A new red-bracted species of Calathea (Marantaceae) from Peru

Calathea schunkei from San Martín Province, Peru, is described as new. This species is distinguished by the red bracts and unusual floral morphology. The petals do not reflex at anthesis, the outer staminode is lacking, and the stigma is blunt rather than the typical “scoop” shape. These unusual floral features are shared withC. timothei, a high altitude Colombian species.     

Biotransformation of cycloalkanones: Controlled oxidative and reductive bioconversions by anAcinetobacter species

Summary Culture conditions have been optimised to enable resting cell cultures ofAcinetobacter calcoaceticus NCIMB 9871 to selectively undertake either oxidative or reductive biotransformations of various bicyclic ketones.     

Protection against acute paraquat toxicity by ambroxol

An expression vector for G-CSF, pASLB3-3, was constructed and introduced into Namalwa KJM-1 cells (Hosoi et al., 1988), and cells resistant to 100 nM of methotrexate (MTX) were obtained. Among them, the highest producer, clone SC57, was selected and the productivity of this clone was further characterized. The maximal production of G-CSF was at the most 1.8 g/ml/day using a 25 cm² tissue culture flask, even though the cell number was above 7×10⁵ cells/ml. The limiting factors at high density were analyzed as the deficiency of nutrients, such as glucose, cysteine and serine, and pH control. The depression of specific G-CSF productivity per cell under the batch culture conditions was overcome by using a perfusion culture system, Biofermenter^TM (Sato, 1983) with modifications of nutrients supplementation by a dialysis membrane and/or dissolved oxygen (DO) supplementation by microsilicone fibers. ITPSGF medium was modified to elevate concentrations of amino acids and glucose by 2.0- and 2.5-times, respectively. Under the control of pH at 7.4 and DO at 3 ppm, the specific G-CSF productivity was not depressed even at high cell density (above 1×10⁷ cells/ml), and the amount of G-CSF reached 41 g/ml. These results indicated the possibility of finding the optimum culture conditions for the production of recombinant proteins by Namalwa KJM-1 cells.Abbreviations ABTS 2,2-Azino-di-(3-ethylbenzothiazoline)-6-sulfonic acid - BSA Bovine Serum Albumin - BSA-PBS Phosphate-buffered Saline without Ca²⁺ and Mg²⁺ containing Bovine Serum Albumin - dhfr Dihydrofolate Reductase - DO Dissolved Oxygen - G-CSF Granulocyte Colony-stimulating Factor - HEPES 4-(2-Hydroxyethyl)-1-piperazineethansulfonic Acid - IFN Interferon - MTX Methotrexate - PBS(-) Phosphate-buffered saline without Ca²⁺ and Mg²⁺ - Tween-PBS Phosphate-buffered saline without Ca²⁺ and Mg²⁺ containing 0.05% of Tween 20     

Adaptations of Goldner's Masson Trichrome Stain for the Study of Undecalcified Plastic Embedded Bone

Specialized adaptations for application of Goldner's Masson trichrome stain to plastic embedded undecalcified bone specimens are presented. This stain can be used successfully on methyl-glycol methacrylate, glycol methacrylate and Spurr embedded bones. The stain affords the advantage of good cellular staining due to the hematoxylin component with concomitant sharp discrimination of mature bone matrix which stains green, immature new bone matrix which stains red, and calcified cartilage which stains very pale green. Use of red filters during photomicrography aids in bone-osteoid discrimination in black and white photographs.     

The induction of sensitivity to gibberellin in aleurone tissue of developing wheat grains

Aleurone tissue from undried immature developing wheat grains (Triticum aestivum L. cv. Sappo), normally insensitive to gibberellic acid, can be made to respond to the hormone by a series of temperature treatments. Incubation of the de-embryoed grains at temperatures above 27° C for at least 8 h causes the tissue to become sensitive. Prolonged incubation at temperatures below 27° C does not effect a change in sensitivity. In addition to the requirement for exposure to an elevated temperature for a period of several hours the tissue must also subsequently be subjected to a period at a lower temperature for just a few seconds for the response to be observed. Once sensitized, the tissue remains responsive to gibberellic acid for substantial periods of time. Exposure of the tissue to temperatures which induce sensitivity to gibberellic acid also results in an increased leakage of amino acids. It is suggested that the increase in sensitivity to gibberellin requires two separate processes to take place. One could be a homeoviscous adaptation of the cell membranes in response to elevated temperature, the other a subsequent, permanent change in conformation of membrane components.     

Equilibrium studies of zinc(II) and cobalt(II) binding to tripeptide analogues of the amino terminus of human serum albumin

Serum albumin is known to bind several divalent metal ions at the amino terminus of the protein. Two peptide analogues for the amino terminus of human albumin, L-aspartyl-L-alanyl-L-histidine-N-Methyl amide (AAHNMA) and glycylglycyl-L-histidine-N-methyl amide (GGHNMA) have been synthesized, and their interactions with Zn(II) and Co(II) ions have been studied using analytical potentiometry. The stability constants of the species and their distribution as a function of pH were determined in 0.16-M KNO₃ at 25°. Comparison of the modes of interaction of the Zn(II) and Co(II) with each of the above peptides indicate that, although Co(II) is a valuable tool for the study of Zn(II) interaction with metalloenzymes, it is considerably less useful as a Zn(II) model with small peptide molecules. The potentiometric properties of the two peptide-Zn(II) systems have been compared to the potentiostatic properties of the albumin-Zn(II) system. The results indicate that AAHNMA is a better analogue for the Zn(II)-HSA interaction than is GGHNMA. The findings suggest that the Zn(II)-HSA binding site is best described as a compound site containing both a histidyl and a neighboring carboxyl group.     

The location of highly repetitious DNA in the somatic chromosomes of Drosophila melanogaster

In situ hybridization of Drosophila melanogaster somatic chromosomes has been used to demonstrate the near exact correspondence between the location of highly repetitious DNA and classically defined constitutive heterochromatin. The Y chromosome, in particular, is heavily labeled even by cRNA transcribed from female (XX) DNA templates (i.e., DNA from female Drosophila with 2 Xs and 2 sets of autosomes). This observation confirms earlier reports that the Y chromosome contains repeated DNA sequences that are shared by other chromosomes. In grain counting experiments the Y chromosome shows significantly heavier label than any other chromosome when hybridized with cRNA from XY DNA templates (i.e., DNA from male Drosophila with 1 X and 1 Y plus 2 sets of autosomes). However, the preferential labeling of the Y is abolished if the cRNA is derived from XX DNA. We interpret these results as indicating the presence of a class of Y chromosome specific repeated DNA in D. melanogaster. The relative inefficiency of the X chromosome in binding cRNA from XY and XYY DNA templates, coupled with its ability to bind XX derived cRNA, may also indicate the presence of an X chromosome specific repeated DNA.     

Tissue fixation and osmium black formation with nonvolatile octavalent osmium compounds

Summary Several compounds of osmium^VIII, including potassium osmiamate and coordination complexes of OsO₄ with ammonia and various heterocyclic nitrogen compounds, have been synthesized and characterized. They have also been evaluated as substitutes for OsO₄ in postfixation of biological specimens and in light and electron microscopic cytochemical methods resulting in osmium black formation.The most useful of these osmic compounds, a molecular addition complex of hexamethylenetetramine (methenamine) with OsO₄, has a negligible vapor pressure of OsO₄. It has the molecular formula C₆H₁₂N₄.2OsO₄ and has been designated osmeth. Although it has only limited solubility, aqueous solutions of the compound (or of OsO₄) can be rapidly prepared by dissolution in a minimal amount of dimethylformamide and subsequent dilution with distilled water or buffer. Although stable in the solid state, the complex in solution undergoes partial dissociation releasing OsO₄, and the odor of OsO₄ becomes apparent.Such solutions of osmeth are (0.25%) considerably less concentrated with respect to OsO₄ than solutions (1–2%) ordinarily employed for ultrastructural preservation or in cytochemical studies. Osmeth has limited value for postosmication after glutaraldehyde fixation because the generation (release) of OsO₄ appears to be slow. Adequate osmication of tissue blocks exists only at the surface, but effective osmication can be achieved throughout tissue sections. In cytochemical reactions resulting in the formation of osmium blacks, the osmeth solutions are as effective as OsO₄ solutions of equivalent concentrations. Our findings indicate that OsO₄ solutions of less than 1% may be satisfactorily utilized in many cytochemical studies.Osmeth is safer and more convenient to handle than OsO₄ because small amounts may be solubilized as needed. It should be considered as a substitute for OsO₄ in ultrastructural cytochemistry.This investigation was supported by NIH research grant number DE 02668 from the National Institute of Dental Research and by NIH grant number RR 05333 from the Division of Research Facilities and ResourcesVisiting Professor, Dental Research Center, University of North Carolina at Chapel Hill, Jan.-May, 1975. Supported in part by USPHS Grant HD 09209     

Fitness of Karyotypes in DROSOPHILA PSEUDOOBSCURA

In the dynamics of the survival of chromosomal polymorphism selection may be operating at the genic level, at the chromosomal level or at the supergene level. Tests designed to distinguish between these levels were run on Drosophila pseudoobscura. There was no evidence for heterosis, a necessary requirement for gene-determined chromosomal polymorphism. A strong chromosmal selection was observed. No evidence was found for the presence within one locality of more than a single superallele for each supergene (= gene order). These results are compared to those found by others.