Gold nanoparticles (GNPs) are claimed as outstanding biomedical tools for cancer diagnostics and photo-thermal therapy, but without enough evidence on their potentially adverse immunological effects. ...Using a model of human dendritic cells (DCs), we showed that 10 nm- and 50 nm-sized GNPs (GNP10 and GNP50, respectively) were internalized predominantly via dynamin-dependent mechanisms, and they both impaired LPS-induced maturation and allostimulatory capacity of DCs, although the effect of GNP10 was more prominent. However, GNP10 inhibited LPS-induced production of IL-12p70 by DCs, and potentiated their Th2 polarization capacity, while GNP50 promoted Th17 polarization. Such effects of GNP10 correlated with a stronger inhibition of LPS-induced changes in Ca2+ oscillations, their higher number per DC, and more frequent extra-endosomal localization, as judged by live-cell imaging, proton, and electron microscopy, respectively. Even when released from heat-killed necrotic HEp-2 cells, GNP10 inhibited the necrotic tumor cell-induced maturation and functions of DCs, potentiated their Th2/Th17 polarization capacity, and thus, impaired the DCs' capacity to induce T cell-mediated anti-tumor cytotoxicity in vitro. Therefore, GNP10 could potentially induce more adverse DC-mediated immunological effects, compared to GNP50.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Porphyromonas gingivalis, an asaccharolytic Gram-negative oral anaerobe, is a major pathogen associated with adult periodontitis, a chronic infective disease that a significant percentage of the ...human population suffers from. It preferentially utilizes dipeptides as its carbon source, suggesting the importance of dipeptidyl peptidase (DPP) types of enzyme for its growth. Until now DPP IV, DPP5, 7 and 11 have been extensively investigated. Here, we report the characterization of DPP III using molecular biology, biochemical, biophysical and computational chemistry methods. In addition to the expected evolutionarily conserved regions of all DPP III family members, PgDPP III possesses a C-terminal extension containing an Armadillo (ARM) type fold similar to the AlkD family of bacterial DNA glycosylases, implicating it in alkylation repair functions. However, complementation assays in a DNA repair-deficient Escherichia coli strain indicated the absence of alkylation repair function for PgDPP III. Biochemical analyses of recombinant PgDPP III revealed activity similar to that of DPP III from Bacteroides thetaiotaomicron, and in the range between activities of human and yeast counterparts. However, the catalytic efficiency of the separately expressed DPP III domain is ~1000-fold weaker. The structure and dynamics of the ligand-free enzyme and its complex with two different diarginyl arylamide substrates was investigated using small angle X-ray scattering, homology modeling, MD simulations and hydrogen/deuterium exchange (HDX). The correlation between the experimental HDX and MD data improved with simulation time, suggesting that the DPP III domain adopts a semi-closed or closed form in solution, similar to that reported for human DPP III. The obtained results reveal an atypical DPP III with increased structural complexity: its superhelical C-terminal domain contributes to peptidase activity and influences DPP III interdomain dynamics. Overall, this research reveals multifunctionality of PgDPP III and opens direction for future research of DPP III family proteins.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Cilj rada je ispitati upoznatost posjetitelja s ponudom turističkih seoskih gospodarstava u Dubrovačko– neretvanskoj županiji (DNŽ) te utvrditi obilježja potražnje i zadovoljstvo posjetitelja ukupnom ...ponudom. Anketno osobno ispitivanje provedeno je na uzorku od 111 posjetitelja turističkih seoskih gospodarstva u DNŽ. Rezultati istraživanja pokazali su da je većina posjetitelja slabo ili djelomično upoznata s ponudom turističkih seoskih gospodarstava pri čemu se većina njih o ponudi informirala putem preporuke prijatelja i rodbine. Većini posjetitelja ovo je bio prvi posjet nekom turističkom seoskom gospodarstvu u zadnjih godinu dana. Kao razloge posjeta turističkim seoskim gospodarstvima većina ispitanika navodi zdravu hranu i uživanje u specijalitetima, druženje s obitelji i prijateljima, odmor i opuštanje, te bijeg od gužve. Ispitanici su zadovoljni ukupnom ponudom turističkih seoskih gospodarstava pri čemu su najzadovoljniji ljubaznošću domaćina, ponudom hrane i pića, te udobnošću smještaja na odabranom gospodarstvu, dok su manje zadovoljni mogućnošću sudjelovanja u poljoprivrednim poslovima i zastupljenosti domaćih životinja na posjećenim gospodarstvima. Rezultati ovog istraživanja daju vrijedne informacije turističkim seoskim gospodarstvima u svrhu planiranja ponude i marketinških aktivnosti.
The aim of this paper is to examine the familiarity of visitors with the offer of tourist family farms in Dubrovnik - Neretva County, to determine the characteristics of demand and visitors satisfaction with the overall offer of tourist family farms. A survey was conducted on a sample of 111 visitors. The results showed that the majority of visitors were poorly or partially familiar with the offer of tourist family farms, where most of them were informed about the offer through the recommendation of friends and relatives. For most visitors, this was their first visit to a tourist farm in the last year. Main reason for visiting tourist family farms was healthy food, having pleasure of eating local specialities, quality time with their friends and family, and an escape from urban crowds. Visitors are satisfied with the overall offer of tourist farms, whereby they were most pleased with the warmth and kindness of the hosts, food and drink variety, the comfort of the accommodation, while they was less satisfied with the possibilities in participating in agricultural activities and the lack of animals on visited farms. The results of this research provide valuable information to tourist family farms for the purpose of supply planning and marketing activities.
Yeast dipeptidyl peptidase III (yDPP III) is a member of the metallopeptidase family M49 involved in intracellular protein catabolism. Elucidation of the yDPP III crystal structure has pinpointed the ...zinc-coordinating residues (two His from H(460)ELLGH(465) motif and the second Glu from E(516)ECRAE(521) motif), and several amino acid residues potentially important for catalytic activity whose roles have not been investigated. Here, three putative catalytic residues of the yDPP III, His578, Arg582 and Lys638 were substituted and the resultant single mutants characterized. The replacement of His578 with an asparagine significantly (122-fold) lowered the catalytic efficiency, kcat/Km, for Arg-Arg-2-naphthylamide (Arg2-2NA) hydrolysis, and affinity for hydroxamate inhibitor Tyr-Phe-NHOH (decline by 14-fold). The R582Q mutant exhibited an order of magnitude higher activity with all four dipeptide derivatives examined, compared to the wild type. The molecular dynamics simulations revealed the change in the H-bond networking in the R582Q variant active-site region. The mutation of Lys638, to Leu, slightly increased the specificity constant for Arg2-2NA hydrolysis. However, the affinity for Tyr-Phe-NHOH, and activity for the substrates with uncharged P2 side chains (Ala-Ala-, Ala-Arg- and Phe-Arg-2NA) were dramatically reduced, indicating the importance of the evolutionary conserved salt bridge Lys(638)-Glu(516) for the modulation of DPP III substrate specificity.
Brassica rapa auxin amidohydrolase (BrILL2) participates in the homeostasis of the plant hormones auxins by hydrolyzing the amino acid conjugates of auxins, thereby releasing the free active form of ...hormones. Herein, the potential role of the two conserved Cys residues of BrILL2 (at sequence positions 139 and 320) has been investigated by using interdisciplinary approaches and methods of molecular biology, biochemistry, biophysics and molecular modelling. The obtained results show that both Cys residues participate in the regulation of enzyme activity. Cys320 located in the satellite domain of the enzyme is mainly responsible for protein stability and regulation of enzyme activity through polymer formation, as has been revealed by enzyme kinetics and differential scanning calorimetry analysis of the BrILL2 wild type and mutants C320S and C139S. Cys139 positioned in the active site of the catalytic domain is involved in the coordination of one Mn(2+) ion of the bimetal center and is crucial for the enzymatic activity. Although the point mutation Cys139 to Ser causes the loss of enzyme activity, it does not affect the metal binding to the BrILL2 enzyme, as has been shown by isothermal titration calorimetry, circular dichroism spectropolarimetry and differential scanning calorimetry data. MD simulations (200 ns) revealed a different active site architecture of the BrILL2C139S mutant in comparison to the wild type enzyme. Additional possible reasons for the inactivity of the BrILL2C139S mutant have been discussed based on MD simulations and MM-PBSA free energy calculations of BrILL2 enzyme complexes (wt and C139S mutant) with IPA-Ala as a substrate.
Poor antimicrobial activity and lack of protection against UV irradiation are weaknesses of cotton textiles. In an attempt to impart superior antimicrobial and UV‐protective properties to cotton, ...layer‐by‐layer deposition of chitosan and magnesium lignosulfonate, and in situ synthesis of silver (Ag) nanoparticles (NPs), was performed. Lignin, in a chitosan/lignin multilayer, simultaneously acts as a UV protective macromolecule and natural reducing and stabilizing agent, allowing formation of Ag NPs. Four bilayers of this coating is sufficient for fabrication of a chitosan/lignin/Ag‐NP textile nanocomposite treatment with 50+ UV protection. 100% reduction of Gram‐negative bacteria Escherichia coli, gram‐positive bacteria Staphylococcus aureus, and yeast Candida albicans can be achieved with a 12‐bilayer coating, when 20 mM silver nitrate solution and sodium borohydride are applied. On the other hand, four bilayers impregnated with Ag NPs, using 10 mM silver nitrate solution, provides sufficient antimicrobial activity independent of an added reducing agent. This treatment exhibits no inhibition of human keratinocyte cells growth on the skin, indicating low cytotoxicity.
Seminoma is a testicular tumor type, routinely diagnosed after orchidectomy. As cfDNA represents a source of minimally invasive seminoma patient management, this study aimed to investigate whether ...cfDNA methylation of six genes from liquid biopsies, have potential as novel seminoma biomarkers.
cfDNA methylation from liquid biopsies was assessed by pyrosequencing and compared with healthy volunteers' samples.
Detailed analysis revealed specific CpGs as possible seminoma biomarkers, but receiver operating characteristic curve analysis showed modest diagnostic performance. In an analysis of panels of statistically significant CpGs, two DNA methylation panels emerged as potential seminoma screening panels, one in blood CpG8/CpG9/CpG10 (
) and the other in seminal plasma CpG1(
)/CpG1(
).
The presented data promote the development of liquid biopsy epigenetic biomarkers in the screening of seminoma patients.
Sparkling wines accounted for 7.7% of the total wine consumption in 2011 in the world. Even though sparkling wine remains a long way behind still wine in total volume of production, world consumption ...of sparkling wine is expected to grow at a faster rate with an 8.5% increase by 2016. Although Croatia is a country with a high residual per capita consumption, the share of sparkling wine consumption compared with total wine consumption amounts only 1.3%. To collect data on Croatian sparkling wine consumers' behavior and attitudes, we performed online research with 273 consumers. The research confirmed the low frequency of sparkling wine consumption. The most important characteristics of sparkling wines for Croatian consumers are their intrinsic characteristics (taste, smell) together with price:quality ratio, while visual appearance, expert reviews, or wine prizes are less important. Consumption of sparkling wines is generally associated with specific celebrations. Due to such perception of exclusive wine use, there is a need to break down this prejudice by better sparkling wine promotion through wine-tasting events, seminars, and gastronomy offers, especially for younger consumers.
The purpose of this study was to collect all available randomized controlled trials (RCT) on the treatment of open tibial fractures with an external fixator (EF) and intramedullary nailing (IMN) for ...meta-analysis to provide reliable evidence-based data for clinical decision-making.
The systematic review was undertaken in accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) and AMSTAR (Assessing the Methodological Quality of Systematic Review). An electronic search of PubMed, Cochrane Library, and Web of Science was performed until 1 March 2023 to identify RCTs which compared either IMN or EF to fix the open tibial fracture. Outcome measures were: postoperative superficial and deep infection, time to union, delayed union, malunion, nonunion and hardware failure. In addition, pain and health-related quality of life were evaluated after 3 and 12 months of follow-up.
Sixteen publications comprising 1011 patients were included in the meta-analysis. The pooled results suggested that the IMN technique had a lower postoperative superficial infection and malunion rate (RR = 3.56, 95%CI = 2.56-4.95 and RR = 1.96, 95%CI = 1.12-3.44, respectively), but higher hardware failure occurrence in contrast to EF (RR = 0.30; 95%CI = 0.13-0.69). No significant differences were found in the union time, delayed union or nonunion rate, and postoperative deep infection rate between the treatments. Lower levels of pain were found in the EF group (RR = 0.05, 95%CI = 0.02-0.17,
< 0.001). A difference in quality of life favoring IMN after 3 months was found (RR = -0.04, 95%CI = -0.05-0.03,
< 0.001), however, no statistical difference was found after 12 months (RR = 0.03, 95%CI = -0.05-0.11,
= 0.44).
Meta-analysis presented reduced incidence rates of superficial infection, malunion, and health-related quality of life 3 months after treatment in IMN. However, EF led to a significant reduction in pain and incidence rate of hardware failure. Postoperative deep infection, delayed union, nonunion and health-related quality of life 12 months following therapy were similar between groups. More high-quality RCTs should be conducted to provide reliable evidence-based data for clinical decision-making.
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