Serotonergic mechanisms are involved in the hemostatic action of erythropoietin in uremic patients

Summary Recombinant human erythropoietin was administered to 12 patients with end-stage renal failure on long-term hemodialysis. They responded to the therapy with a shortening of the prolonged bleeding time, starting from the 1st week of therapy, before a significant increase in hemoglobin concentration was achieved. We also observed an increase in the activity of tissue plasminogen activator and a decrease in the activity of its inhibitor. There were no changes in platelet count but a significant increase in blood and platelet serotonin concentration was found. The shortening of the prolonged bleeding time before the correction of the anemia correlated with the rise in blood and platelet serotomin concentration during erythropoietin therapy. We suggest the possible involvement of an serotonergic mechanism in the hemostatic action of recombinant human erythropoietin.     

Analgesic, antioedematous and antioxidant activity of γ-butyrolactone derivatives in rodents

In this paper, the analgesic, antioedematous, motor-impairing and antioxidant properties of four γ-butyrolactone derivatives (BM113, BM113A, BM138 and BM138A) are described. Pain was induced by thermal (hot-plate test), chemical (writhing test) or mechanical (Randall-Selitto model) stimulation. All in-vivo assays were carried out in mice pretreated intraperitoneally with the test compounds, except for the evaluation of anti-inflammatory and analgesic activities in the carrageenan-induced paw oedema model, in which rats were pretreated orally with these compounds. In the hot-plate assay, BM113A and BM138A dose dependently prolonged the latency of the nociceptive reaction. Their analgesic activity, measured as a median effective dose (ED(50)=4.7 mg/kg), was similar to that of morphine (2.4 mg/kg). In the writhing test, all four compounds, in particular BM113A and BM138A, showed higher potency than the reference drug acetylsalicylic acid (the ED(50) values were 3.7, 2.3 and 46.1 mg/kg, respectively). BM138 caused a dose-dependent diminution of paw oedema (up to 49%) in the carrageenan model and BM138A at 200 mg/kg reduced mechanical hyperalgesia in the Randall-Selitto test (～30% when compared with the control). None of the γ-butyrolactone derivatives tested at the ED(50) obtained in the hot-plate test influenced the locomotor activity of mice, although in the rotarod test at 24 rpm, BM113A and BM138 at 100 mg/kg showed some motor-impairing properties. In vitro, a concentration-dependent ABTS radical cation-scavenging activity of BM138 and BM138A (up to 80% inhibition of the radical absorbance) was observed. The results of the present study suggest that BM138 and BM138A could be of interest for future investigations as antinociceptive and antioedematous agents with potential free radical-scavenging properties.     

New conjugates of muramyl dipeptide and nor-muramyl dipeptide linked to tuftsin and retro-tuftsin derivatives significantly influence their biological activity

The synthesis and biological activity of new conjugates of muramyl dipeptide (MDP) and nor-muramyl dipeptide (nor-MDP) with tuftsin and retro-tuftsin derivatives containing isopeptide bond between ε-amino group of lysine and carboxyl group of simple amino acids such as Ala, Gly and Val are presented. We presumed, based on the cytokine profile, that the examined conjugates of tuftsin and MDP were capable of activating antibacterial mechanisms by switching on Th1 immune response. The most active were compounds 11, 14 and 19-23.     

Characterization of genes associated with internalization of enteroaggregative Escherichia coli

On animal models enteroaggregative Escherichia coli (EAEC) can cause mild, but significant mucosal damage, suggesting the invasive capability of these strains. In the study we investigated the ability of typical, aggR-positive and atypical, aggR-negative EAEC isolates to enter intestinal epithelial Int407 cells in relation to the distribution of genes encoding the putative invasins described among pathogenic E. coli categories. The results demonstrated that regardless of origin and affiliation to typical and atypical EAEC, most isolates examined were internalized by the epithelial cells to different extent. Although as many as 50 (84.3%) EAEC demonstrated a variety of combinations of the aggB, afaD, ipaH and tia genes determined, there was no correlation between the invasion efficiency of these strains and the presence of any particular gene involved in invasion. Most of EAEC examined belonged to phylogenetic group B2 and D.     

Aggregation-promoted expansion of neuraly committed human umbilical cord blood progenitors in vitro

Human umbilical cord blood (HUCB) has been established as a promising source of hematopoietic as well as various non- hematopoietic stem cell populations. This offers numerous advantages of HUCB stem/progenitor cells for therapies, however in vitro conditions that contribute to long term propagation of proliferating undifferentiated cells have not yet been established. Here we evaluate culture conditions promoting spheroid aggregates/neurospheres formation which, together with serum withdrawal and mitogenes treatments in strictly defined media, maintain population of HUCB progenitor cells in undifferentiated and dividing state exhibiting neurogenic potential in vitro. Our results indicate that formation and maintenance of three-dimensional aggregates enhanced by cell culture rotating motion, is crucial for high and prolonged expression of genes and proteins characteristic for cord blood stem cells and their further neural commitment.     

Migratory capabilities of human umbilical cord blood-derived neural stem cells (HUCB-NSC) in vitro

Many types of neural progenitors from various sources have been evaluated for therapy of CNS disorders. Prerequisite for success in cell therapy is the ability for transplanted cells to reach appropriate target such as stroke lesion. We have established neural stem cell line from human umbilical cord blood neural stem (HUCB-NSC). In the present study we evaluated migratory capabilities of cells (HUCB-NSC) and the presence of various migration-related receptors. Immunocytochemical analysis revealed abundant expression of CXCR4, PDGFR-alpha, PDGFR-beta, c-Met, VEGFR, IGF-1R and PSA-NCAM receptors in non-adherent population of HUCB-NSC cultured in serum free (SF) conditions (SF cells). Biological activity of selected receptors was confirmed by HUCB-NSC in vitro migration towards SDF-1 and IGF-1 ligands. Additionally, rat brain-derived homogenates have been assessed for their chemoattractive activity of HUCB-NSC. Our experiments unveiled that brain tissue was more attracted for HUCB-NSC than single ligands with higher potency of injured than intact brain. Moreover, adherent HUCB-NSC cultured in low serum (LS) conditions (LS cells) were employed to investigate an impact of different extracellular matrix (ECM) proteins on cell motility. It turned out that laminin provided most permissive microenvironment for cell migration, followed by fibronectin and gelatin. Unexpected nuclear localization of CXCR4 in SF cells prompted us to characterize intracellular pattern of this expression in relation to developmental stage of cells cultured in different conditions. Continuous culture of LS cells revealed cytoplasmatic pattern of CXCR4 expression while HUCB-NSC cultured in high serum conditions (HS cells) resulted in gradual translocation of CXCR4 from nucleus to cytoplasm and then to arising processes. Terminal differentiation of HUCB-NSC was followed by CXCR4 expression decline.     

Correlation between fat mass and blood pressure in healthy children

Obesity is a well-known risk factor for arterial hypertension. The aim of this study was to analyze which surrogate marker of adiposity, i.e., body mass index (BMI) or fat mass (FM), as measured by bioimpedance analysis (BIA), best correlated with blood pressure in healthy children. Body weight, height, and casual blood pressure (BP) were measured in 193 healthy children (103 boys), aged 8–16 years. Body composition was determined by BIA. The correlation between BMI and age was linear, whereas the correlation between percentage of FM and age was nonlinear and it was different in boys and girls. Blood pressure standard deviation scores (SDS) correlated with FM SDS (BIA) over the entire normal range (systolic: r = 0.26, p = 0.002; diastolic: r = 0.33, p < 0.01). An evaluation of the children based on BP (three groups: BP < 50th percentile, 50th < BP > 95th percentile; BP > 95th percentile) revealed that hypertensive children had a higher BMI (17.6 vs. 19.4 vs. 26.2 kg/m², respectively) and a greater FM (14.0 vs. 16.8 vs. 30.2%, respectively). In conclusion, the divergence in FM in healthy boys and girls can be determined by BIA but not by BMI. In healthy children, BP within the entire normal range correlated with FM, children with established hypertension presented with a significantly higher FM. The study points to FM as an important determinant of BP pressure in obese and non-obese children.     

ORIGINAL ARTICLE: Prenatal Priming of Cord Blood T Lymphocytes by Microbiota in the Maternal Vagina

Problem In the vagina of women at the reproductive age, more than 170 strains of bacteria and yeasts are found. The effect of vaginal flora on neonatal T cells is yet to be investigated. Method of study We analyzed CD45RA and CD45RO expression on neonatal CD4+ T cells and cytokine production in CBMC cultures (interferon‐γ (IFN‐γ ), interleukin‐4 (IL‐4) and IL‐12) related to vaginal bacteria isolated from a maternal vagina. We collected vaginal swabs from 36 women at the first stage of the delivery and cord blood from their newborns. IFN‐γ, IL‐4, and IL‐12 in stimulated CBMC were measured and the expression of CD45RA/CD45RO on CD4+ T cells was assessed. Results We noted the difference in CD45RO CD4+ expression and IL‐12 levels between the newborns whose mothers were or were not colonized with Lactobacillus in the vagina (newborns whose mothers were colonized with Lactobacillus: CD45RO‐10%±3; IL‐12‐0.2 pg/mL ± 0.05; newborns whose mothers were not colonized with Lactobacillus: CD45RO‐6%±3; IL‐12‐2.0 pg/mL ± 0.7). Conclusion Our results may indicate that lactobacilli in maternal vagina influence the development of neonatal immune system. Yet, more research is needed using specified bacterial antigens.