Changes in photosynthetic activity of the diatom Phaeodactylum tricornutum in a culture of limited volume

Changes in photosynthetic activity of a marine diatom duringalgal growth were studied with a typical culture medium formarine algae, ASP-2 (l). As the algal specimen, Phaeodactylumtricornutum was used for the experiments. Nitzschia closteriumand Chaetoceros sp. were also supplementarily used. Photosynthetic and p-benzoquinone Hill activities remarkablychanged with time during algalgrowth; with maximum activityfound in cells at log phase. A rapid decrease occurred in theinterphase from the log to stationary phase. The activity changewas not accompanied by variation in photosynthetic pigment content. The low concentration of phosphorus source was suggested asthe main cause for the change. On supply of extra inorganicphosphate, the time length for holding high photosynthetic activitybecame longer; or, the activity of the cells at stationary phaserecovered at least partly even in the dark. Dark recovery wasnot accompanied by either algal growth or an increase in thecontent of photosynthetic pigments. Inactivation of photosynthesis in the stationary growth phaseand activation by added phosphate in the dark were inferredto be due to changes in activities of both the CO₂-fixing andelectron transfer systems. The observed activity change maynot be attributable to a deficiency in inorganic phosphate asthe substrate for photophosphorylation. Similar changes in photosynthetic activity were also observedwith Nitzschia closterium and Chaetoceros sp. (Received January 30, 1971; )     

EFFECT OF LIGHT ON AUXIN TRANSPORT AND ELONGATION OF AVENA MESOCOTYL

The present work was undertaken to find if there are relations between light and auxin action on elongation of coleoptilar node and mesocotyl with Avena seedlings. Red light inhibited the elongation of mesocotyl and simultaneously decreased the rate of transport of diffusible auxin through the node. Red light also inhibited the transport of exogenously given IAA through the nodal region. The light inhibition of IAA transport was closely related to the increase of IAA immobilization. As the age proceeds, the ability of IAA immobilization increased with the decrease in the rate of mesocotyl elongation, even if the seedling was grown in complete darkness. The nature of radioactive substances found in the IAA-C¹⁴ treated tissue was examined by paper chromatography. The above results strongly suggested that the increase of IAA immobilization might result in the inhibition of mesocotyl elongation.     

The Fate of L-Tyrosine-UL−14C in Shoot-Forming Tobacco Callus

Tobacco callus fed L-tyrosine-UL^?14C was sampled at 3-day intervals for 15 days, homogenized and studied with respect to distribution of incorporated radioactivity. The supernatant obtained by centrifuging of the homogenates at 270 g contained the bulk of the radioactivity although significant activity was also detected in the pellet. Sucrose density gradient centrifugation of the supernatant showed over 90% of the recovered label to be associated with a fraction designated as “less dense than mitochondria”, with the remainder being found in the fraction identified as “mitochondria”. During tissue culture, virtually all of the radioactivity in the fraction “less dense than mitochondria” was recovered in the supernatant obtained by centrifugation at 100,000 g. From 4 to 18% of the labeling in the 100,000 g supernatant fraction was attributable to tyrosine-containing protein, and the rest to free tyrosine and unidentified anionic constituents. The highest proportions of radio-activity in the 270 g pellet were associated with substances extractable with NaCl, pronase, 4.6 N NaOH, and acetolyzing reagent. Low but substantial labeling characterized the extracts obtained with Triton X-100 and 1 N NaOH. The final unextractable residue contained 20% of the 270 g pellet radioactivity.     

Possible Involvement of ERK 1/2 in UVA‐Induced Melanogenesis in Cultured Normal Human Epidermal Melanocytes

UV‐induced melanogenesis is a well known physiological response of human skin exposed to solar radiation; however, the signaling molecules involved in the stimulation of melanogenesis in melanocytes following UV exposure remain unclear. In this study we induced melanogenesis in vitro in normal human epidermal melanocytes using a single irradiation with UVA at 1 kJ/m² and examined the potential involvement of mitogen‐activated protein kinases (MAPK) as UVA‐responsive signaling molecules in those cells. UVA irradiation did not affect the proliferation of melanocytes, but it did increase tyrosinase mRNA expression, which reached a maximum level 4 hr after UVA irradiation. The amount of tyrosinase protein, as quantitated by immunoblotting, was also increased at 24 hr following UVA irradiation. Among the MAPK examined, extracellular signal‐related kinase (ERK) 1/2 was phosphorylated within 15 min of UVA irradiation, but no such phosphorylation was observed for c‐Jun N‐terminal kinases (JNK) or p38. Accordingly, the activity of ERK1/2 was also increased shortly after UVA irradiation. These responses of ERK1/2 to UVA irradiation were markedly inhibited when cells were pre‐treated with N‐acetyl‐l ‐cysteine, an antioxidant, or with suramin, a tyrosine kinase receptor inhibitor. The formation of (6‐4)photoproducts or cyclobutane pyrimidine dimers was not detected in cellular DNA after UVA irradiation. These findings suggest that a single UVA irradiation‐induced melanogenesis is associated with the activation of ERK1/2 by upstream signals that originate from reactive oxygen species or from activated tyrosine kinase receptors, but not from damaged DNA.     

Quantitation and Visualization of Ultraviolet‐Induced DNA Damage Using Specific Antibodies: Application to Pigment Cell Biology

The major types of DNA damage induced by sunlight in the skin are DNA photoproducts, such as cyclobutane pyrimidine dimers (CPDs), (6‐4)photoproducts (6‐4PPs) and Dewar isomers of 6‐4PPs. A sensitive method for quantitating and visualizing each type of DNA photoproduct induced by biologically relevant doses of ultraviolet (UV) or sunlight is essential to characterize DNA photoproducts and their biological effects. We have established monoclonal antibodies specific for CPDs, 6‐4PPs or Dewar isomers. Those antibodies allow one to quantitate photoproducts in DNA purified from cultured cells or from the skin epidermis using an enzyme‐linked immunosorbent assay. One can also use those specific antibodies with in situ laser cytometry to visualize and measure DNA photoproducts in cultured cells or in the skin, using indirect immunofluorescence and a laser‐scanning confocal microscope. This latter method allows us to reconstruct three‐dimensional images of nuclei containing DNA photoproducts and to simultaneously examine DNA photoproducts and histology in multilayered epidermis. Using those techniques, one can determine the induction and repair of these three distinct types of DNA photoproducts in cultured cells and in the skin exposed to sublethal or suberythematous doses of UV or solar simulated radiation. As examples of the utility of these techniques and antibodies, we describe the DNA repair kinetics following irradiation of human cell nuclei and the photoprotective effect of melanin against DNA photoproducts in cultured pigmented cells and in human epidermis.     

封育对科尔沁沙地小叶锦鸡儿群落植被特征及空间异质性的影响

以科尔沁沙地小叶锦鸡儿群落为对象,分析了放牧和不同封育年限下小叶锦鸡儿群落的植被特征及植被分布的小尺度空间异质性.结果表明:放牧和封育样地内植被均以1年生草本植物为主,物种数没有明显差异;封育6年、封育12年样地的植株密度分别为(124.46±5.22)株·m~(-2)和(203.05±10.38)株·m~2,显著高于放牧样地(P<0.05);封育样地的Shannon-Wiener多样性指数、Simpson多样性指数、Pielou均匀度指数均低于放牧样地,并随着封育年限的增加而减小;封育样地植被分布的小尺度空间异质性小于放牧样地,并且封育年限越长,空间异质性越小.

Abstract：

This paper studied the vegetation characteristics and small-scale spatial heterogeneity of Caragana mirophylla community in Horqin Sandy Land in northeast Inner Mongolia of China under grazing and under 6-and 12 years enclosure, aimed to assess the effects of grazing and enclosure on vegetation restoration. In the sampling plots of grazing and different years enclosure, the species composition of C. mirophylla community all dominated by annual herbaceous plants. The species richness in grazed plot and in the plots enclosed for 6 and 12 years was 22, 19, and 20, respectively, with no significant difference. In the plots enclosed for 6 and 12 years, the plant density was (124.46±5.22) plants·m~(-2) and (203. 05±10. 38) plants·m~(-2), respec- tively, being significantly higher than that in grazed plot, which suggested that enclosure was an effective method to accelerate the vegetation restoration in Sandy Land. The Shannon-Wiener index, Simpson species diversity, and Pielou evenness in enclosed plots were lower than those in grazed plot, and decreased with increasing enclosure duration. The small-scale spatial heteroge-neity of vegetation in enclosed plots was smaller than that in grazed plot. The longer the enclosure duration, the smaller the spatial heterogeneity was.     

Establishment of a Murine Model for Metastasis of Cytokine-Producing Tumor to the Brain

The A375 cell line, derived from human malignant melanoma, has characteristics of interleukin-6 (IL-6) production. By using this cell line, we have investigated a murine metastasis model of IL-6-producing tumors to the brain by injecting A375 cells directly into the left cardiac ventricle. Nude mice were anesthetized with intraperitoneal injection of pentobarbital sodium. Next, A375 cells suspended in phosphate-buffered saline (PBS) were injected into the left cardiac ventricle of mice. An intracardiac injection of 10⁵ cells developed tumor colonies in the brain after 4 to 6 weeks. Metastatic cells were found in every lobe of the brain. An immunocytochemical study revealed IL-6 production by A375 cells at the metastatic sites in the brain. By the transfection of genes encoding proteins into A375 cells, a novel model of protein expression in the brain in vivo could be constructed. Our system does not require great skill. Our experimental model will facilitate future studies of the local effects of proteins in the brain.     

INTRACELLULAR LOCALIZATION OF NATIVE AUXIN IN AVENA COLEOPTILE

Intracellular localization of the native auxin in the Avenacole-optile tip was investigated by separating cellular fractionsby differential centrifugation. Each fraction was extractedwith ether and the auxin activity was measured by the sensitizedAvena curvature test. After the removal of the native free auxin,each fraction was alkaline-hydrolyied, and from these hydrolyzatesthe bound auxin was extracted with ether and its activity wasmeasured. Both the native free auxin and the native bound auxinin these extracts were identified as IAA by paper chromatography.The results show that the native free auxin occurs only in thesupernatant soluble cytoplasm, and that the native bound auxinlocalizes also in the supernatant. The distribution of the externallyapplied IAA was also investigated. (Received February 27, 1962; )     

FORMATION AND CHARACTERIZATION OF THREE TYPES OF YEAST INVERTASE

Three types of invertase (invertase I, II and III) are separatedfrom the soluble and insoluble fractions (4,500xg, 10 min supernatantand pellets of the homogenate, respectively) of baker's yeastby a DEAE cellulose column chromatography. The invertases Iand II are eluted with 0.1 M sodium acetate buffer (pH 3.9)and with 0.1 M sodium acetate buffer (pH 6.2) containing 0.1M NaCl from DEAE cellulose respectively, whereas the invertase-IIIremains adsorbed on the cellulose under these conditions. Theyare present in proportions of 2.5: 1 : 0.06 in the soluble fractionand 1.4: 1 : 0.12 in the insoluble fraction of the fresh baker'syeast cells. While in-vertase-II remains at a constant level,invertases I and III in the soluble fraction increase upon incubationof cells for the formation of invertase under the continuoussupply of sucrose. Invertases I and II differ from each other considerably in theoptimum pH and slightly in the response to (activation and inactivationby) crude papain and are identical with respect to the heatstability and probably to the affinity for sucrose. ¹Present address: Chemical Laboratory, Nippon Medical School,Konodai, Ichikawa-shi, Chiba-ken.