The correlation between N-glycosylation of plasma proteins and age in children with diabetes mellitus type 1

Glikozilacija je jedna od najčešćih posttranslacijskih modifikacija proteina te ima važnu ulogu u mnogim bolestima. Šećerna bolest tipa 1 je među najraširenijim autoimunosnim bolestima današnjice čiji uzrok kod djece nije poznat. S obzirom na razvoj suvremenih analitičkih metoda omogućena je analiza glikozilacije u velikom broju uzoraka te je primijećeno da glikozilacija ovisi o raznim navikama i tjelesnim čimbenicima. Istraživanjima je ustanovljeno da se glikanski profili mijenjaju u raznim bolestima pa je bitno otkriti izvor ovih promjena. Iz navedenih razloga je u ovom radu ispitan utjecaj dobi na glikozilaciju proteina u uzorcima plazme ispitanika dobivenih iz registra uzoraka dječjeg dijabetesa u Danskoj (engl. The Danish Society for Childhood Diabetes Registry – DanDiabKids). Analiza N-glikana u navedenim uzorcima provedena je HILIC-UPLC metodom, a dobiveni podaci su statistički obrađeni u Excel-u i MedCalc-u Spearmanovim testom korelacije. Usporedbom dobi ispitanika s dobivenim rezultatima analize glikana primjećena je korelacija u nekim glikanskim skupinama od kojih je najveći Spearmanov koeficijent korelacije primjećen u populaciji muških ispitanika za glikansku skupinu 6. Također je primjećena i usporedivost dobivenih rezultata bolesne dječje populacije s postojećim podacima o ovisnosti između glikozilacije i dobi zdrave dječje populacije. Glikozilacija kod bolesne dječje populacije se, kao i kod zdrave dječje populacije, značajno razlikuje u usporedbi s glikozilacijom odrasle populacije. Ovim istraživanjem dokazana je povezanost između N-glikozilacije plazmatskih proteina i dobi kod djece sa šećernom bolesti tipa 1.Glycosylation, being one of the most common posttranslational modifications of proteins, plays an important role in many diseases such as diabetes. Diabetes mellitus type 1 is among the most widespread autoimmune diseases of our time, whose cause is unknown. Considering the development of modern analytical methods, glycosylation analysis is enabled in a large number of samples and it is noticed that glycosylation depends on many different habits and physical factors. Studies have shown that glycan profiles are changing in different diseases so it is important to discover the source of these changes. Therefore, this paper tested the impact of age on the glycosylation of proteins in plasma samples of subjects of The Danish Society for Childhood Diabetes Registry – DanDiabKids. N-glycan analysis of the above mentioned samples were done by HILIC-UPLC and the data were statistically processed in Excel and MedCalc using Spearman's correlation test. Comparing age of subjects with glycan analysis results we noticed a correlation in some of the glycan peaks, with the highest Spearman's correlation coefficient being noticed in glycan peak 6 for male subjects. We noticed the comparability between the results of the sick children population and existing data on correlation between glycosylation and the age of healthy children population. Glycosylation in sick children, as in the healthy children population, is significantly different from glycosylation in adults. This research proved the connection between N-glycosylation of plasma proteins and age of children with diabetes mellitus type 1

Costs in the function of business decision making for mushroom production

Gljivarstvo je poljoprivredna grana koja se posljednjih godina sve više razvija u Republici Hrvatskoj. Pri tome se poseban naglasak pridaje razvoju tehnologije koja bi ubrzala proces proizvodnje, povećala prinose i minimalizirala ljudski rad u samoj proizvodnji. Uzgoj šampinjona analiziran je na gospodarstvu s mjesečnom proizvodnjom šampinjona od 15.000 kg, što godišnje čini ukupno 180.000 kg. Izračuni se temelje na razlici troškova ručnog i strojnog rada punjenja uzgojne prostorije. Prag korisnosti ili točka pokrića iznosi 83.414 kg, a prikazuje razinu proizvodnje pri kojoj ručni i strojni način rada imaju jednake troškove. Za proizvodnju iznad navedene količine isplativo je koristiti liniju strojeva za rinfuzno punjenje, dok je za količinu ispod navedenog iznosa ekonomski učinkovitije ručno punjenje uzgojne prostorije. Na analiziranom gospodarstvu smanjenje troškova primjenom strojnog rada iznosi 167.778,00 kn. Ukupna investicija u kupnju stroja iznosi 1.207.500,00 kn, a povrat je 7,2 godine.Mushroom growing, as a branch of agriculture has been rapidly developing in recent years. Emphasis is particularly put on the application of technology that can accelerate the production process to increase yield and to minimize human labor invested in production. Mushroom production was analyzed on a farm with a monthly production of 15,000 kg or an annual production of 180,000 kg. The calculations were based on the defined difference between the costs of manual labor and machine service invested in the mushroom growing room. The break-even point or the coverage point was set at 83,414 kg, referring to the production level with the same manual and machine work costs. Machine refilling for mushroom production above the mentioned amount is more profitable, while manual refilling is economically efficient within the production below the determined amount. On the analyzed farm, applied machine work led to the lowering of costs for 167,778.00 HRK per year. Purchase of machinery requires a total investment of 1,207,500 HRK with the return of 7.2 years

Glycosylation Alterations in Multiple Sclerosis Show Increased Proinflammatory Potential

Multiple sclerosis (MS) is an inflammatory autoimmune disorder affecting the central nervous system (CNS), with unresolved aetiology. Previous studies have implicated N-glycosylation, a highly regulated enzymatic attachment of complex sugars to targeted proteins, in MS pathogenesis. We investigated individual variation in N-glycosylation of the total plasma proteome and of IgG in MS. Both plasma protein and IgG N-glycans were chromatographically profiled and quantified in 83 MS cases and 88 age- and sex-matched controls. Comparing levels of glycosylation features between MS cases and controls revealed that core fucosylation (p = 6.96 × 10−3) and abundance of high-mannose structures (p = 1.48 × 10−2) were the most prominently altered IgG glycosylation traits. Significant changes in plasma protein N-glycome composition were observed for antennary fucosylated, tri- and tetrasialylated, tri- and tetragalactosylated, high-branched N-glycans (p-value range 1.66 × 10−2–4.28 × 10−2). Classification performance of N-glycans was examined by ROC curve analysis, resulting in an AUC of 0.852 for the total plasma N-glycome and 0.798 for IgG N-glycome prediction models. Our results indicate that multiple aspects of protein glycosylation are altered in MS, showing increased proinflammatory potential. N-glycan alterations showed substantial value in classification of the disease status, nonetheless, additional studies are warranted to explore their exact role in MS development and utility as biomarkers

Initial experiences of hemodialysis HeRO graft in University Hospital Dubrava – A report of two cases and review of literature

U Kliničkoj bolnici Dubrava u Zagrebu prvi je put u Hrvatskoj ugrađen novi sustav za krvožilni pristup kod bolesnika koji se liječi kroničnom hemodijalizom. Radi se o kombinaciji centralnoga venskog katetera i grafta engleskog naziva Hemodialysis Reliable Outflow (HeRO) device, skraćenog naziva HeRO-graft. Glavna je indikacija za primjenu ovoga krvožilnog pristupa okluzija ili značajna stenoza centralnih vena. Iz literature se doznaje da ovaj krvožilni pristup ima značajne prednosti u odnosu na tunelirani centralni venski kateter koji bi trebao biti zadnja opcija kod bolesnika koji zahtijevaju trajno nadomještanje izgubljene bubrežne funkcije. Za razliku od tuneliranih katetera ovaj je sustav u potpunosti smješten potkožno, što ga čini otpornijim na infekcije. Do sada su u Kliničkoj bolnici Dubrava ugrađena dva HeRO-grafta za hemodijalizu i prema prvim iskustvima potrebna je detaljna obrada kandidata za ovu proceduru. Potrebno je dobro poznavati indikacije i kontraindikacije te je nužna suradnja više timova: nefrološkog, anesteziološkog, radiološkog i kirurškog. Osobito je potrebno obratiti pažnju na stanje arterijske cirkulacije okrajine koja se koristi za ovaj krvožilni pristup kako bi se izbjegle moguće ishemijske komplikacije. Kod obaju naših bolesnika postupak ugradnje protekao je uredno te smo započeli s korištenjem HeRO-grafta za potrebe hemodijalize.A new vascular access system in patients undergoing chronic hemodialysis was implanted for the first time in Dubrava University Hospital in Zagreb. It is a combination of a central venous catheter and a graft known as Hemodialysis Reliable Outflow device (HeRO graft). The main indication for this vascular access is the occlusion or a significant stenosis of the central veins. Literature supports the significance of this type of vascular access over tunneled central venous catheters which should be the last option for patients requiring permanent renal replacement therapy. Unlike tunneled catheters, this system is completely located subcutaneously which makes it more resistant to infections. So far, two HeRO grafts for hemodialysis have been implanted in Dubrava University Hospital, and according to the first experiences, detailed workup on candidates is required before this procedure. It is necessary to be well aware of the indications and contraindications, and the cooperation of several specialties is required: nephrology, anesthesiology, radiology and surgery. It is especially important to pay attention to the condition of the arterial circulation of the arm used for this vascular access in order to avoid possible ischemic complications. In both of our patients, the implantation procedure had no immediate complications and we started using the HeRO graft

Potencijalni prediktivni biljezi postoperativnih komplikacija kolorektalnog karcinoma temeljeni na omjeru nezrelih granulocita i limfocita, udjelu nezrelih granulocita te IG i IT omjeru

Introduction: The markers of inflammations are a significant predictor of postoperative outcome after colorectal cancer surgery. Along with leukocytes, C-reactive protein (CRP), procalcitonin (PCT) and neutrophil/lymphocyte ratio (NLR) we tried to test the role of immature granulocytes (IG) counts and trends in the postoperative period. Methods: We recorded matched three patient according to their sex, age, and tumor localization who had an uneventful recovery with three patients who had an anastomotic leak required reoperation. We obtained the IG count with every complete blood count and correlated these with CRP, PCT, NLR. For the best prediction we calculated the potential Zagreb score by adding 1 point for elevated NLR ratio for more than 75%, minimally three consecutively elevated % IG, doubling of immature granulocytes relative value to absolute neutrophils count ratio (IG ratio) and immature granulocytes relative value to leucocytes count ratio (IT ratio) at three consecutive time points. Results and Conclusion: Potential Zagreb score seems to predict the need for reoperation in time. Herein we introduce its concept, and we plan the retrospective study to test its feasibility and precision.Uvod: Markeri upale su važni čimbenici u predviđanju poslijeoperacijskog tijeka u kolorektalnoj kirurgiji. Uz leukocite, C reaktivni protein, prokalcitonin i neutrofilno/limfocitni omjer, uvodimo mogućnost korištenja broja i trendova nezrelih granulocita u postoperacijskom periodu. Metode: Zabilježili smo kliničke podatke tri pacijenta koja su imala uredan poslijeopracijski tijek i tri koja su imala dehiscencu anastomoze. Pacijenti su upareni po dobi, spolu, i lokalizaciji karcinoma kolorektuma. Obradili smo podatke o broju nezrelih granulocita koji je integralni dio kompletne krvne slike na analizatoru Sysmex XN 1000, te korelirali podatke sa CRP-om, PCT-om, NLR-om. Najbolje predviđanje smo dobili kada je ustanovljen potencijalni Zagreb score: povećan NLR za više od 75%, tri povećanja % IG zaredom i udvostručenje IG-a and IT-a u tri mjerenja zaredom. Rezultati i zaključak: Potencijalni Zagreb score je predvidio potrebu za reoperacijom na vrijeme. Ova serija slučajeva je uvela koncept i planiramo retrospektivno, na većem uzorku, testirati izvedivost i preciznost potencijalnog Zagreb scorea

Traian and the Danubian Provinces. The political, economic and religious life in the Danubian Provinces: Proceedings of the 4th International Conference on the Roman Danubian Provinces, Zagreb, 15th - 17th November 2017.

The “Department of Archaeology“ of the Faculty of Humanities and Social Sciences of the University of Zagreb gladly hosted the 4th International Conference on the Roman Danubian Provinces in 2017. The Conference was organized with the support of the Faculty of Humanities and Social Sciences of the University of Zagreb, the Croatian Archaeological Society, and the Croatian Science Foundation through the project 6505 Between the Danube and the Mediterranean. Exploring the role of Roman military in the mobility of people and goods in Croatia during the Roman Era. Since it was organized in Zagreb, the Conference remained in Pannonia, not far from Siscia, the capital of Pannonia Savia, and in the territory of the Res publica Andautoniensium. The main topic of the Conference was Traian and the Roman Danubian Provinces or, more specifically, the political, economic and religious life in the Danubian Provinces.(from the Foreword)The “Department of Archaeology“ of the Faculty of Humanities and Social Sciences of the University of Zagreb gladly hosted the 4th International Conference on the Roman Danubian Provinces in 2017. The Conference was organized with the support of the Faculty of Humanities and Social Sciences of the University of Zagreb, the Croatian Archaeological Society, and the Croatian Science Foundation through the project 6505 Between the Danube and the Mediterranean. Exploring the role of Roman military in the mobility of people and goods in Croatia during the Roman Era. Since it was organized in Zagreb, the Conference remained in Pannonia, not far from Siscia, the capital of Pannonia Savia, and in the territory of the Res publica Andautoniensium. The main topic of the Conference was Traian and the Roman Danubian Provinces or, more specifically, the political, economic and religious life in the Danubian Provinces.(from the Foreword

Traian and the Danubian Provinces. The political, economic and religious life in the Danubian Provinces: Proceedings of the 4th International Conference on the Roman Danubian Provinces, Zagreb, 15th - 17th November 2017.

The “Department of Archaeology“ of the Faculty of Humanities and Social Sciences of the University of Zagreb gladly hosted the 4th International Conference on the Roman Danubian Provinces in 2017. The Conference was organized with the support of the Faculty of Humanities and Social Sciences of the University of Zagreb, the Croatian Archaeological Society, and the Croatian Science Foundation through the project 6505 Between the Danube and the Mediterranean. Exploring the role of Roman military in the mobility of people and goods in Croatia during the Roman Era. Since it was organized in Zagreb, the Conference remained in Pannonia, not far from Siscia, the capital of Pannonia Savia, and in the territory of the Res publica Andautoniensium. The main topic of the Conference was Traian and the Roman Danubian Provinces or, more specifically, the political, economic and religious life in the Danubian Provinces.(from the Foreword)The “Department of Archaeology“ of the Faculty of Humanities and Social Sciences of the University of Zagreb gladly hosted the 4th International Conference on the Roman Danubian Provinces in 2017. The Conference was organized with the support of the Faculty of Humanities and Social Sciences of the University of Zagreb, the Croatian Archaeological Society, and the Croatian Science Foundation through the project 6505 Between the Danube and the Mediterranean. Exploring the role of Roman military in the mobility of people and goods in Croatia during the Roman Era. Since it was organized in Zagreb, the Conference remained in Pannonia, not far from Siscia, the capital of Pannonia Savia, and in the territory of the Res publica Andautoniensium. The main topic of the Conference was Traian and the Roman Danubian Provinces or, more specifically, the political, economic and religious life in the Danubian Provinces.(from the Foreword

Stable population structure in Europe since the Iron Age, despite high mobility

Ancient DNA research in the past decade has revealed that European population structure changed dramatically in the prehistoric period (14,000–3000 years before present, YBP), reflecting the widespread introduction of Neolithic farmer and Bronze Age Steppe ancestries. However, little is known about how population structure changed from the historical period onward (3000 YBP - present). To address this, we collected whole genomes from 204 individuals from Europe and the Mediterranean, many of which are the first historical period genomes from their region (e.g. Armenia and France). We found that most regions show remarkable inter-individual heterogeneity. At least 7% of historical individuals carry ancestry uncommon in the region where they were sampled, some indicating cross-Mediterranean contacts. Despite this high level of mobility, overall population structure across western Eurasia is relatively stable through the historical period up to the present, mirroring geography. We show that, under standard population genetics models with local panmixia, the observed level of dispersal would lead to a collapse of population structure. Persistent population structure thus suggests a lower effective migration rate than indicated by the observed dispersal. We hypothesize that this phenomenon can be explained by extensive transient dispersal arising from drastically improved transportation networks and the Roman Empire’s mobilization of people for trade, labor, and military. This work highlights the utility of ancient DNA in elucidating finer scale human population dynamics in recent history

N-glycosylation of plasma proteins as monitoring marker of metabolic syndrome

Metabolički sindrom obuhvaća poremećaje poput visceralne pretilosti te inzulinske rezistencije koja često prethodi šećernoj bolesti tipa 2. Istraživanja su spomenute poremećaje povezala s promjenama N-glikozilacije proteina plazme, jedne od najčešćih i najkompleksnijih post-translacijskih modifikacija. Glavni ciljevi ovog doktorskog rada jesu identificirati N-glikanske strukture prisutne na proteinima plazme koje: a) omogućavaju predikciju i rano prepoznavanje pojedinaca s povećanim rizikom razvoja inzulinske rezistencije i šećerne bolesti tipa 2 i b) su povezane s indeksom tjelesne mase te pretilošću i promjenama indeksa tjelesne mase kod pojedinaca podvrgnutih operaciji želuca. Analizirani su uzorci krvne plazme oko 2 000 ispitanika iz TwinsUK registra sakupljani u više vremenskih točaka tijekom 20 godina pri čemu su bilježeni detaljni podaci o zdravstvenom statusu ispitanika poput dijagnoze inzulinske rezistencije i šećerne bolesti tipa 2, te uzorci krvne plazme 37 pretilih ispitanika okupljenih u Sveučilišnoj bolnici Oxford gdje su bili podvrgnuti restriktivnoj dijeti i operaciji želuca, a krv im je uzrokovana kontinuirano tijekom studije. Izolirani, pročišćeni te fluorescentno obilježeni N-glikani proteina plazme i imunoglobulina G analizirani su HILIC-UPLC metodom. Statistička obrada dobivenih rezultata pokazala je da se N-glikani proteina plazme u pojedinaca koji će razviti šećernu bolest tipa 2 i inzulinsku rezistenciju značajno razlikuju od ispitanika koji će ostati zdravi te se primijećene promjene počinju pojavljivati godinama prije službene dijagnoze što omogućuje uspješno pravovremeno identificiranje pojedinaca visokorizičnih za poremećaj u metabolizmu glukoze. Nadalje, opažene promjene N-glikana proteina plazme i imunoglobulina G u pretilih pojedinaca uslijed mršavljenja rezultat su podvrgavanja restriktivnoj dijeti i operaciji želuca. Iste promjene su primijećene i uslijed longitudinalnog praćenja pojedinaca kojima se mijenjao indeks tjelesne mase. Zaključno, primjenom N-glikana kao biljega ranog izdvajanja visokorizičnih pojedinaca te kao biljega praćenja metaboličkoga i upalnog statusa organizma, može se na vrijeme prevenirati i odgoditi razvoj ozbiljnih komorbiditeta koji se povezuju s metaboličkim sindromom, te čak i spriječiti pojavu samog sindroma, ali i pratiti učinkovitost terapijskog pristupa.Metabolic syndrome includes disorders such as visceral obesity and insulin resistance, which often precedes type 2 diabetes. Studies have linked these disorders to changes in plasma N-glycosylation, one of the most common and complex post-translational modifications. The main objectives of this study are to identify N-glycan structures present on plasma proteins that: a) allow prediction and early identification of individuals at increased risk of developing insulin resistance and type 2 diabetes and b) are associated with body mass index and obesity and body mass index changes in individuals undergoing bariatric surgery (restrictive gastric surgery). Blood plasma samples of about 2000 subjects from the TwinsUK registry collected at multiple time points over 20 years were analysed, recording detailed data on the health status of subjects, as well as blood plasma samples of 37 obese subjects gathered at the Oxford University Hospital where they were subjected to a restrictive diet and bariatric surgery, and their blood was sampled continuously throughout the study. Isolated, purified and fluorescently labelled plasma protein and immunoglobulin G N-glycans were analysed by HILIC-UPLC method. Statistical processing of the obtained results showed that plasma protein N-glycans in individuals to be diagnosed with type 2 diabetes and insulin resistance differ significantly from healthy controls, and that the observed changes begin to appear years before official diagnosis, which allows successful timely identification of individuals at high risk of impaired glucose metabolism. Furthermore, observed significant changes in plasma and immunoglobulin G N-glycans in obese individuals due to weight loss are the result of being subjected to a low-calorie, restrictive diet and bariatric surgery. The same changes were observed due to longitudinal follow-up of individuals with changes in their body mass index. In conclusion, the use of N-glycans as markers of timely identification of high-risk individuals and as monitoring markers of metabolic and inflammatory status of the organism, can prevent and delay the development of serious comorbidities associated with metabolic syndrome, and even prevent the occurrence of the syndrome itself, but also can monitor the efficacy of used therapeutic approach