Long‐term cytological and histological outcomes in women managed with loop excision treatment under local anaesthetic for high‐grade cervical intraepithelial neoplasia

Y. L. Woo, C. Badley, E. Jackson and R. Crawford Long‐term cytological and histological outcomes in women managed with loop excision treatment under local anaesthetic for high‐grade cervical intraepithelial neoplasia Objective: This study examines the impact of excision margin status after large loop excision of the transformation zone (LLETZ) under local anaesthetic for high‐grade cervical intraepithelial neoplasia (HG‐CIN) on the cytological and histological outcomes up to 5 years after treatment. Methods: Prospective cytological and histological data were obtained by examination of the colposcopy database at Addenbrooke’s Hospital, Cambridge, UK. All women aged between 19 and 50 years who underwent treatment for HG‐CIN by LLETZ under local anaesthetic were included in the study. Patients without follow‐up data were excluded from the study. The excision margin status was correlated with the subsequent cytological and histological outcomes. Results: A series of 967 women with CIN2 and CIN3 underwent LLETZ excision under local anaesthetic. Overall, 42% of women had disease present at the excision margin following LLETZ. Women with CIN3 were more likely than those with CIN2 to have an involved excision margin (P < 0.0001). Cytological recurrence was highest at 12 months (16%) and did not correlate with the CIN grade or excision margin status. Histological recurrence/persistence was also highest at 12 months follow‐up (15%) and this correlated with grade of CIN and margin status (P < 0.0001). Conclusions: Histological recurrence/persistence correlates with grade of CIN and excision margin status. Management of HG‐CIN in an outpatient setting under local anaesthetic is safe, cost effective and yields a favourable long‐term outcome.     

Biolistic co-transformation of the nuclear and plastid genomes

Particle gun-mediated (so-called 'biolistic') transformation represents a universal genetic transformation technology that is widely applied in nearly all groups of organisms. The mechanism of how accelerated DNA-coated particles, after their entry into the cell, deliver the foreign DNA to the target compartment is not known. Here we have studied this process in plants by performing co-transformation experiments with vectors targeted to two different cellular compartments, the nucleus and the plastids (chloroplasts). We find that coating of particles with both plastid and nuclear transformation vectors can result in co-transformation of chloroplasts and the nucleus. In contrast, mixing of particles coated individually with the vectors does not produce co-transformed plants. Our data suggest that a single DNA-coated particle can transform more than one compartment of the plant cell, opening up the possibility to generate doubly transgenic plants in one step. Importantly, co-transformation can also be obtained in the absence of selection, thus providing a method to produce marker-free transgenic genomes. In addition, our findings raise the possibility of occasional inadvertent co-transformation of two genomes and, therefore, have important implications for the molecular characterization and regulation of transgenic plants.     

Tn5 transposase-assisted transformation of indica rice

Here, we describe experiments on Tn5 transposase-assisted transformation of indica rice. Transposomes were formed in vitro as a result of hyperactive Tn5 transposase complexing with a transposon that contained a 19-bp tetracycline operator (tetO) sequence. To form modified projectiles for transformation, the Tn10-derived prokaryotic tetracycline repressor (TetR) proteins, which can bind transposomes via the high affinity of TetR for tetO, were immobilized onto the surface of bare gold microscopic particles. These projectiles were introduced into cells of the indica rice cultivar Zhuxian B by particle bombardment. Once projectiles were inside the cell, tetracycline induced an allosteric conformational change in TetR that resulted in the dissociation of TetR from tetO, and thus generated free transposomes. Molecular evidence of transposition was obtained by the cloning of insertion sites from many transgenic plants. We also demonstrated that the introduced foreign DNA was inherited stably over several generations. This technique is a promising transformation method for other plant species as it is species independent.     

Cells from icons to symbols: molecularizing cell biology in the 1980s

Over centuries cells have been the target of optical and electronic microscopes as well as others technologies, with distinctive types of visual output. Whilst optical technologies produce images ‘evident to the eye’, the electronic and especially the molecular create images that are more elusive to conceptualization and assessment. My study applies the semiotic approach to the production of images in cell biology to capture the shift from microscopic images to non-traditional visual technologies around 1980. Here I argue that the visual shift that coincides with the growing dominance of molecular biology involves a change from iconic to symbolic forms.     

Experimental studies on the distribution of the aphid counts

The present paper dealt with the sequential changes of the distribution pattern of apterous females aphid populations, that were artificially settled at the beginning on the experimental barley ‘field'. The aphids were settled at random or even with a fixed denisty per plant. For five or six days after the settling, the number of individuals followed the negative binomial distributions in all cases while the parameters k and p were varying. The estimated values of k were rather small for the first one week after the settling, which may suggest that the number of moving aphids between plants was relatively small and the degree of concentration expressing the intrinsic increase was high. After that, as the number of individuals increased, the number of moving aphids between plants would be considered to be increased. It was found that with the lapse of time the degree of concentration decreased or k became larger. The distribution of aphids per blade in a plant was also described briefly.     

Heterologous expression of Arabidopsis pattern recognition receptor RLP23 increases broad‐spectrum resistance in poplar to fungal pathogens

The pattern recognition receptor AtRLP23 from Arabidopsis thaliana recognizes the epitopes (nlp24s) of necrosis and ethylene‐inducing peptide 1‐like proteins (NLPs) and triggers pattern‐triggered immunity (PTI). Here, we established methods for studying the early events of PTI in the hybrid poplar cultivar Shanxin (Populus davidiana × Populus bolleana) in response to the flagellin epitope. We confirmed that wild‐type Shanxin cannot generate PTI responses on nlp24 treatment. Four NLP homologues were characterized from two common fungal pathogens of Shanxin, namely Marssonina brunnea f. sp. monogermtubi (MbMo) and Elsinoë australis (Ea), which cause black leaf spot and anthracnose disease, respectively, and the nlp24s of three of them could be responded to by Nicotiana benthamiana leaves expressing AtRLP23. We then created AtRLP23 transgenic Shanxin lines and confirmed that the heterologous expression of AtRLP23 conferred on transgenic Shanxin the ability to respond to one nlp24 of each fungal pathogen. Consistently, infection assays with MbMo or Ea showed obviously lower levels of disease symptoms and significantly inhibited the growth of fungi on the transgenic poplar compared with that in wild‐type poplar. Overall, our results indicated that the heterologous expression of AtRLP23 allowed transgenic Shanxin to generate a PTI response to nlp24s, resulting in increased broad‐spectrum fungal disease resistance.     

丛枝菌根真菌对褐土玉米氮素吸收和土壤N2O排放的影响

探究不同氮肥水平下丛枝菌根(AM)真菌对褐土玉米土壤N₂O排放和氮转化功能基因的影响,为阐明AM真菌在褐土N₂O排放中的作用和效应提供理论依据。设置氮肥用量(NⅠ:105 mg/kg;NⅡ:210 mg/kg)、AM真菌(M0:不接种AM真菌;M1:接种根内根孢囊霉(Rhizophagus intraradices);M2:接种摩西斗管囊霉(Funneliformis mosseae);M3:接种Rhizophagus intraradices + Funneliformis mosseae等比例混合)双因素盆栽试验。测定植株地上部全氮含量、土壤铵态氮、硝态氮含量和N₂O排放量,采用实时荧光定量聚合酶链式反应(PCR)法分析土壤硝化功能基因(amoA-AOA和amoA-AOB)和反硝化功能基因(nirS、nirK和nosZ)的丰度。结果表明,两种施氮水平下,接种AM真菌均可显著降低土壤N₂O排放通量和累积排放量,不同AM真菌处理下N₂O累积排放量表现为:M0>M2>M1>M3。相同AM真菌处理的土壤N₂O排放通量和累积排放量在NⅡ施氮水平高于NⅠ施氮水平;相同AM真菌处理的玉米菌根侵染率在NⅡ施氮水平低于NⅠ施氮水平。与M0相比,NⅠ条件下M1、M2和M3处理土壤铵态氮含量分别降低24.5%、20.8%和45.3%,硝态氮含量分别降低19.7%、14.9%和30.2%,植株地上部全氮含量分别增加16.3%、35.2%和59.6%;与M0相比,NⅡ条件下M1、M2和M3处理土壤铵态氮含量分别降低20.9%、24.8%和40.0%,硝态氮含量分别降低36.3%、25.6%和45.2%,植株地上部全氮含量分别增加33.2%、43.9%和95.4%。两种施氮水平下,AM真菌可显著降低土壤硝化功能基因(amoA-AOA和amoA-AOB)丰度,增加反硝化功能基因(nirS、nirK和nosZ)丰度。AM真菌与N₂O排放通量呈极显著负相关。本盆栽试验条件下,接种AM真菌均可增强两种氮肥用量玉米植株氮素吸收能力,调节硝化、反硝化相关功能基因的丰度,减少土壤N₂O气体的排放,且两种AM真菌混合处理的N₂O减排效应强于单一AM真菌接种。     

喀斯特11种典型生态恢复树种凋落叶分解及其对土壤碳排放的激发效应

旨在探究喀斯特地区退化生态系统植被恢复树种凋落叶分解过程及其对土壤碳排放的激发效应,为选择合适的树种进行植被恢复提供数据支持。以中国林科院热带林业实验中心大青山石山树木园11种适应性强、耐干旱贫瘠的优良石山树种为研究对象,利用¹³C自然丰度法区分凋落叶和土壤来源CO₂并量化土壤激发效应,比较不同生态恢复树种凋落叶分解及其激发效应的差异,探讨凋落物分解及其激发效应与凋落物性状之间的关联。结果表明:(1)11个生态恢复树种凋落叶在碳相关化学性质(水溶性碳、半纤维素和单宁含量等)、养分含量(磷和镁含量等)及化学计量特征(碳磷比和氮磷比)等方面均表现出较高程度变异。(2)不同生态恢复树种凋落叶分解及其诱导的土壤激发效应具有极显著差异(P<0.001);在整个培养实验期间,11个生态恢复树种凋落叶平均分解了35.3%,其中海南椴分解最快,达到50%,而青冈栎分解最慢,仅分解16.5%。(3)总体上看,凋落叶处理的土壤呼吸速率(5.1 mg C kg^-1 土壤 d^-1)是对照土壤呼吸速率(2.3 mg C kg^-1土壤d^-1)的2.2倍,凋落叶添加显著促进土壤有机碳分解,平均达到37.6%;其中海南椴、割舌树和任豆凋落叶输入则抑制土壤有机碳分解(抑制程度分别为-13.2%、-6.9%和-22.5%),产生负激发效应。(4)凋落叶分解与非结构性碳(r=0.63,P=0.04)和水溶性碳(r=0.91,P<0.001)呈显著正相关,与叶干物质含量(r=0.64,P=0.03)、纤维素(r=0.62,P=0.04)和锰含量(r=-0.63,P=0.04)呈显著负相关。多元回归分析结果表明,水溶性碳、钾和钙含量相结合可以解释生态恢复树种凋落叶分解变异的98%;然而,凋落叶性状与土壤激发效应强度之间并没有显著相关性。从土壤养分归还角度考虑,喀斯特退化生态系统恢复树种可以选择光皮梾木、海南椴、顶果木和降香黄檀等凋落叶分解较快的树种,以促进土壤养分循环和植被恢复;另一方面,从土壤碳固持角度来看,海南椴、任豆和割舌树等凋落叶输入会抑制土壤有机碳分解,从而有利于提高退化生态系统土壤碳封存能力。     

Plant protoplast technology: Current status

Robust and reproducible protoplast-to-plant systems are crucial for underpinning genetic manipulation technology involving somatic hybridisation and transformation. Novel and effective approaches for maximising the efficiency of such protoplast cultures include supplementation of media with surfactants and artificial gas carriers, such as perfluorochemicals and haemoglobin. Physical parameters, particularly electrostimulation, also enhance the development of protoplasts and protoplast-derived cells in culture. DNA uptake into protoplasts is now a routine and universally accepted procedure in plant biotechnology for introducing and evaluating both short-term (transient) and long-term (stable) expression of genes in cells and regenerated plants. Importantly, protoplast fusion overcomes pre- and post-zygotic sexual incompatibility barriers and generates novel germplasm through new nuclear-cytoplasmic combinations. In this respect, considerable progress has been made in generating somatic hybrid plants, particularly in citrus, brassicas and potato. Isolated protoplasts are also a unique single cell system for evaluating aspects of ultrastructure, genetics and physiology, with potential for the biosynthesis of novel secondary products, including commercially-important recombinant proteins (e.g. antibodies), and as systems in toxicity screening. Recent advances in protoplast technology have benefited from advances in animal and microbial cell culture, with interesting parallels existing between these systems. Further innovations will necessitate the strengthening of interdisciplinary links in these research fields and the requirement for continued dialogue and co-operation between workers with diverse but complementary skills.     

Signal amplification in biological and electrical engineering systems: Universal role of cascades

In this paper we compare the cascade mechanisms of signal amplification in biological and electrical engineering systems, and show that they share the capacity to considerably amplify signals, and respond to signal changes both quickly and completely, which effectively preserves the form of the input signal. For biological systems, these characteristics are crucial for efficient and reliable cellular signaling. We show that this highly-efficient biological mechanism of signal amplification that has naturally evolved is mathematically fully equivalent with some man-developed amplifiers, which indicates parallels between biological evolution and successful technology development.