Morphology,nutrition and physiology of Sphaerotilus discophorus

Summary By means of hay infusion-Fe(OH)₃–MnCO₃ enrichment cultures, 11 pure strains of the filamentous, sheathed bacterium, Sphaerotilus discophorus were isolated from streams, rivers and lakes. The morphology of the colonies, filaments and cells are shown in a series of photographs. The strains grew well in dilute organic media but not in many of the common bacteriological media. All strains required thiamin and biotin for growth. Glucose, mannitol, salicin, raffinose, glycerol and other compounds were suitable energy sources and peptone and casein hydrolysate were satisfactory nitrogen sources for growth. The temperature range for growth was 5–35°C and the pH range 6.0–8.6.The sudanophilic granules of S. discophorus are composed of poly--hydroxybutyric acid which may be as much as 40% of the dry weight of the cells. Growth with iron and manganese salts results in extensive deposition of oxides on these metals on the filaments. Chemical analyses of 60 hr cultures showed that the iron content of the filaments ranged from 4.25–7.10% and manganese from 1.12–1.43%, calculated on a dry weight basis. Resting cell suspensions oxidized a variety of sugars, sugar alcohols, amino acids and other compounds and concomitantly assimilated about 90% of these substrates. The properties of S. discophorus are compared with those of S. natans.Based in part on a thesis presented by the senior author in partial fulfillment of the requirements for the degree of Doctor of Philosophy, Washington State University, 1963.     

Dehydrogenase activity and toxinogenesis of a production strain ofCorynebacterium diphtheriae during submerged cultivation

A simple biochemical procedure was obtained for studying metabolism ofCorynebacterium diphtheriae during submerged cultivation based on the modification of the assay of dehydrogenase activity using 2,3,5-triphenyltetrazolium chloride as redox indicator. Results obtained by the estimation of the dehydrogenase activity using TTC are in a good accordance with oxygen consumption assayed manometrically. By following dehydrogenase activity in submerged cultivations of a production strain ofCorynebacterium diphtheriae PW8-Weissensee we found that a massive toxin production is connected with the decrease of the activity of cells. This fall of activity occurs yet during the exponential phase of growth. Especially a sudden fall of succindehydrogenase activity exactly indicates the beginning of a considerable toxin accumulation in the medium. The presence of inhibitory concentrations of iron ions in the medium not only increases the level of dehydrogenase activity but changes its whole kinetics. A retarded and irregular fall of the activity occurs instead of a sharp one typical for good toxin production.     

Growth factor requirements of two strains ofSphaerotilus discophorus

The relatively complex growth-factor requirements of two strains ofSphaerotilus discophorus, strains 31 and 32, have been elucidated. In addition to thiamin and biotin which are required by other strains ofS. discophorus, the two strains must be supplied also with adenine or guanine and with cyanocobalamin for growth in a glucose — Casamino Acids — mineral salts medium. The cyanocobalamin can be replaced by methionine but only if relatively large amounts of this amino acid, 100 µg or more per ml, are added to the medium. The methionine requirement of the two strains is approximately 3 times greater than that of otherS. discophorus strains.     

Stimulation of poly-β-hydroxybutyrate oxidation in Sphaerotilus discophorus by manganese and magnesium

Summary When grown with glucose, S. discophorus synthesized large amounts of poly--hydroxybutyrate which accumulated intracellularly as sudanophilic granules. The rate of endogenous oxygen consumption by such cells was markedly increased by Mn⁺⁺ and even more by Mg⁺⁺. It has been shown that these inorganic ions stimulate the oxidation of the intracellular poly--hydroxybutyrate.Dedicated by the senior author to Prof. C. B. van Niel on the occasion of his 70th birthday with gratitude for many unforgettable years of association, instruction and stimulation.     

A novel family of potentially mobile DNA elements encoding site-specific gene-integration functions: integrons

A family of novel mobile DNA elements is described, examples of which are found at several independent locations and encode a variety of antibiotic resistance genes. The complete elements consist of two conserved segments separated by a segment of variable length and sequence which includes inserted antibiotic resistance genes. The conserved segment located 3' to the inserted resistance genes was sequenced from Tn21 and R46, and the sequences are identical over a region of 2026 bases, which includes the sulphonamide resistance gene sull, and two further open reading frames of unknown function. The complete sequences of both the 3' and 5' conserved regions of the DNA element have been determined. A 59-base sequence element, found at the junctions of inserted DNA sequences and the conserved 3' segment, is also present at this location in the R46 sequence. A copy of one half of this 59-base element is found at the end of the sull gene, suggesting that sull, though part of the conserved region, was also originally inserted into an ancestral element by site-specific integration. Inverted or direct terminal repeats or short target site duplications, both of which are characteristics of class I and class II transposons, are not found at the outer boundaries of the elements described here. Furthermore, the conserved regions do not encode any proteins related to known transposition proteins, except the DNA integrase encoded by the 5' conserved region which is implicated in the gene insertion process. Mobilization of this element has not been observed experimentally; mobility is implied from the identification of the element in at least four independent locations, in Tn21, R46 (IncN), R388 (IncW) and Tn1696. The definitive features of these novel elements are (i) that they include site-specific integration functions (the integrase and the insertion site); (ii) that they are able to acquire various gene units and act as an expression cassette by supplying the promoter for the inserted genes. As a consequence of acquiring different inserted genes, the element exists in a variety of forms which differ in the number and nature of the inserted genes. This family of elements appears formally distinct from other known mobile DNA elements and we propose the name DNA integration elements, or integrons.     

Three-dimensional osseo-ligamentous model of the thorax representing initiation of scoliosis by asymmetric growth

A biomechanical model of the human thorax was constructed to investigate how asymmetric growth of the thorax might initiate spinal lateral curvature and axial rotation as seen in scoliosis deformities. Geometric data specifying nodal points of the model were taken from stereo-radiographs of an adolescent subject. An initially symmetrical geometry was created by 'mirroring' measurements of a hemi-thorax and spine. Published data provided cross-sectional measurements of the ribs, material properties of tissues and global flexibilities of the intervertebral motion segments. The ribs, sternum, intervertebral motion segments and intercostal ligaments were represented by elastic elements. Model deformations were calculated by the direct stiffness finite element method, with growth represented by an initial strain term in the constitutive law. Non-linear behavior was accommodated by running the model recursively, with updated node locations at each step. Both stress relaxation and stress modulation of growth in the component tissues were simulated. Thoracic growth of 20% with asymmetric growth of the ribs was simulated to give rib length asymmetries of 11%. similar to that observed in a previous study of patients with idiopathic scoliosis. This resulted in the model having a small thoracic scoliosis curvature convex toward the side of the longer ribs. Variations of the model which permitted free motion at the costo-vertebral joints or produced changes in the curvature of the posterior parts of the ribs resulted in axial rotation of the vertebrae similar to that observed clinically. The model supports the idea that growth asymmetry could initiate a small scoliosis during adolescence.     

Rotational and translational movement features of the pelvis and thorax during adult human locomotion

The kinematics of the pelvis and thorax are important in gait studies since their movement patterns are closely related to gait efficiency and 'smoothness' of locomotion. The purpose of this study was to identify features of normal gait patterns for later comparisons with pathological and developmental gait patterns. A two camera SELSPOT system interfaced with an HP1000 minicomputer was used to obtain three-dimensional kinematic/temporal data for the pelvis and thorax. Data from treadmill walking of eight adults were used for within subject (at different speeds) analyses. The analyses revealed a very complex pattern with a set of breakpoints which was consistent over all subjects. Some features were invariant over a range of walking speeds although the total range of motion changed considerably.     

Lamellar stacking in three-dimensional crystals of Ca(2+)-ATPase from sarcoplasmic reticulum.

Electron microscopy of multilamellar crystals of CA(2+)-ATPase currently offers the best opportunity for obtaining a high-resolution structure of this ATP-driven ion pump. Under certain conditions small, wormlike crystals are formed and provide views parallel to the lamellar plane, from which parameters of lamellar stacking can be directly measured. Assuming that molecular packing is the same, data from these views could supplement those obtained by tilting large, thin platelike crystals. However, we were surprised to discover that the lamellar spacing was variable and depended on the amount of glycerol present during crystallization (20% versus 5%). Projection maps (h,0,l) from these womklike crystals suggest different molecular contacts that give rise to the different lamellar spacings. Based on an orthogonal projection map (h,k,0) from collapsed, wormlike crystals and on x-ray powder patterns, we conclude that molecular packing within the lamellar plane is the same as that in thin, platelike crystals and is unaffected by glycerol. Finally, the orientation of molecules in the lamellar plane was characterized from freeze-dried, shadowed crystals. Comparing the profile of molecules in these multilamellar crystals with that previously observed in helical tubes induced by vanadate gives structural evidence of the conformational change that accompanies binding of calcium of Ca(2+)-ATPase.