Seed germination and dormancy traits of forbs and shrubs important for restoration of North American dryland ecosystems

• In degraded dryland systems, native plant community re‐establishment following disturbance is almost exclusively carried out using seeds, but these efforts commonly fail. Much of this failure can be attributed to the limited understanding of seed dormancy and germination traits.
• We undertook a systematic classification of seed dormancy of 26 species of annual and perennial forbs and shrubs that represent key, dominant genera used in restoration of the Great Basin ecosystem in the western United States. We examined germination across a wide thermal profile to depict species‐specific characteristics and assessed the potential of gibberellic acid (GA₃) and karrikinolide (KAR₁) to expand the thermal germination envelope of fresh seeds.
• Of the tested species, 81% produce seeds that are dormant at maturity. The largest proportion (62%) exhibited physiological (PD), followed by physical (PY, 8%), combinational (PY + PD, 8%) and morphophysiological (MPD, 4%) dormancy classes. The effects of chemical stimulants were temperature‐ and species‐mediated. In general, mean germination across the thermal profile was improved by GA₃ and KAR₁ for 11 and five species, respectively. We detected a strong germination response to temperature in freshly collected seeds of 20 species. Temperatures below 10 °C limited the germination of all except Agoseris heterophylla, suggesting that in their dormant state, the majority of these species are thermally restricted.
• Our findings demonstrate the utility of dormancy classification as a foundation for understanding the critical regenerative traits in these ecologically important species and highlight its importance in restoration planning.

     

Oxygen uptake rates and liver-specific functions of hepatocyte and 3T3 fibroblast co-cultures

Bioartificial liver (BAL) devices have been developed to treat patients undergoing acute liver failure. One of the most important parameters to consider in designing these devices is the oxygen consumption rate of the seeded hepatocytes which are known to have oxygen consumption rates 10 times higher than most other cell types. Hepatocytes in various culture configurations have been tested in BAL devices including those formats that involve co-culture of hepatocytes with other cell types. In this study, we investigated, for the first time, oxygen uptake rates (OUR)s of hepatocytes co-cultured with 3T3-J2 fibroblasts at various hepatocyte to fibroblast seeding ratios. OURs were determined by measuring the rate of oxygen disappearance using a ruthenium-coated optical probe after closing and sealing the culture dish. Albumin and urea production rates were measured to assess hepatocyte function. Lower hepatocyte density co-cultures demonstrated significantly higher OURs (2 to 3.5-fold) and liver- specific functions (1.6-fold for albumin and 4.5-fold for urea production) on a per cell basis than those seeded at higher densities. Increases in OUR correlated well with increased liver-specific functions. OURs (V(m)) were modeled by fitting Michaelis-Menten kinetics and the model predictions closely correlated with the experimental data. This study provides useful information for predicting BAL design parameters that will avoid oxygen limitations, as well as maximize metabolic functions.     

Assessing the performance of pea and lentil at different seeding densities as trap crops for the management of wireworms in spring wheat

Wireworms, the larvae of click beetles (Elateridae), are difficult to manage due to their habitats and behaviour. Wireworms pose a major threat to the wheat crop in the north‐western USA. Seed treatment with neonicotinoids, biological control management and some cultural controls are recommended to manage these pests. Trap cropping is an emerging way to manage wireworms. In strawberry and potato crops, trap cropping has been found effective at attracting wireworms away from the principal crop. An earlier study in the Golden Triangle area of Montana found that pea and lentil could be effective trap crops for managing wireworms in spring wheat. In the present study, experiments were conducted at two locations. The effectiveness of peas and lentils as trap crops with wheat at different seeding densities was assessed [pea at 0, 4, 8, 16 seeds/sq.ft. or 0, 43, 86, 172 seeds/sq.m.; lentil at 0, 6, 12, 18 OR 0, 65, 130, 194 seeds/sq.m.; both with wheat at 0, 11, 22, or 28 seeds/sq.ft. or 0, 120, 230, 300 seeds/sq.m.] in a randomized design where all three crops were intercropped. Both trap crops were found to be effective in protecting wheat at standard seeding rates of 8 seeds/sq.ft. for pea and 12 seeds/sq.ft. for lentils. At these seeding rates, higher numbers of wireworms were found to be attracted to the trap crop, resulting in higher yield (7%–10%) of the associated spring wheat plant stands at 22 seeds/sq.ft. To develop an effective trap crop strategy, the pea–wheat and lentil–wheat spatial patterns that are possible need to be assessed in further field trials. Proper design and evaluation of the cost–benefit ratio of pea and lentil as trap crops are likely to produce good results for wheat crops in Montana.     

The Impact of Seeding Method on Diversity and Plant Distribution in Two Restored Grasslands

Previous studies have compared grassland restoration techniques based on resulting species richness and composition. However, none have determined if different techniques generate different plant distributions in space, which may further impact restoration success. This study tests if there are quadrat‐scale (1 m²) differences between paired drilled and broadcast plantings in diversity, composition, and plant distributions. Higher competition intensity in and more contiguous spaces between rows in drill‐seeded restorations were hypothesized to result in larger patches of native grasses and exotic species. Two paired drill‐ and broadcast‐seeded plantings were sampled in June 2007 in Iowa, U.S.A. Within 10 quadrats in each planting, we measured species abundance with point intercept sampling and plant distributions by dividing the quadrat into 64 cells and recording the most abundant species in each cell. Drilled and broadcast plantings at both sites had similar Simpson’s diversity and evenness. However, the effect of planting type on species richness, composition, and plant distribution was site dependent. Native warm‐season grasses in one site, and exotic species in the second, occupied more space and were distributed in larger patches in drilled plantings. Furthermore, drilled canopies consistently captured more light than broadcast canopies. This suggests that initial differences in seed placement can affect resulting plant distributions, resource use, and potentially long‐term species turnover. Mechanisms structuring vegetation in these communities need to be further investigated to determine if this approach can provide more information on long‐term diversity maintenance in restorations than traditional measures.     

Observation of sequence specificity in the seeding of protein amyloid fibrils

It is well established that the rate of formation of fibrils by amyloidogenic proteins is enhanced by the addition of preformed fibrils, a phenomenon known as seeding. We show that the efficiency of seeding fibril formation from solutions of hen lysozyme by a series of other proteins depends strongly on the similarity of their sequences. This observation is consistent with the importance of long-range interactions in stabilizing the core structure of amyloid fibrils and may be associated with the existence of a species barrier observed in the transmissible spongiform encephalopathies. In addition, it is consistent with the observation of a single dominant type of protein in the deposits associated with each form of amyloid disease.     

Oscillating perfusion of cell suspensions through three-dimensional scaffolds enhances cell seeding efficiency and uniformity

We developed a bioreactor for automated cell seeding of three-dimensional scaffolds by continuous perfusion of a cell suspension through the scaffold pores in oscillating directions. Using quantitative biochemical and image analysis techniques, we then evaluated the efficiency and uniformity of perfusion seeding of Polyactive foams as compared to conventional static and spinner flask methods. Finally, we assessed the efficacy of the perfusion seeding technique for different scaffolds and cell types. Perfusion seeding of chondrocytes into Polyactive foams resulted in "viable cell seeding efficiencies," defined as the percentages of initially loaded cells that were seeded and remained viable, that were significantly higher (75 +/- 6%) than those by static (57% +/- 5%) and spinner flask seeding (55% +/- 8%). In addition, as compared to static and spinner flask methods, cells seeded by perfusion were respectively 2.6-fold and 3.8-fold more uniformly distributed and formed more homogeneously sized cell clusters. Chondrocytes seeded by perfusion into Hyaff-11 nonwoven meshes were 26% and 63%, respectively, more uniformly distributed than following static and spinner flask seeding. Bone marrow stromal cells seeded by perfusion into ChronOS porous ceramics were homogeneously distributed throughout the scaffold volume, while following the static method, cells were found only near the top surface of the ceramic. In summary, we demonstrated that our cell seeding perfusion bioreactor generated constructs with remarkably uniform cell distributions at high efficiencies, and was effective for a variety of scaffolds and different mesenchymal cell types.     

Multistep nucleus formation and a separate subunit contribution of the amyloidgenesis of heat-denatured monellin

Monellin (MN) is a sweet-tasting plant protein known to form fibrous aggregates in the heat-denatured state. Here the amyloid-type aggregation process of MN is extensively characterized. The amyloidgenesis was initiated in a highly denatured state of MN. A seeding effect of skipping a lag phase of the amyloid formation kinetics established a nucleation-dependent aggregation mechanism. A finely controlled experimental protocol revealed an additional prenucleus stage preceding the maturation of the nucleus, indicating that the initial lag phase is composed of multiple conformational events. The results obtained for the aggregation properties of the separate A and B subunit chains of MN and a recombinant single-chain MN suggest that the B chain exclusively contributed to the amyloid-type aggregation. These findings suggest a scheme for the amyloidgenesis of MN and their subunits, and provide a unique model of amyloidgenesis that is regulated by the subunit composition of protein.     

经皮椎板间内镜与椎板小开窗术治疗腰椎间盘突出症临床疗效比较

目的：分析和比较椎板间内镜与椎板小开窗术治疗腰椎间盘突出症的临床疗效和安全性指标。方法：使用回顾性分析的方法 对2012-2014 年共计126 例在我科行椎板间内镜手术或椎板小开窗手术的腰椎间盘突出患者进行分析和比较。通过纳入和排除 标准的筛选,经皮椎板间内镜组纳入48例,椎板小开窗组纳入78 例。结合详实的术后随访,对两组患者在花费,住院时间等一般 性指标,疼痛指标,功能指标,并发症等数据进行分析和比较。结果：两组患者在术后均取得明显的治疗疗效,在疼痛、功能等指标 中都有明显的改善。但两组之间并无明显统计学差异(P>0.05)。而椎板间内镜组在住院时间,出血量,切口长度及并发症等方面明 显的优于小开窗组,具有统计学意义(P<0.05)。结论：经皮椎板间内镜手术作为一种脊柱微创手术,治疗效果确切,安全性好,能体 现微创的优势,可作为椎间孔镜技术在治疗椎间盘突出症的有益补充,在临床中进一步的开展和推广。     

Establishment and characterization of a cell line from embryos of the Indianmeal moth,Plodia interpunctella

A cell line, UMN-PIE-1181, initiated in November, 1981, from embryos of a malathion-resistant strain of Indianmeal moth, Plodia interpunctella, was in the 83rd passage on January 28, 1985. The line consists of single, small, fibroblastlike cells that are polyploid with chromosome numbers ranging from 56 to 180. Growth rate is dependent on seeding density, there being no growth at or below seeding densities of $\text{2 ×}\text{10}{}^5\text{5}\text{,}\text{ml}$; optimum growth requires a fetal bovine serum concentration of at least 5%. Twenty-nine isozymes were examined. Five enzymes from the cell lines resolved well and subsequently were compared to enzymes extracted from 4-day-old embryos and other life stages of the insects. Phosphomannose isomerase, malic enzyme, malate dehydrogenase, phosphoglucose isomerase, and glucose-6-phosphate dehydrogenase in extracts from the cultured cells and from the insects had identical patterns. Two bands for glutamate-oxalacetate transaminase, present in the cell line, were not observed in the tissue extracts. Furthermore, lactate dehydrogenase from the cultured cells appeared as four bands but was not detectable in any of the samples run from the various life stages of the insects.     

A power-driven increment borer for sampling high-density tropical wood

High-density hardwood trees with large diameters have been found to damage manually operated increment borers, thus limiting their use in the tropics. Therefore, we herein report a new, low-cost gasoline-powered sampling system for high-density tropical hardwood trees with large diameters. This system provides increment cores 15 mm in diameter and up to 1.35 m in length, allowing minimally invasive sampling of tropical hardwood tree species, which, up to the present, could not be collected by conventional 5 or 10 mm increment borers. This system provides a single core sample with ample amount of wood for multidisciplinary analyses, including ring width, stable isotope and wood anatomical measurements. The borer never gets stuck inside stems, even in hollowed trees, cores will never twist during coring, and the gasoline drill gives ample flexibility in the field. It is anticipated that the dendrochronological community will find our technique very useful in the pursuit of tropical tree ring research.