Spotlight on phytochrome nomenclature

These recommendations for genes encoding phytochromes were developed independently by Quail et al., but are broadly consistent with the Commission's guidelines. Their original article, kindly provided in advance of publication, appeared as a Letter to the Editor inPlant Cell (6:468–471, 1994) and is published with permission of the American Society of Plant Physiologists.     

Cell-substrate adhesion and metastatic potential of cultured mesothelioma cells induced by asbestos

Cell-substrate adhesion was quantified for two cultured mesothelioma cell lines (epitheliomatus and sarcomatous) on glass, fibronectin and laminin substrates. Interference reflection microscopy (IRM) was used to image the adhesion patterns of cells and a grey level analysis was employed to quantify adhesion. Sarcomatous cells demonstrated marked adhesion to glass and fibronectin-coated substrates but not to laminin-coated substrate, with the greatest adhesion occurring on the fibronectin-coated surface. This adhesion was accompanied by cytoplasmic spreading. By contrast, epitheliomatous cells showed little tendency to adhere to any of the substrates and only showed significant spreading when in contact with the laminin substrate (P < 0.01). A bioassay was used to determine the metastatic potential of each of the cell lines. Via the intravenous route, the sarcomatous cells killed the host rats in 24.7 ± 1.5 (S.D.) days compared to 27.3 ± 0.9 (S.D.) days for the epitheliomatous cells (P < 0.01). After subcutaneous inoculation of tumour cells, the sarcomatous cells killed the host rats in 54.7 ± 0.7 (S.D.) days compared to 48.5 ± 0.5 (S.D.) days for the epitheliomatous cells (P < 0.01). We conclude that the results of the metastasis bioassays were consistent with the predicted behavior of these cell lines based on their ability to adhere to substrates in the in vitro adhesion assays.     

Essential trace elements in humans

Serum arsenic concentrations of persons suffering from renal failure and undergoing hemodialysis treatment (n=85) and of healthy controls (n=25) were determined by hydride-generation AAS technique after microwave digestion. The results were evaluated by comparing the values of both groups, considering physiological factors and individual data, as well as comorbid conditions of the hemodialysis (HD) patients. Serum arsenic levels were diminished in the patient group compared with controls (mean values 8.5±1.8 ng/mL vs 10.6±1.3 ng/mL). Furthermore, additional diseases within the hemodialysis group, particularly injuries of the central nervous system (CNS), vascular diseases, and cancer, were correlated to occasionally markedly decreased serum arsenic concentrations. It was concluded that arsenic homeostasis is disturbed by HD treatment and certain additional diseases. Desirable arsenic concentrations in the body seem to be reasonable. This consideration results in the conclusion that arsenic could play an essential role in human health. Thus, reference arsenic concentrations in different human tissues and body fluids should be established in order to recognize not only arsenic intoxication, but also arsenic deficiency. Perhaps arsenic deficiency contributes to the increased death risk of HD patients, and therefore, arsenic supplementations for patients with extremely low serum arsenic concentrations should be taken into account.     

Interpretation of gas residence time distributions in large airlift tower loop reactors

Gas-residence time distribution (RTD) response curves measured in a 23 m high pilot plant airlift tower loop reactor, which consisted of a riser, a special downcomer construction and at the top of the riser a large head. The measurements were evaluated by means of a deterministic dispersion model, which yielded the following particular parameters for the riser, downcomer and the head: Gas-Bo numbers, gas-mean residence times, gas holdups, liquid velocities, gas and liquid circulation times as well as a fraction of the large and small bubbles in a model medium (water) and during cultivation of baker's yeast.List of Symbols A cross section - Bo Bodenstein number - Bo _d (= l _d w _G,d /D _d ) - Bo _h (= l _h w _G,h /D _h ) - Bo _r (= l _r w _G,r /D _r ) - D longitudinal dispersion coefficient - E gas holdup - E(t) RTD-density function - L, l length parameter - q fraction of the gas throughput which is not recirculated (approximately equal to fraction of the large bubbles) - r fraction of the throughput which is recirculated (approximately equal to the fraction of the small bubbles) - t _c circulation time - t _cL liquid circulation time - t _c,L ^* liquid circulation time calculated from the measured w _Ld in the downcomer - V ^h hydrodynamical calculated gas-liquid volume - V _d ^h (=V _d+0.75/2 V _k ) - V _k ^h =(0.25V _k ) - V _r ^h = (V _r+0.75/2 V _k ) - V _L liquid volume - V _G dispersed gas volume - V _G ^* gas throughput at the gas distributor (given in m³/h) under standard conditions, 1 bar and 25°C) - V _G,d ^* gas throughput in downcomer (=V _G ^* ) - V _G,h ^* gas throughput in head (=V _G ^* ) - V _G,r ^* gas throughput in riser (V _G ^* (1+) - w _g gas velocity - w _G,rel relative gas velocity with respect to the liquid velocity w _L - w _G,d gas velocity in the downcomer (=w _G,rel –w _Ld ) - w _G,h gas velocity in the head (=w _G,rel ) (since wLh = o) - w _G,r gas velocity in the riser (=w _G,rel +w _Lr ) - w _L liquid velocity - w _L,d liquid velocity in the downcomer measured with mass flow meter - w _sg ·w _SL superficial gas and liquid velocities - first moment of the response curve - mean residence time Indices d downcomer - G gas phase - h head - L liquid phase - r riser - h hydrodynamic (upper position) Dedicated to the 65th birthday of Proffessor Fritz Wagner.The authors gratefully acknowledge the financial support by the Krupp Industrietechnik, Grevenbroich and the support of Pleser Co, Darmstadt. H. M. Rüffer thanks the Verband der Chemischen Industrie for a Fond der Chemie scholarship, and W. Liwei thanks the government of Lower Saxony for a graduate scholarship.     

Interpretation of the gas residence time distributions in large stirred tank reactors

The behaviour of dispersed gas in large aerated stirred tank reactors is modelled by means of a Markov-process, which distinguishes between small recirculation bubbles with stagnant gas content, large rising bubbles with active gas content and exchange of stagnant and active gas contents, the gas exchange region at the impeller. The measurements of the gas residence time distributions (RTDs) in an 1.5 m³ aerated stirred tank reactor with water and Penicillium chrysogenum cultivation medium are interpreted by this model.List of Symbols CPR CO₂ production rate - OTR oxygen transfer rate - PRS pseudo random signal - RTD residence time distribution - V gas volume - recirculation coefficient - mean gas residence time Indices act active gas - ex gas exchange - stagn stagnant gas - tot total gas Dedicated to the 65th birthday of Professor Fritz Wagner.The authors thank Hoechst AG for the strain and the medium components, the GBF for the support of the experiments and H.M. Rüffer thanks the Verband der Chemischen Industrie for a Fond-der-Chemie scholarship.     

Neutral adaptation of the genetic code to double-strand coding

Summary We lay new foundations to the hypothesis that the genetic code is adapted to evolutionary retention of information in the antisense strands of natural DNA/RNA sequences. In particular, we show that the genetic code exhibits, beyond the neutral replacement patterns of amino acid substitutions, optimal properties by favoring simultaneous evolution of proteins encoded in DNA/RNA sense-antisense strands. This is borne out in the sense-antisense transformations of the codons of every amino acid which target amino acids physicochemically similar to each other. Moreover, silent mutations in the sense strand generate conservative ones in its antisense counterpart and vice versa. Coevolution of proteins coded by complementary strands is shown to be a definite possibility, a result which does not depend on any physical interaction between the coevolving proteins. Likewise, the degree to which the present genetic code is dedicated to evolutionary sense-antisense tolerance is demonstrated by comparison with many randomized codes. Double-strand coding is quantified from an information-theoretical point of view.     

Emulsion liquid membrane extraction of lactic acid from aqueous solutions and fermentation broth

Studies on the batch extraction of lactic acid using an emulsion liquid membrane system are reported. The membrane phase consists of the tertiary amine carrier Alamine 336 and the surfactant Span 80 dissolved in n-heptane/paraffin and aqueous solutions of sodium carbonate in the internal phase. The effects of internal phase reagent, extraction temperature, and initial external phase pH on the extraction efficiency and the emulsion swelling are examined. A statistical factorial experiment on extraction from clarified lactic acid fermentation broth was carried out to obtain knowledge of the performance of the extraction system from a broth. The extraction efficiency from the fermentation broth is found to be lower as compared to aqueous solutions of pure lactic acid. The effect of pH and the presence of other ionic species on selectivity are discussed. (c) 1993 John Wiley & Sons, Inc.     

Fed-batch cultivation of recombinant escherichia coli JM103 and production of the fusion protein SPA::EcoRI in a 60-L working volume airlift tower loop reactor

SPA::EcoRI fusion protein was produced by Escherichia coli JM103 carrying the multicopy expression plasmid pMTC48, the multicopy repressor plasmid pRK248, and the multicopy protection plasmid pEcoR4 in a 60-L working volume airlift tower loop reactor on M9 minimal medium with glucose. Cell mass concentration, total cell count, number of colony-forming units, specific growth rate, yield coefficient, and metabolite (acetate, pyruvate, succinate, lactate, ethanol) concentrations were monitored during the growth phase and gene expression. Gene expression was induced by temperature shift or chemically by isopropyl-thiogalactosidase in the airlift tower loop reactor (ALTR) at constant cultivation time and in a small stirred tank reactor at different cultivation times. During induction, the cultivation medium was supplemented with concentrated Luria-Bertani (LB) medium. The intracellular enzyme activity was evaluated as a function of the time after the start of the induction. It was found that the reduction of the glucose concentration and increase of the dissolved oxygen concentration reduced the acetate produced and increased the intracellular enzyme activity. (c) 1993 John Wiley & Sons, Inc.     

Two early replicated,developmentally controlled genes of Physarum display different patterns of DNA replication by two-dimensional agarose gel electrophoresis

The nature of replication origins in eukaryotic chromosomes has been examined in some detail only in yeast, Drosophila, and mammalian cells. We have used highly synchronous cultures of plasmodia of the myxomycete Physarum and two-dimensional agarose gel electrophoresis to examine replication of two developmentally controlled, early replicated genes over time in S-phase. A single, discrete origin of replication was found within 4.8 kb of the LAV1-5 gene, which encodes a homolog of profilin. In contrast, the LAV1-2 gene appears to be surrounded by several origins. Two origins were identified within a 15 kb chromosomal domain and appear to be inefficiently used. Replication forks collide at preferred sites within this domain. These terminating structures are long lived, persisting for at least 2 h of the 3 h S-phase. Analysis of restriction fragment length polymorphisms (RFLPs) within the LAV1-2 domain indicates that replication of alleles on different parental chromosomes is a highly coordinated process. Our studies of the these two early replicated, plasmodium-specific genes indicate that both a fixed, narrow origin region and a broader zone containing two closely spaced origins of DNA replication occur in Physarum.     

Application of continuous zone electrophoresis to preparative separation of proteins

A comprehensive study of the application of continuous zone electrophoresis to preparative separation of proteins in free solution is presented. First, the influence of electric field strength, buffer residence time in the chamber, sample flow rate, and sample concentration on separation resolution and throughput were studied. Using multiple injections of sample into the electrophoresis chamber, a throughput of 500 mg protein/h was achieved for partially purified model proteins. Experiments on Escherichia coli crude extracts yielded a fivefold purification of beta-galactosidase along with a simultaneous separation of proteins from cell debris in a single step. Experiments correlating the electrophoretic mobility in continuous electrophoresis with the elution behavior in ion-exchange chromatography were performed on more than a dozen proteins which conclusively showed that separation of proteins in continuous zone electrophoresis is governed by net surface charge. Based on these results, the fraction numbers in which the proteins eluted could be correctly predicted. Proteins and enzymes with differences >0.5 M elution molarities in ion-exchange chromatography were separated by continuous zone electrophoresis on a preparative scale (mg/h or g/h) with >90% recovery. This corresponds to a preparative scale separation of proteins and enzymes which differ in apparent electrophoretic mobility by only 0.70 x 10(-5) cm(2)/V . s. (c) 1993 John Wiley & Sons, Inc.