Quantification of antibiotics in honey by liquid chromatography tandem mass spectrometry

U ovom radu korištena je metoda za kvalitativno i kvantitativno određivanje sulfonamida i kloramfenikola u medu kombiniranom tehnikom tekućinske kromatografije visoke djelotvornosti i spektrometrije masa. Optimizirani su uvjeti za istovremeno izdvajanje sulfonamida i kloramfenikola iz meda ekstrakcijom na čvrstoj fazi, te je optimizacijom uvjeta postignuta niža granica kvantifikacije za sulfonamide koja iznosi 5,0 g/kg. Granica detekcije za kloramfenikol iznosi 0,3 g/kg. Uzorci su ukoncentrirani i pročišćeni ekstrakcijom na čvrstoj fazi (SPE, engl. Solid phase extraction). Daljnja analiza provodila se na temelju podataka o zadržavanju analita na C18 kromatografskoj koloni i podataka dobivenih MS/MS analizom pomoću sustava ABSciex Qtrap 4500. Optimizirani su volumeni otapala za eluiranje kolone za ekstrakciju i pH-vrijednost uzorka. Također je ispitana ponovljivost pripreme uzorka na granicama kvantifikacije, te je izražena relativnim standardnim odstupanjem (RSD). Prinosi metode ekstrakcije za kloramfenikol iznosi 84,62 % i za sulfonamide od 80,80 % do 125,00 %. Kao unutarnji standard tijekom kvantitativne analize spomenutih antibiotika koristio se izotopno obilježeni kloramfenikol, sulfametazin i sulfametoksazol. Metoda se primjenjivala za analizu sulfonamida i kloramfenikola u uzorcima meda prikupljenim sa različitih geografskih lokacija.Quantitative and qualitative method was used for determination of sulfonamides and chloramphenicol in honey by liquid chromatography coupled to mass spectrometry. Conditions were optimized for simultaneous extraction of sulfonamides and chloramphenicol from honey in solid state. Lower detection limit was achieved for sulfonamides (5.0 g/kg). Detection limit for chloramphenicol was 0.3 g/kg. Samples were concentrated and refined by sold phase extraction. Further analysis was conducted by data of analyte retention at the C18 column chromatography and by the MS/MS data obtained with ABSciex Qtrap 4500 system. pH value of sample and the volume of elution solvent were optimized. Repeatability of method was determined and expressed as relative standard deviation (RSD). Extraction recoveries were high with values in the range 84.62 % for chloramphenicol and of 80.80 % to 125.00 % for sulfonamides. Isotopically marked chloramphenicol was used as internal standard for quantitative analysis of antibiotics. Method was applied for determination of antibiotic in honey samples obtained from the various geographical locations

Bacteroides fragilis bakterijemija u atipičnoj prezentaciji akutnog apendicitisa

Acute appendicitis is one of the most common surgical emergencies and atypical presentation can delay its diagnosis. We report a case of a patient with Bacteroides fragilis bacteremia with a 5-day fever, vomiting and diarrhea. Blood cultures warned physicians to extend the diagnostics with computed tomography that revealed acute appendicitis.Akutni apendicitis jedno je od najčešćih hitnih kirurških stanja, a atipična prezentacija može odgoditi dijagnozu. Prikazujemo slučaj bolesnika s Bacteroides fragilis bakterijemijom te petodnevnom temperaturom, povraćanjem i proljevom. Hemokulture su upozorile liječnike da prošire dijagnostiku kompjutoriziranom tomografijom koja je pokazala da se radi o akutnom apendicitisu

Liječenje starijih bolesnika s kroničnim hepatitisom C: retrospektivno kohortno istraživanje

The prevalence of chronic hepatitis C increases in elderly patients. The aims of this study were to identify the factors associated with hepatocellular carcinoma (HCC) and end-stage liver disease development and to evaluate the efficacy and safety of pegylated interferon (PEG-IFNα) plus ribavirin (RBV) therapy in elderly patients. A retrospective cohort study included all consecutive patients with hepatitis C virus (HCV) infection treated with PEG-IFNα+RBV between 2003 and 2013. Elderly patients had a higher frequency of poor prognostic factors including genotype 1 infection, high fibrosis, and high fibrosis index based on four factors (FIB-4) score. The sustained virologic response (SVR) rate for genotype 1 was significantly lower (35.8% vs. 57.1%), while the frequency of PEG-IFNα (27.2% vs. 7.8%), RBV dose reduction (19.6% vs. 9.7%) and treatment discontinuation (13.0% vs. 4.1%) was significantly higher in elderly patients. However, age was not associated with SVR in multivariate analysis, and comparable SVR rates were achieved when adjusted for fibrosis score (Ishak ≤3: 66.7% vs. 69.8%). During the follow-up, HCC was diagnosed in 18 elderly patients (3 SVR+, 4 SVR- and 9 untreated patients). In conclusion, selected elderly patients can achieve comparable SVR rates as younger patients, but with a higher rate of side effects. Since complications of HCV infection occur more frequently in elderly patients, they should be given priority for antiviral therapy.Učestalost kroničnog hepatitisa C (KHC) raste u starijim dobnim skupinama. Ciljevi ovoga istraživanja bili su utvrditi čimbenike povezane s razvojem hepatocelularnog karcinoma (HCC) i dekompenzirane jetrene bolesti te procijeniti učinkovitost i sigurnost terapije pegiliranim interferonom (PEG-IFNα) i ribavirinom (RBV) u starijih bolesnika. Retrospektivna kohortna studija je uključila sve bolesnike s KHC koji su liječeni PEG-IFNα + RBV između 2003. i 2013. godine u Klinici za infektivne bolesti “Dr. Fran Mihaljević”. Bolesnici u dobi od >65 godina češće su imali nepovoljne prognostičke čimbenike, tj. HCV-1 genotip, uznapredovali stadij fibroze i viši zbir indeksa fibroze zasnovan na četiri čimbenika (fibrosis index based on four factors, FIB-4). Trajni virusološki odgovor (sustained virologic response, SVR) je bio značajno niži (35,8% prema 57,1%), dok je učestalost smanjenja doze PEG-IFNα (27,2% prema 7,8%), RBV (19,6% prema 9,7%) i prekida liječenja (13,0% prema 4,1%) bila značajno češća u starijih bolesnika. Dob nije bila povezana sa SVR u multivarijatnoj analizi, a stariji bolesnici su imali podjednaki SVR kao i mlađi bolesnici ovisno o stadiju fibroze (Ishak ≤3: 66,7% prema 69,8%). Tijekom praćenja HCC je dijagnosticiran u 18 bolesnika u dobi od >65 godina (3 SVR+, 4 SVR-, 9 neliječenih). Zaključno, stariji bolesnici imaju podjednaku vjerojatnost postizanja SVR kao i mlađi, ali uz češće nuspojave. Budući da se komplikacije infekcije virusom hepatitisa C češće javljaju u ovoj populaciji, stariji bolesnici trebaju imati prednost u primjeni antivirusne terapije

Treatment of chronic hepatitis C in Croatian war veterans: experiences from Croatian reference center for viral hepatitis

Aim To examine the risk factors, comorbidity, severity of liver disease, treatment course, and outcome in Croatian war veterans with chronic hepatitis C, especially those suffering from posttraumatic stress disorder (PTSD). Methods We collected medical records of 170 adult men diagnosed with chronic hepatitis C who started treatment with a combination of pegylated interferon-alpha and ribavirin between January 2003 and June 2009 at the Croatian Reference Centre for Viral Hepatitis. Results Participants’ mean age was 43 ± 9 years. Among 170 participants, there were 37 war veterans (22%). The main risk factor in veteran patients were operative procedures with transfusions (46% vs 5% in non-veterans; P < 0.001) and in non-veteran patients intravenous drug use (42.1% vs 13%; P < 0.001). The average duration of infection was longer in war veterans (14.5 ± 3.4 vs 12.2 ± 7.2 years; P = 0.020). The percentage of PTSD comorbidity in the whole group was 11% (18/170) and in the war veterans group 49% (18/37). The prevalence of sustained virological response in patients with PTSD was 50% and in patients without PTSD 56%. Treatment reduction in patients with PTSD (33%) was higher than in patients without PTSD (12%; P = 0.030). Conclusion Croatian war veterans are a group with high risk of chronic hepatitis C infection because many of them were wounded during the Croatian War 1991-1995. Considerations about PTSD as a contraindication for interferon treatment are unjustified. If treated, patients with PTSD have an equal chance of achieving sustained virological response as patients without PTSD

Treatment of chronic hepatitis C in Croatian war veterans: experiences from Croatian reference center for viral hepatitis

Aim. To examine the risk factors, comorbidity, severity of liver disease, treatment course, and outcome in Croatian war veterans with chronic hepatitis C, especially those suffering from posttraumatic stress disorder (PTSD). ----- Methods. We collected medical records of 170 adult men diagnosed with chronic hepatitis C who started treatment with a combination of pegylated interferon-alpha and ribavirin between January 2003 and June 2009 at the Croatian Reference Centre for Viral Hepatitis. ----- Results. Participants' mean age was 43±9 years. Among 170 participants, there were 37 war veterans (22%). The main risk factor in veteran patients were operative procedures with transfusions (46% vs 5% in non-veterans; P<0.001) and in non-veteran patients intravenous drug use (42.1% vs 13%; P<0.001). The average duration of infection was longer in war veterans (14.5±3.4 vs 12.2±7.2 years; P=0.020). The percentage of PTSD comorbidity in the whole group was 11% (18/170) and in the war veterans group 49% (18/37). The prevalence of sustained virological response in patients with PTSD was 50% and in patients without PTSD 56%. Treatment reduction in patients with PTSD (33%) was higher than in patients without PTSD (12%; P=0.030). ----- Conclusion. Croatian war veterans are a group with high risk of chronic hepatitis C infection because many of them were wounded during the Croatian War 1991-1995. Considerations about PTSD as a contraindication for interferon treatment are unjustified. If treated, patients with PTSD have an equal chance of achieving sustained virological response as patients without PTSD

Sezonska dinamika prisustva culicoides spp u Srbiji u periodu 2015-2016. godine

Genus Culicoides spp. includes small insects 0.5-2 mm in length, usually grey or black and at first glance very similar to mosquitoes. They are strictly hematophagous, feeding by attacking hosts outdoors and indoors (although they are less susceptible to the stationing like Aedes mosquitoes do). The sting is very painful at the injection site and often followed by hypersensibilisation with the consequent formation of allergic dermatitis. In addition, the insects carry and transmit a multitude of diseases, often of a zoonotic character, and therefore are of great epidemiological importance. In our country, continuous monitoring of Culicolides spp. has been carried out and seasonal dynamics of their appearance in the period 2015-2016 is presented in this article. During October 2015, the presence of Culicolides spp. was confirmed in 10.00% of samples; in November, their presence was not established, whereas in December, 2.35% of samples proved positive for the presence of Culicolides spp. During 2016, from January to March, no Culicolides spp. were found in any of the examined samples. During April, their prevalence was 9.63%, in May - 6.74%, in June - 3.70%, in July - 15.78%, in August - 18.07%, in September - 27.27%, and in October - 45.65%. In Serbia, the dominant Culicoides spp. species are Obsoletus complex and Pulicaris complex established in 57.21% and 33.37% of samples, respectively. Other species are present in lesser extent. In Obsoletus complex, the dominant species was Culicoides obsoletus/scoticus. The percentage of Culicoides obsoletus /scoticus males in samples was 25.52%. Non-pigmented (young) females were present in 66.06% of samples; females who took blood in 7.55% and 0.87% were gravid females. In Pulicaris complex, the dominant species was Culicoides pulicaris. Males of Culicoides pulicaris were found in 19.23% of samples, non-pigmented (young) females in 70.96%, females who took blood in 9.08% while 0.73% were gravid females.Rod Culicoides spp. su mali insekti veličine 0,5-2 mm, većinom sivkaste ili crne boje i na prvi pogled veoma slični komarcima. Oni su striktne hematofage. Ubod je veoma bolan, a mesta uboda su često hipersenzibilisana uz posledično nastajanje alergijskih dermatitisa. Uz to oni prenose mnoštvo oboljenja često zoonotskog karaktera tako da su od izuzetnog epidemiološkog značaja. U našoj zemlji se vrši stalni monitoring Culicolides-a i ovde dajemo prikaz sezonske dinamike njihovog pojavljivanja u periodu 2015-2016.godine. Tokom 2015. godine u oktobru je njihovo prisustvo ustanovljeno u 10.,00% uzoraka, u novembru ih nije bilo, a tokom decembra su bili prisutni u 2,35%. Tokom 2016. godine tokom januara, februara i marta nisu nađeni ni u jednom prispelom uzorku. Tokom aprila meseca njihova prevalencija je iznosila 9,63%, u maju je bila 6,74%, u junu 3,70%, u julu 15,78%, u avgustu 18,07%, u septembru 27,27% i u oktobru 45,65%. U Srbiji su dominantne vrste Culicoides spp. iz Obsoletus kompleksa i ustanovljeni su u 57,21%. Culicoides spp. iz Pulicaris kompleksa ustanovljeni su 33,37% dok su ostale vrste manje zastupljene. Kod Culicoides spp. iz Obsoletus kompleksa dominantna vrsta je bila Culicoides obsoletus/scoticus. Mužjaci su nađeni u 25,52%, nepigmentisane (mlade) ženke u 66,06%, ženke koje su uzele krv u 7,55%, a 0,87% su bile gravidne ženke. Kod Culicoides spp. iz Pulicaris kompleksa dominantna vrsta je bila Culicoides pulicaris. Mužjaci su nađeni u 19,23%, nepigmentisane (mlade) ženke u 70,96%, ženke koje su uzele krv u 9,08%, a 0,73% su bile gravidne ženke

Phytochemistry, Toxicology and Therapeutic Value of Petasites hybridus Subsp. Ochroleucus (Common Butterbur) from the Balkans

Petasites hybridus (Common butterbur) is extensively used in traditional medicine, and is currently gaining interest and popularity as a food supplement and for its medicinal properties. It contains a large number of active compounds of potential therapeutic activity, but also toxic pyrrolizidine alkaloids. Science-based information is needed to support the developing modern use of this plant, and to direct continued safe practice in traditional medicine. The present study focused on the essential oils from leaves and rhizomes of the understudied P. hybridus ssp. ochroleucus from the Balkans, and evaluated its phytochemistry and potential therapeutic activities (antimicrobial, antioxidant, anti-cholinesterase and anti-inflammatory), as well its toxicology potential (acute toxicity in insects and mice). We studied the essential oils, which are not commonly used in traditional practices, but have a potential for safe use since the toxic pyrrolizidine alkaloids, which are non-volatiles, are usually not present in the distilled essential oils. Pyrrolizidine alkaloids were indeed not detected in the essential oils; ingestion of the essential oils did not induce toxicity signs in mice, and topical application did not elicit skin irritation in humans. The essential oils had no antimicrobial properties against 20 pathogenic bacterial strains, but demonstrated good local anti-inflammatory activity in a Carrageenan-induced paw edema test. An insect toxicity test demonstrated that the leaf essential oil is an efficient insect repellent, and the demonstrated anti-cholinesterase activity suggests a potential for the treatment of neurological conditions. Isopetasin, a sesquiterpene found in plants of the genus Petasites, known to have anti-inflammatory effects, was present only in the rhizomes essential oil (3.9%), and sesquiterpene lactones concentrations were high, likely contributing to the antioxidant activity

Enhancing Socially Responsible action and Approach to Nature-Based Solutions in Architectural Education within the framework of the Project: Eco-friendly Built Environment [poster]

This event was realized as a joint event of the COST Action CA17133 “Circular City: Implementing nature-based solutions for creating a resourceful circular city” and a 6-year project on climate change adaptation “Coast to Coast Climate Challenge”. Posters on significant European and global scientific projects dealing with a broad spectrum of problems related to climate change were exhibited in parallel with the conference “Done. What’s Next?” More than 320 participants – including professionals in climate adaptation, international researchers, students and 21 stand holders from companies and organizations- participated in this event. In accordance with the thematic importance of the event and the presence of key stakeholders in relation to the topic of the ECOBUILT project, it was of great importance to present it to wither international public. The ECOBUILT project was presented as a significant initiative within the field of education, especially from the aspect of enhancing socially responsible action and approach in architectural education. https://ecobuilt-project.org/ecobuilt-project-presented-at-final-event-of-the-cost-action-ca17133-circular-city

Značaj određivanja koštanih biomarkera u proceni rizika od fraktura i praćenju terapije kod pacijenata sa osteoporozom

Osteoporosis is characterised by reduced bone mineral density (BMD), increased bone fragility and risk of fractures. We investigated clinical significance of bone turnover biomarkers determination for the prediction of fracture risk and response to therapy. We performed three months follow-up study in 48 women with osteoporosis. During the follow-up period 32 patients were on antiresorptive and 16 patients received anabolic therapy. Serum alkaline phosphatase (ALP), osteocalcin, C-terminal telopeptide of collagen type I and vitamin D were measured before and 3 months after initiation of the therapy. Higher osteocalcin and ALP levels were significantly associated with increased risk of BMD loss. Following antiresorptive therapy levels of C-terminal telopeptide (P lt 0.001) and osteocalcin (P lt 0.001) significantly decreased. Anabolic therapy significantly increased osteocalcin (P lt 0.01). At the study entry 25.5% of the patients had optimal levels of 25(OH)D and 17% had severe vitamin D deficiency. Vitamin D supplementation during three months improved vitamin D status of the patients (P lt 0.001). In conclusion, our data indicate that osteocalcin and C-terminal telopeptide determination may be useful for BMD loss prediction and monitoring of osteoporosis therapy. This study confirmed importance of vitamin D testing in osteoporosis in order to detect the patients with deficiency and determine the need for supplementation.Osteoporoza se karakteriše smanjenom mineralnom gustinom kostiju, što povećava fragilnost kostiju i rizik za nastanak fraktura. U ovoj studiji pratili smo 48 pacijentkinja sa osteoporozom tokom tri meseca sa ciljem da utvrdimo značaj određivanja koštanih biomarkera u proceni rizika od fraktura i praćenju terapije osteoporoze. Tokom trajanja studije 32 pacijentkinje su bile na antiresorptivnoj, a 16 na anaboličkoj terapiji. Svim pacijentkinjama odredili smo aktivnost alkalne fosfataze (ALP) i koncentracije osteokalcina, C-terminalnog telopeptida kolagena tip I i vitamina D u serumu pre i tri meseca nakon uvođenja terapije. Sa povećanjem nivoa osteokalcina i ALP rastao je rizik za smanjenje mineralne gustine kostiju (BMD). Antiresorptivna terapija dovela je do značajnog smanjenja koncentracija C-terminalnog telopeptida (P lt 0,001) i osteokalcina (P lt 0,001). Tokom anaboličke terapije nivo osteokalcina je značajno porastao (P lt 0,01). Na početku studije 25,5% pacijentkinja je imalo optimalne koncentracije 25(OH)D u serumu, a tešku deficijenciju 17% žena. Nakon 3 meseca suplementacije većina je poboljšala status vitamina D (P lt 0,001). Naši rezultati ukazuju da određivanje osteokalcina i C-terminalnog telopeptida može biti korisno za predviđanje rizika za smanjenje BMD i praćenje efekata terapije osteoporoze. Takođe, potvrđena je važnost određivanja koncentracije 25(OH)D za procenu statusa vitamina D u osteoporozi, u cilju otkrivanja pacijenata sa insuficijencijom i utvrđivanja potrebe za suplementacijom