Hypothesis: the origin of life in a hydrogel environment

A hypothesis is proposed that the first cell(s) on the Earth assembled in a hydrogel environment. Gel environments are capable of retaining water, oily hydrocarbons, solutes, and gas bubbles, and are capable of carrying out many functions, even in the absence of a membrane. Thus, the gel-like environment may have conferred distinct advantages for the assembly of the first cell(s).     

Conditional lethality involving nuclear and cytoplasmic chlorophyll mutants in soybeans

Summary A conditionally lethal phenotype occurred when a nuclear chlorophyll mutant (y ₂₀-k ₂) was present with a cytoplasmic chlorophyll mutant (cyt-Y ₂) in soybean (Glycine max [L.] Merr.). Nuclear mutant y ₂₀-k ₂, Genetic Type Collection Number T253, has yellow foliage, tan-saddle-pattern seed and is viable. The y ₂₀-k ₂ mutant cannot be separated by classical genetic tests into two separate components, y ₂₀ (yellow foliage) and k ₂ (tan-saddle-pattern seed). Mutant cyt-Y ₂, T275, is inherited cytoplasmically, has yellow foliage, and is viable. The genotype cyt-Y ₂ y ₂₀-k ₂/ y ₂₀-k ₂ is a conditional lethal; the genotype is lethal under field conditions, but plants survive under greenhouse conditions. This interaction is unique to y ₂₀-k ₂. This conditionally lethal genotype may be useful in molecular studies on the interaction between nuclear and plastid genomes.This is a joint contribution of North Central Region, USDA ARS, and Journal Paper No. J-11429 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa, and the Agriculture Experiment Station, Univ. of Puerto Rico, Mayaguez Campus, Mayaguez, Puerto Rico 00708. Projects 2471 and 2475. The research was supported in part by the Iowa Soybean Promotion Board.     

Biofunctionalized nanoneedles for the direct and site-selective delivery of probes into living cells

Background

Accessing the interior of live cells with minimal intrusiveness for visualizing, probing, and interrogating biological processes has been the ultimate goal of much of the biological experimental development.

Scope of review

The recent development and use of the biofunctionalized nanoneedles for local and spatially controlled intracellular delivery brings in exciting new opportunities in accessing the interior of living cells. Here we review the technical aspect of this relatively new intracellular delivery method and the related demonstrations and studies and provide our perspectives on the potential wide applications of this new nanotechnology-based tool in the biological field, especially on its use for high-resolution studies of biological processes in living cells.

Major conclusions

Different from the traditional micropipette-based needles for intracellular injection, a nanoneedle deploys a sub-100-nm-diameter solid nanowire as a needle to penetrate a cell membrane and to transfer and deliver the biological cargo conjugated onto its surface to the target regions inside a cell. Although the traditional micropipette-based needles can be more efficient in delivery biological cargoes, a nanoneedle-based delivery system offers an efficient introduction of biomolecules into living cells with high spatiotemporal resolution but minimal intrusion and damage. It offers a potential solution to quantitatively address biological processes at the nanoscale.

General significance

The nanoneedle-based cell delivery system provides new possibilities for efficient, specific, and precise introduction of biomolecules into living cells for high-resolution studies of biological processes, and it has potential application in addressing broad biological questions.This article is part of a Special Issue entitled Nanotechnologies - Emerging Applications in Biomedicine.     

Restriction fragment patterns of chloroplast and mitochondrial DNA of Dasypyvum villosum (L.) candargy and wheats

To elucidate the phylogenetic relationships and cytoplasmic types, restriction endonuclease fragment patterns of chloroplast (cp) and mitochondrial (mt) DNAs isolated from two different accessions of Dasypyrum villosum (L.) candargy were compared with those of tetraploid wheat (Triticum durum Desf., PI265007), hexaploid wheat (Triticum aestivum L., cv Chinese Spring), Aegilops longissimum (S. and M., in Muschli) Bowden and Hordeum vulgare L. T. aestivum and T. durum had identical restriction patterns for their cp and mtDNAs in digestions with four different enzymes. Likewise, no differences were found between the restriction fragment patterns of two accessions of D. villosum. But, there were distinct differences in chloroplast and mitochondrial DNA restriction fragment patterns between D. villosum and tetraploid and hexaploid wheats. A. longissimum (G609) showed a similar pattern to those wheats for PstI digestion of cpDNA. Organellar DNA from Hordeum vulgare (cv Himalaya) showed a distinctly different restriction pattern from those of wheat and D. villosum. These results suggest that D. villosum is unlikely to be the donor of cytoplasm to wheats, and its cytoplasmic organelles were also different from those of A. longissimum.Contribution No. 92-522-J from the Kansas Agricultural Experiment Station; Kansas State University, Manhattan, Kansas, USA     

Measurement of grouped intracellular solute osmotic virial coefficients

Models of cellular osmotic behaviour depend on thermodynamic solution theories to calculate chemical potentials in the solutions inside and outside the cell. These solutions are generally thermodynamically non-ideal under cryobiological conditions. The molality-based Elliott et al. form of the multi-solute osmotic virial equation is a solution theory which has been demonstrated to provide accurate predictions in cryobiological solutions, accounting for the non-ideality of these solutions using solute-specific thermodynamic parameters called osmotic virial coefficients. However, this solution theory requires as inputs the exact concentration of every solute in the solution being modeled, which poses a problem for the cytoplasm, where such detailed information is rarely available. This problem can be overcome by using a grouped solute approach for modeling the cytoplasm, where all the non-permeating intracellular solutes are treated as a single non-permeating “grouped” intracellular solute. We have recently shown (Zielinski et al., J Physical Chemistry B, 2017) that such a grouped solute approach is theoretically valid when used with the Elliott et al. model, and Ross-Rodriguez et al. (Biopreservation and Biobanking, 2012) have previously developed a method for measuring the cell type-specific osmotic virial coefficients of the grouped intracellular solute. However, the Ross-Rodriguez et al. method suffers from a lack of precision, which—as we demonstrate in this work—can severely impact the accuracy of osmotic model predictions under certain conditions. Thus, we herein develop a novel method for measuring grouped intracellular solute osmotic virial coefficients which yields more precise values than the existing method and then apply this new method to measure these coefficients for human umbilical vein endothelial cells.     

Control of DNA Replication by the Nucleus/Cytoplasm Ratio in Xenopus

The nucleus/cytoplasm (N/C) ratio controls S phase dynamics in many biological systems, most notably the abrupt remodeling of the cell cycle that occurs at the midblastula transition in early Xenopus laevis embryos. After an initial series of rapid cleavage cycles consisting only of S and M phases, a critical N/C ratio is reached, which causes a sharp increase in the length of S phase as the cell cycle is reconfigured to resemble somatic cell cycles. How the N/C ratio determines the length of S phase has been a longstanding problem in developmental biology. Using Xenopus egg extracts, we show that DNA replication at high N/C ratio is restricted by one or more limiting substances. We report here that the protein phosphatase PP2A, in conjunction with its B55α regulatory subunit, becomes limiting for replication origin firing at high N/C ratio, and this in turn leads to reduced origin activation and an increase in the time required to complete S phase. Increasing the levels of PP2A catalytic subunit or B55α experimentally restores rapid DNA synthesis at high N/C ratio. Inversely, reduction of PP2A or B55α levels sharply extends S phase even in low N/C extracts. These results identify PP2A-B55α as a link between DNA replication and N/C ratio in egg extracts and suggest a mechanism that may influence the onset of the midblastula transition in vivo.     

Cytoplasmic/nuclear localization of tuberin in different cell lines

Summary. Tuberous sclerosis (TSC) is an autosomal dominantly inherited disease affecting 1 in 6000 individuals. The TSC gene products, hamartin and tuberin, form a complex, of which tuberin is assumed to be the functional component being involved in a wide variety of different cellular processes. Tuberin has been demonstrated to be localized to both, the cytoplasm and the nucleus. The cytoplasmic/nuclear localization of tuberin is known to be regulated by the serine/threonine protein kinase Akt. Akt also regulates the cytoplasmic/nuclear localization of the cyclin-dependent kinase inhibitor p27. In this study the localization of these two Akt-regulated proteins was analysed in different cell lines.     

大葱小孢子母细胞至二胞早期花粉发育的超微结构观察

用电镜观察了章丘大葱 (AlliumfistulosumL .)从小孢子母细胞至二胞早期花粉发育的超微结构。终变期的花粉母细胞 ,胼胝壁外方的相邻初生壁间及胞间隙内 ,存在胞间物质 ,四分体期 ,此物质尚部分存在。小孢子母细胞减数分裂前 ,细胞质内含有脂滴 ,小孢子有丝分裂以后 ,脂滴增多增大。小孢子分裂后期 ,质体已积累淀粉粒 1至多个。二胞早期花粉之营养细胞质内 ,有些含淀粉质体亦含脂滴。各发育期 ,核糖体及多聚合糖体丰富 ,并有很多的粗面内质网、高尔基体及小泡、线粒体 ,显示蛋白质、糖类及其它物质合成及运输作用的活跃。小孢子缺中央大液泡。有丝分裂后期 ,细胞器集中于未来的营养细胞极。小孢子胞质分裂期 ,有些内质网贴近或与花粉质膜相连 ,它们或有可能互相融合 ,扩大质膜面积而适应花粉的生长。还讨论了不同时期高尔基体小泡的作用。     

沙门菌CWDMs脂代谢检测

采用毛地黄皂苷敏感试验和菌细胞胆固醇、甘油三脂及胆碱酯酶定性与定量分析法,检测伤寒杆菌和甲型副伤寒杆菌经L 型变异后形成的细胞壁缺陷突变株(CW DM )的脂类代谢活性,了解这些CW DM 变异的性质和探讨细菌细胞壁缺陷突变与细菌演变的关系。结果表明,沙门菌CW DM s 具有显著的胆固醇和甘油三脂代谢活性、对毛地黄皂苷高度敏感并且还具有与白色念珠菌相似的胆固醇和甘油三脂的含量,但未能检出胆碱酯酶活性。CW DM s返祖菌丧失了脂类代谢酶类和胞浆膜不含胆固醇,恢复了与其亲代细菌型相似的代谢特征。提示在沙门菌天然即存在有与脂类及胆固醇代谢相关的基因,细胞壁的缺陷导致这些脂类及胆固醇代谢基因活化,以致 CW DM s 能够表达固醇和甘油三脂代谢活性和胞浆膜含有胆固醇