Pea extracellular Cu/Zn-superoxide dismutase responsive to signal molecules from a fungal pathogen

We previously reported that the release of O₂ ⁻ from isolated pea cell walls was enhanced by a 70-kDa glycoprotein elicitor but was suppressed by mucin-type glycopeptide suppressors (supprescins A and B) prepared from pycnospore germination fluid of Mycosphaerella pinodes, causal agent of Mycosphaerella blight of pea. Here, we show that superoxide dismutase (SOD) in the apoplast fluid/cell wall of pea seedlings responds to the fungal elicitor and suppressor molecules. In a pharmacological study and with internal amino acid sequencing, the apoplastic SOD in a pea cultivar Midoriusui was found to be a Cu/Zn type SOD. We cloned a full-length cDNA of the Cu/Zn-SOD and designated it as PsCu/Zn-SOD1. An increase in PsCu/Zn-SOD1 mRNA and the PsCu/Zn-SOD1 protein was induced by treatment with the elicitor more intensively than by wounding. Such induction by the elicitor or wounding, however, was inhibited by the concomitant presence of supprescins. The SOD activity of recombinant PsCu/Zn-SOD1 was regulated directly by these signal molecules in a manner similar to their effect on the SOD activity in the apoplastic fluid and in the cell wall-bound proteins. Based on these findings, we discuss a role for PsCu/Zn-SOD1 in the pea defense response. The nucleotide sequence data of PsCu/Zn-SOD1 reported are available in the DDBJ/EMBL/GenBank databases under accession number AB189165.     

Expression of receptor tyrosine kinases VEGFR-1 (FLT-1), VEGFR-2 (KDR), EGFR-1, PDGFRa and c-Met in canine primary brain tumours

Inhibition of tumour growth and angiogenesis by targeting key growth factor receptors is a promising therapeutic strategy for central nervous system tumours. Characterization of these growth factor receptors in canine primary brain tumours has not been done. Using quantitative real‐time TaqMan polymerase chain reaction (PCR), we evaluated the expression of messenger RNA (mRNA) for five tyrosine kinase growth factor receptors (vascular endothelial growth factor receptor [VEGFR]‐1, VEGFR‐2, endothelial growth factor receptor [EGFR]‐1, platelet‐derived growth factor receptor a [PDGFRa], and c‐Met) relative to normal cerebral cortex in 66 spontaneous canine primary brain tumours. Increased expression of VEGFR‐1 and VEGFR‐2 mRNA was greatest in grade IV astrocytomas (glioblastoma multiforme) and grade III (anaplastic) oligodendrogliomas. EGFR‐1 mRNA expression was more consistently increased than the other receptors in all tumour types, while increased PDGFRa mRNA expression was mostly restricted to oligodendrogliomas. The similarities in increased expression of these tyrosine kinase growth factor receptors in these canine tumours, as compared to data from their human counterparts, suggest that common molecular mechanisms may be present.     

Implantation study on the potential of murine epidermal stem cell differentiation

AIM: To investigate the potential of murine epidermal stem cell (ESC) differentiation after seeded in a biodegradable carrier and implanted subcutaneously into syngeneic recipient mice. METHODS: ES cells were induced in vitro to differentiate into ESCs. After stained with a fluorescent dye Hoechst 33342, these ESCs were seeded into a polyglycolic acid (PGA) net containing collagen gel, functioning as a cell carrier, and implanted subcutaneously into 129/J mice, which were syngeneic to these stem cells. RESULTS: The ESCs kept alive in the implant when observed under a fluorescent microscopy 3 weeks or longer after implantation, and could differentiate into hair follicle-like structure, glandular structure, and gave rise to additional structures displaying features resembling native dermis. No apparent rejection or severe side effects were observed at least 10 weeks post-implantation. CONCLUSION: It is feasible to use these ESCs as seed cells in the study to fabricate dermal equivalent having the potential to develop dermal appendages.     

荔枝果皮对外源钙和蔗糖吸收及向细胞壁沉着的研究

利用放射性同位素示踪技术研究荔枝果皮对外源钙和蔗糖的吸收及细胞壁构建规律。结果表明,进入果皮的蔗糖主要被用以构建细胞壁,但随果实发育,蔗糖用于构建细胞壁比例减少,更多分布于果皮组织的可溶性成分中;果梗向果实运输钙的效率远远低于蔗糖运输效率;NAA处理果实短期内可明显促进蔗糖向果实运输,但不能促进果梗的钙进入果实,说明蔗糖和钙向果实运输有不同调控机制;施于果实表面的钙虽可被吸收并成为细胞壁的结构钙,但比例不到千分之一;硝酸(根)离子和NAA可一定程度促进外源钙向细胞壁沉着;抗裂的怀枝果皮细胞壁的钙含量比易裂的糯米糍高,其细胞壁结合外源钙的能力也强于后者,说明前者细胞壁中果胶半乳糖醛酸残基含量高于后者,也是怀枝具有较强抗裂性的物质基础之一;2品种14C-蔗糖向果皮细胞壁沉着均在果实发育初期最活跃,怀枝细胞壁构建也只是在果实发育初期比糯米糍更活跃,这就意味着抗裂性形成的关键时期是在果皮发育的初期。     

3类生境下脐橙果实细胞壁酶、瓤囊壁超微结构和品质变化的研究

为改进脐橙果实质地和科学生态区划与高品质栽培技术提供理论依据,研究了四川3类不同生境下脐橙果实果胶甲酯酶(PE)、多聚半乳糖醛酸酶(PG)、纤维素酶(CX)、瓤囊壁超微结构及主要品质指标的变化。结果表明:果实生长发育期细胞壁酶活性逐渐升高,但不同生境类型的变化趋势不同。超微结构中,南亚热带攀枝花脐橙的线粒体明显,中胶层清晰,细胞壁形态结构较完整;中亚热带江安和中北亚热带雅安脐橙细胞壁的中胶层明显裂解,线粒体膜分解,细胞器空泡化。不同生境SSC和可溶性糖差异显著。PG酶活性与≥10℃积温、1月份平均气温、年平均气温显著负相关,与年降雨量显著正相关。固酸比与≥10℃积温、年平均气温、1月均气温极显著正相关,与日照时数极显著正相关,与降雨量显著负相关。     

Comparative sensitivity of barley (Hordeum vulgare L.) to insecticide and fungicide on different stages of cell cycle

In this study, the comparison of cytogenetic effects of insecticide and fungicide in different phases of cell cycle was investigated in the root tip cells of barley (Hordeum vulgare L.). The seeds of H. vulgare L. Var. Karan 16 were treated with different concentrations (0.05%, 0.1% and 0.5%) of insecticide Profenophos (PF) and fungicide Mancozeb (MZ) for 6 h after presoaking durations of 7, 17 and 27 h.The different presoaking durations were used to bring the cells in various phases of cell cycle. Negative control was run parallel in distilled water. Cytogenetic examinations of root meristems exposed to the PF and MZ showed significant inhibition of mitotic index (MI) as well as significant increase in chromosomal aberrations (CAs). These parameters were dependent on the concentrations of insecticide and fungicide. The present study shows that PF and MZ both caused more damage in S phase of cell cycle which indicates that S phase is more sensitive in comparison to other phases.     

Recombinant hFGF-10 adenovirus promotes proliferation of kerotinocytes

AIM:To construct the recombinant adenoviral vector containing human fibroblast growth factor 10 (hFGF-10) gene, and to study the effect of the recombinant adenovirus on the proliferation of kerotinocytes. METHODS:HFGF-10 gene was amplified by PCR and ligated with shuttle vector pAdTrack-CMV to get the recombinant plasmid pAdTrack-CMV-hFGF-10, which was linearized with PmeI and transferred into Escherichia coli BJ5183 containing the adenoviral bone plasmid pAdEasy-1 for homologous recombination to obtain the recombinant adenoviral plasmid pAdEasy-hFGF-10. The recombinant adenoviral plasmid was then transfected into HEK-293 cell line to package and amplify the recombinant adenovirus. The expression of hFGF-10 in HaCat cells infected with the recombinant adenovirus was detected by Western blotting. The influence of the recombinant adenovirus on the proliferation of kerotinocytes was checked by MTT. RESULTS:The recombinant adenovirus containing hFGF-10 gene was successfully constructed, which effectively infected HaCat cells. The result of Western blotting showed that a protein in culture media of the infected HaCat cells reacted with hFGF-10 antibody. The recombinant adenovirus stimulated the proliferation of kerotinocytes. CONCLUSION:HaCat cells infected with the recombinant adenovirus expresses and secrets hFGF-10 protein, which promotes the proliferation of HaCat cells.     

Effects of cinnamyl aldehyde on c-Fos and c-Myc expression in NIH3T3 cells

AIM: Cinnamyl aldehyde (CA) is one alcohol ingredient derived from Cinnamomum cassia,which is widely used in treating chronic skin wound in Chinese medicine with the curative effect of ‘rescuing YANG’.The purpose of the present study was to investigate the expression of c-Fos,c-Myc proteins at different time points in NIH3T3 treated with CA and explore the possible mechanism of promoting cell proliferation by CA.METHODS: MTT assay was used for observing cell proliferation.Expression of c-Fos and c-Myc proteins in NIH3T3 cells were assessed by immunocytochemistry assay.RESULTS: The cell proliferation was promoted obviously when CA concentration was between 8.8×10^-2 μg/L and 8.8×10 μg/L.CA at concentration of 5.5 μg/L significantly induced expression of c-Fos,c-Myc proteins at 2-3 h after the stimulation compared with control group (P<0.01).CONCLUSION: CA increases expression of c-Fos and c-Myc proteins,which may be one of mechanisms for CA to promote NIH3T3 cell proliferation.