Induction of apoptosis in estrogen dependent and independent breast cancer cells by the marine terpenoid dehydrothyrsiferol

Breast cancer (BCA) represents the highest incidence of death in 35- to 60-year-old women. Above all, hormone unresponsive BCA is still associated with poorer prognosis than hormone receptor expressing malign, mammary tumors. There is a consistent need for effective compounds to treat especially the first variant of this disease. Therefore, we investigated the cytotoxic effects of the marine polyether triterpenoid dehydrothyrsiferol (DT) in four BCA cell lines. Annexin V labeling revealed higher rates of DT-induced apoptosis in hormone insensitive than in estrogen receptor expressing cells. Flow cytometric analysis of combined DNA fragmentation and total DNA labeling allowed us to ascribe apoptotic cells to their cell cycle stage. Although, high cell mortality was detected in mitogen dependent G(1)-phase, time, concentration, and cell line dependent populations of apoptotic cells were also found to be of S-phase and G(2)/M-phase origin. These results suggest that the induction of apoptosis by DT might be transduced through more than one effector pathway. Cell cycle distributions and 5-bromo-2'-deoxyuridine incorporation varied in a treatment dependent manner and differed from control experiments with colchicine and doxorubicin which exclude that DT functions as a mitosis inhibitor. In summary, we propose that DT might be an interesting candidate for an antitumor drug development regimen.     

The effect of electrical stimulation on leg muscle pump activity in spinal cord-injured and able-bodied individuals

The purpose of this study was to examine the difference in: (1) effective muscle pump activity (MPA) between voluntary and electrically (ES) induced contractions in able-bodied subjects (ABS); and (2) ES-induced MPA between spinal cord-injured (SCI) individuals and ABS. MPA was measured as relative volume changes in the calf using strain-gauge plethysmography during repeated muscle contractions in the supine position while venous outflow was impeded by a thigh cuff inflated to a range of pressures. Ten SCI individuals and ten ABS participated in this study. ABS showed no significant difference between voluntary and electrically induced MPA [58.1 (18.4)% versus 67.7 (8.7)%, respectively]. SCI individuals showed a significantly lower ES-induced MPA than ABS [21.5 (15.9)% versus 67.7 (8.7)%, respectively]. The low MPA in SCI individuals may be explained by: (1) extensive leg muscle atrophy and/or (2) an “atrophic” vascular system in the legs. The electrical current level seemed to influence MPA (43 mA, 21.5% versus 60 mA, 30.8%) for SCI individuals, whereas no influence of muscle contraction rate on MPA was observed in ABS. The results of this study demonstrate that although ES-induced leg muscle contractions result in adequate MPA in ABS, it leads to significantly less effective MPA in SCI individuals. Accepted: 21 March 2000     

Factors influencing the generation of murine dendritic cells from bone marrow: The special role of fetal calf serum

Since the generation of dendritic cells (DC) from myeloid precursor cells within the bone marrow (BM) was first reported, it has developed to one of the standard tools for studying DC biology in rodent systems. However, antigen-specific effects of such DC on polyclonal T cell populations may be influenced by the additional presentation of bystander antigens derived from fetal calf sera (FCS). This may lead to misinterpretations of the antigen-specific effects on the T cell response. In addition, many factors, including FCS, also influence the quantity and quality of BM-DC already during culture. Here, we review the factors that have an impact on the generation of BM-DC, including criteria regarding the mice, but also technical parameters and the different growth factors. The critical role of FCS in BM-DC cultures and alternative methods using murine sera or serum-free media are discussed.     

Hypotensive and peripheral vascular effects in healthy volunteers of repeated oral administration of pinacidil

Summary The hypotensive and peripheral vascular effects of two different oral formulations of pinacidil were investigated in twelve healthy male volunteers.Arterial blood pressure, heart rate and calf blood flow were measured on Days 1 and 7. The vasodilator activity of both formulations was confirmed by a significant increase in calf blood flow on both days, which was correlated with a significant decrease in diastolic blood pressure.There was no differences between the tablet and the pellet.     

小牛脾提取物注射液对家兔白细胞计数的影响

目的探讨小牛脾提取物注射液肌肉注射后对家兔白细胞计数的影响。方法选取健康合格家兔,随机分成3组:肌肉注射1次组、肌肉注射2次组、生理氯化钠溶液对照组。结果肌肉注射1次组,与0h相比,注射后6~12h内,83%的家兔白细胞升高达峰值(P<0.05),但随后便开始下降,至24h白细胞已降至正常水平;肌肉注射2次组,与0h相比,注射后6~12h内100%家兔白细胞显著性升高(P<0.05),至24h仍有83%的家兔白细胞显著升高(P<0.05);肌肉注射1次和2次组在6、12h时间点与对照组相应时间点的白细胞计数均有显著差异(P<0.01);肌肉注射2次组在24h时白细胞计数与对照组相应时间点相比有显著差异(P<0.01)。结论小牛脾提取物注射液肌肉注射2次/d,升高白细胞的效果优于肌肉注射1次/d的效果。     

Photodynamic sterilization of red cells and its effect on contaminating white cells: viability and mechanism of cell death

BACKGROUND: Phthalocyanines are useful sensitizers for photodynamic sterilization of red cell concentrates. Various lipid-enveloped viruses can be inactivated with only limited red cell damage. Because white cells are involved in the immunomodulatory effects of blood transfusions, the study of the effect of photodynamic treatment on these cells is imperative. STUDY DESIGN AND METHODS: White cell-enriched red cell suspensions were photodynamically treated with either the hydrophobic Pc4 (HOSiPcOSi-(CH3)2(CH2)3N(CH3)2) or water-soluble aluminum phthalocyanine tetrasulfonate (AIPCS4) under virucidal conditions. Viability of white cell subpopulations on Days 0, 1, and 4 after treatment was determined by fluorescence-activated cell sorting by flow cytometric analysis of propidium iodide uptake. Apoptosis induction was studied by DNA ladder formation and staining for an early marker of apoptosis (annexin V). RESULTS: Treatment with Pc4 causes a significant decrease in cell viability of all white cells, as shown by prodidium iodide uptake. Monocytes and granulocytes are the most sensitive, and lymphocytes are relatively more resistant. Some of the cells die by apoptosis, which is induced within 30 minutes after treatment. Treatment with AIPCS4 damages only monocytes; other cell populations are not affected. CONCLUSIONS: Physicochemical properties of the photosensitizers partly determine their effect on white cells. Differences in intracellular localization are likely to be responsible for the effects observed.     

Serum supplement, inoculum cell density, and accessory cell effects are dependent on the cytokine combination selected to expand human HPCs ex vivo

BACKGROUND: The prolonged periods of pancytopenia associated with cord blood transplants suggest that in some cases cell numbers may be limiting. The possibility that limiting cell numbers may be overcome and prolonged periods of pancytopenia abrogated by the transplantation of human umbilical cord blood cells expanded ex vivo has led to efforts to define optimal culture conditions for these cells. STUDY DESIGN AND METHODS: Cord blood CD34+ cells were cultured with three cytokine combinations: SCF+G-CSF+GM-CSF+MGDF (SGGM); IL-6+ SCF+MGDF+Flt3-ligand (6SMF); and IL-1+IL-3+IL-6+G-CSF+GM-CSF+SCF+Epo (GFmix). Serum effects, inoculum concentration (cells/mL) seeding density (cell/cm(2)) and accessory cell effects on the expansion of CD34+ cells were determined. RESULTS: Cellular outputs were significantly higher with fetal calf serum (FCS) than with cord blood serum (CBS) or adult group AB serum (ABS) in the presence of 6SMF, however, CBS was as effective as FCS. The best seeding concentrations varied for each of the cytokine combinations, and inoculum densities exceeding 1000 cells per cm(2) proved detrimental for cultures containing GFmix and SGGM. Accessory cell studies indicated that populations expressing the CD33 antigen inhibited the expansion of purified CD34+ cells in the presence of GFmix or SGGM, but not in the presence of 6SMF. CONCLUSION: Serum supplement, inoculum cell concentration, seeding densities, and accessory cell effects are dependent upon the cytokine combination selected to expand cord blood HPCs ex vivo. Thus, each of these measures should be assessed to establish reproducible and reliable conditions for the selection of different cytokine combinations to culture cord blood HPCs.     

在乙型肝炎表面抗原破坏试验中对ELISA法检验结果影响因素的研究

试验表明，在ＨＢｓＡｇ破坏试验中，用ＥＬＩＳＡ法检测时，样品含小牛血清量愈多检出ＨＢｓＡｇ的灵敏度愈高。某些阴离子表面活性剂使检测结果呈假阴性，加适宜中和剂可将之克服。新洁尔灭与脂肪醇聚氧乙烯醚可使检测结果呈假阳性。所试中和剂中，仅亚硫酸钠与半胱氨酸可使测得的Ｓ／Ｎ值下降，其余无明显影响。     

Are adipose‐derived stem cells cultivated in human platelet lysate suitable for heart valve tissue engineering?

Tissue‐engineered heart valves represent a promising strategy for the growing need for valve replacements in cardiovascular medicine. Recent studies have shown that adipose‐derived stem cells (ADSC) are a viable cell source, as they are readily available in both the young and the elderly, show diverse differentiation potential and adapt their extracellular matrix (ECM) to a varying mechanical load. In vitro culture medium is usually enriched with fetal calf serum (FCS). However, a promising substitute has recently been found in human platelet lysate (HPL), which is superior in terms of proliferation speed and allogenicity. This study sought to elucidate the suitability of ADSC and HPL for heart valve tissue engineering (TE). ADSC harvested from five healthy individuals were cultured in both FCS and HPL. The cells were observed for differentiation potential, proliferation speed and immunophenotype, using immunohistochemistry and FACS analysis. Neotissue was assessed for ECM composition, human collagen I (hColl1) formation, histomorphology and mechanical stiffness under uniaxial tensile stress. Neotissue cultured in HPL was found to be significantly inferior in mechanical rigidity; it showed a three‐fold higher proliferation rate and a more dense ECM, but also a more heterogeneous hColl1 distribution. ECM analysis showed significantly higher amounts of DNA and glycosaminoglycans (GAG) in HPL‐cultured tissue. No significant differences were observed for differentiation potential and immunophenotype, apart from a lower CD166 expression in HPL. The mechanical inferiority of neotissue cultured in HPL represents a limitation to the use of HPL‐enriched media for heart valve TE with ADSC. This result concurs with data published about HPL and myofibroblasts derived from the venous wall. Similarly, the mechanical inferiority is not rooted in a difference in ECM composition, but rather in hColl1 architecture. Stem cell properties, as documented in the literature, are retained with HPL. A possible connection between the mechanical inferiority and the observed decrease in CD166 needs further investigation. Copyright © 2016 John Wiley & Sons, Ltd.