Selective solubilization and purification of cell-surface concanavalin A-binding glycoproteins from Novikoff hepatocellular carcinoma cells

Novikoff hepatocellular carcinoma cells possess cell-surface glycoproteins that bind the lectin, concanavalin A. A subset of Con A-binding plasma membrane glycoproteins was solubilized by addition of n-butanol to a suspension of Novikoff cells. Glycoproteins solubilized into the n-butanol-saturated aqueous phase of the two-phase mixture were purified by sequential chromatography on DEAE-cellulose and Sepharose-conjugated concanavalin A. Glycoproteins specifically bound to the Sepharose-conjugated Con A exhibited apparent M_r = 72,000 to 125,000. The plasma membrane localization of these components was inferred by their isolation from cells surface labeled with NaIO₄/ NaB³H₄. A xenoantiserum, raised against glycoproteins specifically bound to Sepharose-conjugated concanavalin A was employed to identify reactive components in nonionic detergent extracts of Novikoff tumor cells or rat hepatocytes surface labeled using lactoperoxidase-catalyzed iodination (¹²⁵I). Major reactive peptides in extracts of Novikoff cells exhibited apparent M_r = 74,000, 82, 000, 110,000, and 135,000, while those in extracts of hepatocytes possessed apparent M_r = 98,000 and 105,000. The reactivity of the antiserum with extracts of ¹²⁵I-labeled Novikoff cells was abolished by absorption of the antiserum with hepatocytes, indicating that the qualitative differences observed may result from structural modification of one or more cell-surface glycoproteins, rather than the expression of new or inappropriate glycoproteins. This antiserum will provide a useful probe to investigate alterations in the expression or structure of glycoproteins that occur as a consequence of malignant transformation or adaptation of malignant cells to growth in the ascitic form.     

Rapid selective stimulation by growth factors of the incorporation by BALB/C 3T3 cells of [35S]methionine into a glycoprotein and five superinducible proteins

Within four hours of adding fibroblast growth factor, epidermal growth factor, prostaglandin F_2α, or serum to quiescent $\text{Balb}\text{c}$ 3T3 cells we observe selective increases in the incorporation of [³⁵S]methionine into six proteins; “major excreted protein” (MEP) and five “superinducible proteins” (SIPs). The mechanisms regulating the extracellular expression of MEP and the SIPs differ. 1) The levels of MEP but not SIPs are increased by NH₄Cl; and 2) Cycloheximide increase SIP and decreases MEP production. These results suggest that production of MEP and the SIPs are controlled by other proteins; MEP by a positive, and the SIPs by a negative effector.     

1,6-Diamino-2,5-anhydro-1,6-dideoxy-dl-glucitol and some derivatives thereof

1,6-Diamino-2,5-anhydro-1,6-dideoxy-dl-glucitol dihydrochloride and some derivatives were synthesized from 3,5-di-O-acetyl-1,6-dibromo-1,6-dideoxy-d-mannitol. Introduction of the 2,5-anhydro ring and subsequent replacement of the terminal bromine atoms by azide gave low yields of the diazide; therefore, a reverse reaction-sequence was applied. The azido groups were reduced with hydrogen sulfide-pyridine, and the amino groups formed were methylated by using formaldehyde-formic acid and subsequently treating with borohydride. According to ¹³C-n.m.r. investigations, the symmetrically substituted, 2,5-anhydroglucitol derivatives are present mainly in the ⁴T₃ [“north” (N) type of twist] conformation, whereas the analogous l-iditol derivatives mainly adopt the ³T₄ [“south” (S)] type. The different quaternary salts obtained on methylation of the corresponding 1,6-bis(dimethylamino)derivatives with methyl iodide (aiming at the structure of epi-muscarine) showed no muscarine-like, biological activity.     

Surface antigen changes occurring in short-term cultures of activated human T lymphocytes: analysis by flow cytometry

Surface antigens of activated and cultured human T cells were studied using peripheral blood lymphocytes activated with conditioned medium from phytohemagglutinin-activated leukocytes and maintained in liquid culture for 2 weeks with conditioned medium containing Interleukin 2. The ensuing cell population was tested for kinetic changes in cell size and for the expression of surface antigens by immunofluorescence staining with a panel of monoclonal antibodies and analysis by flow cytometry. Upon activation, the cell population progressively increased in size to large blasts, with the rapid appearance on all of the large dividing cells of the antigen recognized by OKT9, the transferrin receptor. Cells within the population continued to express the common peripheral T-cell antigens bound by OKT3 and UCHT1, and also the antigen bound by 3A1, but never the antigen bound by OKT6, a thymic cell marker. From the time of activation an increasing proportion of the T cells, up to 80%, expressed the antigen detected with OKIa and FMC4, which recognise nonpolymorphic Ia determinants. This sequence of events was followed by a general decrease in size of the cell population, a process accompanied by further phenotypic changes. The percentage of cells expressing Ia antigens decreased, but most striking was the rapid change in the OKT4:OKT8 ratio of cells within the population, from 60:40 to 40:60. Thereafter the proportions of OKT4⁺ to OKT8⁺ cells within the cultures remained relatively stable and it is suggested that these data provide evidence for a possible change in phenotype of cultured human T lymphoblasts, from OKT4 to OKT8.     

兰属(Cymbidium)5种植物叶结构特征

采用光学显微镜和体视镜,对兰属5种植物（春兰、蕙兰、建兰、寒兰、墨兰）的叶结构特征（叶脉结构、叶表皮结构、解剖结构）进行了观察和测量。结果显示,兰属5种植物的叶脉结构相似,叶表皮结构和解剖结构存在较大差异,其中最主要的差异是皮下纤维束的多少和边缘角质层锯齿形态。依据叶结构特征聚类分析结果,5种兰可分为春兰组、建兰组和墨兰组3组。本研究表明叶结构特征对兰属5种植物的分类具有重要意义。     

Enucleation eliminates a differentiation-specific surface marker from normal and leukemic murine erythroid cells

Mammalian erythropoiesis includes a step in which the nucleus is extruded through the cell membrane. We have investigated the relationship between concanavalinA (conA) plasma membrane receptors, which are known to leave the incipient reticulocyte during enucleation, and regions of the plasma membrane which bind merocyanine 540, a differentiation-specific marker of hematopoietic cells. The distribution of these two fluorescent probes was examined on living cells from the spleens of neonatal mice and on erythroleukemia cells induced to enucleate in culture. In both cases, the region of the membrane extruded with the nucleus preferentially binds conA and merocyanine 540, whereas the plasma membrane which is left behind retains the capacity to bind another lectin, wheat germ agglutinin (WGA). The implications of these findings are discussed with respect to the mechanism by which markers are eliminated from the erythrocyte cell surface.     

木姜子叶精油的化学成分研究

采用气相色谱-质谱联用仪对产于秦岭太白山的木姜子叶的挥发油成分进行分析鉴定,从分离出的79个峰中鉴定了32种成分,其含量占挥发油总量的90.59％,主要成分为1,3,3-三甲基-乙-氧杂双环[2,2,2]辛烷(59.96％),1,8-桉油醇(8.96％),香茅醛(6.86％),2-甲基-5-(1-甲基乙烯基)环己酮(4.34％)和澄花醇乙酯(3.19％)。     

麦草栽培姬菇配方的数学模型研究

借助二次通用旋转组合设计法研究了麦草栽培姬菇配方的数学模型,以探讨利用麦草替代棉籽壳的新配方.试验结果得出了配料的取值范围及最佳栽培配方,即麦草79％,麸皮15％,玉米粉4％,过磷酸钙1％,黄豆饼粉1％.在实际生产中很有应用价值.     

杉木人工林水量平衡和蒸散的研究

 本文根据我们严密设计的小集水区径流场连续6年的水文测定数据,进行了杉木人工林水量平衡和蒸散的研究。结果表明：集水区年平均降雨量1065.5mm, 在林冠作用面降雨量的分配中,林冠截留雨量264.6mm,截留率24.8%;穿透过林冠层的雨量799.82mm,树干径流量1.08mm,分别占降雨量的75.1%和0.1%。林内降水到达林地时,在枯枝落叶层这个作用面上净降水进行再分配,其中,地表径流量9.27mm,地下径流量203.00mm,总径流系数0.199。土壤蓄水量月变化较大,但年变化很小,占降雨量的1.2%。系统水量最大的输出是蒸散,每年以气态形式返回大气的水量866.03mm,占降雨量81.3%。在蒸散的水量中,林冠截留雨量的直接物理蒸发量为264.6mm,占总蒸散量的31.6%。