The role of apoptosis in mineralizing murine versus avian micromass culture systems

Chondrocyte apoptosis is thought to be an important step in the calcification of cartilage in vivo; however, there are conflicting reports as to whether or not this apoptosis is a necessary precursor to mineralization. The goal of this study was to determine whether or not apoptosis is necessary for mineralization in an in vitro murine micromass model of endochondral ossification. C3H10T1/2 murine mesenchymal stem cells were plated in micromass culture in the presence of 4 mM inorganic phosphate with the addition of the apoptogens, camptothecin, or staurosporine, to induce apoptosis. The rate and total accumulation of mineralization was measured with ⁴⁵Ca uptake. In these studies, both apoptogens increased the rate of mineralization, with staurosporine increasing ⁴⁵Ca accumulation by about 2.5 times that of controls and camptothecin increasing total amounts of mineralization about 1.5 times that of controls. Inhibiting cell apoptosis with the caspase inhibitor, ZVAD‐fmk, to prevent apoptosis, caused slower rates of ⁴⁵Ca uptake; however, total amounts of ⁴⁵Ca accumulation reached the same values by day 30 of culture. FTIR data showed mineralization in all samples treated with 4 mM inorganic phosphate, with the highest mineral to matrix ratios in the camptothecin treated samples. J. Cell. Biochem. 111: 653–658, 2010. © 2010 Wiley‐Liss, Inc.     

Estimation of contaminant depletion in unsaturated soils using a reduced-order biodegradation model and carbon dioxide measurement

The objective of this study was to develop a reduced-order model of biodegradation in unsaturated soils that allows the estimation of contaminant depletion, using available on-line measurements. Hexadecane was chosen as a model compound for petroleum hydrocarbons. A two-compartment model was developed, decoupling the intrinsic biodegradation kinetics from limiting factors imposed by field conditions, such as oxygen transfer and contaminant bioavailability. Two new experimental protocols (one for the liquid phase and the other for the solid phase) were developed to monitor hexadecane depletion, hexadecane mineralization, total mineralization, and evolution of the degraders. Using the liquid-phase experiment, parameters of a Haldane kinetic model and yield coefficients were identified and used in the complete model of biodegradation in soil. Using the carbon dioxide production curve, a biocontact kinetic model was identified so that, despite the high sensitivity of the model outputs to variations in the parameters, hexadecane depletion could be correctly predicted with an average error on the entire time trajectory of about 8%. Moreover, the ratio between hexadecane mineralization and total mineralization remained constant after a brief transient period, indicating that hexadecane mineralization could be deduced from the total carbon dioxide measurement. Finally, the new model developed in this study allows real-time monitoring of contaminant biodegradation, using on-line carbon dioxide measurement.     

A comparison of double vial to serum bottle radiorespirometry to measure microbial mineralization in soils

Two methods of measuring mineralization rates were compared for their ability to quantify the microbiol mineralization of organic compounds in soils. In each of three soils used, the serum bottles gave higher yields for the mineralization of both [¹⁴C]glucose and [¹⁴C]cellulose (lignocellulose) than the double vials. In two of the soils, the serum bottles also showed less variation between replicates than the double vials. Furthermore, the mineralization of glucose in the serum bottles fit a first order rate model, whereas the mineralization of glucose in the double vials showed best fit to a linear model. Results using different amounts of soils indicated that container geometry may have placed unfavorable restrictions upon the double vial system, lowering the yield of ¹⁴CO₂ from the soils. Therefore, the serum bottle method was found to be the better method for measuring mineralization rates in soils.     

Involvement of type VI collagen in interleukin-4-induced mineralization by human osteoblast-like cells in vitro

We recently showed that interleukin-4 (IL-4) enhanced collagen and osteocalcin accumulation and caused mineralization in human periosteal osteoblast-like (SaM-1) cells. At that time, the expression of alpha1(VI) collagen mRNA was induced. In the present study, the possible role of IL-4-induced type VI collagen in the in vitro mineralization in osteoblasts was investigated. Addition of IL-4 in the early stage (for the first 10 days) was essential for the mineralization. The mRNA levels of alpha1(VI) and alpha2(VI) collagen and protein level of type VI collagen were transiently increased by IL-4 treatment up to day 5, whereas the alpha1(I) procollagen mRNA level was greater at day 10 than at day 5. Addition of anti-type VI collagen antibody remarkably reduced the extracellular accumulations of calcium and hydroxyproline induced by IL-4. Furthermore, the transfection of antisense oligonucleotides of alpha1(VI) to SaM-1 cells in the presence of IL-4 partially inhibited IL-4-induced type I collagen accumulation. These results demonstrated that type VI collagen played important roles for IL-4-induced mineralization and hydroxyproline accumulation mostly type I collagen accumulation, in human periosteal osteoblast-like cells.     

Source of the soybean N credit in maize production

Nitrogen response trials throughout the United States Corn Belt show that economic optimum rates of N fertilization are usually less for maize (Zea mays L.) following soybean (Glycine max L.) than for maize following maize; however, the cause of this rotation effect is not fully understood. The objective of this study was to investigate the source of the apparent N contribution from soybean to maize (soybean N credit) by comparing soil N mineralization rates in field plots of unfertilized maize that had either nodulated soybean, non-nodulated soybean, or maize as the previous crop. Crop yields, plant N accumulation, soil inorganic N, and net soil mineralization were measured. Both grain yield (6.3 vs. 2.8 Mg ha^–1) and above-ground N accumulation (97 vs. 71 kg ha^–1) were greatly increased when maize followed nodulated soybean compared with maize following maize. A partial benefit to yield and N accumulation was also observed for maize following non-nodulated soybean. Cumulative net soil N mineralization following nodulated soybean, non-nodulated soybean, and maize was 112, 92 and 79 kg N ha^–1, respectively. Net mineralization of soil N appeared to be influenced by both quality (C:N ratio) and quantity of residue from the previous crop. In addition to an increase in plant available N from mineralization, the amount of soil inorganic N (especially in soil 5 cm from the row) was greater following nodulated soybean than non-nodulated soybean or maize. Based on these data, the soybean N credit appears to result from a combination of a decrease in net soil mineralization in continuous maize production and an increase in residual soil N from symbiotic fixation.     

重金属污染对土壤有机质积累的影响

采用田间采样分析与室内培养试验相结合的方法,研究了不同重金属污染土壤中有机质积累的差异及重金属污染强度对土壤有机质矿化动态变化的影响.结果表明：污染土壤中重金属的大量积累可减弱有机物质的矿化速率,增加土壤有机质的积累.土壤中颗粒态有机质及其占总有机碳的比例随重金属积累的增加而增加;而微生物生物量碳占总碳的比例却随土壤重金属污染水平的提高而下降.污染土壤中颗粒态有机质对重金属有显著的富集,这可能是影响土壤有机物质进一步矿化的原因之一.重金属污染可改变土壤有机质的矿化速率,影响土壤有机质的积累与分配.     

铲草和施肥对降香黄檀与印度檀香混交林土壤氮素矿化淋溶的影响

珍贵树种降香黄檀与印度檀香混交种植是当前华南地区人工林发展的一种重要模式.本研究设置对照(不做处理)、铲草、施肥、铲草+施肥4个处理,研究抚育措施对林地土壤净矿化速率、净硝化速率、净铵化速率和氮素淋溶速率的影响.结果表明:4个处理0~10 cm土层在春、秋季有最大净氮矿化速率,分别为18.92、18.13 mg·kg^-1·month^-1;在春、秋季有最大硝化速率,分别为20.35、18.85 mg·kg^-1·month^-1;夏、冬季有最大铵化速率,分别为0.22、0.26 mg·kg^-1·month^-1;秋季的氮素淋溶最严重,为15.98 mg·kg^-1·month^-1,全年最大淋溶为86.69 mg·kg^-1.铲草、施肥、铲草+施肥都在一定程度上抑制了土壤氮的净矿化和硝化速率,铲草、施肥、铲草+施肥处理年氮矿化量分别下降26.2%、16.1%、6.3%,年氮硝化量分别下降17.1%、16.6%、1.4%,同时也抑制了土壤铵态氮的累积.铲草、施肥、铲草+施肥处理全年氮素淋溶量依次减少25.2%、8.6%、6.1%.相对于铲草、施肥、铲草+施肥抚育措施,季节因素对土壤氮素矿化和淋溶过程的影响更显著.铲草、施肥、铲草+施肥措施在一定程度上抑制了土壤氮素硝化和铵化过程,减少了土壤氮素的矿化和淋溶损失量,有利于土壤肥力的保存和氮素的累积.     

In situ examination of the time-course for secondary mineralization of Haversian bone using synchrotron Fourier transform infrared microspectroscopy.

At the tissue level it is well established that the rate of remodeling is related to the degree of mineralization. However, it is unknown how long it takes for an individual bone structural unit (BSU) to become fully mineralized during secondary mineralization. Using synchrotron Fourier transform infrared microspectroscopy (FTIRM) we examined the time required for newly formed bone matrix to reach a physiological mineralization limit. Twenty-six, four-month old female New Zealand white rabbits were administered up to four different fluorochrome labels at specific time points to evaluate the chemical composition of labeled osteons from the tibial diaphysis that had mineralized for 1, 8, 18, 35, 70, 105, 140, 175, 210, 245, 280, 315, 350, and 385 days. Interstitial bone from 505 day old rabbits was used as a reference value for the physiological limit to which bone mineralizes. Using synchrotron FTIRM, area integrations were carried out on protein (Amide I: 1688-1623 cm(-1)), carbonate (v(2)CO(3)(2-): 905-825 cm(-1)), and phosphate (v(4)PO(4)(3-): 650-500 cm(-1)) IR bands. IR spectral data are presented as ratios of phosphate/protein (overall matrix mineralization) and carbonate/protein. The rate of mineralization of osteonal bone proceeded rapidly between day 1 and 18, reaching 67% of interstitial bone levels. This was followed by a slower, more progressive accumulation of mineral up to day 350. By 350 days the rate of increase plateaued. The ratio of carbonate/protein also increased rapidly during the first 18 days, reaching 73% of interstitial bone levels. The ratio of carbonate/protein plateaued by day 315, reaching levels not significantly different to interstitial bone levels. In conclusion, our data demonstrate that bone accumulates mineral rapidly during the first 18 days (primary mineralization), followed by a more gradual increase in the accumulation of mineral (secondary mineralization) which we found to be completed in 350 days.     

天山林区土壤总氮矿化过程对季节性冻融的响应

森林土壤总氮矿化对冻融过程的响应机制尚不明确,氮矿化速率和转化情况尚缺乏定量刻画。通过土壤原位法与室内培养分析相结合,利用~(15)N同位素稀释技术,研究冻融期间天山林区乔木林地、灌丛、草地3种群落类型土壤总氮矿化及转化累积量的动态,分析土壤总氮矿化速率与土壤温度、含水率及微生物量氮(MBN)的相互关系。结果表明:(1)冻融过程及群落类型对总氮矿化速率和MBN含量有极显著的影响(P0.01),秋、春季冻融期的总氮矿化速率相比冻结期更高;(2)季节性冻融期间,乔木林地土壤总氨化累积量在3种群落类型中最高(163.9 kg N hm~(-2) a~(-1)),秋、春冻融期占整个时期的比值约为66%;而总硝化累积量在3种群落类型中相差较小,秋、春冻融期占比均约为77.4%;(3)土壤温度和含水率显著影响总氮矿化速率、净氮矿化速率和MBN速率,随土壤温度增加,总氨化速率(林地和灌丛)显著升高(P0.05);随土壤含水率增加,净氨化速率(灌丛)和净硝化速率(灌丛)显著降低(P0.05)。通过揭示天山林区土壤总氮矿化速率(总氨化速率和总硝化速率)及转化累积量对冻融过程的响应情况,本研究为天山森林土壤中氮的生物地球化学过程提供了有价值的基础数据。     

Cartilage calcification in the human thoracic column

A histological and microradiological study of the cartilage calcification processes in the human thoracic column of an old man has been performed. Two different types of cartilage mineralization have been identified. The first corresponds to a calcification of the hyaline cartilage ground substance where chondrocytes are apparently intact. The second is a real mineralization of the chondrocyte lacunae in an uncalcified matrix, which we have called cartilaginous necrosis.     

Effects of chemical composition on nitrogen mineralization from green manures of seven tropical leguminous trees

Green manures from seven tropical leguminous trees were incubated with soil to determine the rates and controls of net nitrogen release. Fresh green manure (leaves and succulent twigs) was mixed with moist soil and incubated in polyethylene bags. Net N mineralization from green manures was estimated by the accumulation of extractable ammonium and nitrate minus the accumulation in soil alone. Patterns of N mineralization were complex, differed among species, and at 12 weeks ranged from 10 to 65 percent of original green-manure N. Cumulative net N mineralization was negatively correlated with initial soluble polyphenol content in the early phases of decomposition (1 through 8 weeks) and with initial lignin content in later phases (4 through 12 weeks). Neither initial percent N nor lignin: N ratio were strongly correlated with N mineralization. The best chemical index of N release was the initial polyphenol: N ratio. This study confirms previous findings that N mineralization from tropical legumes is controlled more by soluble polyphenols than by lignin or N content.     

Sites of mineral deposition in metal-accumulating cells

The basophil cells of the hepatopancreas of gastropods contain intracellular granules which act as the sites of accumulation or detoxification of environmentally available metals. A technique is described which results in the loss of these concretions and allows an ultrastructural study of their formation. Manganese ions are used as a probe for this mineralization process which is shown to occur on the inner surface of a vesicular membrane.     

不同化学性质叶凋落物添加对土壤有机碳矿化及激发效应的影响

凋落物输入可显著影响土壤有机碳(SOC)矿化速率,但添加不同化学性质叶凋落物对土壤有机碳矿化释放CO₂及激发效应的影响及其机理仍不清楚。本研究将亚热带6种树种¹³C标记的叶凋落物添加至天然次生林0～10 cm原位土柱中,比较不同树种叶凋落物添加对土壤总CO₂、外源凋落物和土壤来源CO₂释放速率和累积量以及激发效应的影响,并量化叶凋落物化学性质与土壤CO₂释放累积量、激发效应的相关关系。结果表明: 添加叶凋落物能够显著提高土壤总CO₂和土壤来源CO₂释放量,存在显著正激发效应,激发效应值为68%～128%。不同树种叶凋落物添加对土壤有机碳矿化和激发效应的影响存在显著差异。Pearson相关分析和逐步多元线性回归分析发现,凋落物来源CO₂释放累积量与叶凋落物C、P和纤维素含量呈显著负相关,而土壤来源CO₂释放量与叶凋落物C:N和木质素:N呈显著正相关。综上,不同化学性质的叶凋落物对土壤有机碳矿化和激发效应的影响存在异质性,在亚热带地区森林类型转变过程中营造具有高质量叶凋落物的人工林将有助于减少森林土壤碳损失。     

Simulation of nitrogen mineralization and transport in loess-parabrownearthes: Plot experiments

Summary In order to simulate the Nmin distributions in field plot experiments, a first order kinetic model for mineralization is combined with a transport model described earlier in this journal. Comparison of simulated and experimental figures shows satisfying agreement. The main advantages of these models are: i. only few and easy to measure input data are required and ii. both model equations, for mineralization and transport of Nmin, are simple to solve rendering short computing times even using a microcomputer.Aqualfs and Udalfs, according to the 7th approximation     

Mechanisms of glucocorticoid-induced growth retardation: impairment of cartilage mineralization

Immature female mice were treated with daily doses of triamcinolone diacetate. Using histochemical and fluorescent methods, the mineralization pattern of the proximal epiphyseal plate of the humerus was studied at various intervals. Following 1 injection, a noticeable decrease was noted in the oxytetracycline incorporation at the mineralizing zone of the growth plate. By the 10th injection, the mineralization process was almost totally arrested. Marked accumulation of phospholipids appeared in experimental plates concomitantly with a lack of acidic glycosaminoglycans' synthesis as well as degradation. Chondroclastic activity was also inhibited resulting in an increased number of hypertrophic chondrocytes at the mineralization zone. It is suggested that the hormone's antianabolic effect led to an impairment in the activity of the chondrocytes' hydrolytic enzymes and thereby interfered with the maturation of the matrix, a prerequisite phase for normal mineralization.     

松嫩平原碱化草地植物-环境系统的仿真模拟

 本研究以系统仿真的方法对松嫩平原碱化草地植物—环境系统进行模拟。模型的系统变量包括植物种的地上、地下生物量,土壤水分、有机质。可溶性和交换性Na+和Ca++浓度和植物的凋落物生物量。模型所考虑的过程有：不同土壤碱化条件下的植物生长季节动态;土壤水、盐运动;植物的蒸腾作用;土壤表面的蒸发;凋落物的积累和分解;土壤有机质的积累和矿化;地下生物量对土壤持水和导水特性的控制作用,以及收获强度对系统平衡的影响等。模型成功地解释了植物生物量形成过程与环境之间的动态耦合、相关作用。模拟结果说明：与地下生物量密切相关的土壤非毛管孔隙度与土壤的碱化和脱碱过程有极强的相关作用,这种作用是通过改变土壤的饱和持水量来实现的。非毛管孔隙度随地下生物量增加,导致饱和含水量增加和脱碱作用加强。收获强度过大导致地下生物量的减少、非毛管孔隙度的减少和碱化作用的加强。     

Age-related changes in porosity and mineralization and in-service damage accumulation

It has been proposed that bone damageability (i.e. bone's susceptibility to formation of damage) increases with the elevation or suppression of bone turnover. Suppression of turnover via bisphosphonates increases local bone mineralization, which theoretically should increase the susceptibility of bone to microcrack formation. Elevation of bone turnover has also been proposed to increase bone microdamage through an increase in bone intracortical porosity and local stresses and strains. The goal of this paper was to investigate the above proposals, i.e., whether or not increases to mineral content and porosity increase bone in-service damageability. To do this, we measured in vivo diffuse damage area (Df.Dm.Ar, %) and microcrack density (Cr.Dn) (cracks/mm(2)) in the same specimen from human cortical bone of the midshaft of the proximal femur obtained from cadavers with an age range of eight decades and examined their relationships with porosity, mineralization and age. Results of this study showed that Cr.Dn and Df.Dm.Ar increased with a decrease in bulk mineralization. This finding does not appear to support the proposal that damage accumulation increases with low bone turnover that results in increases mineralization. It was proposed however that the negative correlation between damage accumulation and mineralization may be attributed to highly mineralized regions of bone existing with under-mineralized regions resulting in an overall decrease in average bone mineralization. It was also found that microdamage accumulates with increasing porosity which does appear to support the proposal that elevated bone turnover that results in increased porosity can accelerate microdamage accumulation. Finally, it was shown that linear microcracks and Df.Dm.Ar accumulate with age differently, but because they correlate with each other, one may be the precursor for the other.     

Correlation between loss of alkaline phosphatase activity and accumulation of calcium during matrix vesicle-mediated mineralization

Activity of the bone/liver/kidney isozyme of alkaline phosphatase (AP) is known to be critical for mineralization in developing bone, although its role is unclear. The work now reported explores changes in the activity of this Zn2+-containing enzyme that occur during Ca2+ accumulation by matrix vesicles (MV). A marked loss (up to 65-70%) in AP activity was found to accompany Ca2+ accumulation by MV. These two events were highly correlated, both temporally and quantitatively. Investigation into possible causes revealed that the decline in AP activity during Ca2+ uptake was not due to action of proteases but rather resulted from interaction with the developing mineral phase, loss of metal ions (Zn2+ and Mg2+) from the active site of the enzyme, and concomitant irreversible denaturation of the enzyme. Protease inhibitors did not protect AP from loss of activity during mineralization; in contrast, protease treatments, which progressively destroyed the ability of MV to accumulate Ca2+ actually reduced loss of AP activity. These findings clearly demonstrate that AP is present at the site of MV mineralization and that its catalytic activity is profoundly reduced by the mineralization process.     

松嫩平原碱化草地植物－环境系统的仿真模拟

本研究以系统仿真的方法对松嫩平原碱化草地植物－环境系统进行模拟。模型的系统变量包括植物种的地上、地下生物量,土壤水分、有机质、可溶性和交换性Ｎａ￣＋和Ｃａ￣＋＋浓度和植物的凋落物生物量。模型所考虑的过程有：不同土壤碱化条件下的植物生长季节动态;土壤水、盐运动;植物的蒸腾作用;土壤表面的蒸发;凋落物的积累和分解;土壤有机质的积累和矿化;地下生物量对土壤持水和导水特性的控制作用,以及收获强度对系统平衡的影响等。模型成功地解释了植物生物量形成过程与环境之间的动态耦合、相关作用。模拟结果说明：与地下生物量密切相关的土壤非毛管孔隙度与土壤的碱化和脱碱过程有极强的相关作用,这种作用是通过改变土壤的饱和持水量来实现的。非毛管孔隙度随地下生物量增加．导致饱和含水量增加和脱碱作用加强。收获强度过大导致地下生物量的减少、非毛管孔隙度的减少和碱化作用的加强。     

Hierarchical modeling of the elastic properties of bone at submicron scales: the role of extrafibrillar mineralization

We model the elastic properties of bone at the level of mineralized collagen fibrils via step-by-step homogenization from the staggered arrangement of collagen molecules up to an array of parallel mineralized fibrils. A new model for extrafibrillar mineralization is proposed, assuming that the extrafibrillar minerals are mechanically equivalent to reinforcing rings coating each individual fibril. Our modeling suggests that no more than 30% of the total mineral content is extrafibrillar and the fraction of extrafibrillar minerals grows linearly with the overall degree of mineralization. It is shown that the extrafibrillar mineralization considerably reinforces the fibrils’ mechanical properties in the transverse directions and the fibrils’ shear moduli. The model predictions for the elastic moduli and constants are found to be in a good agreement with the experimental data reported in the literature.