3D Cell Technology in Biomedical Research

Traditional two dimensional cell culture has enabled great strides in biomedicine but needs to be improved to be able to keep up with the demands of modern biomedical research. 2D monolayer culture cannot replicate tissue responses and needs to be supplemented with extensive animal research. Growing cells in three dimensional scaffolds provides a more functional model for biomedical research than traditional monolayer culture. Depending on the needs and the complexity of the model there are several ways that 3D models can be initiated. Simple spheroids can be grown in low adherence plates and in hanging drops while larger spheroids and co-cultured ones need to be grown in systems with greater support such as hydro gels. The system that offers the greatest flexibility is the magnetic levitation approach. In the paper we offer a brief resume to various 3D methods and their characteristics to ease the choice of methods for implementing 3D cell culture techniques

Urolithiasis from a molecular perspective

Urolitijaza se javlja u relativno velikom udjelu europskoga stanovništva, ali unatoč povećanom zanimanju za temeljna istraživanja i sve većoj prisutnosti molekularne biologije u medicini, njena molekularna osnova i uzroci nisu otkriveni. U radu se pokušalo dati uopćeni pregled glavnih interesnih područja molekularnih biologa koji se bave istraživanjem urolitijaze, uz napomenu da još podosta toga ostaje istražiti i daje to tek početni korak.Urolithiasis affects relatively large part of the European population, but in spite of the increased interest in basic research and increasing molecular approach in medicine, molecular causes for urolithiasis remain yet to be discovered. This paper attempts to give a general overview of the main areas of interest of molecular biologists, who have been engaged in the research of molecular urolithiasis, while emphasizing that there is still a lot left for research and that this is just the firs

Influence of pH, Temperature and Common Ion on Magnesium Hydrogenurate Octahydrate Solubility

Physico-chemical investigation of urolithiasis today is mostly focused on applying physico-chemical description of precipitation processes to the stone formation with the aim to distinguish between stone formers and nonstone formers. This is done by calculating supersaturation for different solid phases which can be formed in urine using data obtained by urine analysis and existing solubility product, dissociation and complexation constants. In order for this approach to succeed it is of utmost importance that system is described as detailed as possible, i.e. that all species that can be formed are taken into account. Magnesium hydrogenurate octahydrate, Mg(C5H3N4O3)2·8H2O (Mg(HU)2·8H2O), is among species which can precipitate in the urine and for which solubility data doesn’t exist. In order to fill this void crystals of Mg(C5H3N4O3)2·8H2O phase I and phase II have been prepared and characterized. Solubility product constant of Mg(C5H3N4O3)2·8H2O phase I in water at 37 °C and phase II at different temperatures, pH and in different solvents have been determined by measuring total concentration of uric acid and magnesium ions in solutions at different time periods. Results show that in water at 37 °C thermodynamically less stable phase I is more soluble (Ksp=(5.64±0.20).10–9 mol3 dm–9) than phase II (Ksp=(1.66±0.13).10–9 mol3dm–9). Solubility of Mg(HU)2·8H2O phase II increases with temperature. At equilibrium the solubility of phase II is the lowest in the presence of excess of magnesium ions, while solubility in the presence of uric acid is comparable with the one obtained in water

Rizični čimbenici i koštana masa u bolesnika s recidivirajućom urolitijazom: presječno istraživanje na 144 ispitanika

Patients with urolithiasis, particularly hypercalciuria, may have reduced bone mineral density (BMD). There are numerous risk factors contributing to reduction of BMD such as advanced age, sedentary life-style, smoking, low calcium intake, etc. The aim of our study was to investigate the association of lifestyle risk factors and daily intake of milk and dairy products with determinants of BMD in a group of recurrent calcium stone formers (RSF) compared with healthy subjects (HS). The study was carried out at the De-partment of Mineral Research, Faculty of Medicine in Osijek, Croatia. The study included 144 subjects, i.e. 56 RSF and 78 HS. BMD was assessed by dual-energy x-ray absorptiometry. A standard self-reported questionnaire was used to collect data on lifestyle risk factors. Current dietary intake was assessed by personal interview that included questions about milk and dairy product intake. Low BMD was observed in 44.64% of RSF and 35.90% of HS. RSF consumed significantly less milk and dairy products than HS. Calcium restriction in dietary recommendations might be unnecessary due to the impact on bone mineral loss in RSF and dual-energy x-ray absorptiometry should be included in the routine evaluation of RSF.Bolesnici s urolitijazom, osobito oni s hiperkalciurijom, imaju smanjenu koštanu mineralnu gustoću (bone mineral density, BMD). Rizični čimbenici gubitka koštane mase su uznapredovala dob, sjedilački način života, pušenje i smanjen unos kalcija. Cilj našeg istraživanja bio je ustanoviti povezanost rizičnih čimbenika i dnevnog unosa mlije-ka i mliječnih prerađevina s odrednicama BMD u bolesnika s recidivirajućom kalcijskom urolitijazom te ih usporedi-ti sa zdravim ispitanicima. Istraživanje je provedeno na Zavodu za mineralni metabolizam Medicinskog fakulteta u Osijeku. U istraživanju su sudjelovala 144 ispitanika, od čega 56 bolesnika s recidivirajućom kalcijskom urolitija-zom i 78 zdravih ispitanika. BMD je određen metodom dvoenergetske apsorpciometrije X zraka (DXA). Podatci o čimbenicima rizika dobiveni su anketnim upitnikom, a unos hrane je ocijenjen osobnim intervjuom koji je uključi-vao pitanja o unosu mlijeka i mliječnih proizvoda. Snižen BMD zabilježen je u 44,64% bolesnika s recidivirajućom urolitijazom i u 35,90% zdravih ispitanika. Bolesnici s recidivirajućom urolitijazom konzumirali su znatno manje mlijeka i mliječnih proizvoda u odnosu na zdrave ispitanike. Nepotrebna je preporuka smanjenog unosa kalcija bolesnicima s recidivirajućom urolitijazom zbog utjecaja na gubitak koštane mase, a DXA treba biti dio rutinske procjene bolesnika s recidivirajućom urolitijazom

Characterization of the TCRβ repertoire of peripheral MR1-restricted MAIT cells in psoriasis vulgaris patients

Abstract Psoriasis vulgaris (PV) is an inflammatory skin disease largely driven by aberrant αβT cells. Mucosal-associated invariant T (MAIT) cells, which constitute the largest circulating innate-like αβT cell community in human adults, are characterized by a semi-invariant TCRVα7.2 receptor and MR1-restricted affinity toward microbial metabolites. Limited MAIT TCRα diversity is complemented by a more variable TCRβ repertoire, but its footprint in the MAIT repertoire of PV patients has never been tested. Here, we used bulk TCRSeq, MiXCR, VDJTools, and Immunarch pipelines to decipher and compare TCRβ clonotypes from flow-sorted, peripheral TCRVα7.2+MR1-5-OP-RU-tet+MAIT cells from 10 PV patients and 10 healthy, matched controls. The resulting TCRβ collections were highly private and individually unique, with small public clonotype content and high CDR3β amino acid length variability in both groups. The age-related increase in the ‘hyperexpanded’ clonotype compartment was observed in PV, but not in healthy MAIT repertoires. The TCRβ repertoires of PV patients were also marked by skewed TRBV/TRBJ pairing, and the emergence of PV-specific, public CDR3β peptide sequences closely matching the published CDR3β record from psoriatic skin. Overall, our study provides preliminary insight into the peripheral MAIT TCRβ repertoire in psoriasis and warrants further evaluation of its diagnostic and clinical significance