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Recreational SCUBA (rSCUBA) diving is a specific type of underwater activity preformed in extreme environmental conditions. However, information on molecular events underlying (patho)physiological events that follow exposure to such conditions is still largely unexplored. To contribute to better understanding cardiac and immune system adaptation mechanisms triggered by repeated rSCUBA diving we measured copeptin, IgA, IgG, and IgM, complement components C3 and C4, and differential blood count parameters, including neutrophil­to­lymphocyte ratio (NLR), N­glycosylation of IgG and total plasma proteins (TPP), as well as cell­free DNA in blood samples of fourteen recreational male divers who performed five dives, one per week, on the depth 20­30 m for 30 min, after non­dive period of 5 months. Changes in measured parameters suggested that first dive triggered stress response that was abolished with repetition of diving since its beneficial effects on cardiac system (copeptin level) and immune status (NLR, immunoglobulins level, as well as IgG and TPP N­glycosylation pattern towards anti­inflammatory status) were observed after the following dives. In conclusion, rSCUBA diving practiced on a regular basis induce cardiac adaptive response and promotes anti­ inflammatory status of the organism thus conferring cardioprotection as well as multiple health benefits.

Additional file 1: Table S1. Abbreviations, major structures and descriptions of all glycans complementary to every plasma glycan peak. Table S2. Comparability of self-sampling methods with standard plasma method. Figure S1. Representative chromatogram with 39 separated glycan peaks acquired from plasma sample. Figure S2. Representative chromatogram with 39 separated glycan peaks acquired from Noviplex card (plasma separation disk). Figure S3. Representative chromatogram with 39 separated glycan peaks acquired from Mitra sticks (Neoterxy). Figure S4. Representative chromatogram with 39 separated glycan peaks acquired from DBS.

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.

Experimental protocol for for ultra performance liquid chromatography-mass spectrometry N-glycan analysis of the native and desialylated human apo-transferrin. Supplemental information for the published paper: Friganovi��, Tomislav; Toma��i��, Antonela; ��eba, Tino; Biru��, Ivan; Kerep, Robert; Borko, Valentina; ��aki��, Davor; Gabri��evi��, Mario; Weitner, Tin: Low-pressure chromatographic separation and UV/Vis spectrophotometric characterization of the native and desialylated human apo-transferrin // Heliyon, 7 (2021), 9; e08030, 8 doi:10.1016/j.heliyon.2021.e08030, This work was supported by funding from the Croatian Science Foundation grant UIP-2017- 05-9537 - Glycosylation as a factor in the iron transport mechanism of human serum transferrin (GlyMech), as well as the European Structural and Investments funding for the "Croatian National Centre of Research Excellence in Personalized Healthcare" (contract #KK.01.1.1.01.0010) and "Centre of Competences in Molecular Diagnostics" (contract #KK.01.2.2.03.0006).

Multiple sclerosis (MS), a disease affecting the central nervous system, is characterized by demyelination and inflammation. Mechanism underlying the development of MS is not yet fully understood, but it is considered to be influenced by both genetic and environmental factors. N- glycosylation is one of the most complex co- and post-translational modifications and a highly regulated process. N-glycans do not only increase protein structural diversity, but can modify their function as well. Multiple studies have shown the importance of N-glycan changes in different autoimmune and inflammatory diseases, which highlights N-glycans as potential diagnostic and/or prognostic biomarkers. Even though plasma protein and immunoglobulin G (IgG) N-glycosylation changes are well investigated in many diseases, the studies on N-glycan changes in multiple sclerosis are scarce. Hence, we aimed to investigate N-glycosylation patterns of plasma proteins and IgG in multiple sclerosis. We have analysed plasma and IgG N-glycosylation profiles in 83 multiple sclerosis cases, 88 age- and sex- matched controls and 85 sex-matched controls, but who passed the majority of lifetime risk for MS development (>70 years of age). Core fucosylation showed to be the most prominently altered IgG glycosylation trait, as it was significantly decreased in MS subjects compared to the healthy controls (adjusted p=0.0069). Among all generated plasma glycosylation traits, the most significant changes were observed in antennary fucosylated and high branched N-glycans, which were both increased in the MS group (adjusted p=0.0067 and 0.0219, respectively). On the other hand, low branched N-glycans were significantly decreased in MS subjects (adjusted p=0.0167). Our results demonstrated that plasma protein and IgG N- glycosylation markedly changes in multiple sclerosis. However, additional studies are needed to determine the background of these changes, their role in multiple sclerosis development and their potential use as diagnostic biomarkers.

Genetic studies associated certain N- glycosyltransferase loci with type 1 diabetes development. However, individual variation in plasma N-glycosylation has mostly been studied in the context of diabetes complications, and its role in type 1 diabetes onset is mainly unknown. In this study the aim was to explore N-glycome and genome interplay at the onset of type 1 diabetes. Plasma protein and IgG N-glycans were chromatographically analysed in a study population comprising 1917 children and adolescents (0.6-19.1 years) with recent-onset type 1 diabetes from the DanDiabKids registry. In the follow-up study results for 188 of these participants were compared with those for their 244 unaffected siblings. A total of 1105 new-onset type 1 diabetes patients were genotyped at 183, 546 genetic markers, testing these for genetic association with 24 IgG and 39 plasma protein N- glycans. Significant genetic associations were validated in 455 samples. Our study showed that onset of type 1 diabetes was characterised by an increase in plasma and IgG high-mannose and bisecting GlcNAc structures, a decrease in monogalactosylation, and an increase in IgG disialylation. Models including age, gender and N- glycans yielded significant discriminative power between children with type 1 diabetes and their unaffected siblings, with AUCs of 0.915 and 0.869 for inclusion of plasma and IgG N-glycans, respectively. This study identified novel associations that were not previously reported for the general European population: 1) novel genetic associations of IgG glycans were identified for SNPs on chromosome 22 close to candidate gene MGAT3 ; these include core fucosylated digalactosylated disialylated IgG N-glycan with bisecting GlcNAc and its asialylated version ; 2) novel genetic locus was associated with plasma protein N-glycosylation, the complement C3 gene (C3). C3 variants reside on chromosome 19 within the protein coding region, and the associated high-mannose Man9 glycan is attached on a domain involved in pathogen binding of the complement component C3, whereas identified C3 variants are often co-inherited with another type 1 diabetes risk-associated variant. This study identified distinct N-glycosylation of plasma proteins and IgG at type 1 diabetes onset, novel genetic variants driving these changes and potential underlying molecular mechanisms, which are still unknown for many identified type 1 diabetes risk- associated variants.

Type 1 diabetes (T1D) is a chronic disease characterized by the autoimmune destruction of pancreatic insulin-producing beta cells with the unknown cause. Alterations in glycosylation have been linked with various diseases and are emerging as potential biomarkers of a disease development or progress. In order to search for glycomic changes in T1D that could be associated with the disease onset, plasma samples of 1105 children and adolescents (0-18 years) diagnosed with T1D were collected within three months of diagnosis through the Danish Registry of Childhood and Adolescent Diabetes. Samples were previously genotyped for 183, 546 single nucleotide polymorphisms (SNPs) on the Immunochip. HILIC-UPLC was applied for quantification of the N-glycans of both total plasma proteins and IgG. Genome-wide association study identified five genome-wide significant loci associated with total plasma proteins and/or IgG N-glycans. All of the identified loci, except for the complement C3 gene (C3) locus, were previously associated with N-glycosylation. SNPs within the exon regions of the C3 gene are associated with the oligomannose N-glycan levels. C3 protein glycosylation changes within the T1D population and their influence on complement activation should be further investigated. In the follow- up experiments, N-glycans from 187 children with T1D and their 244 unaffected siblings were quantified. Our results showed that children with a recent diagnosis of T1D had significantly higher levels of mannose and GlcNAc terminal residues on total plasma proteins and IgG, respectively, when compared to their unaffected siblings. This suggests that the complement system could be activated in T1D through the mannose-binding lectin pathway.

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.

Female cancer patients are almost two times more vulnerable to depression than men. 16% of them report clinical levels of depression, compared to 8% of men. This is consistent with the comparably higher prevalence of depression in female population in general. We adopted this premise of our work from the observational, epidemiological perspective although we do respect the doubts expressed by Parker and Brotchie 2010. Anyhow, younger breast cancer patients are at higher risk for depression, and this observation is partially inconsistent with the state in the general population. Primary objective was to find out whether the risk for depression decreases with patients’ age. If it were so, our second objective was to check if there are any moderating effects of age to the association of biological variables and depression. We performed this cross-sectional study at Department of Medical Oncology, University Hospital for Tumors Zagreb, University Hospital Center "Sestre milosrdnice" Zagreb, Croatia during Spring 2015. Targeted population was the general population of patients diagnosed with breast cancer. We included a consecutive sample of patients. Depression was measured by Beck Depression Inventory II (BDI-II). The analysis was performed by “PROCESS” Release 2.16.2 and executed from within IBM SPSS version 23. We included 153 breast cancer patients. Correlation of age and BDI-II result was indeed a negative one, but it was not significant (Spearman’s ρ=-0.05 ; p=0.572). However, interaction between age and tumor size was significant (p=0.036). In patients younger than 47 years (what was 23% of all patients), association of tumor size and BDI- II was significant and positive: larger the tumor, more severe the depression. In patients older than 61, the association of tumor size and BDI-II become negative, but it was not significant. We have not observed significant interaction of depression, age and any other biological variable. Breast cancer patients’ perception of tumor size and it’s potential as stressor responsible for the increased risk for depression is moderated by patients’ age.

V nalogi uvodoma ugotavljamo, da so idejna razhajanja znotraj slovenskega katoliškega tabora med obema vojnama posegla tudi na področje katoliškega šolstva. Raziskali smo, kakšen vpliv je imel na omenjena razhajanja francoski personalizem. Pokazali smo, da so temu idejnemu toku sledili katoličani, ki so izhajali predvsem iz križarskega gibanja, medtem ko mu tradicionalni katoliški tabor ni bil naklonjen. V nalogi, ki temelji na zgodovinski metodi pedagoškega raziskovanja, smo izpostavili Stanka Cajnkarja, Stanka Gogalo in Jakoba Šolarja (vsi so bili pripadniki križarskega gibanja), ki so ideje francoskega personalizma aplicirali na področje svojega pedagoškega udejstvovanja. V raziskavi smo poskušali potrditi hipotezo, da je bil francoski personalizem v medvojnem obdobju eden izmed idejnih tokov, ki so ga Gogala, Šolar ter Cajnkar vključevali v svoj pedagoški koncept, hkrati pa je bil vzrok za odstopanje od uradne pedagoške doktrine Katoliške cerkve. Pokazali smo, da ideje francoskega personalizma, ki so jih izpostavljeni slovenski avtorji aplicirali na področje pedagogike, sovpadajo z idejami francoskega personalista in pedagoga Jacquesa Maritaina. Čeprav je vpliv Maritaina in ostalih francoskih personalistov na slovenske avtorje slabo raziskan, je analiza pedagoških idej obravnavanih slovenskih avtorjev vendarle pokazala, da so Maritaina poznali in upoštevali. In the first part of the study we have shown, that the ideological differentiation within Slovene Catholic community during the two world wars intervened in the area of Catholic education. We explore, what influence French personalism had on those discrepancies. We have shown, that this idea is the course followed by Catholics, which came mainly from the Cross movement, while within traditional Catholic Church was not favorable. In the study, based on the historical method of pedagogic research, we exposed Stanko Gogala, Jakob Šolar and Stanko Cajnkar (all were members of the Cross movement), who applied the principles of personalism on the pedagogy. In this study we tried to verify the hypothesis, that in the interwar period the French personalism was one of the ideological currents that Šolar, Gogala and Cajnkar had included in their educational concept, but in the same time they distances themselves from the official Catholic Church. We have shown, that the ideas of French personalism, which have exposed Slovene pedagogues applied on the pedagogy, coincide with ideas of Catholic philosopher and a representative of french personalism, Jacques Maritain. Although the influence of Maritain and others French personalists on the Sloven authors is poorly explored, the analysis of educational ideas, discussed by the Sloveneian author, however, showed, that they knew and respected Maritain.