Evaluation of human umbilical cord blood as a source of embryonic stem cells

Human umbilical cord blood (HUCB) has been poorly characterised as a source of embryonic stem cells (ESCs). The aim of this study, therefore, was to evaluate HUCB as source of mesenchymal stem cells (MSCs) with embryonic characteristics. HUCB was collected from consenting women undergoing elective caesarean sections. HUCB was meticulously explanted into MesenCult media and incubated. Qualitative and quantitative immunophenotyping of cells was achieved using fluorescein isothiocyanate (FITC) labelled antibodies (CD34, CD45, CD29, CD44, CD73 and CD105) phenotypic markers. Immunocytochemistry was carried out for the human ESC markers CD9, stage-specific embryonic antigen-1 and 4 (SSEA-1 and SSEA-4), E-cadherin, Podocalyxin (PODXL), sex-determining region Y-box 2 (SOX2), NANOG and Octamer (OCT3/4). MSCs were cultured to induce differentiation into adipogenic, osteogenic, chondrogenic and neurogenic cells. Immunocytochemistry was used to identify fatty acid binding protein-4 (FABP-4), osteocalcin, aggrecan, SOX2 and oligodendrocyte-4 (Olig-4) markers. The cells were strongly positive for the MSC markers CD29, CD44, CD73 and CD105; these cells also expressed the ESC markers CD9, SSEA-1 and SSEA-4, E-cadherin, PODXL, SOX2, NANOG and OCT3/4. Additionally, the MSCs expressed the adipogenic FABP-4, osteogenic osteocalcin, chondrogenic aggrecan and neural Olig-4 and SOX2 markers after differentiation. Therefore, HUCB is a rich source for MSCs with embryonic characteristics

Integrative analysis of copy number and gene expression in breast cancer using formalin-fixed paraffin-embedded core biopsy tissue: a feasibility study.

BACKGROUND: An absence of reliable molecular markers has hampered individualised breast cancer treatments, and a major limitation for translational research is the lack of fresh tissue. There are, however, abundant banks of formalin-fixed paraffin-embedded (FFPE) tissue. This study evaluated two platforms available for the analysis of DNA copy number and gene expression using FFPE samples. METHODS: The cDNA-mediated annealing, selection, extension, and ligation assay (DASL™) has been developed for gene expression analysis and the Molecular Inversion Probes assay (Oncoscan™), were used for copy number analysis using FFPE tissues. Gene expression and copy number were evaluated in core-biopsy samples from patients with breast cancer undergoing neoadjuvant chemotherapy (NAC). RESULTS: Forty-three core-biopsies were evaluated and characteristic copy number changes in breast cancers, gains in 1q, 8q, 11q, 17q and 20q and losses in 6q, 8p, 13q and 16q, were confirmed. Regions that frequently exhibited gains in tumours showing a pathological complete response (pCR) to NAC were 1q (55%), 8q (40%) and 17q (40%), whereas 11q11 (37%) gain was the most frequent change in non-pCR tumours. Gains associated with poor survival were 11q13 (62%), 8q24 (54%) and 20q (47%). Gene expression assessed by DASL correlated with immunohistochemistry (IHC) analysis for oestrogen receptor (ER) [area under the curve (AUC) = 0.95], progesterone receptor (PR)(AUC = 0.90) and human epidermal growth factor type-2 receptor (HER-2) (AUC = 0.96). Differential expression analysis between ER+ and ER- cancers identified over-expression of TTF1, LAF-4 and C-MYB (p ≤ 0.05), and between pCR vs non-pCRs, over-expression of CXCL9, AREG, B-MYB and under-expression of ABCG2. CONCLUSION: This study was an integrative analysis of copy number and gene expression using FFPE core biopsies and showed that molecular marker data from FFPE tissues were consistent with those in previous studies using fresh-frozen samples. FFPE tissue can provide reliable information and will be a useful tool in molecular marker studies. TRIAL REGISTRATION: Trial registration number ISRCTN09184069 and registered retrospectively on 02/06/2010

The differential contribution of the innate immune system to a good pathological response in the breast and axillary lymph nodes induced by neoadjuvant chemotherapy in women with large and locally advanced breast cancers

The tumour microenvironment consists of malignant cells, stroma, and immune cells. The role of adaptive immunity in inducing a pathological complete response (pCR) in breast cancer with neoadjuvant chemotherapy (NAC) is well studied. The contribution of innate immunity, however, is poorly documented. Breast tumours and axillary lymph nodes (ALNs) from 33 women with large and locally advanced breast cancers (LLABCs) undergoing NAC were immunohistochemically assessed for tumour-infiltrating macrophages (TIMs: M1 and M2), neutrophils (TINs), and dendritic cells (TIDCs) using labelled antibodies and semiquantitative methods. Patients’ blood neutrophils (n = 108), DCs (mDC1 and pDC), and their costimulatory molecules (n = 30) were also studied. Pathological results were classified as pCR, good (GPR) or poor (PRR). In breast and metastatic ALNs, high levels of CD163+ TIMs were significantly associated with a pCR. In blood, high levels of neutrophils were significantly associated with pCR in metastatic ALNs, whilst the % of mDC1 and pDC and expression of HLA-DR, mDC1 CD40, and CD83 were significantly reduced. NAC significantly reduced tumour DCs but increased blood DCs. PPRs to NAC had significantly reduced HLA-DR, CD40, and CD86 expression. Our study demonstrated novel findings documenting the differential but important contributions of innate immunity to pCRs in patients with LLABCs undergoing NAC

Semi-Analytical Perturbative Approaches to Third Body Resonant Trajectories

In the framework of multi-body dynamics, successive encounters with a third body, even if well outside of its sphere of influence, can noticeably alter the trajectory of a spacecraft. Examples of these effects have already been exploited by past missions such as SMART-1, as well as are proposed to benefit future missions to Jupiter, Saturn or Neptune, and disposal strategies from Earth's High Eccentric or Libration Point Orbits. This paper revises three totally different descriptions of the effects of the third body gravitational perturbation. These are the averaged dynamics of the classical third body perturbing function, the Opik's close encounter theory and the Keplerian map approach. The first two techniques have respectively been applied to the cases of a spacecraft either always remaining very far or occasionally experiencing extremely close approaches to the third body. However, the paper also seeks solutions for trajectories that undergo one or more close approaches at distances in the order of the sphere of influence of the third body. The paper attempts to gain insight into the accuracy of these different perturbative techniques into each of these scenarios, as compared with the motion in the Circular Restricted Three Body Problem.Comment: In: 66th International Astronautical Congress, Jerusalem, 12-16 October 201

M1 macrophages evoke an increase in polymeric immunoglobulin receptor (PIGR) expression in MDA-MB468 breast cancer cells through secretion of interleukin-1β

High expression of polymeric immunoglobulin receptor (PIGR) in breast cancer is associated with increased 5-year survival rate. However, the factors influencing PIGR expression in breast cancer have not been elucidated. The aim of this study was to determine the role of macrophages and cytokines affecting expression of PIGR in two breast cancer cell lines. M1, M2 macrophage conditioned media (CM) and recombinant human cytokines were used to determine factors which increased PIGR expression in MCF7 (HTB-22) and MDA-MB468 (HTB-132) breast cancer cell lines. The level of PIGR expression in the cells and PIGR secretory component were evaluated by real-time quantitative polymerase chain reaction and Western blotting. M1 macrophage CM induced a dose-dependent increase in PIGR mRNA expression in MDA-MB468 cells, up to 20-fold. The level of PIGR expression in MCF7 cells was very low and not affected by M1 and M2 CM. Interferon gamma (IFN-γ) and interleukin (IL)-1β also increased PIGR expression in MDA-MB468 and MCF7 cells. However, IL-1β was demonstrated to increase in M1 macrophages, while IFN-γ was not. The role of IL-1β secreted from M1 macrophages in increasing expression of PIGR was confirmed by IL-1 receptor blockade, indicating that IL-1β was the major M1 macrophage-derived cytokine that enhanced PIGR expression. Elevated PIGR expression in breast cancer in vivo may reflect the polarization state of tumor-associated immune cells

Assessment of natural density of distribution for fish by the catches at light stations

Algorithm for calculation of illumination produced by shipboard lights at the sea surface and underwater is developed and implemented with Microsoft© Office Excel software. The surface illumination is calculated for the fishing vessel STR-420 with the light equipment for saury fishery. Irregularity of the lighting around the vessel is shown that depends on the lamps arrangement. Optimal scheme of the lamps arrangement is proposed for a new saury trap with the catch volume > 7000 m3. The optimal arrangement provides more regular lighting and therefore larger illuminated area for attraction of fish with positive phototaxis, as saury. Relationship between natural density of saury distribution and its catch per trap is determined for this trap

Modulation of T regulatory and dendritic cell phenotypes following ingestion of Bifidobacterium longum, active hexose correlated compound (AHCC®) and azithromycin in healthy individuals

The probiotic Bifidus BB536 (BB536), which contains Bifidobacterium longum, has been shown to have enhanced probiotic effects when given together with a standardized extract of cultured Lentinula edodes mycelia (AHCC®, Amino Up Co. Ltd., Sapporo, Japan). BB536 and AHCC® may modulate T cell and dendritic cell (DC) phenotypes, and cytokine profiles to favour anti-inflammatory responses following antibiotic ingestion. We tested the hypothesis that orally administered BB536 and/or AHCC®, results in modulation of immune effector cells with polarisation towards anti-inflammatory responses following antibiotic usage. Forty healthy male volunteers divided into 4 equal groups were randomised to receive either placebo, BB536, AHCC® or a combination for 12 days in a double-blind manner. After 7 days volunteers also received 250 mg azithromycin for 5 days. Cytokine profiles from purified CD3+ T cells stimulated with PDB-ionomycin were assessed. CD4+ CD25+ forkhead box P3 (Foxp3) expression and peripheral blood DC subsets were assessed prior to treatment and subsequently at 7 and 13 days. There was no difference in cytokine secretion from stimulated CD3+ T cells between treatment groups. Compared with baseline, Foxp3 expression (0.45 ± 0.1 vs. 1.3 ± 0.4; p = 0.002) and interferon-gamma/interleukin-4 (IFN-γ/IL-4) ratios were increased post-treatment in volunteers receiving BB536 (p = 0.031), although differences between groups were not significant. For volunteers receiving combination BB536 and AHCC®, there was an increase in myeloid dendritic cells (mDC) compared with plasmacytoid DC (pDC) counts (80% vs. 61%; p = 0.006) at post treatment time points. mDC2 phenotypes were more prevalent, compared with baseline, following combination treatment (0.16% vs. 0.05%; p = 0.002). Oral intake of AHCC® and BB536 may modulate T regulatory and DC phenotypes to favour anti-inflammatory responses following antibiotic usage

Anchor system for installation of fishing gears on soft grounds

Mushroom anchors are proposed for positioning of vessels and fishing gears on soft grounds, as sands and silts. Designs of the anchors of original shape are presented. Their holding force is determined for certain navigational conditions, and the forces acting on a floating object are calculated for the cases of anchoring perpendicular and parallel to the water flow. Weight of the mushroom anchors necessary for anchoring of a floating object is calculated and scheme of the anchorage is presented for the depth 250 m. The mushroom anchors using for rescue operations in the Okhotsk Sea and for shrimp pot fishery in the Japan Sea is discussed, in particular the episode of positioning the vessel of deadweight 2433 t in conditions of 3-points roughness and the current velocity 2 knots at the sea surface and 4 knots at the sea bottom with using two stern and one bow mushroom anchors. Using of the compact mushroom anchor for shrimp pot fishery allows to reduce the traps moving along the bottom during their hauling. Possibly, the mushroom anchors could be used for other gears installation, as well, as stationary nets, crab traps, bottom longlines, etc

Effects on quality of life, anti-cancer responses, breast conserving surgery and survival with neoadjuvant docetaxel: a randomised study of sequential weekly versus three-weekly docetaxel following neoadjuvant doxorubicin and cyclophosphamide in women with primary breast cancer

<p>Abstract</p> <p>Background</p> <p>Weekly docetaxel has occasionally been used in the neoadjuvant to downstage breast cancer to reduce toxicity and possibly enhance quality of life. However, no studies have compared the standard three weekly regimen to the weekly regimen in terms of quality of life. The primary aim of our study was to compare the effects on QoL of weekly versus 3-weekly sequential neoadjuvant docetaxel. Secondary aims were to determine the clinical and pathological responses, incidence of Breast Conserving Surgery (BCS), Disease Free Survival (DFS) and Overall Survival (OS).</p> <p>Methods</p> <p>Eighty-nine patients receiving four cycles of doxorubicin and cyclophosphamide were randomised to receive twelve cycles of weekly docetaxel (33 mg/m²) or four cycles of 3-weekly docetaxel (100 mg/m²). The Functional Assessment of Cancer Therapy-Breast and psychosocial questionnaires were completed.</p> <p>Results</p> <p>At a median follow-up of 71.5 months, there was no difference in the Trial Outcome Index scores between treatment groups. During weekly docetaxel, patients experienced less constipation, nail problems, neuropathy, tiredness, distress, depressed mood, and unhappiness. There were no differences in overall clinical response (93% vs. 90%), pathological complete response (20% vs. 27%), and breast-conserving surgery (BCS) rates (49% vs. 42%). Disease-free survival and overall survival were similar between treatment groups.</p> <p>Conclusions</p> <p>Weekly docetaxel is well-tolerated and has less distressing side-effects, without compromising therapeutic responses, Breast Conserving Surgery (BCS) or survival outcomes in the neoadjuvant setting.</p> <p>Trial registration</p> <p>ISRCTN: <a href="http://www.controlled-trials.com/ISRCTN09184069">ISRCTN09184069</a></p