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Recently, we purified from adult murine bone marrow (BM) a population of CXCR4(+), Oct-4(+) SSEA-1(+), Sca-1(+) lin(-) CD45(-) very small embryonic-like (VSEL) stem cells and hypothesized that similar cells could be also present in human cord blood (CB). Here, we report that by employing a novel two-step isolation procedure -- removal of erythrocytes by hypotonic lysis combined with multiparameter sorting -- we could isolate from CB a population of human cells that are similar to murine BM-derived VSELs, described previously by us. These CB-isolated VSELs (CB-VSEL) are very small (3-5 micro m) and highly enriched in a population of CXCR4(+)AC133(+)CD34(+)lin(-) CD45(-) CB mononuclear cells, possess large nuclei containing unorganized euchromatin and express nuclear embryonic transcription factors Oct-4 and Nanog and surface embryonic antigen SSEA-4. Further studies are needed to see if human CB-isolated VSELs similar to their murine BM-derived counterparts are endowed with pluripotent stem cell properties.

Bovine colostrum is considered to provide anti-infective protection. Here, we present the first randomized controlled trial (RCT) aimed at assessing the preventive use of colostrum against upper respiratory tract infections (URTIs) in healthy pre-school children. We analyzed 57 children—35 in the colostrum (COL—dried bovine colostrum) and 22 in the placebo (PBO—dried whey) group, who received these substances as follows: first 15 days 2 × 500 mg and then 30 days 1 × 500 mg. The reporting on the children’s health status, specifically on the frequency and gravity of URTI symptoms and abdominal side effects, was performed via an online survey. The influence of colostrum on the frequency of days with URTI symptoms remained significant until the 20th week of observation and reached 31% of median reduction. The median reduction reached 37% when the gravity of symptoms was analyzed. When we grouped symptomatic days into episodes of second gravity level, the reduction in their frequency was even larger (50%) and lasted until the end of the trial (21 weeks). No significant side effects, especially abdominal, were reported during the trial. Colostrum supplementation in pre-school children is well tolerated, safe and provides protection from frequency of URTIs and their gravity.

Colostrum supplementation has been confirmed to protect from upper respiratory tract infections (URTIs) in athletes. Our trial was designed to find out whether other young adults who have potentially been exposed to increased risk of developing URTIs can also benefit. Homogenous population of medical (MED) students (at risk) and health science (HSci) peers were supplemented with a relatively low dose (0.5–1.0 g/day) of bovine colostrum (COL) or placebo (PBO) over 45 days and then once again over 7 days starting at day 87. The trial lasted 107 days. Subjects were monitored solely by them filling out online daily questionnaires containing questions about frequency and severity of URTIs symptoms, well-being, and potential gastrointestinal side-effects. A significant level of protection from URTIs was observed as expressed by dropping frequency of symptomatic days in COL vs. PBO group among MED vs. HSci students. The same effect was also recorded for severity of symptoms, as well as general well-being perception. Overall, it can be concluded that although young healthy people seem to have sufficient defenses from URTIs, COL supplementation can provide significant support in such protection among those at higher infectious risk because of exposure to a heavy workload and increased contact with infectious agents.

The aim of this prospective study is to compare the sensitivity and specificity of the liver shear wave elastography to the golden standard liver biopsy in staging liver fibrosis. Ninety-five patients were included in this study. These patients were sent for liver biopsy as a possible living liver donor or because of different pathologies including viral and autoimmune hepatitis and congenital liver diseases. A shear wave elastography and US-guided liver biopsy were done at the same setting by one experienced radiologist. One experienced histopathologist, blinded to SWE results, read the specimens. We included 95 patients in the study with a mean age of 30 years (range 3-65 years). We had 15/95 (16%) patients with hepatitis B/C, 61/95 (64%) patients with another liver disease, and 19/95 (20%) were donors. The mean of liver stiffness measured by elastography in patients was 6.5±0.19 kPa. The mean liver stiffness measured by elastography in patients with F0-F1 fibrosis was 5.39 ± 0.62 kPa, F2 was 7.32 ± 0.41, at stage F3 was 8.46 ± 0.33, and in the F4 stage, it was 11.42 ± 2.8 kPa. We found a significant difference in the mean level of liver stiffness in different degrees of fibrosis (p = 0.0001). The shear wave elastography could be used to assess liver fibrosis regardless of the cause.

Increased intestinal permeability has been implicated in various pathologies, has various causes, and can develop during vigorous athletic training. Colostrum bovinum is a natural supplement with a wide range of supposed positive health effects, including reduction of intestine permeability. We assessed influence of colostrum supplementation on intestinal permeability related parameters in a group of 16 athletes during peak training for competition. This double-blind placebo-controlled study compared supplementation for 20 days with 500 mg of colostrum bovinum or placebo (whey). Gut permeability status was assayed by differential absorption of lactulose and mannitol (L/M test) and stool zonulin concentration. Baseline L/M tests found that six of the participants (75%) in the colostrum group had increased intestinal permeability. After supplementation, the test values were within the normal range and were significantly lower than at baseline. The colostrum group Δ values produced by comparing the post-intervention and baseline results were also significantly lower than the placebo group Δ values. The differences in stool zonulin concentration were smaller than those in the L/M test, but were significant when the Δ values due to intervention were compared between the colostrum group and the placebo group. Colostrum bovinum supplementation was safe and effective in decreasing of intestinal permeability in this series of athletes at increased risk of its elevation.

Background and objective: The health supplement bovine colostrum reportedly improves immunity and regulates intestinal homeostasis. Reliable assessment methods are needed to ensure the satisfactory biological activity of all marketed colostrum products. Of the well-established effects of colostrum use, the restoration of appropriate intestinal permeability assessed with the lactulose/mannitol (L/M) differential sugar absorption test upon supplementation with colostrum has been consistently observed. Milking time after delivery is one of the factors that influences the composition of bovine colostrum, which causes a rapid decrease in bioactive components. Materials and methods: We use the L/M test to evaluate the intestinal permeability reduction upon supplementation with colostrum (2 × 500 mg) harvested at various times after delivery (2, 24, and 72 h) or a placebo (whey). In our randomized, double-blind placebo-controlled (DBPC) trial, 31 healthy athletes were divided into four groups and assessed at baseline and after the intervention. Results: The trial revealed that only colostrum collected after 2 h and 24 h caused a significant reduction of intestinal permeability. The comparison of post-intervention vs. baseline Δ values produced statistically significant results for 2 h colostrum versus the placebo and 72 h colostrum groups. Conclusions: We conclude that the change of bovine colostrum composition over the first three days of lactation is accompanied by a decrease in its biological activity as measured with the L/M test. This test may offer a biological quality measure for colostrum.

This paper describes a simple hydrothermal procedure for high-yield synthesis of single-crystalline ZnO hexagonal nanoplates in a surfactant-free system at 70 °C. The structures and morphologies of the synthesized ZnO nanoplates are derived from characterisation by X-ray diffraction, and scanning and transmission electron microscopy. Their optical properties are recorded by Raman and photoluminescence spectroscopy. These ZnO hexagonal nanoplates exhibit the enhanced photocatalytic activity of phenol photodecomposition, suggesting that they could be served as an active system for the treatment of the waste water, in addition to their common applications.

The transplantation of hematopoietic stem and progenitor cells (HSPC) is an established lifesaving therapy. Bone marrow (BM), harvested from heparinized cadaveric organ donors, peripheral blood (PB) and cord blood (CB), are important sources of hematopoietic stem cells. HSPCs, which are used for transplantation purposes, are routinely evaluated in terms of number of mononuclear cells (MNCs), CD34+ MNCs count and viability. The efficacy of grafting is determined additionally in clonogenic tests in vitro. These tests deliver important information about the number of HSPCs and their proliferative potential. Unfortunately, they do not give a possibility to evaluate the functional HSPC chemotactic reactivity in the SDF-1 gradient, which is probably the key phenomenon for HSPC homing after transplantation procedure. Thus, the aim of our study was to optimize HSPC isolation according to their chemotactic reactivity in SDF-1 gradient. Using multiparameter cell sorter (FACS Aria, BD) we examined the HSPCs attracted by SDF-1 on a single cell level. The population of cells which participated in the chemotactic process was highly enriched in CXCR4+lin-AC133+CD45+ cells (referred as hematopoietic stem cells) and to our surprise in CXCR4+lin-AC133+CD45- cells (referred as pluripotent stem cells) in quantitative amounts. Since reactivity of HSPCs may depend on various factors involved in the protocol of their isolation and short-term storage, we tested the most commonly used anticoagulants (ACD, CPDA-1, EDTA and Heparin) and culture media (DME, IMDM, RPMI). HSPCs, harvested from CB, PB and BM, were subsequently investigated for clonogenic growth of CFU-GM in methylcellulose cultures and for the level of apoptosis by employing annexin V staining. Evaluating clonogenic potential, ability of chemotactic reactivity in SDF-1 gradient and intensification of apoptosis of HSPC as the most safe anticoagulant and medium were selected. This study has proved that chemotactic reactivity of HSPCs is a new but very important parameter which should be included in the procedure of their isolation.

Histone deacetylase inhibitors (HDIs) are a group of compounds that exhibit anticancer activity, but their significance and usefulness in breast cancer (BC) treatment are still controversial. The ability of cancer cells to invade and migrate is augmented by the acquisition of a mesenchymal phenotype - a process known as epithelial-to-mesenchymal transition (EMT). Changes in the expression level of different cadherins, so-called cadherin switches, have been used to monitor the EMT process in development and tumor progression, in particular migration and invasion potential. The aim of this study was to analyze the influence of two HDIs - valproic acid (VPA) and vorinostat (SAHA) - on the migration potential of different BC cell types, as well as on EMT, or its reverse process - mesenchymal-to-epithelial transition, progression by means of shift in epithelial and mesenchymal marker expression. HDI treatment-induced expression of E- and N-cadherin at the mRNA and protein levels was evaluated by qPCR, Western blotting and immunostaining methods, respectively. BC cell proliferation and migration were assessed by BrdU, xCELLigence system and wound-healing assay. VPA and SAHA inhibited the proliferation and migration in a dose- and time-dependent manner, regardless of the BC cell type. Unawares, BC cells having a more mesenchymal phenotype (MDA-MB-468) were found to overexpress N-cadherin, whereas BC lines having an epithelial phenotype (T47D, MCF7) responded to HDI treatment by a significant increase of E-cadherin expression. We suggest that HDAC inhibition results in a more relaxed chromatin concomitant to an increase in the expression of already expressing genes. By using multiple cancer cell lines, we conclude that HDI induction or reversal of EMT is not a universal mechanism, yet inhibition of cell migration is, and thus EMT should not be considered as the only measurement for tumor aggressiveness.

Studies on mixed chimerism are currently focused primarily on obtaining less toxic conditioning protocols. With these issues in mind, we have undertaken the attempt to optimize the procedure of mixed chimerism induction in mice. In order to reduce toxicity, we used decreasing doses of total body irradiation (TBI) together with combination of blocking antibodies. We also tried to eliminate immunosuppression (cyclophosphamide - CP) treatment after bone marrow transplantation. B6.SJL-PtprcaPep3b mice were injected with 20-30 x 106 bone marrow cells from Balb C mice. Mice were treated with TBI (3 - 1.5 - 0 Gy) on \"-1\" day of the experiment and blocking antibodies against CD40L (\"0\", and \"4\" days) and additionally anti-CD8 (\"-2\" day) and/or anti-NK1.1 (\"-3\" day). Mice in certain groups also received CP (175 mg/kg) on \"2\" day. Presence of mixed chimerism was assessed in peripheral blood cells by flow cytometry on the 1st, 2nd, 3rd, 4th, 6th and 8th weeks of the experiment by detecting of CD45.1 (characteristic for B6.SJL-PtprcaPep3b strain) and CD45.2 (characteristic for Balb C strain) antigens expression. We also analyzed the percentage of peripheral blood CD8 T-cells (CD3e/CD8a) and NK cells (Ly-49D/NK1.1). We found that reduction of TBI dose and elimination of CP decrease the rate of mixed chimerism formation. The highest percentage of donor cells was obtained in the group of animals treated with 3 Gy of TBI, CP and combination of anti-CD40L, anti-CD8, and anti-NK1.1 antibodies. The 3 Gy TBI was necessary to induce stable mixed chimerism, but it could be obtained without the CP use. The percentage of CD3e/CD8a and Ly-49D/NK1.1 cells was significantly lower in the groups of mice treated by corresponding antibodies. Moreover, we observed the lowest number of peripheral blood Ly-49D/NK1.1 cells in the group of animals with highest mixed chimerism. Our experiments in mice model can help in better understanding of mixed chimerism phenomenon and in selecting the method of mixed chimerism induction with lowest possible toxicity. This also might improve the protocols of stable mixed chimerism induction in humans, and in the future, the effectiveness of vascularized organ transplantation.