Methylation of KvDMR1 involved in regulating the imprinting of CDKN1C gene in cattle

The CDKN1C gene encodes a cyclin‐dependent kinase inhibitor and is one of the key genes involved in the development of Beckwith–Wiedemann syndrome and cancer. In this study, using a direct sequencing approach based on a single nucleotide polymorphism (SNP) at genomic DNA and cDNA levels, we show that CDKN1C exhibits monoallelic expression in all seven studied organs (heart, liver, spleen, lung, kidney, muscle and subcutaneous fat) in cattle. To investigate how methylation regulates imprinting of CDKN1C in cattle, allele‐specific methylation patterns in two putative differential methylation regions (DMRs), the CDKN1C DMR and KvDMR1, were analyzed in three tissues (liver, spleen and lung) using bisulfite sequencing PCR. Our results show that in the CDKN1C DMR both parental alleles were unmethylated in all three analyzed tissues. In contrast, KvDMR1 was differentially methylated between the two parental alleles in the same tissues. Statistical analysis showed that there is a significant difference in the methylation level between the two parental alleles (P < 0.01), confirming that this region is the DMR of KvDMR1 and that it may be correlated with CDKN1C imprinting.     

M?ssbauer effect studies on some bioinorganic complexes of europium.

The bioinorganic complexes of europium with N-acetyl-DL-alanine, N-acetyl-DL-valine, and DL-alanyl-DL-alanine have been synthesized and the M?ssbauer spectra at room temperature have been measured for these solid state complexes. The M?ssbauer parameters indicate that the water molecules in these complexes are not directly linked to the central europium ion and are outside the coordination sphere of europium and biological ligands, and that the chemical bond between the europium ion and the ligands may be predominantly ionic in character, with the possibility of partial covalent contribution.     

Formation of lymphocyte colonies under serum-free culture conditions in normal individuals and patients with chronic lymphocytic leukemia

This paper describes a culture system which supports the formation of B cell and some T cell colonies under serum-free conditions in peripheral blood samples of normal individuals and patients with chronic lymphocytic leukemia (CLL) of B cell type. In this system, serum is replaced by bovine serum albumin, transferrin, cholesterol, insulin and catalase or horseradish peroxidase. In addition, it is necessary to add staphylococcus protein A, mitomycin-treated T cells as feeders and phytohemagglutinin leukocyte-conditioned medium as a source of growth factors. The plating efficiency is greatly enhanced when normal cells are incubated with galactose oxidase prior to plating and when CLL cells are exposed sequentially to neuraminidase and galactose oxidase.     

休眠与萌发过程中苹果种子的呼吸代谢

“秦冠”苹果种子的总呼吸强度和EMP途径的呼吸强度在2℃条件下层积之后逐渐增加,解除休眠临界期大体在层积后第30天发生,此后增长速度较快。HMP途径和TCAC在临界期之前变化极慢,而后急剧增加。种子萌发后子叶和胚轴的总呼吸强度、EMP和TCAC初期呈急剧上升趋势,大约当胚轴长度达2cm时迅速下降,HMP在萌发过程中始终呈上升趋势。细胞透性层积之后逐渐降低,解除休眠至萌发阶段呈上升趋势。用不同浓度呼吸抑制剂处理解除休眠种子的试验结果表明,除1mM碘乙酸能明显刺激萌发和生长外,NaCN、SHAM和丙酮酸都表现为抑制作用。     

Conformation of the second disulfide loop in human transforming growth factor-alpha studied by two-dimensional NMR spectroscopy

The determination of the conformation of a cyclic heptadecapeptide derived from the second loop of human transforming growth factor-α, [Ala²¹]-hTGF-α-(16–32), is described. Two-dimensional homonuclear Hartmann–Hahn and rotating-frame cross-relaxation spectroscopy in H₂O were used to obtain the complete proton resonance assignments and the necessary distance constraints between nonbonded hydrogen atoms to derive a conformation without involving any energy minimization. The result is an ellipsoidal-shaped structure with a turn at Gly19 and a bend formed by residues 26–29, Gln-Glu-Asp-Lys. Comparison is made with the second loop of human epidermal growth factor and the results are discussed in terms of receptor binding and biological activity.     

小鼠肝脏免疫抑制蛋白质(LISP)的分离纯化和鉴定

用硫酸铵分段盐析及DEAE-Sephadex A-50、羟磷灰石和CM纤维素等多种柱层析方法,从正常小鼠肝浸液中分离纯化出一种免疫抑制蛋白质(LISP)。在体外用微量该蛋白质就能强烈抑制小鼠T、B淋巴细胞对促有丝分裂原和同种异型抗原的增生反应。纯化的蛋白质在聚丙烯酰胺凝胶电泳(PACE)和等电聚焦(IEF)鉴定时均显示为一条区带,其等电点(pI)值在7.5—7.8范围。沉降系数利S_(20),w为5.39。Sephadex G-100凝胶层析测得LISP的分子量为78,000道尔顿。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)提示LISP是由二个相同的亚基组成,亚基分子量为38,500道尔顿。LISP是一种既非糖蛋白又非脂蛋白的碱性蛋白质,对它的氨基酸组成也作了分析。