The effect of nitrogen containing bisphosphonates,zoledronate and alendronate,on the production of pro-angiogenic factors by osteoblastic cells

Bisphosphonates (BPs) have been shown to influence angiogenesis. This may contribute to BP-associated side-effects such as osteonecrosis of the jaw (ONJ) or atypical femoral fractures (AFF). The effect of BPs on the production of angiogenic factors by osteoblasts is unclear. The aims were to investigate the effect of (1) alendronate on circulating angiogenic factors; vascular endothelial growth factor (VEGF) and angiopoietin-1 (ANG-1) in vivo and (2) zoledronate and alendronate on the production of VEGF and ANG-1 by osteoblasts in vitro. We studied 18 post-menopausal women with T score ⩽ −2 randomized to calcium/vitamin D only (control arm, n = 8) or calcium/vitamin D and alendronate 70 mg weekly (treatment arm, n = 10). Circulating concentrations of VEGF and ANG-1 were measured at baseline, 3, 6 and 12 months. Two human osteoblastic cell lines (MG-63 and HCC1) and a murine osteocytic cell line (MLO-Y4) were treated with zoledronate or alendronate at concentrations of 10⁻¹²–10⁻⁶ M. VEGF and ANG-1 were measured in the cell culture supernatant. We observed a trend towards a decline in VEGF and ANG-1 at 6 and 12 months following treatment with alendronate (p = 0.08). Production of VEGF and ANG-1 by the MG-63 and HCC1 cells decreased significantly by 34–39% (p < 0.01) following treatment with zoledronate (10⁻⁹–10⁻⁶ M). Treatment of the MG-63 cells with alendronate (10⁻⁷ and 10⁻⁶) led to a smaller decrease (25–28%) in VEGF (p < 0.05). Zoledronate (10⁻¹⁰–10⁻⁶ M) suppressed the production of ANG-1 by MG-63 cells with a decrease of 43–49% (p < 0.01). Co-treatment with calcitriol (10⁻⁸ M) partially reversed this zoledronate-induced inhibition. BPs suppress osteoblastic production of angiogenic factors. This may explain, in part, the pathogenesis of the BP-associated side-effects.     

Factors shaping the tolerance of local Tibetan herders toward snow leopards

Snow leopards (Panthera uncia) have long co-existed with livestock herding people across Asia’s high mountains. Multiple use landscapes however imply potential competition for shared resources, livestock predation, and the risk of retaliatory killing of predators. Community-based conservation is a central pillar for supporting people’s livelihoods and safeguarding predators and their habitat. Based on the theory of planned behavior, we examined the factors that shape herders’ tolerance of snow leopards. Our questionnaire-based study was conducted in the Sanjiangyuan Region, China, encompassing four communities with varying livelihoods, experiences with livestock depredation and levels of exposure to community conservation interventions. Our results showed that respondents generally held positive views towards snow leopards, although women tended to have relatively more negative views towards snow leopards compared with men. Current household income was largely dependent on caterpillar fungus rather than livestock. Social norms around religion and the role of community leaders in our study area seemed to be the main determinants of the generally benign association of people with wildlife, overshadowing potential influences of community-based conservation interventions. Our work suggests that conservations programs will be aided through collaborations with communities and religious institutions, and that conservationists must proactively engage with women as significant actors in conservation.     

The effect of local dominance and reciprocal tolerance on feeding aggregations of ocellated antbirds

We studied ocellated antbirds (Phaenostictus mcleannani) to test the hypothesis that reciprocal tolerance between dominant individuals can favour feeding in aggregations. Mated pairs hold large non-exclusive feeding ranges, but roost and nest in a small portion of this range (‘roosting area’); adjacent roosting neighbours are unrelated. Ocellated antbirds congregate to feed on arthropods fleeing from nomadic swarms of army ants that move across the ranges of many pairs. We used playback experiments to simulate acoustic challenges, and results showed that males responded aggressively to other males only in their roosting areas. Responses to adjacent neighbours were less aggressive than to non-neighbours (i.e. the ‘dear enemy’ effect). Prey intake rates were higher when birds fed in their own roosting area or in that of adjacent neighbours compared with more distant sites. Males tolerated adjacent neighbours at swarm fronts where prey are most dense, but more distant neighbours were displaced. Despite small samples for some analyses, our results suggest that reciprocal tolerance between adjacent unrelated neighbours can ameliorate intraspecific competition within ephemeral feeding aggregations.     

Rapid evaluation of sorghum for aluminum tolerance

Two screening methods tested in this study were based on the observation that roots of freshly germinated seeds exhibit their relative tolerance to Al during the first 48 hours of growth. Sorghum (Sorghum bicolor L. Moench) varieties were evaluated using toxic/nontoxic soil pairs in petri dishes and toxic/nontoxic solution pairs in a flowing aqueous system. The soils had 0, 61, and 72% Al saturation and the solutions had 0, 1.85 and 3.70 M l^–1 Al from AlCl₃ and 0.25 mM l^–1 Ca from Ca(NO₃)₂. Relative root lengths in both systems correctly differentiated 13 genotypes of known Al tolerance into tolerant and intolerant groups. Twenty six other sorghums genotypes were also screened using genotypes of known Al reactions as checks. The soil with 72% Al saturation and the 1.85 M l^–1 Al solution gave the highest correlation between the two systems and both effectively arrayed sorghums of known and unknown tolerance. For routine screening the authors prefer the soil system for its simplicity, efficiency, and use of a natural growth medium.Journal paper 11637. Purdue University Agricultural Experiment Station, Lafayette, Indiana. Contribution from the Agronomy Department.     

The histopathological characterization of oralCandida leukoplakias

Sixty four biopsy specimens of oralCandida leukoplakias were examined. Histological sections were stained with periodic acid-Schiff (PAS) reagent and by Grocott’s silver method.C. albicans isolates were cultivated from all the patients but fungal hyphae were proved histologically only in 23 of them. PAS-positive and Grocott-positive elements were exclusively observed in the superficial epithelial layers,e.g., the cornified layer and the stratum grnulosum. The inflammatory reaction was characterized by an early phase during which polymorphonuclear cells predominated. The skeletal muscle fascicles immediately adjacent to the infected epithelium showed striking degeneration and atrophy associated with a marked infiltrate of chronic inflammatory cells.     

Number of gammaglobulin containing cells after the single local administration of antigen

The number of cells containing gammaglobulin in the regional lymphatic nodes of rabbits during the response to the single local dose of diphtheria toxoid was demonstrated using immunofluorescent technique. Antigen-dependent increase of the number of IgG—containing cells in regional lymph nodes was found. The possible source of the gammaglobulin without demonstrable specificity to the administered antigen is discussed. Dedicated to Academician Ivan Málek on the occasion of his 60th birthday     

Effects of systemic versus local administration of corticosteroids on mucosal tolerance

Respiratory exposure to allergen induces T cell tolerance and protection against the development of airway hyperactivity in animal models of asthma. Whereas systemic administration of dexamethasone during the delivery of respiratory Ag has been suggested to prevent the development of mucosal tolerance, the effects of local administration of corticosteroids, first-line treatment for patients with bronchial asthma, on mucosal tolerance remain unknown. To analyze the effects of systemic versus local administration of different types of corticosteroids on the development of mucosal tolerance, mice were exposed to respiratory allergen to induce mucosal tolerance with or without systemic or intranasal application of different doses of dexamethasone or prednisolone. After the induction of mucosal tolerance, proliferation of T cells was inhibited in tolerized mice, whereas systemic applications of corticosteroids restored T cell proliferation and secretion of Th2 cytokines. In contrast, inhaled corticosteroids showed no effect on both T cell proliferation and cytokine secretion. In addition, mice systemically treated with corticosteroids showed an increased airway hyperactivity with a significant lung inflammation, but also an increased T effector cells/regulatory T cells ratio in the second lymphoid organs when compared with mice that receive corticosteroids by inhalation. These results demonstrate that local administration of corticosteroids has no effect on the development of immune tolerance in contrast to systemically applied corticosteroids. Furthermore, although different concentrations of corticosteroids are administered to patients, our results demonstrated that the route of administration rather than the doses affects the effect of corticosteroids on respiratory tolerance induction. Considering the broad application of corticosteroids in patients with allergic disease and asthma, the route of administration of steroid substances seems crucial in terms of treatment and potential side effects. These findings may help elucidate the apparently contradicting results of corticosteroid treatment in allergic diseases.     

Transgenic tall fescue containing the Agrobacterium tumefaciens ipt gene shows enhanced cold tolerance

Embryogenic calli of Festuca arundinacea were transformed with the Agrobacterium tumefaciens isopentenyl transferase (ipt) gene driven by a maize ubiquitin promoter. Tillering ability, levels of chlorophyll a and b, and cold tolerance were greatly increased in the transgenic turfgrass, which resulted in the plants remaining more vigorous and staying green longer under lower temperatures.     

Visual detection of saikosaponins by on-membrane immunoassay and estimation of traditional Chinese medicines containing Bupleuri radix

The purpose of this study was to describe the simple, rapid, and environmental-cost effective determination method for saikosaponins in complicated samples like Bupleuri radix and traditional Chinese medicines (TCM). Saikosaponin standards, extracts of Bupleuri radix and TCM, were applied to a polyethersulphone (PES) membrane and developed by acetonitrile-water (1:4, by volume). Saikosaponin a (SSa), SSc, and SSd were visually detected by an immunostaining method (called Eastern blotting technique) using a monoclonal antibody (MAb) against SSa. At least 62.5 ng of SSa, SSc, and SSd were clearly detectable individually. These coloring spot areas of saikosaponins on PES membrane were calculated by using the NIH Imaging software and three saikosaponins can be analyzed quantitatively between 62.5 ng and 1.0 microg. Saikosaponins in Bupleuri radix and TCM were determined and these results of SSa and total saikosaponin concentrations were in good agreement with those from the ELISA analysis.     

Epidemiologic analysis of prenatal exposure to cough medicines containing dextromethorphan: no evidence of human teratogenicity

BACKGROUND: In a recent experimental study on chick embryos, Andaloro et al. (1998, Pediatr. Res. 43:1-7) observed a relationship between neural crest/neural tube defects (NTDs) and prenatal exposure to dextromethorphan. These authors made an extrapolation of their results to the human embryo, indicating that this drug could produce NTDs in humans. Rosenquist (1999, Teratology 60:58-60) based on the results of the epidemiologic study performed by Ferencz et al. (1997, Perspect. Pediatr. Cardiol. 5:50-162), concluded that dextromethorphan could cause congenital heart defects in humans. Because this drug is an over-the-counter drug, the suggestion of those authors has led to great controversy and public concern about the possible teratogenic effect of this drug on the human embryo. METHODS: We present the results of a case-control study by using logistic regression analyses on the effect of prenatal exposure to drugs containing dextromethorphan only, or in combination with other drugs, on human development. We mostly analyzed dextromethorphan. The study was designed in part as hypothesis confirmation for NTDs and congenital heart defects and, in part, as hypotheses generation by testing the association with many other congenital defects. RESULTS: The results do not show a relation between the occurrence of NTDs and heart defects or other defects with exposure to drugs containing dextromethorphan. CONCLUSIONS: The usual use of dextromethorphan in cough medications during pregnancy does not increase the risk for congenital defects.     

Fluoro-Jade B evidence of induced ischemic tolerance in the rat spinal cord ischemia: physiological, neurological and histopathological consequences

Fluoro-Jade B, a marker of degenerating neurons, was used to label histopathological changes in the rat spinal cord after transient ischemia and ischemic preconditioning (IPC). To characterize postischemic neurodegenerations and consequent neurological changes, a particular attention was paid to the standardization of ischemic conditions in animals of both groups. 1. The control ischemic rats were submitted to a reversible occlusion of descending aorta by insertion and subsequent inflation of a 2F Fogarty catheter for 12 min. 2. In the IPC rats, an episode of short 3 min occlusion and 30 min reperfusion preceded the 12 min ischemia. Postischemic motor function testing (ambulation and stepping) was provided repeatedly for evaluation of neurological status 2 h and 24 h after surgery and at the end of postischemic survival, i.e. after 48 h. Fluoro-Jade B staining was used to demonstrate degenerated neurons. In the control rats, neurological consequences of histopathological changes in lumbosacral spinal cord, manifested as paraplegia, were present after 12 min ischemia. Thus, numbers of degenerated Fluoro-Jade B positive cells were visible in gray matter of the most injured L(4)-S(2) spinal cord segments. Slight motor function impairment, consequential from significant decreasing in Fluoro-Jade B-positivity in the L(4)-S(2) spinal cord segments of the IPC rats, was considered the pathomorpfological evidence that IPC induces spinal cord tolerance to ischemia. Our results are consistent with the previously published silver impregnation method for histopathological demonstration of ischemic degeneration.     

The effect of the bisphosphonate alendronate on viability of canine osteosarcoma cells in vitro

The objective of this study was to determine the effect of alendronate on the viability of canine osteosarcoma cells and nonneoplastic canine cells. The sample population was composed of canine osteosarcoma tumor cells. Osteosarcoma cells and canine fibroblasts were maintained in culture under standard conditions. The MTT assay for cell viability was performed after 24, 48, and 72 h of incubation with alendronate (0.001 to 1000 microM) or no drug (control). Plates were set up so that each concentration and the control had a sample number of 8. The optical density (OD) of each well was measured at 540 nm using an enzyme-linked immunosorbent assay microplate reader. The percent viability was determined for each concentration and for each incubation time. After 24 h of incubation of POS (parent osteosarcoma) and HMPOS cells with alendronate, there was no significant difference in mean OD at any drug concentration when compared with control samples. A significant concentration- and time-dependent reduction in mean OD of osteosarcoma cells was observed after 48 and 72 h of incubation, with alendronate concentrations ranging from 10 to 1000 microM. The lowest percent cell viability observed in treated cells was 35%. Conversely, alendronate did not significantly affect mean OD in fibroblasts, and the lowest percent cell viability observed was 76%. Our data indicate that alendronate may have the potential to inhibit canine osteosarcoma tumor growth. It will be important to determine the clinical relevance of these in vitro findings. If similar findings are observed in vivo, use of alendronate may also be indicated as an adjuvant to existing chemotherapeutic protocols.     

内毒素耐受对核苷酸结合寡聚化结构域2信号通路的影响

为研究内毒素耐受对核苷酸结合寡聚化结构域2(nucleotide-binding oligomerization domain containing 2,NOD2)信号通路的影响,将小鼠单核-巨噬细胞RAW264.7分为两组,分别给予小剂量脂多糖(lipopolysaccharide,LPS)(100ng/mL)或磷酸盐缓冲液(phosphate buffered saline,PBS)预处理20h,建立内毒素耐受组和对照组。每组细胞分别给予大剂量LPS(1 000ng/mL)或热灭活烟曲霉孢子刺激,于刺激后0、2、6、12、24h采用定量聚合酶链反应(polymerase chain reaction,PCR)检测细胞NOD2、受体相互作用蛋白2(receptor-interacting protein 2,RIP2)和肿瘤坏死因子α(tumor necrosis factorα,TNF-α)mRNA表达;采用酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测细胞上清液中白细胞介素8(interleukin 8,IL-8)和TNF-α浓度。结果显示,内毒素耐受组无论是大剂量LPS还是热灭活烟曲霉孢子刺激均不能增加NOD2、RIP2和TNF-αmRNA表达及细胞上清液中IL-8、TNF-α浓度;而对照组大剂量LPS和热灭活烟曲霉孢子刺激均可提高NOD2、RIP2和TNF-αmRNA表达及细胞上清液中IL-8、TNF-α浓度,尤以刺激后12h增加显著,与刺激前(0h)比较,差异有统计学意义(P0.05)。结果提示,内毒素耐受可能对NOD2信号通路有抑制作用。     

Experimental evaluation of local wave speed in the presence of reflected waves

Wave speed (also called pulse wave velocity) is the speed by which disturbance travels along the medium and it depends on the mechanical and geometrical properties of the vessel and on the density of the blood. Wave speed is a parameter of clinical relevance because it is an indicator of arterial stiffness and cardiovascular diseases.