Identifikacija bioaktivnih proteina gljive Ophiocordyceps sinensis i određivanje njihovog antioksidacijskog i citotoksičnog učinka pomoću shotgun analize proteoma

Research background. Ophiocordyceps sinensis, a highly valued medicinal fungus, is close to extinction due to overexploitation. Successful cultivation of O. sinensis fruiting body (OCS02®) shows that the cultivar has a promising nutritional value and numerous bioactive compounds. Antioxidant and antiproliferative properties and biologically active proteins of the OCS02® are investigated for possible development into nutraceuticals. Experimental approach. The chemical composition of the OCS02® cold water extract was determined, and the antioxidant activities were examined using ferric reducing, DPPH• and O2•- scavenging assays. Tetrazolium dye (MTT) cytotoxic assay was performed to assess the antiproliferative activity of the extract. Bioactive proteins in the active fraction of the extract were identified using liquid chromatography (LC) and tandem-mass spectrometry (MS/MS). Results and conclusions. The OCS02® extract exhibited strong O2•- scavenging (expressed as Trolox equivalents (18.4±1.1) mol/g) and potent cytotoxic activities against adenocarcinomic human alveolar basal epithelial (A549) cells (IC50=(58.2±6.8) µg/mL). High molecular mass polysaccharides, proteins and protein-polysaccharide complexes could have contributed to the antioxidant and cytotoxic selectivity of the OCS02®. LC-MS/MS analysis identified several potential cytotoxic proteases and an oxalate decarboxylase protein which may exhibit protection effects on kidneys. Novelty and scientific contribution. The findings demonstrate the potential of OCS02® to be developed into functional food due to its promising superoxide anion radical scavenging capacity, cytotoxic effect and presence of biopharmaceutically active proteins.Pozadina istraživanja. Vrlo cijenjena medicinska gljiva Ophiocordyceps sinensis je na rubu izumiranja zbog njezine prekomjerne eksploatacije. Uspješnim uzgojem plodišta gljive O. sinensis (OCS02®) potvrđeno je da taj kultivar ima obećavajuća hranjiva svojstva te sadržava brojne bioaktivne spojeve. Ispitana su njegova antioksidacijska i antiproliferacijska svojstva te sastav biološki aktivnih proteina, s ciljem mogućeg razvoja nutraceutika. Eksperimentalni pristup. Utvrđen je kemijski sastav ekstrakta gljive u hladnoj vodi, a antioksidacijska je aktivnost ispitana pomoću FRAP metode te metodama uklanjanja DPPH˙ i O2 radikala. Citotoksičnost odnosno antiproliferacijska aktivnost ekstrakta ispitana je testom na osnovi tetrazolija (MTT test). Bioaktivni proteini su identificirani u aktivnoj frakciji ekstrakta pomoću tekućinske kromatografije i tandemske spektrometrije masa. Rezultati i zaključci. Ekstrakt gljive OCS02® imao je izrazito jako svojstvo uklanjanja superoksid radikala (izraženo u ekvivalentima Troloxa (18,4±1,1) mol/g) i snažan citotoksični učinak (IC50=(58,2±6,8) µg/mL) na humane epitelne stanice adenokarcinoma pluća (A549). Moguće je da polisaharidi, proteini i kompleksi proteina s polisaharidima velike molekularne mase pridonose antioksidacijskoj i citotoksičnoj selektivnosti gljive OCS02®. Tekućinskom kromatografijom i tandemskom spektrometrijom masa identificirano je nekoliko potencijalno citotoksičnih proteaza te protein oksalat dekarboksilaza koji bi mogli imati zaštitni učinak na bubrege. Novina i znanstveni doprinos. Dobiveni rezultati pokazuju da se gljiva OCS02® može upotrijebiti u proizvodnji funkcionalne hrane zbog njezine obećavajuće sposobnosti uklanjanja superoksidnih aniona, citotoksičnog učinka te prisutnosti biofarmaceutski aktivnih proteina

Association between polymorphisms 3' of ApoE and susceptibility to schizophrenia and its subtypes

This study investigated the effects of rs439401 and three other polymorphisms (rs5112, +7203 C/T, rs7247227) located within or close to regulatory elements in the 3' region downstream of the ApoE gene on susceptibility to Schizophrenia and on cognitive performance in affected and normal individuals. Association analysis was conducted in a case-control sample of 427 schizophrenia cases and 208 controls derived from the Western Australian Family Study of Schizophrenia (WAFSS). The clinically affected individuals were divided into two distinct subtypes: cognitive deficit (CD; n=185)/ cognitive spared (CS; n=154). Additional analysis was also performed in a sample of 563 men (aged≥65; representative of the general elderly population) obtained from the Health in Men Study (HIMS). rs439401 was the only SNR found to be associated with increased risk of schizophrenia (P=0.0176) and cognitive deficit (P=0.0049). Within Schizophrenia subjects, significant association was found between the minor allele of rs439401 and decline in general cognitive ability (P=0.0131) and verbal memory (P=0.0489). In the HIMS cohort, the minor allele of rs439401 was associated with verbal memory decline (P=0.0202), with its effects independent of the ApoE ε alleles. rs439401 is located close to a peroxisome proliferator-activated receptor-y (PPAR-y) response element (PPRE) in the ApoE/ApoCI intergenic region. Expression analysis, using lymphoblastoid cell lines (LCLs) as a model, revealed no difference in APOE levels between Schizophrenia and controls, and between rs439401 genotypes. Overall, the results point to the contribution of rs439401 and possibly APOE to increased susceptibility of schizophrenia and decline in cognitive function. rs439401 may affect PPRE function, which consequently alters APOE expression. APOE distribution may influence schizophrenia risk through its effects on neurite outgrowth. neuronal plasticity, CNS glucose and cholesterol homeostasis, or tau phosphorylation

Leptin and soluble leptin receptor in association with gestational diabetes: a prospective case–control study

Purpose: To assess the association of serum leptin and its receptor (SLeptinR) with the risk of gestational diabetes mellitus (GDM) and to evaluate the longitudinal circulation of these peptides in pregnancy. Methods: This study consisted of 53 subjects diagnosed with GDM and 43 normal glucose tolerance (NGT) pregnant women. Serum leptin and SLeptinR were measured at 24–28 weeks, prior and after delivery, and post-puerperium. Results: Lower levels of leptin and SLeptinR were observed in GDM compared to NGT. Leptin [OR 0.97 (95% CI 0.94–1.0)] and SLeptinR [OR 0.86 (95% CI 0.79–0.93]) were inversely associated with GDM. Participants in the lowest tertile for leptin and SLeptinR had a 2.8-fold (95% CI 1.0–7.6) and a 5.7-fold (95% CI 1.9–17.3) higher risk of developing GDM compared with the highest tertile, respectively. These relationships were attenuated after adjustment for covariates. In both the groups, peak leptin was observed at 24–28 weeks, decreasing continuously during pregnancy (p > 0.05) and after delivery (p < 0.017). SLeptinR level increased (p < 0.001) during pregnancy and decreased (p < 0.005) after delivery in GDM, however, levels remained the same in NGT. In GDM, leptin and SLeptinR was positively and inversely correlated with BMI and HOMA-IR at 24–28 weeks and post-puerperium, respectively. The cord levels of both leptin and SLeptinR were lower than maternal levels. There were no significant differences in serum cord leptin and SLeptinR levels between the groups. Conclusion: Leptin and SLeptinR are independently and inversely associated with GDM. Lower levels of these peptides may play an important role in the pathophysiology of GDM and pre-diabetic state in post-puerperium

Subversion of immunoproteasome subunit expression in dengue virus serotype 2-infected HepG2 cells

Abstract: INTRODUCTION: Infection with all serotypes of dengue virus (DV) results in augmented antigen presentation by MHC class I molecules. However, the upregulation of immunoproteasome subunits only results from infection with two serotypes. This study aims to elucidate changes in the expression of immunoproteasome subunits resulting from infection with DV, particularly DV serotype 2 (DV2). METHODS: HepG2 cells were grown in various culture milieu. Total cellular RNA and proteins were extracted and quantified. RESULTS: Results demonstrated sequestration of immunoproteasome subunits LMP2 and LMP7 in DV2-infected cells. CONCLUSIONS: This study provides insights into the mechanisms underlying immune evasion by DV