Luminescence Induced in Intact Leaves and Isolated Chloroplasts by Alcohols and Other Substances

Elodea leaves and spinach chloroplasts emit red light when treated with alcohols or certain other solvents for chlorophyll. The intensity of the light, the lag phase and the threshold concentration vary considerably between different alcohols. Light emission from a leaf starts a few seconds to a few minutes after the addition of alcohol to the medium, reaches a maximum after 2–45 minutes (sometimes more) and then continues for many hours. Despite the faintness of the glow, the total number of photons given off from a leaf after addition of alcohol may exceed the number of photons given off from the same sample as long-lived afterglow after saturating irradiation with far-red light. The maximum yield of photons per chlorophyll molecule is a little more than 10^–5. The alcohol-induced luminescence is not influenced by a decrease in the oxygen tension to one fifth of the normal. Electron micrographs of treated leaves reveal that the thylakoid lipids contract to drops at the edges of the grana. Treatment of isolated chloroplasts with ethanol results in light emission when the concentration is high enough to dissolve the chlorophyll. It is estimated that the surface free energy in the thylakoid lipid-aqueous interface, due to ordinary interfacial tension, is large enough to account for the light emission observed when the interfaces contract or disappear.     

Factors Affecting Cell Fusion Induced by Sendai Virus

Cell fusion mediated by exogenous Sendai virus appears to occur in four temperature-dependent stages. The first two, which include viral adsorption, are pH dependent and can be inhibited by viral antibody. Viral envelope constituents remain detectable on the cell surface during the third stage and disappear only when cell-to-cell fusion supervenes. The relationship of these interactions to possible mechanisms of cell fusion are discussed.     

Factors Affecting the Rate of Photophosphorylation by Isolated Chloroplasts of Euglena gracilis*

SYNOPSIS The rate of photophosphorylation by Euglena chloroplasts depends not only on the physiologic stage of cell growth but also on the stage of development of chloroplasts in these cells at the time of harvesting. Both of these processes can be markedly influenced by a number of environmental factors; they are affected neither in a parallel manner nor completely independent of each other. In addition, the rate of photophosphorylation of chloroplasts can also be greatly affected by the conditions employed for their isolation. After investigating the various environmental factors both during cell growth and chloroplast isolation, we have developed a procedure which increased the photophosphorylation rate of our chloroplast preparations more than 5-fold, giving a specific activity in the range of 100-150 μmoles ATP/mg chlorophyll/hr routinely. The procedure is simple, needs no special equipment and requires only 2 or 3 days for cell growth.     

Induction Kinetics of Millisecond-Delayed Luminescence in Intact Bryopsis Chloroplasts

The induction curve of delayed luminescence emitted from 0.5to 2.5 ms after excitation of dark-adapted intact chloroplastsof the green alga, Bryopsis maxima, showed three transient peaks,L₁, L₂ and L₃ (in order of appearance), at about 0.1, 1 and5 s after theonset of intermittent illumination. Intact chloroplastswere needed for L₂ to appear, whereas L₁ and L₃ were presentin hypotonically treated chloroplasts. L₁ and L₂ are related to the electric field generated acrossthe thylakoid membranesbecause the two peaks parallelled theappearance of the first and second peaks of electrochromic absorptionchanges at 560 nm and they were totally abolished by valinomycinand CCCP. A smaller contribution to the L₁ and L₂ of the protonactivity gradient across the membranes, or of pH changes insideor outside the membranes, was suggested by the partial suppressionof the transient by NH₄CI. L₃ is related to the proton gradient or pH changes because thetransient was inhibited by NH₄CI and CCCP but enhanced by N,N'-dicyclohexylcarbodiimide.In the presenceof valinomycin, which somewhat lowered the peakheight of L₃, the kinetics of delayed luminescence parallelledthat of fluorescence. Electrogenic reactions which occur sequentiallyduring the dark to light transition of the photosynthetic machineryin intact chloroplasts is discussed in connection with transientchanges in delayed luminescence. (Received November 8, 1982; Accepted May 21, 1983)     

Factors Affecting the Enhancement of Oxidative Stress Tolerance in Transgenic Tobacco Overexpressing Manganese Superoxide Dismutase in the Chloroplasts

Two varieties of tobacco (Nicotiana tabacum var PBD6 and var SR1) were used to generate transgenic lines overexpressing Mn-superoxide dismutase (MnSOD) in the chloroplasts. The overexpressed MnSOD suppresses the activity of those SODs (endogenous MnSOD and chloroplastic and cytosolic Cu/ZnSOD) that are prominent in young leaves but disappear largely or completely during aging of the leaves. The transgenic and control plants were grown at different light intensities and were then assayed for oxygen radical stress tolerance in leaf disc assays and for abundance of antioxidant enzymes and substrates in leaves. Transgenic plants had an enhanced resistance to methylviologen (MV), compared with control plants, only after growth at high light intensities. In both varieties the activities of FeSOD, ascorbate peroxidase, dehydroascorbate reductase, and monodehydroascorbate reductase and the concentrations of glutathione and ascorbate (all expressed on a chlorophyll basis) increased with increasing light intensity during growth. Most of these components were correlated with MV tolerance. It is argued that SOD overexpression leads to enhancement of the tolerance to MV-dependent oxidative stress only if one or more of these components is also present at high levels. Furthermore, the results suggest that in var SR1 the overexpressed MnSOD enhances primarily the stromal antioxidant system.     

Iron Deficiency Induced Changes on Electron Transport Rate in Pisum Sativum Chloroplasts

Iron deficiency induced decrease in the rate of whole electron transport chain in chloroplasts of pea (Pisum sativum L.). Such reduction was mainly due to the loss of photosystem (PS) 2 activity. The same result was obtained when the ratio of variable to maximum chlorophyll fluorescence (F_v/F_m) was evaluated. The loss in PS 2 activity was primarily due to a loss of 33, 23 and 17 kDa polypeptides. In contrast, iron deficiency induced the synthesis of 28 and 29 kDa polypeptides.     

Factors Affecting Egg-ratio in Planktonic Rotifers

Edmondson’s egg ratio (number of amictic eggs per female) is an important life history variable, which has been in wide use to understand and predict patterns of population growth in planktonic rotifers under field conditions. It is also useful as an indicator of the health of rotifers under culture conditions. Generally, an inverse relationship exists between the egg ratio and the density of females in a population. A number of abiotic and biotic factors influence the egg ratio. For example, temperature can cause marked changes in the egg ratio by influencing the frequency of egg production and the hatching times of parthenogenetic eggs. Also, preferential feeding on ovigerous females of rotifers by invertebrate predators such as Asplanchna will lower the egg ratios of the population. The easy detachment of eggs, as may be the case in some members of the Brachionidae especially during enhanced reproduction when food levels are high, may also cause an underestimation of the egg ratio. In this review, we discuss the egg ratio of selected rotifer species in relation to the role of diel changes in egg production, the frequency or the intensity of feeding, the problems of distinguishing between different egg types and the negative effect of stressors such as toxicants and diet quality.     

Factors Affecting Grain Enlargement in Wheat

When most of the grains were removed from wheat ears the remaininggrains developed increased numbers of aleurone and endospermcells, and the increased grain volume was accompanied by anenlarged endosperm cavity. The giant grains usually synthesizedstarch more rapidly than normal grains. Flag leaf photosynthesiswas unaffected and additional sugar was retained in the culmand glumes. Total nitrogen and free amino acid content increased.Gibberellin and auxin concentrations were greater in the glumesof the partly degrained ears, and auxin in the grains was alsogreater than in equivalent grains from intact ears.     

Factors Affecting Transfection in Bacillus stearothermophilus

The conditions for the infection of Bacillus stearothermophilus 4S with TP-1C phage deoxyribonucleic acid (DNA) are described. Cells from log-phase cultures are the most susceptible to phage DNA infection (transfection). A cellular component (competence factor) which enhances transfection is released into the culture medium during the transition period between the log and stationary phase of growth. Transfection is stimulated in the order of decreasing effectiveness, by Fe(3+), Mn(2+), and Mg(2+). The efficiency of transfection is the highest in cells growing at 60.5 C and does not occur in cells growing at 67 C although the cells are growing normally. A cellular component (competence factor) of this organism, which is released into the culture medium, advances by 40 min some step in the uptake of phage DNA.     

Seasonal and Experimentally Induced Changes in the Ultrastructure of Chloroplasts of Pinus silvestris

The ultrastructure of chloroplasts in mesophyll cells of Pinus silvesris was examined under the electron microscope. Secondary needles were regularly sampled from a tree in a natural stand for one year. Primary needles from one-year-old seedlings exposed to frost hardening and dehardening conditions in a controlled environment chamber were also studied. These seedlings were exposed to 8 or 55 W m^-2. All needles were put in fixative at the different sampling dates and stored in a refrigerator until they were prepared for electron microscopy at the end of the experimental period. During the summer the choroplasts were symmetrically shaped and heavily loaded with starch. The membrane systems were well developed and consisted of both grana and stroma thylakoids. In autumn and during early artificial frost hardening the starch content was reduced, the chloroplasts appeared amoeboid and membrane-free stroma regions were seen. Later the chloroplasts became swollen and aggregated in one part of the cell. Starch was lost and the chloroplasts aggregated earlier at 8 W m^-2 than at 55 W m^-2. During winter the stroma thylakoids were first reduced in number and later even the grana thylakoids were damaged, resulting in mostly disorganized single membranes. Also the chloroplast envelope disappeared. In spring and early summer the chloroplasts migrated to the proximity of the cell walls. The membrane systems were reorganized and starch accumulated. During the first days of artificial dehardening the photosynthetic membranes were severely damaged, especially at 55 W m^-2, but soon new membranes were formed. Starch accumulated earlier at 55 than at 8 W m^-2. The reported ultrastructural variations are discussed in relation to functional and biochemical fluctuations caused by the season or by artificial variations in the climate as demonstrated earlier.     

Factors Affecting Chloroplast Replication in Spinach

Chloroplast replication has been studied in discs cut from thebase of young spinach leaves and cultured on sterile nutrientagar. In discs grown in a growth cabinet chloroplast numbersper cell increased logarithmically with time over a 7-day cultureperiod. Chloroplast replication proceeds in a similar way incultured discs and in intact leaves. Cytokinins do not affect chloroplast replication in this systembut they stimulate the fresh-weight growth of discs. Chloroplastreplication is temperature dependent, having an optimum at 25°C. By contrast chloroplast size is at a maximum in discscultured at 12 °C. Light stimulates chloroplast replication, a linear relationshipoccurring between chloroplast number per cell and the dailyquantity of light given to discs up to a saturating value of250 J d^–1. Daylength does not affect chloroplast formationin spinach. In a number of experiments a general relationship was establishedbetween chloroplast number per cell and cell size but no evidenceis available to suggest that this correlation is causal. Theresults of experiments in which discs were transferred fromdark to light suggest that some of the events which precedechloroplast replication may occur at similar rates in both lightand dark.     

乙醇诱导杂交瘤细胞凋亡的检测

为建立杂交瘤细胞凋亡检测模型,在乙醇诱导下,采用荧光染色、MTT等方法检测H18杂交瘤细胞凋亡时的形态学及增殖活性变化,并用流式细胞仪对其进行定量分析,夹心ELISA检测IgG抗体分泌的变化情况。结果发现5lOmmol／L乙醇作用5～6h的凋亡诱导效果最为明显,在诱导条件下,杂交瘤细胞的活细胞数显著下降,凋亡细胞比例较高。抗体浓度明显下降,与乙醇浓度呈负相关,但与细胞增殖活性无明显的线性关系。故以此为凋亡检测模型,可为后续抗凋亡细胞株的建立与筛选研究提供初步的实验基础,并为乙醇对IgG型抗体分泌的影响研究提供了可能的实验模型。     

ART中影响新生儿性别比的相关因素分析

辅助生殖技术(assisted reproductive technology,ART)在给广大不孕不育患者带来福音的同时,也带来了男女出生性别比的失衡,相关影响因素有待研究分析。通过收集并分析2013年1月至2014年12月于广州医科大学附属第三医院接受ART治疗并成功分娩4 635个新生儿的3 462个周期患者的临床资料发现,ICSI组与IVF组比较,活产男婴比例没有统计学差异(P=0.07),但PESA/TESA-ICSI组出生男婴比例较IVF组显著降低(P=0.036);同时,移植囊胚期胚胎出生的男婴比例要显著高于移植卵裂期胚胎的男婴比例(P=0.005)。进一步的分层卡方检验结果显示,除去授精方式的影响,移植胚胎期别仍然可以影响出生婴儿的性别比(P=0.021)。但多因素logistic回归分析提示,以上因素均不能构成独立预测因素。上述结果表明在体外受精-胚胎移植周期中,移植囊胚期胚胎可显著增加男婴出生比例,而PESA/TESA-ICSI授精方式则会显著降低男婴出生比例。     

Long-Term Ethanol Alcoholization Decreases Levodopa- Induced Aggressive Behavior in Rats

One of the old but still unsolved problems in psychiatry is the treatment of patients with a dual diagnosis of schizophrenia and alcoholism. Numerous clinical studies failed to explain unambiguously the mechanisms of formation of these comorbid disorders. Experimental modeling of dual disorders was carried out using a model of schizophrenia based on the impaired development and dopamine hypothesis of schizophrenia. To model comorbid experimental schizophrenia and alcohol abuse, the dopamine precursors, Levodopa/Carbidopa (LC), were used in combination with 15% ethanol. The aim of this study was to investigate social behavior of rats receiving LC and exposed to intermittent semi-compulsory alcoholization. After three months of the experiment, there were significantly more interactions (including aggressive–attacks, fights, bites) and submissive postures in the group of rats that received LC. Combined LC/ethanol administration did not alter rat social behavior in contrast to ethanol administered alone.     

影响耶尔森氏鼠疫杆菌基因组密码子使用的因素分析

基因组密码子使用的影响因素分析有助于发现影响密码子使用的进化动力学 ,对发现和预测进化的方向和模式有重要的作用。同时 ,分析完整的基因组可以发现特定基因组中密码子的使用模式 ,从而重新设计高效的PCR引物和外源导入基因 ,促进外源基因在特定生物体中的高效率表达。导致瘟疫等外源性感染疾病的耶尔森氏鼠疫杆菌完整基因组序列已经测序公布。为了对鼠疫杆菌的同义密码子使用的进化模式有更加深入的了解 ,详细的研究分析鼠疫杆菌的基因组密码子的使用模式和影响密码子使用的因素。结果发现 ,尽管鼠疫杆菌基因组序列中“G” “C”含量相对较低 (4 7.6 4 % ) ,高水平表达基因的密码子第三位碱基使用胞嘧啶 (C)的频率比表达水平低的基因使用胞嘧啶 (C)有显著的提高 ,表达水平较低的基因在密码子的第三位碱基更趋向使用鸟嘌呤 (G)。在表达水平高低的两组基因中 ,对密码子的第三位碱基使用腺嘌呤 (A)和胸腺嘧啶 (T)总体上趋于随机使用。基因的表达水平与对应分析的第一条向量轴呈高度相关 (R =0 .6 3,P <0 .0 0 0 1)。通过分析比较表达水平高低两组基因的密码子使用模式发现 ,基因的表达水平对于密码子使用有显著的影响。GC skew分析结果显示 ,复制转录阶段的选择对密码子使用有一定的影响。在不同长度     

Analysis of a Modern Hybrid and an Ancient Sugarcane Implicates a Complex Interplay of Factors in Affecting Recalcitrance to Cellulosic Ethanol Production

Abundant evidence exists to support a role for lignin as an important element in biomass recalcitrance. However, several independent studies have also shown that factors apart from lignin are also relevant and overall, the relative importance of different recalcitrance traits remains in dispute. In this study we used two genetically distant sugarcane genotypes, and performed a correlational study with the variation in anatomical parameters, cell wall composition, and recalcitrance factors between these genotypes. In addition we also tracked alterations in these characteristics in internodes at different stages of development. Significant differences in the development of the culm between the genotypes were associated with clear differential distributions of lignin content and composition that were not correlated with saccharification and fermentation yield. Given the strong influence of the environment on lignin content and composition, we hypothesized that sampling within a single plant could allow us to more easily interpret recalcitrance and changes in lignin biosynthesis than analysing variations between different genotypes with extensive changes in plant morphology and culm anatomy. The syringyl/guaiacyl (S/G) ratio was higher in the oldest internode of the modern genotype, but S/G ratio was not correlated with enzymatic hydrolysis yield nor fermentation efficiency. Curiously we observed a strong positive correlation between ferulate ester level and cellulose conversion efficiency. Together, these data support the hypothesis that biomass enzymatic hydrolysis recalcitrance is governed by a quantitative heritage rather than a single trait.