Presence of nadph-cytochrome P-450 reductase in rat liver golgi membranes: Evidence obtained by immunoadsorption method

Light Golgi fractions (GF(1+2)) prepared from rat liver homogenates by a modification of the Ehrenreich et al. procedure (J. Cell Biol. 59:45) had significant NADPH-cytochrome P(450) reductase (NADPH-cyt c reductase) activity if assayed immediately after their isolation. An antibody raised in rabbits against purified microsomal and Golgi fractions. To find out whether this activity is located in bona fide Golgi elements or in contaminating microsomal vesicles, we used the following 3-step immunoadsorption procedure: (a) antirabbit IgG (raised in goats) was conjugated to small (2-5 μm) polycrylamide (PA) beads; (b) rabbit anti NADPH-cyt c reductase was immunoadsorbed to the antibody-coated beads; and (c) GF(1+2) was reacted with the beads carrying the two successive layers of antibodies. The beads were then recovered by centrifugation, and were washed, fixed, embedded in agarose, and processed for transmission electromicroscopy. Antireductase- coated beads absorbed 60 percent of the NADPH-cyt c reductase (and comparable fractions of NADH-cyt c reductase and glucose-6-phosphatase) but only 20 percent of the galactosyltransferase activity of the input GF(1+2). Differential vesicle counts showed that approximately 72 percent of the immunoadsorbed vesicles were morphologically recognizable Golgi elements (vesicles with very low density lipoprotein [VLDL] clusters or Golgi cisternae); vesicles with single VLDL and smooth surfaced microsome-like vesicles were too few (approximately 25 percent) to account for the activity. It is concluded that NADPH-cytochrome P(450) reductase is a Golgi membrane enzyme of probably uneven distribution among the elements of the Golgi complex.     

THE IN VITRO DIFFERENTIATION OF DENSITY SUB-POPULATIONS OF COLONY-FORMING CELLS UNDER THE INFLUENCE OF DIFFERENT TYPES OF COLONY-STIMULATING FACTOR

The in vitro proliferation and differentiation of myeloid progenitor cells (CFU-c) in agar culture from CBA/Ca mouse bone marrow cells was studied. Density sub-populations of marrow cells were obtained by equilibrium centrifugation in continuous albumin density gradients. The formation of colonies of granulocytes and/or macrophages was studied under the influence of three types of colony-stimulating factor (CSF) from mouse lung conditioned medium CSF_MLCM), post-endotoxin mouse serum (CSF_ES) and from human urine (CSF_Hu). The effect of the sulphydryl reagent mercaptoethanol on colony development was also examined. The density distribution of CFU-c was dependent on the type of CSF. Functional heterogeneity was found among CFU-c with partial discrimination between progenitor cells forming pure granulocytic colonies and those forming pure macro-phage colonies. Mercaptoethanol increased colony incidence but had no apparent effect on colony morphology or the density distribution of CFU-c.     

In vitro flowering in cassava (Manihot esculenta Crantz)

Meristem-derived plantlets of cassava (Manihot esculenta Crantz) were induced to flower in vitro. Five genotypes out of 13 consistently responded to our culture conditions giving rise to male or female flowers. Male flowers contained anthers in which meiosis occurred and apparently normal pollen grains were formed.     

Red Light-Independent Instability of Oat Phytochrome mRNA in Vivo

Phytochrome A (phyA) mRNA abundance decreased rapidly in total RNA samples isolated from 4-day-old etiolated oat seedlings following a red light pulse. Putative in vivo phyA mRNA degradation products were detectable both before and after red light treatment. Cordycepin-treated coleoptiles were unable to accumulate the chlorophyll a/b-binding protein mRNA in response to red light, indicating that cordycepin effectively inhibited mRNA synthesis. In cordycepin-treated coleoptiles, phyA mRNA rapidly decreased in abundance, consistent with the hypothesis that phyA mRNA is inherently unstable, rather than being destabilized after red light treatment of etiolated oat seedlings.     

Routes to chaos in a model of a bursting neuron.

Chaotic regimens have been observed experimentally in neurons as well as in deterministic neuronal models. The R15 bursting cell in the abdominal ganglion of Aplysia has been the subject of extensive mathematical modeling. Previously, the model of Plant and Kim has been shown to exhibit both bursting and beating modes of electrical activity. In this report, we demonstrate (a) that a chaotic regime exists between the bursting and beating modes of the model, and (b) that the model approaches chaos from both modes by a period doubling cascade. The bifurcation parameter employed is the external stimulus current. In addition to the period doubling observed in the model-generated trajectories, a period three "window" was observed, power spectra that demonstrate the approaches to chaos were generated, and the Lyaponov exponents and the fractal dimension of the chaotic attractors were calculated. Chaotic regimes have been observed in several similar models, which suggests that they are a general characteristic of cells that exhibit both bursting and beating modes.     

Clostridium difficile in association with sporadic diarrhoea.

A total of 154 patients admitted to an infectious diseases unit were included in a year''s prospective survey of sporadic diarrhoeal disease. Stools from 19 of them yielded Clostridium difficile, generally on more than one occasion. Twelve of these patients were assessed as having a severe or moderately severe gastrointestinal illness: Cl difficile was the only pathogen isolated from 10 of them, and two had an associated salmonella infection. Seven had had a recent course of antibiotics, but five had not taken antibiotics. Faeces from seven patients with moderate or mild gastrointestinal illness yielded Cl difficile, and two of these patients also had an associated salmonella infection. Two patients in this group had no antibiotic history. From these findings, the occurrence of C difficile in faeces could not be described as antibiotic-associated. Faecal Cl difficile cytotoxin was detected in only six patients, and generally at low levels. In such patients a more relevant pathogenic index might take account of the numbers of Cl difficile present and of their toxigenic potential.     

温度对干旱、盐胁迫下两种黄芪属种子萌发和幼苗生长的影响

为探讨温度对干旱、盐胁迫下黄芪属种子萌发和幼苗生长特性的影响,以黄芪属蒙古黄芪和扁茎黄芪2种种子为研究对象,纯净水处理为对照组,NaCl、PEG处理为实验组,设置4个渗透势水平(0、-0.1、-0.3、-0.5 MPa),置于5种不同的温度(10、15、20、25、30 ℃)下,每日观察并记录两种种子萌发和幼苗生长情况。结果表明：旱盐胁迫下蒙古黄芪和扁茎黄芪种子萌发最适宜的温度分别为25和20 ℃左右;蒙古黄芪耐高温不耐低温,而扁茎黄芪恰恰相反;但25和20 ℃均适宜两种幼苗生长,包括胚根、胚轴和子叶的生长。蒙古黄芪各处理组(除未发芽的种子)的平均发芽时间都比扁茎黄芪长;NaCl胁迫程度的增加使得两种种子的最终发芽率降低,但蒙古黄芪的耐盐性高于扁茎黄芪;随着PEG胁迫程度的增加,二者的发芽均受到抑制,甚至会出现完全不萌发,但扁茎黄芪的耐旱性高于蒙古黄芪;在相同的渗透势时,尤其是-0.5 MPa,PEG比NaCl对两种种子的影响大;交互胁迫作用下,随着渗透势的增加两种幼苗的鲜重、干重以及胚根、胚轴、子叶的长和宽变化较大;利用Design Expert软件预测发现：温度25 ℃、NaCl渗透势为-0.1 MPa,温度24 ℃、PEG渗透势为-0.04 MPa的处理是蒙古黄芪种子萌发和幼苗生长达到最优化的组合;而扁茎黄芪最优化的组合则为23 ℃下NaCl渗透势为-0.07 MPa的处理,20 ℃下PEG渗透势为-0.13 MPa的处理。     

Freeze-etch and thin section studies ofNeurospora crassa ascospores

Summary Mature, dormant ascospores derived from crosses of wild type stocks of the fungusNeurospora crassa were studied using both thin section and freeze-etch techniques for electron microscopy. The spore wall is composed of three major layers with, perhaps, a fourth distinct layer covering these layers. The spore contains several nuclei, many discrete mitochondria, lipid bodies, vacuoles and small pieces of endoplasmic reticulum. Freeze-etch fracture faces of several membranes are described in this paper. Included is a diagram illustrating various fracture faces and surfaces seen following freeze-etching. The thin section results are less than satisfactory but serve to generally confirm the freeze-etch observations and suggest that further work could lead to the development of techniques which will result in high quality thin sections of ascospores.     

Rat Sertoli cells in culture undergo metabolic co-operation with each other and with fibroblasts

Metabolic co-operation between Sertoli cells from adult rats was detected by adding one group of cells, which were the recipients, to a second group of cells, which had been labelled for 3 h with [3H]uridine and were the donors. Metabolic co-operation also was studied by co-culturing Sertoli cells, which were the donors, with human or Chinese hamster HGPRT- fibroblasts (recipients) in the presence of [3H]hypoxanthine. With both techniques the recipients in contact with donors had significantly more radioactive grains than did the recipients alone. In all cases the proportion of interactions that were positive for metabolic co-operation was greater than 80%.