Complexity untwined: The structure and function of cucumber (Cucumis sativus L.) shoot phloem

Cucurbit phloem is complex, with large sieve tubes on both sides of the xylem (bicollateral phloem), and extrafascicular elements that form an intricate web linking the rest of the vasculature. Little is known of the physical interconnections between these networks or their functional specialization, largely because the extrafascicular phloem strands branch and turn at irregular angles. Here, export in the phloem from specific regions of the lamina of cucumber (Cucumis sativus L.) was mapped using carboxyfluorescein and ¹⁴C as mobile tracers. We also mapped vascular architecture by conventional microscopy and X-ray computed tomography using optimized whole-tissue staining procedures. Differential gene expression in the internal (IP) and external phloem (EP) was analyzed by laser-capture microdissection followed by RNA-sequencing. The vascular bundles of the lamina form a nexus at the petiole junction, emerging in a predictable pattern, each bundle conducting photoassimilate from a specific region of the blade. The vascular bundles of the stem interconnect at the node, facilitating lateral transport around the stem. Elements of the extrafascicular phloem traverse the stem and petiole obliquely, joining the IP and EP of adjacent bundles. Using pairwise comparisons and weighted gene coexpression network analysis, we found differences in gene expression patterns between the petiole and stem and between IP and EP, and we identified hub genes of tissue-specific modules. Genes related to transport were expressed primarily in the EP while those involved in cell differentiation and development as well as amino acid transport and metabolism were expressed mainly in the IP.     

Induction of Apoptosis in Cardiac Myocytes by an A3Adenosine Receptor Agonist

The effects of the selective adenosine (ADO) A₃receptor agonist IB-MECA (N⁶-(3-iodobenzyl)adenosine-5′-N-methylcarboxamide) on cultured newborn rat cardiomyocytes were examined in comparison with ADO, the ADO A₁receptor-selective agonistR-PIA (N⁶-R-phenylisopropyladenosine), or the ADO A₃selective antagonist MRS 1191 (3-ethyl-5-benzyl-2-methyl-6-phenyl-4-phenylethynyl-1,4-(±)-dihydropyridine-3,5 dicarboxylate), using digital image analysis of Feulgen-stained nuclei. At high concentration, IB-MECA (10 μM ) and ADO (200 μM) induced apoptosis; however,R-PIA or MRS 1191 did not have any detectable effects on cardiac cells. In addition, DNA breaks in cardiomyocytes undergoing apoptosis following treatment by IB-MECA were identifiedin situusing the nick end labeling of DNA (“TUNEL”-like) assay. In the presence of 10 μM IB-MECA, disorder in the contraction waves appeared, and a decrease in the frequency of beats was observed. Analysis with light microscopy revealed that the number of contracting cells decreased in a concentration-dependent manner. The A₃receptor agonist IB-MECA caused an increase in intracellular free calcium concentration ([Ca²⁺]_i). The drug produced a rapid rise followed by a sustained increase in [Ca²⁺]_i, which lasted for 40–60 s. Finally, cessation of beating and Ca²⁺transients were observed. Full recovery of contractile activity and rhythmical Ca²⁺transients were observed 15–20 min after IB-MECA treatment. The induction of apoptosis in the cardiocytes by IB-MECA led to the appearance of features of apoptotic nuclei: the onset of condensation, compacting, and margination of nuclear chromatin. These effects were accompanied by the disintegration of the structural framework of the nucleus and nuclear breakdown. The results suggest that activation of the A₃adenosine receptor may participate in the process of apoptosis in cardiomyocytes.     

Cell cycle-dependent disruption of microtubules by methyl jasmonate in tobacco BY-2 cells

Summary Methyl jasmonate, a growth-regulating substance that is ubiquitous in the plant kingdom, was found to disrupt cortical microtubules in tobacco cultured cells. It exerted a microtubule-disrupting effect only in cells at the S phase of the cell cycle. Neither microtubules in preprophase bands, spindles and phragmoplasts nor cortical microtubules at stages of the cell cycle other than the S phase were disrupted by methyl jasmonate. Jasmonic acid was as effective as methyl jasmonate in disrupting cortical microtubules.Abbreviations BUdR 5-bromo-2-deoxyuridine - 2,4-D 2,4-dichlorophenoxyacetic acid - DMSO dimethyl sulfoxide - EGTA ethylene glycol bis(2-aminoethyl ether)-tetraacetic acid - FITC fluorescein isothiocyanate - FUdR 5-fluoro-2-deoxyuridine - JA jasmonic acid - JA-Me methyl jasmonate - PBS phosphate-buffered saline - PMSF phenylmethylsulfonyl fluoride     

Morphological and Neurochemical Effects of Diazepam and Phenobarbital on Selective Culture of Neurons from Fetal Rat Brain

The responses to diazepam (DZ) and phenobarbital (PhB) were studied in enriched neuronal primary cultures from rat embryo hemispheres. Cells were grown in chemically defined medium and the drugs were added for 3 days to cultures, at pharmacologically active concentrations. Following exposure to DZ or to PhB, morphological changes, such as less prominent neuronal processes, were observed in neurons. It was also shown that each drug reduced the specific uptake of 2-deoxy-D-glucose by the cells and interfered with protein and RNA metabolism. It was concluded that both DZ and PhB might affect, at least transiently, the normal growth of neurons in culture.     

培养心肌细胞内的SDH图象分析——大豆磷脂脂质体对缺血再灌心肌膜损伤的影响

琥珀酸脱氢酶(SDH)的活性在判断心肌缺血,再灌性损伤中起重要作用。本文通过IBAS图象分析系统对缺血再灌的培养心肌细胞内的SDH定量评估,以求探讨大豆磷脂脂质体对心肌缺血再灌性损伤的保护作用。结果表明,在缺血及再灌组的心肌细胞内出现较强的SDH损伤性反应,大豆磷脂脂质体能明显减轻这种反应,提示大豆磷脂脂质体对缺血及缺血/再灌心肌具有明显的保护作用。     

Evidence for De Novo rearrangements of Drosophila transposable elements induced by the passage to the cell culture

The genomic distribution and the number of elements of eleven transposon families have been compared by the Southern technique between permanent cultured cells, larval salivary glands and the brains and whole flies of an inbred Drosophila line (inb-c) from which the cells were established. In cultured cells, changes in restriction patterns consistent with various types of rearrangements such as amplification, transposition and excision of the elements of copia, 1731, 412, 297 and mdg-4 transposon families are detected whereas B 104, G and blood elements appear stable. In previous reports these rearrangements were not detected among individuals of the inb-c line or among samples of somatic tissues, or in samples spanning years of maintenance of cultured cells. Hence, we believe that they have been induced de novo during the passage to the cell culture.     

The relationship between cytotoxic effect and binding to mammalian cultured cells of Clostridium perfringens enterotoxin

Abstract The relationship between the cytotoxic effect and binding to different cell lines of Clostridium perfringens enterotoxin was investigated. The enterotoxin released ⁵¹Cr from Vero and MDCK cells labeled with Na₂-⁵¹CrO₄. The effect varied depending upon the dose of enterotoxin and the duration and temperature of the interaction. The enterotoxin gave no effect on FL, KB, or L-929 cells. [¹²⁵I]Enterotoxin bound specifically to Vero and MDCK cells via a binding site of distinct nature, but not to FL, KB, or L-929 cells. The number of the binding sites located on one MDCK cell (1.98 × 10⁶ sites/cell) was three times that on one Vero cell (5.64 × 10⁵ sites/cell), although the binding affinity of MDCK cell ( K _a/ 3.76 × 10⁷ M⁻¹) was 0.1 that of Vero cells ( K _a/ 3.23 × 10⁸ M⁻¹). Binding of the enterotoxin to susceptible cells was temperature-independent.     

Studies on oxygen and volume restrictions in cultured cardiac cells I. A model for ischemia and anoxia with a new approach

Summary A novel incubation unit is described that is highly suitable for thorough studies of oxygen deprivation states. Its application with cultured heart cells is experimentally demonstrated. The release of enzymes, taken as a marker for cell damage, has clearly shown that restriction of the volume of extracellular medium combined with oxygen plus glucose deprivation caused greatest cellular damage. It may be considered as an experimental ischemia-like state. Furthermore, the onset of cellular damage followed a time table very much like that occurring in vivo under similar conditions, more so than any other previously described studies. A time lag between the release of cytoplasmic enzymes and lysosomal enzymes and other observations made in the present study suggests a sequential order of events in which the release of cytoplasmic enzymes occurs at a stage of reversible damage due to oxygen deprivation, whereas the release of lysosomal enzymes may point at irrepairable damage. Supported by grants from The Chief Scientist, Ministry of Health, State of Israel; The Ministry of Education and Sciences, State of Niedersachsen (FRG); and The Foundation for Heart Research from Mr. and Mrs. D. Vidal-Madjar, Paris, France. This study was done as partial fulfilment of Vemuri's Ph.D. thesis in biochemistry.