Intercellular adhesion strengthening as studied through simulated stress by organic acid molecules in potato (Solanum tuberosum L.) tuber parenchyma

Intercellular adhesion in some parenchyma becomes strengthened in response to stress. The present study provides an approach to investigate this phenomenon (usually attributed to pectin methyl esterase and binding of Ca(2+) and/or rhamnogalacturonan-II-borate) through reliable stress simulation by probing organic acid molecules in potato tuber parenchyma. Short-chain monocarboxylic acids induce consistent intercellular adhesion strengthening (3.8-5.3 newton) at pH >or= 3 < pK(a), where pectin methyl esterase activity and Ca(2+) or borate binding are limited, and vice versa at pH > pK(a) with a strength of 1.4-2.0 newton as compared to 0.3-0.4 newton for the nonincubated control. Strengthening of intercellular adhesion is characterized by prominent staining of pectin and protein and immunogold labeling of pectin in the cell wall and the middle lamellar complex, particularly after boiling. Pectin confers strengthening to the primary cell wall, as reflected by: (i) prominent immunogold labeling following boiling; and (ii) puncturing macerated cells by starch gelatinization pressure after enzymatic pectin removal.     

New paradigms of signaling in the vasculature: ephrins and metalloproteases

As our understanding of the control of vasculogenesis and angiogenesis continues to grow, we will be confronted with an increasing number of interacting and intersecting receptor-mediated signaling pathways. If we are to be successful in developing new and novel effective therapeutic reagents that can function as stimulators or inhibitors of these critically important processes, we will have to develop a sophisticated, full understanding of the complex interactions associated with ephrin-based and metalloprotease-based signaling pathways.     

Heparanase induces endothelial cell migration via protein kinase B/Akt activation

Heparanase is a mammalian endoglycosidase that degrades heparan sulfate (HS) at specific intra-chain sites. Blood-borne neutrophils, macrophages, mast cells, and platelets exhibit heparanase activity that is thought to be stored in specific granules. The degranulated heparanase is implicated in extravasation of metastatic tumor cells and activated cells of the immune system. Degranulation and heparanase release in response to an inflammatory stimulus or platelet activation would facilitate cellular extravasation directly, by altering the composition and structural integrity of the extracellular matrix, or indirectly, by releasing HS-bound proinflammatory cytokines and chemokines. We hypothesized that in addition to such indirect effect, the released heparanase may also locally affect and activate neighboring cells such as endothelial cells. Here, we provide evidence that addition of the 65-kDa latent heparanase to endothelial cells enhances Akt signaling. Heparanase-mediated Akt phosphorylation was independent of its enzymatic activity or the presence of cell membrane HS proteoglycans and was augmented by heparin. Moreover, addition of heparanase stimulated phosphatidylinositol 3-kinase-dependent endothelial cell migration and invasion. These results suggest, for the first time, that heparanase activates endothelial cells and elicits angiogenic responses directly. This effect appears to be mediated by as yet unidentified heparanase receptor.     

Identification and characterization of phytoplasmal genes, employing a novel method of isolating phytoplasmal genomic DNA

Phytoplasmas are unculturable, insect-transmissible plant pathogens belonging to the class Mollicutes. To be transmitted, the phytoplasmas replicate in the insect body and are delivered to the insect's salivary glands, from where they are injected into the recipient plant. Because phytoplasmas cannot be cultured, any attempt to recover phytoplasmal DNA from infected plants or insects has resulted in preparations with a large background of host DNA. Thus, studies of the phytoplasmal genome have been greatly hampered, and aside from the rRNA genes, only a few genes have hitherto been isolated and characterized. We developed a unique method to obtain host-free phytoplasmal genomic DNA from the insect vector's saliva, and we demonstrated the feasibility of this method by isolating and characterizing 78 new putative phytoplasmal open reading frames and their deduced proteins. Based on the newly accumulated information on phytoplasmal genes, preliminary characteristics of the phytoplasmal genome are discussed.     

Abnormal `wrinkled' cell walls and retarded development of transgenic Arabidopsis thaliana plants expressing endo-1,4-β-glucanase (cell) antisense

Transgenic Arabidopsis thaliana plants expressing cell antisense exhibit reduced levels of cell mRNA and protein compared with wild-type plants. The former display significant alterations in their phenotype. cell antisense plants have shorter stems and roots and are mechanically weaker than their wild-type counterparts. In cell antisense plants, the cell wall structure is markedly disrupted: both fluorescent confocal microscopy and scanning electron microscopy revealed `wrinkled' cell walls, thus indicating that CEL1 plays an important role in cell wall relaxation during cell growth and expansion. In cell antisense plants, the number of xylem elements per bundle is smaller than in the wild-type. In addition, both xylem elements and interfascicular fibers are significantly less lignified in the former. It is suggested that in A. thaliana, abnormal cell wall deposition affected by CEL1 depletion is associated not only with cell growth, but also with the differentiation process in the vascular and supporting tissues.Equal contributors     

Quantitative traits in plants: beyond the QTL

Phenotypic variation for quantitative traits results from segregation at multiple quantitative trait loci (QTL), the effects of which are modified by the internal and external environments. Because of their favorable genetic attributes (e.g. short generation time, large families and tolerance to inbreeding), plants are often used to test new concepts in quantitative trait analysis. Thus far, the molecular basis underlying allelic variation at QTL is similar to the identified variation for simple mendelian loci; namely, alterations in gene expression or protein function. Further comprehensive dissection of complex phenotypes will depend on our ability to link genetic components of the QTL variation to genomic databases.     

Quantitative microscopy of human DNA topoisomerase II-alpha expression in transitional cell carcinoma of the bladder

OBJECTIVE: To compare the nuclear size of various grades of transitional cell carcinoma (TCC) stained immunohistochemically with the nuclear enzyme topoisomerase II-alpha (topo II-alpha) in bladder urothelial neoplasms. STUDY DESIGN: Histologic sections from 53 consecutive papillary bladder neoplasms were stained immunohistochemically for topo II-alpha expression. There were 18 (33.9%) urothelial neoplasms of low malignant potential (UNLMP), 18 (33.9%) low grade urothelial carcinoma (LGUCa), and 17 (32%) with high grade urothelial carcinoma (HGUCa). The histologic slides were photographed at 400 x magnification and then projected on a screen, and the area with stained nuclei was measured. RESULTS: The cells and nuclei in HGUCa were significantly larger than in LGUCa (P < .05) and UNLMP (P < .01). CONCLUSION: Calculation of the area fraction of nuclei in TCC of the bladder stained with topo II-alpha is an additional method of establishing the grade of these tumors.     

Factors affecting <Emphasis Type="Italic">Sarcocystis</Emphasis> infection of rats on small tropical islands

The purpose of our research was to explore the limits of rat-python-Sarcocystis distribution among rats on the offshore tropical islands of Singapore, and to examine the effect of island size, insular isolation, landscape peculiarities and anthropogenic disturbance. Commensal rats (Rattus rattus) inhabited all of these islands, regardless of the islands size, proximity to the mainland, biogeographic features, and/or degree of anthropogenic interference. Rats caught on Sakijang Pelepah Island had well deliminated white bellies that are similar to those of sylvatic or feral rats. The prevalence of Sarcocystis spp. on individual islands ranged from 57 to 100%. This is consistent with the range found in forested habitats on Singapore Island. It also exhibited a similar diversity to Sarcocystis spp. and the predominance of Sarcocystis singaporensis. On Sakijang Pelepah Island, one rat (white bellied) was exceptionally heavily infected with both Sarcocystis villivilosi and Sarcocystis sulawesiensis. The muscles of the rats from nearly all of the islands contained immature sarcocysts, which implies that active transmission is taking place on these islands. This suggests that reticulated pythons (Python reticulatus), which are the definitive hosts of rat Sarcocystis, might have been established or frequented in all the islands of Singapore. Our study shows that the Sarcocystis infection load of the rats was negatively correlated with human disturbance, hinting that human disturbance restricts the pythons mobility and thus, reduces infection of Sarcocytis in the islands rats.