Mendelian randomization in health research: Using appropriate genetic variants and avoiding biased estimates

Mendelian randomization methods, which use genetic variants as instrumental variables for exposures of interest to overcome problems of confounding and reverse causality, are becoming widespread for assessing causal relationships in epidemiological studies. The main purpose of this paper is to demonstrate how results can be biased if researchers select genetic variants on the basis of their association with the exposure in their own dataset, as often happens in candidate gene analyses. This can lead to estimates that indicate apparent “causal” relationships, despite there being no true effect of the exposure. In addition, we discuss the potential bias in estimates of magnitudes of effect from Mendelian randomization analyses when the measured exposure is a poor proxy for the true underlying exposure. We illustrate these points with specific reference to tobacco research.     

Activities of hydrolytic enzymes in callus cultures of tobacco during organogenesis

Starch, total sugars, reducing sugars and protein contents and the specific activities of hydrolytic enzymes such as amylase, Phosphorylase, soluble acid invertase, wall-bound acid invertase, sucrose synthetase, acid and alkaline phosphatases and ribonuclease were determined in root forming, shoot forming and non-organ-forming callus cultures of tobacco. Organ-forming cultures not only showed higher amounts of the above metabolites but also higher enzyme activities compared to non-organ-forming cultures. The activities of these enzymes in relation to organogenesis is discussed.     

Antiviral properties and interaction of novel chalcone derivatives containing a purine and benzenesulfonamide moiety

A new concise and facile method was explored to synthesize a series of novel chalcone derivatives containing a purine and benzenesulfonamide moiety and their antiviral properties were evaluated against TMV and CMV. Biological assays indicated that several of the derivatives exhibited significant anti-TMV and anti-CMV activities in vivo. In particular, compound d2 displayed excellent inactivating activity against TMV, with the EC₅₀ value of 51.65?μg/mL, which was better than that of ribavirin (150.45?μg/mL). Molecular docking showed that there are four hydrogen bonds between compound d2 and TMV coat protein (TMV-CP). Compound d2 demonstrated strong binding capacity to TMV-CP with K_a?=?1.58?×?10⁵?L/mol and K_d?=?12.16?μM. These findings indicated that chalcone derivatives are worthy of further research and development as templates for new antiviral agents.     

Properties of the Coat Protein of a New Tobacco Mosaic Virus Coat Protein ts-Mutant

Amino acid substitutions in a majority of tobacco mosaic virus (TMV) coat protein (CP) ts-mutants have previously been mapped to the same region of the CP molecule tertiary structure, located at a distance of about 70 Å from TMV virion axis. In the present work some properties of a new TMV CP ts-mutant ts21-66 (two substitutions I21 T and D66 G, both in the 70-Å region) were studied. Thermal inactivation characteristics, sedimentation properties, circular dichroism spectra, and modification by a lysine-specific reagent, trinitrobenzensulfonic acid, of ts21–66 CP were compared with those of wild-type (U1) TMV CP. It is concluded that the 70-Å region represents the most labile portion of the TMV CP molecule. Partial disordering of this region in the mutant CP at permissive temperatures leads to loss of the capacity to form two-layer aggregates of the cylindrical type, while further disordering induced by mild heating results also in the loss of the ability to form ordered helical aggregates.     

人防御素—1转基因烟草的获得及其抗TMV的初步观察

人中性粒细胞防御素是一类具有广谱杀伤病原微生物活力的小分子多肽,有作者设想将其导入作物用于抗病育种。我们前期的工作通过RT-PCR建立了人中性粒细胞防御素-1(HNP-1)的cDNA克隆;接着通过农杆菌介导的方法,将HNP-1 cDNA片段导入烟草,尝试了HNP-1在烟草植株水平的表达,并初步证明获得的转基因烟草对低浓度烟草花叶病毒(TMV)侵染有一定抵抗力。     

Sites of microtubule reorganization in tobacco BY-2 cells during cell-cycle progression

Summary The sites of microtubule (MT) reorganization were examined in synchronized tobacco BY-2 cells. The MTs of these cells were completely destroyed by a combined cold and drug treatment at 0 °C with 100 M propyzamide for 3 h. After the cells were washed and cultured at 30 °C, the reorganization of MTs was observed in detail. Sites for MT reorganization at each stage of the cell cycle were identified on the cell cortex and nuclei, the mitotic apparatus, the nuclei (or the nuclei and cell cortex), and the cell cortex in the S-G₂ phase, M phase, M/G₁ interface, and g₁ phase, respectively. The polypeptide synthesis elongation factor (EF)-1 is co-localized with these sites of MT reorganization. At some stages, microfilaments (MFs) were found to be involved in the reorganization of MTs. Based on these results, the mode of MT reorganization during cell cycle progression is discussed.Abbreviations EF-1 elongation factor 1 - MAP microtubule-associated protein - MF microfilament - MIs mitotic indices - MT microtubule     

Production of mouse monoclonal antibody with galactose-extended sugar chain by suspension cultured tobacco BY2 cells expressing human beta(1,4)-galactosyltransferase

Previously, we developed a transgenic tobacco BY2 cell line (GT6) in which glycosylation was modified by expressing human beta(1,4)-galactosyltransferase (betaGalT). In this study, we produced a mouse monoclonal antibody in GT6 cells, and determined the sugar chain structures of plant-produced antibodies. Galactose-extended N-linked glycans comprised 16.7%, and high-mannose-type and complex-type glycans comprised 38.5% and 35.0% of the total number of glycans, respectively. N-linked glycans with the plant-specific sugars beta(1,2)-xylose and alpha(1,3)-fucose comprised 9.8%. The introduction of human betaGalT into suspension cultured tobacco cells resulted in the production of recombinant proteins with galactose-extended glycans and decreased contents of beta(1,2)-xylose and alpha(1,3)-fucose.     

Elicitation of the Hypersensitive Responses in Tabacco by a10.6 kD Proteinaceous Elicitor from Phytophthora palmi

A 10.6 kD heat resistant, proteinacious elicitor was purified from the culture filtrate of Phytophthora palmivora Butler but not from P. melonis Katsura. The 10.6 kD elicitor is a holoprotein devoid of glycoside. It can cause hypersensitive necrosis of the detached tobacco ( Nicotiatna tabacum L. ) leaves 48 h post-inoculation with dosages of above 40 μg. Four cell types were investigated by using Confocal microscopy, and 2' ,7'-dichlorodihydrofluorescein diacetate (DCFDA) as a probe of H2O2 production. It was showed that oxidative burst occurred in cultured suspension cells as well as in mesophyll cells, epidermal cells and guard cells within 10 min upon the elicitor treatment. The hypersensitive cell death appeared 6 h after the treatment when inoculated with fluorescein diacetate (FDA) as indicator of cell viability. These results suggest that H2O2 accumulation was the main cause of the hypersensitive cell death in tobacco induced by the 10.6 kD elicitor. This 10.6 kD elicitor may belong to the family of elicitins.     

从感染黄瓜花叶病毒的烟草叶筛选抗病毒细胞突变体

用绿岛法从感染CMV的普通烟草(G140)叶筛选到抗病毒细胞突变体R~CMV2、6,其当代无性系群体病情指数较对照降低41.5-45.2%。 Abstract:The 2 CMV-resistant-cell mutants,RCMV-2、6,were selected by using the method of culturing dark green island from CMV infected tobacco(N.tabacum cv G140)leaves.The disease in dex of RCMV-2、6′s was decreased by 41.5~45.2% in comparison with that of the control(G140).