Ispitivanje fenolnog profila i antioksidacijskog učinka meda od metvice (Mentha spp.)

Research background. The composition of honey is influenced by the botanical source and geographical area of the nectar from which it is derived. Unifloral honeys reach higher market value than multifloral honeys due to their specific aromas, which result from volatile and phenolic compounds. Experimental approach. The aim of our study is to characterize the phenolic composition of a rare unifloral variety of honey – mint (Mentha spp.) honey. For this purpose, we performed standard physicochemical analyses, pollen analysis, determined total phenolic and flavonoid content, analysed antioxidant activity and performed qualitative and quantitative analyses of phenolic compounds for five mint honeys. Results and conclusions. Our results indicate that mint honey samples have high phenolic content, expressed in gallic acid equivalents, from (76.7±0.6) to (90.1±1.1) mg/100 g, and flavonoid content, expressed as quercetin equivalents, from (6.7±0.6) to (12.5±0.8) mg/100 g. These honey samples also exhibit strong antioxidant activity, expressed as Trolox equivalents, from (33.6±2.8) to (51.3±1.2) mg/100 g and from (14.4±0.8) to (55.1±2.4) mg/100 g when analysed with DPPH and ABTS assays, respectively. Quantitative LC-MS/MS analysis revealed that the most abundant phenols in all samples were chrysin, apigenin and p-coumaric acid. Qualitative LC-MS/MS analysis identified the presence of kaempferide, diosmetin, acacetin and several caffeic acid derivatives. Novelty and scientific contribution. Our study indicates that mint honey contains unique phenolic profiles, which likely contribute to its distinctive aroma and strong antioxidant activity. A detailed description of the rare honey varieties gives beekeepers greater visibility and easier access to the demanding natural product market.Pozadina istraživanja. Na sastav meda utječu botanički izvor i geografsko podrijetlo nektara iz kojeg je dobiven. Uniflorni medovi postižu veću tržišnu vrijednost od multiflornih zbog svoje specifične arome, koja je rezultat prisustva hlapljivih i fenolnih spojeva. Eksperimentalni pristup. Svrha je našeg rada bila okarakterizirati fenolni sastav rijetke uniflorne sorte meda – meda od metvice (Mentha spp.). U tu svrhu smo proveli standardne fizikalno-kemijske analize i analizu peludi, odredili ukupne udjele fenola i flavonoida, ispitali antioksidacijsku aktivnost te kvalitativno i kvantitativno ispitali fenolne spojeve u pet uzoraka meda od metvice. Rezultati i zaključci. Rezultati pokazuju da uzorci meda od metvice imaju velik udjel fenola, izražen u ekvivalentima galne kiseline, od (76,7±0,6) do (90,1±1,1) mg/100 g, te udjel flavonoida, izražen u ekvivalentima kvercetina, od (6,7±0,6) do (12,5±0,8) mg/100 g. Uzorci su također imali snažnu antioksidacijsku aktivnost, izraženu u Trolox ekvivalentima, i to prema metodi DPPH od (33,6±2,8) do (51,3±1,2) mg/100 g, a prema metodi ABTS od (14,4±0,8) do (55,1±2,4) mg/100 g. Kvantitativnim ispitivanjem pomoću spregnutog sustava tekućinske kromatografije i tandemske spektrometrije masa utvrđeno je da su najzastupljeniji fenoli u svim uzorcima bili krizin, apigenin i p-kumarinska kiselina, dok je kvalitativnom analizom pomoću istog sustava utvrđena prisutnost kemferida, diosmetina, akacetina i nekoliko derivata kafeinske kiseline. Novost i znanstveni doprinos. U istraživanju smo pokazali da med od metvice ima jedinstven fenolni profil, koji pridonosi njegovoj prepoznatljivoj aromi i snažnom antioksidativnom djelovanju. Detaljan opis rijetkih sorti meda omogućava pčelarima veću vidljivost i lakši pristup zahtjevnom tržištu prirodnih proizvoda

Anti-androgens act jointly in suppressing spiggin concentrations in androgen-primed female three-spined sticklebacks - Prediction of combined effects by concentration addition

This is the post-print version of the final paper published in Aquatic Toxicology. The published article is available from the link below. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. Copyright @ 2013 Elsevier B.V.Increasing attention is being directed at the role played by anti-androgenic chemicals in endocrine disruption of wildlife within the aquatic environment. The co-occurrence of multiple contaminants with anti-androgenic activity highlights a need for the predictive assessment of combined effects, but information about anti-androgen mixture effects on wildlife is lacking. This study evaluated the suitability of the androgenised female stickleback screen (AFSS), in which inhibition of androgen-induced spiggin production provides a quantitative assessment of anti-androgenic activity, for predicting the effect of a four component mixture of anti-androgens. The anti-androgenic activity of four known anti-androgens (vinclozolin, fenitrothion, flutamide, linuron) was evaluated from individual concentration-response data and used to design a mixture containing each chemical at equipotent concentrations. Across a 100-fold concentration range, a concentration addition approach was used to predict the response of fish to the mixture. Two studies were conducted independently at each of two laboratories. By using a novel method to adjust for differences between nominal and measured concentrations, good agreement was obtained between the actual outcome of the mixture exposure and the predicted outcome. This demonstrated for the first time that androgen receptor antagonists act in concert in an additive fashion in fish and that existing mixture methodology is effective in predicting the outcome, based on concentration-response data for individual chemicals. The sensitivity range of the AFSS assay lies within the range of anti-androgenicity reported in rivers across many locations internationally. The approach taken in our study lays the foundations for understanding how androgen receptor antagonists work together in fish and is essential in informing risk assessment methods for complex anti-androgenic mixtures in the aquatic environment.European Commission and Natural Environment Research Council

Direct Effects, Compensation, and Recovery in Female Fathead Minnows Exposed to a Model Aromatase Inhibitor

BackgroundSeveral chemicals in the environment have the potential to inhibit aromatase, an enzyme critical to estrogen synthesis.ObjectivesThe objective of this study was to provide a detailed characterization of molecular and biochemical responses of female fathead minnows to a model aromatase inhibitor, fadrozole (FAD).MethodsFish were exposed via water to 0, 3, or 30 microg FAD/L for 8 days and then held in clean water for 8 days, with samples collected at four time points during each 8-day period. We quantified ex vivo steroid production, plasma steroids, and plasma vitellogenin (Vtg) concentrations and analyzed relative transcript abundance of 10 key regulatory genes in ovaries and 3 in pituitary tissue by real-time polymerase chain reaction.ResultsEx vivo 17beta-estradiol (E2) production and plasma E2 and Vtg concentrations were significantly reduced after a single day of exposure to 3 microg or 30 microg FAD/L. However, plasma E2 concentrations recovered by the eighth day of exposure in the 3-microg/L group and within 1 day of cessation of exposure in the 30-microg/L group, indicating concentration- and time-dependent physiologic compensation and recovery. Concentration-dependent increases in transcripts coding for aromatase (A isoform), cytochrome P450 side-chain cleavage, steroidogenic acute regulatory protein, and follicle-stimulating hormone receptor all coincided with increased E2 production and recovery of plasma E2 concentrations.ConclusionsResults of this research highlight the need to consider compensation/adaptation and recovery when developing and interpreting short-term bioassays or biomarkers or when trying to predict the effects of chemical exposures based on mode of action

Support for collective action against refugees: The role of national, European, and global identifications, and autochthony beliefs

To understand recent anti-refugee protests in Europe, we examined how different levels of inclusiveness of group identities (national, European, and global) are related to intentions to protest among native Europeans. We focused on the mediating role of autochthony (a belief that the first inhabitants of a territory are more entitled) and the moderating role of threat. Survey data from 11 European countries (N=1909) showed that national identification was positively associated with autochthony, and therefore, with the intention to protest against refugees. In contrast, global identification was related to lower protest intentions via lower autochthony. These paths were found only among Europeans who perceived refugees as a threat. European identification was not related to the endorsement of autochthony or to collective action. These findings indicate why and when majority members are willing to participate in collective action against refugees, and underscore the importance of global identification in the acceptance of refugees

Application of peritoneal dialysis in the therapy of acute exogenous poisonings

Iznesena su vlastita iskustva primjene peritonealne dijalize kao terapijske metode u 24 slučaja teških akutnih trovanja. Potpuni uspjeh je postignut u svih 10 bolesnika otrovanih lijekovima. Međutim, kada su u pitanju bili gljivini otrovi efekat ove terapijske metode je bio znatno slabiji. Od 14 teških trovanja gljivama 6 bolesnika je umrlo. Ovo se ne može pripisati slaboj efikasnosti peritonealne dijalize jer su nam bolesnici došli na odjeljenje sa jasnim znacima teškog oštećenja jetre, bubrega i drugih organa u organizmu.The authors describe own experience with the application of peritoneal dialysis as a therapeutic method in 24 cases of very serious exogenous poisoning by drugs and mushrooms. In 10 cases of drug poisoning a complete recovery took place, but in cases of mushroom poisoning this method was not very successful. Six patients (42.8) out of the 14 seriously poisoned by mushrooms died. This cannot be attributed to the ineffectiveness of peritoneal dialysis, because all patients were admitted to hospital with clear signs of serious damages of the liver, kidneys and other organs

Oral squamous cell carcinoma detection by salivary biomarkers in a Serbian population

Early detection of oral squamous cell cancer (OSCC) is the key to improve the low 5-year survival rate. Using proteomic and genomic technologies we have previously discovered and validated salivary OSCC markers in American patients. The question arises whether these biomarkers are discriminatory in cohorts of different ethnic background. Six transcriptome (DUSP1, IL8, IL1B, OAZ1, SAT1, and S100P) and three proteome (IL1B, IL8, and M2BP) biomarkers were tested on 18 early and 17 late stage OSCC patients and 51 healthy controls with quantitative PCR and ELISA. Four transcriptome (IL8, IL1B, SAT1, and S100P) and all proteome biomarkers were significantly elevated (p lt 0.05) in OSCC patients. The combination of markers yielded an AUC of 0.86, 0.85 and 0.88 for OSCC total, T1-T2, and T3-T4, respectively. The sensitivity/specificity for OSCC total was 0.89/0.78, for T1-T2 0.67/0.96, and for T3-T4 0.82/0.84. In conclusion, seven of the nine salivary biomarkers (three proteins and four mRNAs) were validated and performed strongest in late stage cancer. Patient-based salivary diagnostics is a highly promising approach for OSCC detection. This study shows that previously discovered and validated salivary OSCC biomarkers are discriminatory and reproducible in a different ethnic cohort. These findings support the feasibility to implement multi-center, multi-ethnicity clinical trials towards the pivotal validation of salivary biomarkers for OSCC detection

Effects of single orally administrated different sized microplastic particles – acute toxicity study in adult male rats

Introduction: As a result of human activity, the amount of plastic waste has increased immensely in the environment. While the main concern has been orientated toward visible litter, more recently fragments of plastic less than 5 mm in diameter, called microplastic particles (MP), raised the attention due to their ability to induce toxic effects in organisms. The toxicity of MP increases with a decrease in size since smaller sized particles are absorbed systematically and accumulated in different organs/tissues, including brain. Aim: Present study is designed to compare possible toxic acute effects of different sized MP, originating from polyethylene-terephthalate bottles, on toxicity signs, sensory-motor functions and relative brain weight in young male Wistar rats. Methods: Rats (n = 24) were divided in four experimental groups: (I) intact (controls); Q that orally received a 2.5 ml of Milli-Q; P1/P2 that were respectively orally administrated with 35 mg/kg of MP in median diameter of either 40 or 70 μm dissolved in a 2.5 ml of Milli-Q. 24 h after treatments, using toxicity clinical test, we assessed the agitation, convulsion, piloerection, sleepiness and lethargy. Parameters were valued as: – (no effect), + (mild effect), ++ (moderate effect) and +++ (major effect). Then, we evaluated sensory-motor functions by monitoring: consciousness (scored 0/1 point), respiration (scored 0/1), spontaneous activity (scored 0/1/2/3), forepaw outstretching (scored 0/1/2), climbing (scored 0/1), visual placing (scored 0/1), cage grasp (scored 0/1), gait posture (scored 0/1/2), geotaxis (scored 0/1), hearing (scored 0/1) and pacing/circling (scored 0/1). Total score was expressed as the sum of average scores of all investigated parameters and graded on the scale from 0 to 15. Then, rats were weighed and sacrificed, while their brains were carefully removed and weighed. The relative brain weights were calculated as the ratio of brain (wet tissue, mg) weight to body weight (g). For toxicity clinical testing a descriptive analysis was used while other data were analysed by One-way analysis of variance followed by Tukey’s posthoc test. Results were considered significant at p < 0.05. Results: Clinical toxicity signs were similar between groups while results of sensory-motor test point to unaltered sensory-motor functions upon MP treatments compared to controls (F (3, 20) = 0.9538; P = 0.4336). The absence of changes might be attributed to acute treatment, brief evaluation period and lack of cumulative effect upon MP treatments. Nonetheless, only smaller sized MP led to an increase in relative brain weight compared to controls (F (3, 20) = 3.425; P = 0.0369; p < 0.05) which might indicate the capability of smaller sized MP to cross blood-brain barrier and be accumulated in the brain inducing oxidative stress and inflammatory response. Although larger sized MP did not cause changes in relative brain weight, the possible modulations of additional parameters that were not the focus of this study should not be omitted. Conclusion: According to the presented results, even a smaller sized MP applied in a single dose could be harmful to health, thus, greater control of plastic usage and disposal is essential for humanity

Rubbery taproot disease of sugar beet in Serbia associated with 'Candidatus phytoplasma solani'

Rubbery taproot disease (RTD) of sugar beet was observed in Serbia for the first time in the 1960s. The disease was already described in neighboring Bulgaria and Romania at the time but it was associated with abiotic factors. In this study on RTD of sugar beet in its main growing area of Serbia, we provide evidence of the association between 'Candidatus Phytoplasma solani' (stolbur phytoplasma) infection and the occurrence of typical RTD symptomatology. 'Ca. P. solani' was identified by PCR and the sequence analyses of 16S ribosomal RNA, tuf, secY, and stamp genes. In contrast, the causative agent of the syndrome “basses richesses” of sugar beet-namely, 'Ca. Arsenophonus phytopathogenicus'-was not detected. Sequence analysis of the stolbur strain's tuf gene confirmed a previously reported and a new, distinct tuf stolbur genotype (named 'tuf d') that is prevalent in sugar beet. The sequence signatures of the tuf gene as well as the one of stamp both correlate with the epidemiological cycle and reservoir plant host. This study provides knowledge that, for the first time, enables the differentiation of stolbur strains associated with RTD of sugar beet from closely related strains, thereby providing necessary information for further epidemiological work seeking to identify insect vectors and reservoir plant hosts. The results of this study indicate that there are differences in hybrid susceptibility. Clarifying the etiology of RTD as a long-known and economically important disease is certainly the first step toward disease management in Serbia and neighboring countries.This is the peer reviewed version of the following article: Ćurčić Ž., Stepanović J., Zübert C., Taški-Ajduković K., Kosovac A., Rekanović E., Kube M., Duduk B. Rubbery taproot disease of sugar beet in Serbia associated with 'Candidatus phytoplasma solani'. Plant Disease 2021, 105 (2), 255 – 263. [https://doi.org/10.1094/PDIS-07-20-1602-RE]

Effects of chronic oral D-galactose treatment on general health status in male Wistar rats

D-galactose (d-gal) is an important physiological nutrient. According to the widely accepted aging metabolic theory d-gal at high levels can be converted into aldose and hydroperoxide, resulting in the overproduction of reactive oxygen species (ROS). Increased ROS levels may subsequently cause oxidative stress, inflammation, mitochondrial dysfunction, and apoptosis which are hallmarks of natural senescence as well as various pathological conditions. We investigated the effects of chronic oral d-gal intake (200 mg/kg and 500 mg/kg for 6 weeks) on physiological, neurological and toxicity parameters in 3 months old male Wistar rats. The obtained results indicate that body weight, food intake, serum glucose, neurological and toxicity status remained unaffected while urine proteins were significantly increased in d-gal treated rats. Although there was no effect on the general health status of the animals, our findings suggest that chronic oral d-gal administration may lead to renal dysfunction