Phenolic Compounds in Centaurea rupestris Tissues and Their Antiphytoviral Activity

Quantitative and qualitative analysis of phenolic compounds in tissue extracts from Centaurea rupestris L. as well as their antiphytoviral activity against Tomato bushy stunt virus was performed. Extracts of flowers, leaves and roots from C. rupestris growing in the wild, as well as extracts from shoots, tissue consisting of callus and shoots and undifferentiated callus grown in vitro, were tested. Between tested extracts predominantly quantitative and only several qualitative differences in phenolics were detected by high performance liquid chromatography. The highest amounts of quercetagetin 3’-methylether-7-O-ß-D-glucopyranoside and quercetin were detected in flowers while in leaves the high-est amounts of luteolin, caffeic and p-coumaric acid were detected. Except for roots, antiphytoviral activi-ty of all other extracts was high, inducing virus inhibition, ranging from 43 to 90 %. Simultaneous appli-cation of quercetin, caffeic or p-coumaric acid with virus decreased the number of lesions indicating that these substances contribute to the antiphytoviral activity of C. rupestris extracts. (doi: 10.5562/cca2272

Proizvodnja ružmarinske kiseline u različitim tkivima ukrasne koprive (Coleus blumei)

Rosmarinic acid is one of the main active components of Coleus blumei and is known to have numerous health benefi ts. The pharmacological significance of rosmarinic acid and its production through in vitro culture has been the subject of numerous studies. Here, the ability of different tissues to accumulate rosmarinic acid and sustainability in production over long cultivation have been tested. Calli, tumours, normal roots and hairy roots were established routinely by application of plant growth regulators or by transformation with agrobacteria. The differences among the established tumour lines were highly heterogeneous. Hairy root lines showed the highest mean growth rate and consistency in rosmarinic acid production. Although some tumour lines produced more rosmarinic acid than the hairy root lines, over a long cultivation period their productivity was unstable and decreased. Further, the effects of plant growth regulators on growth and rosmarinic acid accumulation were tested. 2,4-Dichlorophenoxyacetic acid significantly reduced tumour growth and rosmarinic acid production. 1-Naphthaleneacetic acid strongly stimulated hairy root growth whilst abscisic acid strongly enhanced rosmarinic acid production. Hairy roots cultured in an airlift bioreactor exhibited the highest potential for mass production of rosmarinic acid.Ružmarinska kiselina je fenolni sekundarni metabolit poznat po brojnim zdravstvenim učincima. Sintetiziraju je mnoge ljekovite biljke i glavni je aktivni sastojak ukrasne koprive (Coleus blumei). Farmakološki značaj ružmarinske kiseline, kao i njezina proizvodnja u kulturi in vitro, predmetom je brojnih istraživanja. U ovom je radu ispitana sposobnost akumulacije ružmarinske kiseline u različitim tkivima, te održivost proizvodnje tijekom duljeg vremena uzgoja. Ispitani kalusi, tumori, normalno i kosmato korijenje rutinski su uspostavljeni primjenom biljnih regulatora rasta ili transformacijom pomoću agrobakterija. Razlike između uspostavljenih linija tumora bile su vrlo heterogene, a kosmato korijenje imalo je najveću prosječnu stopu rasta te stabilnost u proizvodnji ružmarinske kiseline. Iako su neke tumorske linije proizvodile znatno više ružmarinske kiseline u odnosu na linije kosmatog korijenja, njihova se produktivnost tijekom duljeg vremena uzgoja smanjila. Nadalje, ispitivani su i učinci biljnih regulatora rasta na rast tkiva i akumulaciju ružmarinske kiseline. 2,4-Diklorfenoksioctena kiselina znatno je smanjila rast tumora i proizvodnju ružmarinske kiseline. 1-Naftalenoctena kiselina snažno je stimulirala rast kosmatog korijenja, dok je abscizinska kiselina snažno potakla proizvodnju ružmarinske kiseline. Utvrđeno je da uzgoj kosmatog korijenja u „airlift“ bioreaktoru ima najveći potencijal za masovnu proizvodnju ružmarinske kiseline

Transvaginal mesh in repair of pelvic organs prolapse as a minimally invasive surgical procedure

Background/Aim. Prolapse of genital organs with or without urinary stress incontinention is the most often health problem in the elderly female population tending to increase with ageing. The aim of this study was to assess the perioperative complications and short-term outcomes of prolaps repair using transvaginal polypropylene mesh (Prolift system, Gynecare, Ethicon, USA). Methods. A retrospective study was conducted evaluating 96 women from September 2006 to January 2010 who undewent vaginal repair with implatation of a soft mesh manufactured by Gynecare, Ethicon, USA. Results. All the patients had a stage 3 or stage 4 prolapse according to the POP-Q system of ICS. Total mesh was used in 12 (13%) patients isolated anterior mesh in 52 (54%) patients and isolated posterior mesh in 32 (33%) patients. We reported one intra-operative bladder injury and no other serious complications. At 3 months, all 96 patients were available for follow-up. Vaginal erosion occured in 9 (9.3%) patients, shrinkage of mesh in 6 (6.2%) patients and de novo urinary incontinence in 5 (5.2%) patients. Failure rate was 6.25% (recurrent prolapse stage 3 or 4 even asymptomatic). Conclusion. Our study suggests that transvaginal polypropylene mesh applied with a tensionfree technique is a safe and effective method with low intraoperative complications and low morbidity rates. However, some complications are serious and require highly specialised management

First evidence of the presence of Multixenobiotic Resistance Mechanism activity in freshwater invasive species, signal crayfish Pacifastacus leniusculus (Dana, 1852)

Background and Purpose: The signal crayfish Pacifastacus leniusculus (Dana, 1852) is one of the most successful invasive species of crayfish in European freshwaters, an extremely diverse though endangered group of ecosystems. The main goal of this study was to functionally characterize multixenobiotic resistance (MXR) mechanism defense activity in P. leniusculus tissues for the first time. MXR mechanism protects the cell from a wide variety of toxic compounds, and it is mediated by the transport activity of ATP-binding cassette (ABC) proteins. Materials and Methods: MXR transporter activity dye assay was performed by using fluorescent model substrate rhodamine B (RB) in combination with inhibitors of MXR efflux pumps: MK571 and Verapamil, known to inhibit multidrug resistance-associated proteins (MRP) and P-glycoprotein (P-gp), respectively. In this assay, the increase in intracellular fluorescence of the substrate dye, indicates inhibition of MXR efflux protein pumps. The assay was performed in three different tissues (gills, hepatopancreas, tail muscle). Additionally, tissues were exposed to selected heavy metals – mercury (HgCl2) and zinc (ZnCl2), known to occur in open freshwaters as pollutants. Results: Optimal time for RB accumulation in gills and hepatopancreas was determined to be 30 minutes. RB efflux in gills was inhibited by MK571 and in hepatopancreas by Verapamil, suggesting that multidrug resistanceassociated proteins are dominant in gills of P. leniusculus, and P-glycoprotein in hepatopancreas. Finally, inhibitory effect of mercury (HgCl2: 10 and 20 μM) and zinc (ZnCl2: 5–20 μM) on multixenobiotic resistance mechanism activity in gills, and only mercury in hepatopancreas, was detected. Conclusions: The results for the first time demonstrate the presence of multixenobiotic resistance mechanism efflux activity as an important tissue specific defense mechanism in P. leniusculus and provide the basis for future molecular and toxicological studies of this invasive and adaptable species

Sastav hranjivih tvari i antioksidacijska aktivnost kore, plodova i sjemenki oskoruše (Sorbus domestica L.)

The aim of this work is to assess the nutritional value of service tree (Sorbus domestica L.) bark, fruit exocarp and mesocarp, and seeds by establishing the levels of macro- and microelements, total phenolics, flavonoids and tannins. Our results revealed that all of the tested service tree samples were rich in potassium. Bark was the best source of calcium and zinc, while seeds were the best source of magnesium. Compared to the bark and seeds, fruit exocarp and mesocarp contained significantly lower amounts of these three elements. Immature exocarp and bark contained the highest amounts of total phenolics and showed the highest antioxidant activity. Maturation significantly decreased the amount of total phenolics in fruits, as well as the antioxidant activity of total phenolics and total tannins from exocarp, but not from mesocarp. Exocarp was the richest in total flavonoids. Based on the obtained data, we have concluded that the under-utilised species S. domestica L. could serve as an important source of mineral elements and antioxidants in the human diet.Svrha je ovoga rada bila procijeniti prehrambenu vrijednost kore, egzokarpa, mezokarpa i sjemenki oskoruše (Sorbus domestica L.) na osnovi udjela makroelemenata i mikroelemenata, ukupnih fenolnih spojeva, flavonoida i tanina. Rezultati su pokazali da su svi ispitani uzorci oskoruše bogati kalijem. Kora je najbolji izvor kalcija i cinka, a sjemenke magnezija. U usporedbi s korom i sjemenkama, egzokarp i mezokarp sadržavaju znatno manje udjele ovih triju elemenata. Najveći udjel ukupnih fenolnih spojeva i najveća antioksidacijska aktivnost izmjereni su u ekstraktima nezrelog egzokarpa i kore. Dozrijevanje je značajno smanjilo ukupan sadržaj fenola u plodovima, kao i antioksidacijsku aktivnost fenolnih spojeva i tanina izoliranih iz egzokarpa, no ne i onih iz mezokarpa. Najveći udjel ukupnih flavonoida izmjeren je u egzokarpu. Na osnovi dobivenih podataka zaključili smo da bi oskoruša (S. domestica L.), kao dosad nedovoljno iskorištena biljna vrsta, mogla biti važan izvor mineralnih tvari i antioksidacijskih spojeva u ljudskoj prehrani

Atypical proliferating mucinous tumors of gigantic dimensions

Background. Ovarian tumors of low malignant potential (LMP) are also known as atypically proliferating tumors. Ovarian tumors of LPM account for approximately 15% of all epithelial ovarian cancers. Mean age of occurrence is 40 years and they are 15-20 cm in diameter. Case report. A 32-year-old female patient was hospitalized as an urgent case with a large tumor mass that filled the entire abdomen. Cyst was 100 × 70 cm dimensions belonging to the right ovary and filled with 18 liters of content. Right adnexectomy, resection of the second ovary, as well as biopsy of the omentum were performed. Lymphadenectomy of the right iliac and obturator area was also performed. After receiving definitive histopathological results it was decided to perform a radical reoperation. On the 10th postoperative day relaparotomy, total hysterectomy and left adnexectomy were performed. The patient was released on the 6th postoperative day. She used to come to regular examinations up to date. Conclusion. This case is a proof that LMP tumors have low malignant potential, they grow slowly and can reach great proportions

Characterisation of Sorbus domestica L. Bark, Fruits and Seeds: Nutrient Composition and Antioxidant Activity

The aim of this work is to assess the nutritional value of service tree (Sorbus domestica L.) bark, fruit exocarp and mesocarp, and seeds by establishing the levels of macro- and microelements, total phenolics, flavonoids and tannins. Our results revealed that all of the tested service tree samples were rich in potassium. Bark was the best source of calcium and zinc, while seeds were the best source of magnesium. Compared to the bark and seeds, fruit exocarp and mesocarp contained significantly lower amounts of these three elements. Immature exocarp and bark contained the highest amounts of total phenolics and showed the highest antioxidant activity. Maturation significantly decreased the amount of total phenolics in fruits, as well as the antioxidant activity of total phenolics and total tannins from exocarp, but not from mesocarp. Exocarp was the richest in total flavonoids. Based on the obtained data, we have concluded that the under-utilised species S. domestica L. could serve as an important source of mineral elements and antioxidants in the human diet

Causative agents of bloodstream infections in two Croatian hospitals and their resistance mechanisms

Blood samples were collected alongside with routine blood cultures (BC) from patients with suspected sepsis, to evaluate the prevalence of different causative agents in patients with bacteraemia. Among 667 blood samples, there were 122 positive BC (18%). Haemoglobin content, platelet number, and systolic blood pressure values were significantly lower in patients with positive BC, whereas serum lactate levels, CRP, creatinine and urea content were significantly higher in patients with positive BC. The rate of multidrug (MDR) or extensively drug resistant (XDR) bacteria was 24% (n = 29): Klebsiella pneumoniae (9), Pseudomonas aeruginosa (9), Acinetobacter baumannii (4), Escherichia coli (1), vancomycin resistant Enterococcus spp (VRE) (3), and methicillin-resistant Staphylococcus aureus MRSA (3). The dominant resistance mechanisms were the production of extended-spectrum β-lactamases, OXA-48 carbapenemase, and colistin resistance in K. pneumoniae, VIM metallo-β-lactamases in P. aeruginosa and OXA-23-like oxacillinases in A. baumannii. The study revealed high rate of MDR strains among positive BCs in Zagreb, Croatia

Diffusion of OXA-48 carbapenemase among urinary isolates of Klebsiella pneumoniae in non-hospitalized elderly patients

Background: Recently, a dramatic increase of Klebsiella pneumoniae positive for OXA-48 β-lactamases was observed first in the hospital setting and later in the long-term care facilities (LTCFs) and community in the Zagreb County, particularly, in urinary isolates. The aim of the study was to analyse the epidemiology and the mechanisms of antibiotic resistance of OXA-48 carbapenemase producing K. pneumoniae strains isolated from urine of non-hospitalized elderly patients. ------ Results: The isolates were classified into two groups: one originated from the LTCFs and the other from the community. Extended-spectrum β-lactamases (ESBLs) were detected by double disk-synergy (DDST) and combined disk tests in 55% of the isolates (51/92). The ESBL-positive isolates exhibited resistance to expanded-spectrum cephalosporins (ESC) and in majority of cases to gentamicin. LTCFs isolates showed a significantly lower rate of additional ESBLs and consequential resistance to ESC and a lower gentamicin resistance rate compared to the community isolates, similarly to hospital isolates in Zagreb, pointing out to the possible transmission from hospitals.ESBL production was associated with group 1 of CTX-M or SHV-12 β-lactamases. Ertapenem resistance was transferable from only 12 isolates. blaOXA-48 genes were carried by IncL plasmid in 42 isolates. In addition IncFII and IncFIB were identified in 18 and 2 isolates, respectively. Two new sequence types were reported: ST4870 and ST4781. ----- Conclusions: This study showed eruptive and extensive diffusion of OXA-48 carbapenemase to LTCFs and community population in Zagreb County, particularly affecting patients with UTIs and urinary catheters. On the basis of susceptibility testing, β-lactamase production, conjugation experiments, MLST and plasmid characterization it can be concluded that there was horizontal gene transfer between unrelated isolates, responsible for epidemic spread of OXA-48 carbapenemase in the LTCFs and the community The rapid spread of OXA-48 producing K. pneumoniae points out to the shortcomings in the infection control measures