Effect of cytokinins on plastid development and photosynthetic polypeptides during organogenesis ofPinus ponderosa Dougl. cotyledons culturedin vitro

InPinus ponderosa Dougl., application of the cytokinins, benzyladenine and 2-isopentenyl adenine, to excised cotyledons, promoted thein vitro formation of meristematic centers which led to bud and shoot production. Meristematic cells showed plastids with poorly developed thylakoid membranes and rudimentary grana, whereas cells in non-meristematic tissues and in growth regulator free medium, had chloroplasts with well developed inner membranes, and more thylakoid membranes and grana than plastids of meristematic cells. Chlorophyll and six polypeptides associated with photosynthesis were present in lower concentrations in cytokinin-treated cotyledons than in those cultured in growth regulator free medium. Both benzyladenine and 2-isopentenyl adenine are effective in inhibiting the accumulation of at least two photosynthetic polypeptides in the first 24 h in culture. The ability of cotyledons to respond in this way to cytokinins is lost after three days in culture in growth regulator free medium prior to treatment with cytokinin.     

In vitro characterisation of a monovalent and bivalent form of a fully human anti Ep-CAM phage antibody

 Antibodies to tumour-associated antigens are increasingly being used as targeting vehicles for the visualisation and for therapy of human solid tumours. The epithelial cell adhesion molecule (Ep-CAM) is an antigen that is overexpressed on a variety of human solid tumours and constitutes an attractive target for immunotargeting. We set out to obtain fully human antibodies to this antigen by selecting from a large antibody repertoire displayed on bacteriophages. Two single-chain variable antibody fragments (scFv) were identified that specifically bound recombinant antigen in vitro. One of the selected antibodies (VEL-1) cross-reacted with extracellular matrix components in immunohistochemistry of colon carcinoma, whereas the other scFv (VEL-2) specifically recognised colon cancer cells. The latter antibody was further characterised with respect to epitope specificity and kinetics of antigen-binding. It showed no competition with the well-characterised anti Ep-CAM MOC-31 monoclonal antibody and had an off-rate of 5 × 10⁻² s⁻¹. To obtain an antibody format more suitable for in vivo tumour targeting and to increase the apparent affinity through avidity, the genes of scFv VEL-2 were re-formatted by fusion to a human (γ1) hinge region and CH3 domain. This “minibody” was expressed in Escherichia coli, specifically bound the Ep-CAM antigen and showed a 20-fold reduced off-rate in surface plasmon resonance analysis. These results show that phage antibody selection, combined with antibody engineering, may result in fully human antibody molecules with promising characteristics for in vivo use in tumour targeting. Received: 13 July 2000 / Accepted: 12 October 2000     

Identification of MK-5710 ((8aS)-8a-methyl-1,3-dioxo-2-[(1S,2R)-2-phenylcyclo- propyl]-N-(1-phenyl-1H-pyrazol-5-yl)hexahydro-imidazo[1,5-a]pyrazine-7(1H)-carboxamide), a potent smoothened antagonist for use in Hedgehog pathway dependent malignancies, part 2

The Hedgehog (Hh-) signaling pathway is a key developmental pathway which gets reactivated in many human tumors, and smoothened (Smo) antagonists are emerging as novel agents for the treatment of malignancies dependent on the Hh-pathway, with the most advanced compounds demonstrating encouraging results in initial clinical trials. A novel series of potent bicyclic hydantoin Smo antagonists was reported in the preceding article, these have been resolved, and optimized to identify potent homochiral derivatives with clean off-target profiles and good pharmacokinetic properties in preclinical species. While showing in vivo efficacy in mouse allograft models, unsubstituted bicyclic tetrahydroimidazo[1,5-a]pyrazine-1,3(2H,5H)-diones were shown to epimerize in plasma. Alkylation of the C-8 position blocks this epimerization, resulting in the identification of MK-5710 (47) which was selected for further development.     

Smog nitrogen and the rapid acidification of forest soil, San Bernardino Mountains, southern California

We report the rapid acidification of forest soils in the San Bernardino Mountains of southern California. After 30 years, soil to a depth of 25 cm has decreased from a pH (measured in 0.01 M CaCl2) of 4.8 to 3.1. At the 50-cm depth, it has changed from a pH of 4.8 to 4.2. We attribute this rapid change in soil reactivity to very high rates of anthropogenic atmospheric nitrogen (N) added to the soil surface (72 kg ha(-1) year(-1)) from wet, dry, and fog deposition under a Mediterranean climate. Our research suggests that a soil textural discontinuity, related to a buried ancient landsurface, contributes to this rapid acidification by controlling the spatial and temporal movement of precipitation into the landsurface. As a result, the depth to which dissolved anthropogenic N as nitrate (NO3) is leached early in the winter wet season is limited to within the top approximately 130 cm of soil where it accumulates and increases soil acidity.     

Towards the Generation of B-Cell Receptor Retrogenic Mice

Transgenic expression of B- and T-cell receptors (BCRs and TCRs, respectively) has been a standard tool to study lymphocyte development and function in vivo. The generation of transgenic mice is time-consuming and, therefore, a faster method to study the biology of defined lymphocyte receptors in vivo would be highly welcome. Using 2A peptide-linked multicistronic retroviral vectors to transduce stem cells, TCRs can be expressed rapidly in mice of any background. We aimed at adopting this retrogenic technology to the in vivo expression of BCRs. Using a well characterised BCR specific for hen egg lysozyme (HEL), we achieved surface expression of the retrogenically encoded BCR in a Rag-deficient pro B-cell line in vitro. In vivo, retrogenic BCRs were detectable only intracellularly but not on the surface of B cells from wild type or Rag2-deficient mice. This data, together with the fact that no BCR retrogenic mouse model has been published in the 7 years since the method was originally published for TCRs, strongly suggests that achieving BCR-expression in vivo with retrogenic technology is highly challenging if not impossible.     

遗传学课程思政教育的探索与实践

课程思政对高校落实立德树人、实现人才培养目标具有重要的意义。在专业课中进行课程思政改革有助于探究高校专业课程中育人功能的发挥。遗传学是生命科学专业重要的基础课程,在生命科学知识体系的构建中具有重要作用。立足内蒙古农业大学示范课程改革的教学实践,我们重塑了教学目标,改革教学方法,挖掘遗传学课程中蕴含的思政元素,使德育教育深入教师心、学生心,贯穿于教学、教评全过程,使知识传授、能力培养与价值引领同向同行,以提升学生的综合素质。     

Tit for tat? A mycorrhizal fungus accumulates phosphorus under low plant carbon availability

The exchange of carbohydrates and mineral nutrients in the arbuscular mycorrhizal (AM) symbiosis must be controlled by both partners in order to sustain an evolutionarily stable mutualism. Plants downregulate their carbon (C) flow to the fungus when nutrient levels are sufficient, while the mechanism controlling fungal nutrient transfer is unknown. Here, we show that the fungus accumulates nutrients when connected to a host that is of less benefit to the fungus, indicating a potential of the fungus to control the transfer of nutrients. We used a monoxenic in vitro model of root organ cultures associated with Glomus intraradices, in which we manipulated the C availability to the plant. We found that G. intraradices accumulated up to seven times more nutrients in its spores, and up to nine times more in its hyphae, when the C pool available to the associated roots was halved. The strongest effect was found for phosphorus (P), considered to be the most important nutrient in the AM symbiosis. Other elements such as potassium and chorine were also accumulated, but to a lesser extent, while no accumulation of iron or manganese was found. Our results suggest a functional linkage between C and P exchange.     

Embryologische und histogenetische Studien an „ monokotylen Dikotylen“ III.Anemone apennina L.

Zusammenfassung Anemone apennina gehört in die kleine Grappe der einkeimblättrigen Dikotylen-Arten.Der einzige Kotyledo wird lateral angelegt, drängt aber den Sproßvegetationspunkt später etwas zur Seite. Von der Anlage eines zweiten Kotyledos fehlt jede Spur.Weder der Embryogenese noch der Entwicklungsgeschichte und der Anatomie der Keimpflanzen sind Hinweise für das Vorliegen einer kongenitalen Verwachsung zweier Kotyledonen (Synkotylie) zu entnehmen.Der Kotyledo ist in gerader Linie über dem einen Pol der Xylemplatte der diarchen Keimachsenstele inseriert. Der Kotyledonarstiel wird von einem einfachen Leitstrang durchzogen, der im basalen Teil als diarche Stele und im apikalen als kollaterales Bündel ausgebildet ist. Die Nervatur der zweiteiligen Kotyledonarspreite macht wahrscheinlich, daß es sich um ein asymmetrisch fiederteiliges Blatt handelt.Daraus ergibt sich, daß bei A.apennina eine echte Einkeimblättrigkeit vorliegt, in dem Sinne, daß das einzige Keimblatt einem der beiden Keimblätter verwandter Arten homolog ist.Wir danken der Deutschen Forschungsgemeinschaft für die Unterstützung unserer Arbeiten.     

STAT3小分子抑制剂FLLL31对卵白蛋白致敏小鼠气道炎症治疗活性的初步研究

目的:研究新结构化合物FLLL31抑制卵白蛋白(OVA)致敏引发的小鼠气道炎症的活性,探讨FLLL31治疗哮喘的初步疗效。方法:雄性BALB/c小鼠(18~20 g)随机分为4组,每组10只,包括正常对照组(Control组)、模型对照组(OVA组)、地塞米松组(1 mg/kg,腹腔注射)和FLLL31组(15 mg/kg,口服灌胃)。各组小鼠分别于第0 d和第14 d致敏,每只鼠腹腔注射20μg OVA和2.25 mg Al(OH)3凝胶,FLLL31自致敏第24 d开始给药,给药周期为7 d;于第28、29、30 d以气管滴入OVA进行攻击,攻击前1 h给予受试化合物FLLL31及阳性药地塞米松。结果:小鼠肺灌流液炎症细胞分类计数、肺组织病理分析等实验结果证明,FLLL31以15 mg/kg连续给药7 d后,与模型组相比,抑制了小鼠肺部气道中炎症细胞的浸润,明显改善OVA致敏引发的小鼠气道炎症症状;ELISA结果证明FLLL31降低小鼠肺灌流液中炎症因子白细胞介素6的含量;免疫组化结果显示FLLL31降低OVA致敏小鼠肺部白细胞介素6受体浓度,减少中性粒细胞标志物淋巴细胞抗原6G(Ly6G)的表达(P0.05)。结论:特异抑制STAT3的新结构化合物FLLL31在体内对OVA致敏小鼠模型气道炎症有较好的改善和免疫调节活性。