Radioimmunoassay for fibroblast growth factor (FGF): release by the bovine anterior pituitary in vitro

A radioimmunoassay (RIA) was developed to measure fibroblast growth factor (FGF) using antiserum generated against a synthetic replicate of [Tyr¹⁰]FGF(1–10). The antisera, previously shown to be capable of inhibiting the biological action of FGF on bovine aortic arch endothelial cells in vitro [1], are highly specific for the amino-terminus of FGF. In the RIA, the antisera recognize the decapeptide antigen [Tyr¹⁰]FGF(1–10) and the intact mitogen on an equimolar basis and show less than 0.01% cross-reactivity with N-acetyl-[Tyr¹⁰]FGF(1–10).

Bovine adenohypophysial cells maintained in primary monolayer culture release and ir-FGF which is indistinguishable from the intact mitogen in as much as it is retained on heparin-Sepharose affinity columns and shows a dose-dependent and parallel displacement in RIA. The release of ir-FGF by the bovine adenohypophysis can be increased with forskolin (10⁻⁵ M) or KCl (50 mM). Preincubation of pituitary cells with 17β-estradiol has no measurable effects on basal ir-FGF, but increases the release after KCl treatment 2–3-fold. These results show that ir-FGF can be released by the bovine adenohypophysis in vitro and lend credence to the hypothesis that FGF plays a physiological role in the homeostatic mechanisms regulating mesoderm-derived cell growth.     

An HRP-study of the frequency-place map of the horseshoe bat cochlea: Morphological correlates of the sharp tuning to a narrow frequency band

Summary The frequency-place map of the horseshoe bat cochlea was studied with the horseradish peroxidase (HRP) technique involving focal injections into various, physiologically defined regions of cochlear nucleus (CN). The locations of labeled spiral ganglion cells and their termination sites on inner hair cells of the organ of Corti from injections into CN-regions responsive to different frequencies were analyzed in three dimensional reconstructions of the cochlea. Horseshoe bats from different geographical populations were investigated. They emit orientation calls with constant frequency (CF) components around 77 kHz (Rhinolophus rouxi from Ceylon) and 84 kHz (Rhinolophus rouxi from India) and their auditory systems are sharply tuned to the respective CF-components.The HRP-map shows that in both populations: (i) the frequency range around the CF-component of the echolocation signal is processed in the second half-turn of the cochlea, where basilar membrane (BM) is not thickened, secondary spiral lamina (LSS) is still present and innervation density is maximal; (ii) frequencies more than 5 kHz above the CF-component are processed in the first halfturn, where the thickened BM is accompanied by LSS and innervation density is low; (iii) frequencies below the spectral content of the orientation call are represented in apical turns showing no morphological specializations. The data demonstrate that the cochlea of horseshoe bats is normalized to the frequency of the individual specific CF-component of the echolocation call.The HRP-map can account for the overrepresentation of neurons sharply tuned to the CF-signal found in the central auditory system. A comparison of the HRP-map with a map derived with the swollen nuclei technique following loud sound exposure (Bruns 1976b) reveals that the latter is shifted towards cochlear base by about 4 mm. This discrepancy warrants a new interpretation of the functional role of specialized morphological structures of the cochlea within the mechanisms giving rise to the exceptionally high frequency selectivity of the auditory system.Abbreviations AVCN anteroventral CN - BF best frequency - BM basilar membrane - CF constant frequency - CN cochlear nucleus - DCN dorsal CN - FM frequency modulated - HRP horseradish peroxidase - IHC inner hair cell - LSS secondary spiral lamina - OHC outer hair cell - PVCN posteroventral CN - RF resting frequency - RRc Rhinolophus rouxi from Ceylon - RRi Rhinolophus rouxi from India     

花椰菜(Brassica oleracea var botrytis)下胚轴培养过程中过氧化物酶活性及同工酶谱的变化

花椰菜下胚轴外植体在MS+6BA 5 ppm的培养基上能分化出芽,在MS+2,4-D2ppm的培养基上能脱分化而形成愈伤组织。用3种不同的酚类物质(咖啡酸、阿魏酸、愈创木酚及联苯胺)作氢供体发现分化过程中的过氧化物酶活性高于脱分化过程,其中以咖啡酸作氢供体显示的活性最高,阿魏酸及愈创木酚次之,而联苯胺最小。用聚丙烯酰胺凝胶电泳分离阴极向及阳极向过氧化物酶同工酶,在分化及脱分化培养过程中均不断出现新的酶带,前者有13条,后者为11条,两者的差别主要在阴极向酶带,在分化过程中多了两条酶带(C_1和C_3),同时C_2带活性也比脱分化的高。阳极向酶带也有差别,A_2和A_2两条酶带在分化过程中逐渐加强,但是在脱分化过程中却逐渐消失。反映了两个过程生理上的差别。     

新城疫病毒单克隆抗体的特性及应用

建立了8个分泌抗新城疫病毒(NDV)特异性单克隆抗体(McAb)的杂交瘤细胞株,根据它们的免疫生物学特性可以分为三种类型:(1)具有FA和ELISA特性(FN1、FN4、FN29、FN30、FN35、FNl22);(2)具有FA、ELISA和HI特性(FN7);(3)具有ELISA、HI特性和中和能力(FN106),根据FN30和FN106的ELISA试验,可将11个NDV毒株分为二种不同的抗原群,应用FN4-FITC,FN7-FITC和FN29-HRP试剂,对人工感染NDV和野外送检病例检测结果表明,单抗试剂的DFA阳性率(92.3％)高于病毒分离阳性率(87.2％),两种方法的符合率89.7％,这些单抗试剂用于临床诊断敏感性和特异性高,且方法快速、简便。     

沙雷氏杆菌(Serratia marcescens)红素缩合酶的提取与稳定性的研究

 <正> 红素(Prodigiosin)是某些沙雷氏杆菌(Serratia marcescens)菌株所合成的一种三吡咯衍生物的红色色素,它对某些细菌,真菌和原虫具有抗生作用,因而引起人们的注意。在活菌体内,它由两个中间前体:2-甲基-3-戊基吡咯(MAP)和4-甲氧基-2,2’-双吡咯-5-甲醛(MBC)缩合而成。催化这一反应的酶,我们称之为红素缩合酶。除MAP外,某些人     

大鼠非致瘤四倍体细胞系的建立及细胞遗传学和酶学特征

从大鼠自然诱发的肉瘤细胞中,我们建立了一个四倍体细胞系(4n=84),命名为RC(ratcell)。它具有典型的成纤维细胞外形,能在玻璃表面贴壁生长,但不能生长在琼脂半固体培养基中。该细胞在含15％小牛血清的RPMI 1640培养基中生长良好,至今已连续繁殖112世代,细胞群体倍增时间约为15小时。染色体G-带分析表明,RC为整四倍体细胞,它的1条X染色体在第32至34区为均染区。RC细胞核仁组织者(NORs)活性显然比大鼠二倍体细胞NORs活性的加倍还高(P<0.001)。这个具有非常高NORs活性的RC细胞系对于研究细胞18S+28S rRNA基因转录活性的调控、基因表达与基因剂量关系有一定的意义。RC细胞还有异常高的磷酸酯酶活性,而且它的同工酶谱也与大鼠肌肉细胞明显不同。体内接种实验和扫描电镜的观察表明,RC是非致瘤细胞。RC细胞各号染色体的C-带图样与大鼠二倍体细胞无明显的差异。     

A simple purification of calmodulin by reversed phase chromatography with hydrophobic polymer 3520

A new adsorption chromatography procedure for the purification of calmodulin from bovine brain was developed using polymeric adsorbent 3520. Calmodulin was first isolated by DEAE-Cellulose column chromatography and further purified to apparent homogeneity following elution with 50% ethanol from the adsorbent column. Polyacrylamide gel electrophoresis showed one band either in the presence of Ca2+ or EGTA. The polymeric adsorbent 3520 is a non-polar polymer lacking exchangeable groups. The selective adsorption of calmodulin is based on hydrophobic interaction within the matrix, and is Ca2+ independent. Neither high salt (0.5 M NaC1) nor EGTA (5 mM) was able to elute the CaM from the adsorption column whereas ethanol (50%) eluted it completely. This method is simple to use and it provides highly purified calmodulin with high yield.     

In vitro release of growth hormone-releasing factor from rat hypothalamus: effect of insulin-like growth factor-1

The release of growth hormone-releasing factor (GHRF) from rat hypothalamus was investigated in vitro. After 60 min preincubation the released GHRF from sliced rat hypothalamic fragments during 60 min incubation was detected by a highly specific and sensitive radioimmunoassay for rat GHRF. The release of GHRF was Ca2+-dependent and enhanced by high concentration of K+. Insulin-like growth factor-1 (IGF-1) significantly decreased GHRF release to 65% and 84% of the control at concentrations of 10(-8) M and 10(-7) M, respectively. These results suggest that this in vitro system is useful for the investigation of the mechanism of GHRF release from the hypothalamus and that IGF-1 is probably involved in the feedback inhibition of growth hormone secretion by attenuating GHRF release from the hypothalamus besides countering the effect of GHRF on the pituitary.     

重楼属植物的免疫血清学研究

以海南重楼(Paris dunniana),花叶重楼(P.marmorata),多叶重楼(P.polyphylla),凌云重楼(P.cronquistii),五指莲(P.axialis),滇重楼(Paris polyphylla var.yunnanensis)等六种重楼属植物的根茎为材料,提取分离其球蛋白组份作为抗原,免疫家兔得到相应的抗血清。通过免疫双扩散,聚丙烯酰胺凝胶电泳、免疫电泳、免疫吸收试验以及酶标免疫等方法,研究了重楼属上述六个种和变种的血清学行为以及他们之间的相互关系。在此基础上,用不加权算术平均对群法(UPGMA)经成聚运算,得到基于平均吸收相似系数(Sabs.mean.)和Jaccard's结合系数的两种表相图,这两种表相图的结果是一致的。花叶重楼与多叶重楼有较大的血清反应相似性,五指莲与滇重楼有最大的血清反应相似性。多型种P.polyphylla在形态上的较大变异与血清反应分析的结果是一致的,而且血清反应相似性分析支持了从形态分析得出的假设:Paris dunniana在重楼属的系统发生中是一比较原始的种。