Increased initial cement–bone interlock correlates with reduced total knee arthroplasty micro-motion following in vivo service

Aseptic loosening of cemented tibial components in total knee arthroplasty (TKA) has been related to inadequate cement penetration into the trabecular bone bed during implantation. Recent postmortem retrieval work has also shown there is loss of interlock between cement and bone by resorption of trabeculae at the interface. The goal of this study was to determine if TKAs with more initial interlock between cement and bone would maintain more interlock with in vivo service (in the face of resorbing trabeculae) and have less micro-motion at the cement–bone interface. The initial (created at surgery) and current (after in vivo service) cement–bone interlock morphologies of sagittal implant sections from postmortem retrieved tibial tray constructs were measured. The implant sections were then functionally loaded in compression and the micro-motion across the cement–bone interface was quantified. Implant sections with less initial interdigitation between cement and bone and more time in service had less current cement–bone interdigitation (r²=0.86, p=0.0002). Implant sections with greater initial interdigitation also had less micro-motion after in vivo service (r²=0.36, p=0.0062). This work provides direct evidence that greater initial interlock between cement and bone in tibial components of TKA results in more stable constructs with less micro-motion with in vivo service.     

不同杂种优势群玉米籽粒脱水速率分析

研究不同杂种优势群玉米自交系籽粒脱水速率的特性,筛选脱水速率快的自交系,为选育适应机械化作业的玉米杂交种提供借鉴。本试验采用烘干测定173份玉米自交系的籽粒的脱水速率及其相关性状,利用覆盖玉米全基因组的210对SSR标记对实验材料进行全基因组扫描,通过Structure V2.3.4 软件揭示其群体结构。对不同杂种优势群平均籽粒脱水速率进行方差分析,并筛选出各个群中籽粒灌浆速率快的自交系。研究结果如下：籽粒脱水速率在不同自交系间存在显著差异,与苞叶、穗轴及籽粒的含水率等性状间存在显著相关性。试验共筛选到脱水速率大于1%的自交系20个;授粉后40天时籽粒的含水率低于21%的自交系10个。参试自交系分成P、旅大红骨、瑞德、兰卡斯特和塘四平头5个杂种优势群;授粉后40天脱水速率依次是Reid群0.92%、Lancaster群0.85%、旅大红骨群0.82%、混合群0.80%、P群0.76%、塘四平头群0.56%。     

Synthesis and anti-proliferative activity of a novel 1,2,3-triazole tethered chalcone acetamide derivatives

A new series of 1,2,3-triazole tethered chalcone acetamide derivatives (7a-c & 8a-r) have been synthesized in excellent yields and their structures were determined by analytical and spectral (FT-IR, ¹H NMR, ¹³C NMR & HRMS) studies. The newly synthesized derivatives were evaluated for their cytotoxic activity against four human cancer cell lines, such as HeLa (Human cervical cancer), A549 (Human alveolar adenocarcinoma), MCF-7 (Human breast adenocarcinoma) and SKNSH (Human brain cancer). Among them, compound 7c exhibited good anti-proliferation activity with HeLa (IC₅₀ 7.41 + 0.8 μM), SKNSH (IC₅₀ 8.68 + 1.1 μM), MCF-7 (IC₅₀ 9.76 + 1.3 μM) and MDA-MB-231, while compounds 7a and 7b showed promising anti-proliferation against above four human cancer cell lines with IC₅₀ 7.95–11.62 μM, respectively, compared with the standard drug Doxorubicin. We explored the probable key active site and binding mode interactions in HDAC8 (PDB ID:3SFH) and EHMT2 (PDB ID:3K5K) proteins. The docking results are complementary to the experimental observations.     

Batch production and growth kinetics of hybridomas

Batch cultures of mouse-mouse hybridoma cell lines were carried out and their growth and production kinetics investigated. Three main cell specific production patterns (expressed as pg IgG/cell x hour) were found, which can be used as a classification system for hybridoma cell lines (groups I–III). Cells showing the highest IgG-production at the beginning of the batch culture (during the lag and the onset of the log-phase) were classified as either group I and II. The difference was that cell lines of group II showed a second high cell specific production at the onset of the stationary and death phases. Cell lines of group III had a quite constant production of antibodies during their growth; but IgG secretion completely stopped after the beginning of the stationary phase. The implications of these three production patterns on the design of a production process are discussed.     

Metabolism of polyamines in NaCl-resistant cell lines from Nicotiana sylvestris

The polyamine titers in three cell lines of Nicotiana sylvestris were compared: Type 1, rapidly adapting to NaCl; Type 2, constantly resistant to NaCl; Type 3, a saltsensitive wild strain. During short-term cultivation in MS medium in the presence of 170 mM NaCl (1 passage, 14 d) the changes in polyamine titer in cell suspensions of type 1 (in a slightly adapted state) and non-adapted wild strain (type 3) showed a considerable increase in spermidine and spermine and a decrease in putrescine. After prolonged adaptation to NaCl (20 passages) the putrescine content in the cells of type 1 and of type 2 was increased at the expense of the polyamines. This suggests that the pattern of polyamine titer varies under short- and long-term adaptation to NaCl. The inverse ratio between growth processes and changes in polyamine and proline level indicates that polyamines fulfil primarily a protective and osmorepulatory function in plant cells under NaCl stress.     

Identification of differentially expressed genes in three-pistil mutation in wheat using annealing control primer system

The common wheat line three-pistil (TP) is a valuable mutant for wheat breeding. The TP mutation has normal spike morphology; however, it only produces three pistils per floret. Therefore, it has potential to increase the grain number per spike. In order to determine the underlying molecular mechanism, an annealing control primer system was used to identify the different expressed genes in three-pistil mutation. Using 120 arbitrary ACP primers, we identified three differentially expressed genes in young spikes between two near-isogenic lines (i.e., Chuanmai 28 TP and Chinese Spring TP) and their recurrent parents. We tentatively designated the three differentially expressed genes as DETP-1, DETP-2, and DETP-3. DETP-1 showed similar function with maize cytoplasmic membrane protein, which is involved in cell division in bacteria. DETP-3 is homologous to maize endo-1, 4-beta-glucanase (EGases), which is associated with plant development, cell wall loosening, stem flowering, and root expansion. DETP-2 showed no significant hit with any sequence found in the database and translates unknown protein. These genes would likely play an important role in determining the three pistils trait in wheat.     

Transgene flow to hybrid rice and its male-sterile lines

Gene flow from genetically modified (GM) crops to the same species or wild relatives is a major concern in risk assessment. Transgenic rice with insect and/or disease resistance, herbicide, salt and/or drought tolerance and improved quality has been successfully developed. However, data on rice gene flow from environmental risk assessment studies are currently insufficient for the large-scale commercialization of GM rice. We have provided data on the gene flow frequency at 17 distances between a GM japonica line containing the bar gene as a pollen donor and two indica hybrid rice varieties and four male-sterile (ms) lines. The GM line was planted in a 640m² in an isolated experimental plot (2.4 ha), which simulates actual conditions of rice production with pollen competition. Results showed that: (1) under parallel plantation at the 0-m zone, the transgene flow frequency to the ms lines ranged from 3.145 to 36.116% and was significantly higher than that to hybrid rice cultivars (0.037–0.045%). (2) Gene flow frequency decreased as the distance increased, with a sharp cutoff point at about 1–2 m; (3) The maximum distance of transgene flow was 30–40 m to rice cultivars and 40–150 m to ms lines. We believe that these data will be useful for the risk assessment and management of transgenic rice lines, especially in Asia where 90% of world's rice is produced and hybrid rice varieties are extensively used. Shirong Jia, Feng Wang and Lei Shi contributed equally to this investigation.     

Derivation and characterization of cholesterol-independent non-GS NS0 cell lines for production of recombinant antibodies

Presented is an antibody production platform based on the fed-batch culture of recombinant NS0-derived cell lines. NS0 host cells, obtained from the European Collection of Cell Cultures (ECACC, Salisbury, UK, Part No. 85110503), were first adapted to grow in a protein-free, cholesterol-free medium. The resulting host cell line was designated NS0-PFCF (protein-free, cholesterol-free). The five production cell lines presented here were generated using a common protocol consisting of transfection by electroporation and subcloning. The NS0-PFCF host cell line was transfected using a single expression vector containing the Escherichia coli xanthine-guanine phosphoribosyl transferase gene (gpt), and the antibody heavy and light chain genes driven by the CMV promoter. The five cell lines were chosen after one to three rounds of iterative subcloning, which resulted in a 19-64% increase in antibody productivity when four mother-daughter cell pairs were cultured in a fed-batch bioreactor process. The production cell lines were genetically characterized to determine antibody gene integrity, nucleotide sequences, copy number, and the number of insertion sites in the NS0 cell genome. Genetic characterization data indicate that each of the five production cell lines has a single stably integrated copy of the antibody expression vector, and that the antibody genes are correctly expressed. Stability of antibody production was evaluated for three of the five cell lines by comparing the early stage seed bank with the Working Cell Bank (WCB). Antibody productivity was shown to be stable in two of three cell lines evaluated, while one of the cell lines exhibited a 20% drop in productivity after passaging for approximately 4 weeks. These five NS0-derived production cell lines were successfully cultured to produce antibodies with acceptable product quality attributes in a standardized fed-batch bioreactor process, consistently achieving an average specific productivity of 20-60 pg/cell-day, and a volumetric productivity exceeding 120 mg/L-day (Burky et al., 2006). In contrast to the commonly available NS0 host cell line, which requires serum and cholesterol for growth, and the commonly used expression vector system, which uses a proprietary glutamine synthetase selection marker (GS-NS0), these NS0 cells are cholesterol-independent, grow well in a protein-free medium, use a non-proprietary selection marker, and do not require gene amplification for productivity improvement. These characteristics are advantageous for use of this NS0 cell line platform for manufacturing therapeutic antibodies.     

普通小麦—提莫菲维小麦白粉病抗性渐渗系中渗入片段的准确鉴定

用小麦（ Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍ Ｌ．） 第二部分同源群的３６ 个探针,对携带抗白粉病（ Ｅｒｙｓｉｐｈｅ ｇｒａｍｉｎｉｓｆ．ｓｐ ｔｒｉｔｉｃｉ） 基因Ｐｍ６ 的普通小麦提莫菲维小麦（ Ｔ．ｔｉｍｏｐｈｅｅｖｉ Ｚｈｕｋ ．） 渐渗系进行检测,通过这些渐渗系与受体亲本“Ｐｒｉｎｓ”之间杂交谱带多态性的比较,发现所测出多态性标记位点均位于这５ 个渐渗系的２Ｂ 染色体上,但其渗入片段的位置与大小有明显区别。ＩＧＶ１＿４５６ 和ＩＧＶ１＿４５８ 被鉴定为２Ｂ 染色体从短臂标记Ｘｃｄｏ４０５ 到长臂标记Ｘｂｃｄ１３５ 之间的区域已被提莫菲维２Ｇ 染色体区段所代替;而ＩＧＶ１＿４６３ 则在２Ｂ 染色体长臂Ｘｂｃｄ３０７ 到Ｘｃｄｏ６７８ 标记之间的区域被提莫菲维渗入成分所代替;ＩＧＶ１＿４６４ 、ＩＧＶ１＿４６５ ２ＢＬ染色体上的渗入片段更小,即ＩＧＶ１＿４６４ 在２ＢＬ染色体上标记Ｘｐｓｒ９３４ 与Ｘｂｃｄ１３５ 之间的区域有提莫菲维２Ｇ 染色质成分渗入,ＩＧＶ１＿４６５ 仅在Ｘｂｃｄ１３５ 附近有更小的外源成分渗入。根据５个渐渗系渗入成分重叠区段比较可把Ｐｍ６ 定位于２ＢＬ染色体上Ｘｂｃｄ１３５ ２ＢＬ标记的邻近区域内