Bone marrow-derived mesenchymal stromal cells from patients with end-stage renal disease are suitable for autologous therapy

Background aimsMesenchymal stromal cells (MSCs) are pluripotent cells that have immunosuppressive and reparative properties in vitro and in vivo. Although autologous bone marrow (BM)-derived MSCs are already clinically tested in transplant recipients, it is unclear whether these BM cells are affected by renal disease. We assessed whether renal failure affected the function and therapeutic potential of BM-MSCs.MethodsMSCs from 10 adults with end-stage renal disease (ESRD) and 10 age-matched healthy controls were expanded from BM aspirates and tested for phenotype and functionality in vitro.ResultsMSCs from ESRD patients were >90% positive for CD73, CD90 and CD105 and negative for CD34 and CD45 and showed a similar morphology and differentiation capacity as MSCs from healthy controls. Of importance for their clinical utility, growth characteristics were similar in both groups, and sufficient numbers of MSCs were obtained within 4 weeks. Messenger RNA expression levels of self-renewal genes and factors involved in repair and inflammation were also comparable between both groups. Likewise, microRNA expression profiling showed a broad overlap between ESRD and healthy donor MSCs. ESRD MSCs displayed the same immunosuppressive capacities as healthy control MSCs, demonstrated by a similar dose-dependent inhibition of peripheral blood mononuclear cell proliferation, similar inhibition of proinflammatory cytokines tumor necrosis factor-α and interferon-γ production and a concomitant increase in the production of interleukin-10.ConclusionsExpanded BM-MSCs procured from ESRD patients and healthy controls are both phenotypically and functionally similar. These findings are important for the potential autologous clinical application of BM-MSCs in transplant recipients.     

A Pilot Clinical Study of Hyperacute Serum Treatment in Osteoarthritic Knee Joint: Cytokine Changes and Clinical Effects

The serum fraction of platelet-rich fibrin (hyperacute serum) has been shown to improve cartilage cell proliferation in in vitro osteoarthritic knee joint models. We hypothesize that hyperacute serum may be a potential regenerative therapeutic for osteoarthritic knees. In this study, the cytokine milieu at the synovial fluid of osteoarthritic knee joints exposed to hyperacute serum intraarticular injections was investigated. Patients with knee osteoarthritis received three injections of autologous hyperacute serum; synovial fluid was harvested before each injection and clinical monitoring was followed-up for 6 months. Forty osteoarthritic-related cytokines, growth factors and structural proteins from synovial fluid were quantified and analysed by Multivariate Factor Analysis. Hyperacute serum provided symptomatic relief regarding pain and joint stability for OA patients. Both patients “with” and “without effusion knees” had improved VAS, KOOS and Lysholm-Tegner scores 6 months after of hyperacute serum treatment. Synovial fluid analysis revealed two main clusters of proteins reacting together as a group, showing strong and significant correlations with their fluctuation patterns after hyperacute serum treatment. In conclusion, hyperacute serum has a positive effect in alleviating symptoms of osteoarthritic knees. Moreover, identified protein clusters may allow the prediction of protein expression, reducing the number of investigated proteins in future studies.     

Variability in the Effect of 5-HTTLPR on Depression in a Large European Population: The Role of Age,Symptom Profile,Type and Intensity of Life Stressors

Background

Although 5-HTTLPR has been shown to influence the risk of life stress-induced depression in the majority of studies, others have produced contradictory results, possibly due to weak effects and/or sample heterogeneity.

Methods

In the present study we investigated how age, type and intensity of life-stressors modulate the effect of 5-HTTLPR on depression and anxiety in a European population cohort of over 2300 subjects. Recent negative life events (RLE), childhood adversity (CHA), lifetime depression, Brief Symptoms Inventory (BSI) depression and anxiety scores were determined in each subject. Besides traditional statistical analysis we calculated Bayesian effect strength and relevance of 5-HTTLPR genotypes in specified models.

Results

The short (s) low expressing allele showed association with increased risk of depression related phenotypes, but all nominally significant effects would turn to non-significant after correction for multiple testing in the traditional analysis. Bayesian effect strength and relevance analysis, however, confirmed the role of 5-HTTLPR. Regarding current (BSI) and lifetime depression 5-HTTLPR-by-RLE interactions were confirmed. Main effect, with other words direct association, was supported with BSI anxiety. With more frequent RLE the prevalence or symptoms of depression increased in ss carriers. Although CHA failed to show an interaction with 5-HTTLPR, in young subjects CHA sensitized towards the depression promoting effect of even mild RLE. Furthermore, the direct association of anxiety with the s allele was driven by young (≤30) individuals.

Limitations

Our study is cross-sectional and applies self-report questionnaires.

Conclusions

Albeit 5-HTTLPR has only weak/moderate effects, the s allele is directly associated with anxiety and modulates development of depression in homogeneous subgroups.     

FcRn overexpression in transgenic mice results in augmented APC activity and robust immune response with increased diversity of induced antibodies

Our previous studies have shown that overexpression of bovine FcRn (bFcRn) in transgenic (Tg) mice leads to an increase in the humoral immune response, characterized by larger numbers of Ag-specific B cells and other immune cells in secondary lymphoid organs and higher levels of circulating Ag-specific antibodies (Abs). To gain additional insights into the mechanisms underlying this increase in humoral immune response, we further characterized the bFcRn Tg mice. Our Western blot analysis showed strong expression of the bFcRn transgene in peritoneal macrophages and bone marrow derived dendritic cells; and a quantitative PCR analysis demonstrated that the expression ratios of the bFcRn to mFcRn were 2.6- and 10-fold in these cells, respectively. We also found that overexpression of bFcRn enhances the phagocytosis of Ag-IgG immune complexes (ICs) by both macrophages and dendritic cells and significantly improves Ag presentation by dendritic cells. Finally, we determined that immunized bFcRn mice produce a much greater diversity of Ag-specific IgM, whereas only the levels, but not the diversity, of IgG is increased by overexpression of bFcRn. We suggest that the increase in diversity of IgG in Tg mice is prevented by a selective bias towards immunodominant epitopes of ovalbumin, which was used in this study as a model antigen. These results are also in line with our previous reports describing a substantial increase in the levels of Ag-specific IgG in FcRn Tg mice immunized with Ags that are weakly immunogenic and, therefore, not affected by immunodominance.     

The effect of storage and active ingredient properties on the drug release profile of poly(ethylene oxide) matrix tablets

Different molecular weight forms of poly(ethylene oxide) can be used successfully in controlled release drug delivery due to their excellent matrix forming properties. Drug release of these materials follows nearly zero order kinetics, and is mainly governed by polymer swelling and erosion and diffusion of drug molecules. Because of its partly amorphous structure, poly(ethylene oxide) undergoes structural changes caused by elevated temperature and relative humidity of the storage medium resulting in an increased drug release. This physical process can be highly influenced by the structure of different drug molecules, such as polymer-binding ability and hydration tendency. These properties of two basic drugs embedded into poly(ethylene oxide) matrices were characterized by molecular modelling and an attempt was made to reveal their effect on the change of drug release stability, a prerequisite of the marketing authorization of dosage forms. The findings suggest that both the hydration properties of the active ingredient and the molecular weight of the polymer influence the effect of physical ageing of poly(ethylene oxide) on the drug release properties of the matrix.     

Molecular and morphological divergence in a stygobiont gastropod lineage (Truncatelloidea,Moitessieriidae, Paladilhiopsis) within an isolated karstic area in the Mecsek Mountains (Hungary)

Truncatelloid gastropods are one of the most species‐rich subterranean invertebrate groups. Their current taxonomy is based on morphological characters. However, this is not a comprehensive approach and does not take into account the degree of phylogenetic divergence between stygobiont populations inhabiting hydrologically isolated but geographically close caves. We studied two Paladilhiopsis populations of a small and isolated karstic area (Mecsek Mountains, Hungary) with two hydrologically separated cave systems, investigating morphological (shell morphometrics) and genetic (COI, 16S) divergence together. The populations differed both morphologically and genetically: we found strong divergence in the relative width of the shell (best described by the variable “shell angle”) and a 6.4% divergence in COI. This provides strong support for the presence of two distinct taxa; however, it is still doubtful whether they differ at the species or the subspecies level. In one of the caves, we found representatives of both haplotypes (and phenotypes), which can be explained by secondary contact after an allopatric divergence.     

DNA methylation-associated colonic mucosal immune and defense responses in treatment-na?ve pediatric ulcerative colitis

Inflammatory bowel diseases (IBD) are emerging globally, indicating that environmental factors may be important in their pathogenesis. Colonic mucosal epigenetic changes, such as DNA methylation, can occur in response to the environment and have been implicated in IBD pathology. However, mucosal DNA methylation has not been examined in treatment-naïve patients. We studied DNA methylation in untreated, left sided colonic biopsy specimens using the Infinium HumanMethylation450 BeadChip array. We analyzed 22 control (C) patients, 15 untreated Crohn’s disease (CD) patients, and 9 untreated ulcerative colitis (UC) patients from two cohorts. Samples obtained at the time of clinical remission from two of the treatment-naïve UC patients were also included into the analysis. UC-specific gene expression was interrogated in a subset of adjacent samples (5 C and 5 UC) using the Affymetrix GeneChip PrimeView Human Gene Expression Arrays. Only treatment-naïve UC separated from control. One-hundred-and-twenty genes with significant expression change in UC (> 2-fold, P < 0.05) were associated with differentially methylated regions (DMRs). Epigenetically associated gene expression changes (including gene expression changes in the IFITM1, ITGB2, S100A9, SLPI, SAA1, and STAT3 genes) were linked to colonic mucosal immune and defense responses. These findings underscore the relationship between epigenetic changes and inflammation in pediatric treatment-naïve UC and may have potential etiologic, diagnostic, and therapeutic relevance for IBD.     

Tracking of the kinetic stability of 2 types of total nutrient admixtures containing different lipid emulsions

The physical stability of 2 types of total nutrient admixtures was studied as a function of storage time and temperature. One of them contained only structured triglycerides and the other exclusively long-chain triglycerides as lipid components. To evaluate the possible changes in the kinetic stability of the emulsions and in the surface characteristics of the droplets during storage, particle size analysis, zeta potential, and dynamic surface tension measurements were performed. To follow any chemical decomposition processes that occurred during storage, the pH of the emulsions was also monitored. The mean droplet size of emulsions prepared with lipids containing exclusively long-chain triglycerides showed a remarkable increase after 4 days of storage, in contrast with that of the mixtures containing structured lipids. A combination of size distribution, zeta potential, and dynamic surface tension measurements proved to be useful for an adequate tracking of the kinetic stability of total nutrient admixtures. Structured triglycerides not only provide advantageous metabolic effects but improve the physical stability of total parenteral nutrition admixtures.     

The Production of Nominal and Verbal Inflection in an Agglutinative Language: Evidence from Hungarian

The contrast between regular and irregular inflectional morphology has been useful in investigating the functional and neural architecture of language. However, most studies have examined the regular/irregular distinction in non-agglutinative Indo-European languages (primarily English) with relatively simple morphology. Additionally, the majority of research has focused on verbal rather than nominal inflectional morphology. The present study attempts to address these gaps by introducing both plural and past tense production tasks in Hungarian, an agglutinative non-Indo-European language with complex morphology. Here we report results on these tasks from healthy Hungarian native-speaking adults, in whom we examine regular and irregular nominal and verbal inflection in a within-subjects design. Regular and irregular nouns and verbs were stem on frequency, word length, and phonological structure, and both accuracy and response times were acquired. The results revealed that the regular/irregular contrast yields similar patterns in Hungarian, for both nominal and verbal inflection, as in previous studies of non-agglutinative Indo-European languages: the production of irregular inflected forms was both less accurate and slower than of regular forms, both for plural and past-tense inflection. The results replicate and extend previous findings to an agglutinative language with complex morphology. Together with previous studies, the evidence suggests that the regular/irregular distinction yields a basic behavioral pattern that holds across language families and linguistic typologies. Finally, the study sets the stage for further research examining the neurocognitive substrates of regular and irregular morphology in an agglutinative non-Indo-European language.