Characterization of zebrafish mutants with defects in bone calcification during development

Using the fluorescent dyes calcein and alcian blue, we stained the F3 generation of chemically (ENU) mutagenized zebrafish embryos and larvae, and screened for mutants with defects in bone development. We identified a mutant line, bone calcification slow (bcs), which showed delayed axial vertebra calcification during development. Before 4–5 days post-fertilization (dpf), the bcs embryos did not display obvious abnormalities in bone development (i.e., normal number, size and shape of cartilage and vertebrae). At 5–6 dpf, when vertebrae calcification starts, bcs embryos began to show defects. At 7 dpf, for example, in most of the bcs embryos examined, calcein staining revealed no signals of vertebrae mineralization, whereas during the same developmental stages, 2–14 mineralized vertebrae were observed in wild-type animals. Decreases in the number of calcified vertebrae were also observed in bcs mutants when examined at 9 and 11 dpf, respectively. Interestingly, by 13 dpf the defects in bcs mutants were no longer evident. There were no significant differences in the number of calcified vertebrae between wild-type and mutant animals. We examined the expression of bone development marker genes (e.g., Sox9b, Bmp2b, and Cyp26b1, which play important roles in bone formation and calcification). In mutant fish, we observed slight increases in Sox9b expression, no alterations in Bmp2b expression, but significant increases in Cyp26b1 expression. Together, the data suggest that bcs delays axial skeletal calcification, but does not affect bone formation and maturation.     

Performance of oak,beech and spruce beams after more than 100 years in service

The aim of this study was to investigate differences in the mechanical and fungicidal properties of three different wood species (English oak (Quercus sp.), common beech (Fagus sylvatica) and Norway spruce (Picea abies)) that had been in indoor use for several decades, compared to control specimens of freshly cut timber. The collected material was cut into smaller samples prior to further analysis. Extractive content, mechanical, fungicidal and sorption properties were determined according to standard procedures. The obtained results showed that the mechanical properties of oak wood do not deteriorate over the investigated time frame. On the other hand, the resistance of oak wood against fungi decreases over time. The reason for this is yet to be confirmed; it may be due to degradation of secondary metabolites. Similar results have been reported for spruce wood. There were no statistically significant differences in the mechanical properties of old and new spruce wood. In contrast to oak wood, there were also no significant differences in fungicidal properties, bearing in mind that spruce wood has lower durability than oak wood. Aging of beech wood resulted in a considerable decrease in the tested mechanical properties but showed no significant differences in fungicidal properties. Old beech wood specimens were moderately deteriorated by insects and fungi, which was the reason for the loss of bending and compressive strength. Our results confirm that most of the relevant properties do not deteriorate with time and that wood can be reused for a variety of other applications even after decades in service.     

光照强度对大花旋蒴苣苔叶形态和生理指标的影响

以复苏植物大花旋蒴苣苔(Boea clarkeana)为盆栽实验材料,通过梯度遮光处理,研究不同光照强度(100%、80%、60%、40%和20%透光率)和不同处理时间(30、60、90、120和150d)对大花旋蒴苣苔叶生长指标和生理指标的影响。结果表明:随着光照强度的减弱,大花旋蒴苣苔叶的各生长指标(叶片长、叶片宽、叶柄长、叶柄直径以及叶面积)呈现先增大后减小的趋势,透光率为60%时达到最大值;全光照(100%透光率)时,大花旋蒴苣苔表现出明显的外伤症状,透光率为80%和20%时,分别表现出中度伤害和轻度伤害,透光率为40%和60%时,未出现明显的外伤症状;过氧化氢酶(CAT)、超氧化物歧化酶(SOD)和过氧化物酶(POD)3种抗氧化酶活性均较低且随着光强减弱呈现先降低后升高的趋势,随着处理时间的延长呈先上升后下降的趋势,全光照时抗氧化酶的活性最大,透光率60%处理下酶活性相对较低;丙二醛(MDA)含量、脯氨酸(Pro)含量和质膜透性随着光强减弱呈先降低后升高的趋势,随着处理时间的延长显著升高;叶绿素含量随着光强减弱呈逐渐上升的趋势,随着处理时间的延长先降低后升高。分析表明,40%～60%的透光率有利于大花旋蒴苣苔的生长,强光和弱光对大花旋蒴苣苔的生长均不利。     

Ostéomyélite de l’apophyse odontoïde : intérêt de la TEP/TDM au 18F-FDG dans le bilan d’extension infectieux à distance

Odontoid process is an atypical and very rare localization of osteomyelitis. We reported the case of a 72-year-old hemodialysed man with methicillin-sensitive Staphylococcus aureus osteomyelitis of the odontoid process. Osteomyelitis was diagnosed at MRI and ¹⁸F-FDG PET/CT. While clinical examination and conventional radiographs were non contributive, ¹⁸F-FDG PET/CT also allowed the diagnosis of right foot osteomylitis and multiple vertebral septic localizations. ¹⁸F-FDG PET/CT done at month 3 demonstrated a regression of the odontoid and foot hypermetabolic activity. This case illustrates the atypical presentation of this septic localization and the usefulness of ¹⁸F-FDG PET/CT to perform whole body screening and detect septic metastasis.     

Micro-CT检测中等强度跑台运动对去卵巢大鼠腰椎微结构的影响

目的用micro-CT方法,评估中等强度跑台运动对去卵巢大鼠腰椎微结构的影响。方法将30只3月龄雌性SD大鼠按体重分层后随机分为假手术、去卵巢静止和去卵巢运动三个组。运动组每周进行4次45min、速度18 m/min、坡度5°的跑台训练。正式运动处理14周时,取第2腰椎检测骨密度,取第4腰椎行micro-CT分析及三维结构重建;取第3腰椎椎体进行椎体压缩实验。结果去卵巢运动组第2腰椎骨密度、第3腰椎最大载荷、最大应力和弹性模量以及第4腰椎骨小梁体积和骨小梁数目显著高于去卵巢静止组,骨小梁分离度显著低于去卵巢静止组,而骨小梁厚度无显著变化。结论中等强度跑台运动能改善去卵巢大鼠腰椎的微结构。     

椎弓根钉棒系统加植骨融合治疗胸腰椎爆裂性骨折并脊髓损伤的 临床研究

目的:探讨椎弓根钉棒系统加植骨融合治疗胸腰椎爆裂性骨折并脊髓损伤的疗效。方法:回顾性分析2010年3月至2014年12月,采用椎弓根钉棒系统加植骨融合治疗93例胸腰椎爆裂性骨折并脊髓损伤患者的资料,男56例,女37例。致伤原因:交通事故伤27例,高处坠落伤47例,重物压伤19例。骨折节段:L1骨折22例,L2骨折16例,L3骨折6例,T11骨折15例,T12骨折34例。结果:本研究共93例患者,所有患者经过一般12个月的随访,其中平均随访13.8月(10-16月)。与术前相比,患者术后6个月伤椎前缘高度比值明显增加,Cobb角值和椎管占位率明显降低(t=6.167,7.241,7.143,P0.05)。术后12个月伤椎前缘高度比值明显增加,Cobb角值和椎管占位率明显降低(t=9.345,11.541,11.263,P0.05)。且患者术后12个月与术后6个月在伤椎前缘高度比值、Cobb角、椎管占位率上比较,差异具有统计学意义(t=9.632,8.154,7.415,P0.05),根据Frankel神经分级,术后大部分患者神经功能有所恢复。其中,Frankel分级为A的患者有35例恢复,术后有效恢复率为87.5%;B级患者有25例恢复,术后有效恢复率89.3%。C级和D级患者术后有效恢复率均为100%,B,C,D级患者与A级患者有效恢复率相比,差异没有统计学意义(x~2=0.051,2.196,1.253,P0.05),随访12个月期间,2例患者术后5个月出现内固定物松动,1例术后12个月发生螺钉断裂,其余患者无伤口感染、肺部感染、深静脉血栓等并发症发生。结论:椎弓根钉棒系统加植骨融合能有效治疗胸腰椎爆裂性骨折并脊髓神经损伤,术后患者神经功能恢复较好,并发症较少,值得临床推广使用。     

Mathematical modeling of cell growth in a 3D scaffold and validation of static and dynamic cultures

Tissue engineering, an immensely important field in contemporary clinical practices, aims at the repair or replacement of damaged tissues. The mathematical model proposed herein shows the distribution and growth of cells in their characteristic time in a 3D scaffold model. This study contributes to the progress of simulation techniques in static and dynamic cultures of bone tissue. Brinkman, nutrient transport, and cell growth equations are brought together to quantify the growth behavior of cells. However, when a static culture is being studied, the Brinkman equation is eliminated. The model was validated by experimental cell culture using 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay and scanning electron microscopy. Then, static and dynamic cultures were compared to assess the cell density and cell distribution in the scaffold. Cell counting after 21 days of cell culture showed that the number of cells increased 42‐fold in static and 53.5‐fold in dynamic cultures, which was in good agreement with our model estimations (37‐fold increase in the number of cells in static and 49‐fold increase in dynamic cultures). In conclusion, our mathematical model could predict cell distribution and growth in the scaffold.     

Does intraspecific competition promote variation? A test via synthesis

Competitive diversification, that is, when increasing intraspecific competition promotes population niche expansion, is commonly invoked in evolutionary studies and currently plays a central role in how we conceptualize the process of adaptive diversification. Despite the frequency with which this idea is cited, the empirical evidence for the process is somewhat limited, and the findings of these studies have yet to be weighed objectively through synthesis. Here, we sought to fill this gap by reviewing the existing literature and collecting the data necessary to assess the evidence for competition as a diversifying force. Additionally, we sought to test a more recent hypothesis, which suggests that competition can act to both promote and inhibit dietary diversification depending on the degree to which a consumer depletes its resources. The surprising result of this synthesis was that increasing competition did not have a mean positive effect on population‐level diet breadth or the degree of individual specialization. Instead, we found that increasing intraspecific competition had a restricting effect on population‐level diet breadth in as many cases as it had a diversifying effect. This wide disparity in the effect of competition on consumer diet variation was negatively related to a metric for consumer resource depletion. Altogether, these findings call into question a long‐standing assumption of basic evolutionary models and lend some support to recent theoretical predictions. Specifically, these findings support the idea that competition is primarily diversifying for species with a small effect (per unit biomass) on their resources and that resource depletion limits the diversifying effect of competition for consumers with larger ecological effects.     

Proteomic analysis of bone proteins adsorbed onto the surface of titanium dioxide

Osseointegration is the structural and functional connection between bone tissues and implants such as titanium dioxide (TiO₂). The bone-TiO₂ interface is thought to contain proteoglycans. However, exhaustive analysis of the proteins in this layer has not been performed. In this study, we evaluated the bone protein adhered on the surface of TiO₂ comprehensively. Pig bone protein was extracted by sequential elutions with guanidine, 0.1 M EDTA, and again with guanidine. The proteins obtained from these extractions were allowed to adhere to an HPLC column packed with TiO₂ and were eluted with 0.2 M NaOH. The eluted proteins were identified by LC/MS/MS and included not only proteoglycans but also other proteins such as extracellular matrix proteins, enzymes, and growth factors. Calcium depositions were observed on TiO₂ particles incubated with bone proteins, guanidine-extracted proteins adhered to TiO₂ displayed significantly high amounts of calcium depositions.     

Elastin‐like‐polypeptide based fusion proteins for osteogenic factor delivery in bone healing

Modern treatments of bone injuries and diseases are becoming increasingly dependent on the usage of growth factors to stimulate bone growth. Bone morphogenetic protein‐2 (BMP‐2), a potent osteogenic inductive protein, exhibits promising results in treatment models, but recently has had its practical efficacy questioned due to the lack of local retention, ectopic bone formation, and potentially lethal inflammation. Where a new delivery technique of the BMP‐2 is necessary, here we demonstrate the viability of an elastin‐like peptide (ELP) fusion protein containing BMP‐2 for delivery of the BMP‐2. This fusion protein retains the performance characteristics of both the BMP‐2 and ELP. The fusion protein was found to induce osteogenic differentiation of mesenchymal stem cells as evidenced by the production of alkaline phosphatase and extracellular calcium deposits in response to treatment by the fusion protein. Retention of the ELPs inverse phase transition property has allowed for expression of the fusion protein within a bacterial host (such as Escherichia coli) and easy and rapid purification using inverse transition cycling. The fusion protein formed self‐aggregating nanoparticles at human‐body temperature. The data collected suggests the viability of these fusion protein nanoparticles as a dosage‐efficient and location‐precise noncytotoxic delivery vehicle for BMP‐2 in bone treatment. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1029–1037, 2016