Interconvertible Kinetic States of t-Butylbicycloorthobenzoate Binding Sites of the γ-Aminobutyric AcidA Ionophores

The kinetics of t-[³H]butylbicycloorthobenzoate (TBOB) binding to the convulsant sites of the γ-aminobutyric acid_A (GABA_A) receptor-ionophore complex were examined in synaptosomal membrane preparations of rat brain. On and off rates of TBOB binding were accelerated by 1 μM GABA and decelerated by 1 μM bicuculline methochloride, a GABA_A antagonist. The presence of GABA and bicuculline methochloride created rapid and slow phases of dissociation, respectively. The three groups of rate constants distinguished for the dissociation of 4 nM and 30 nM [³H]TBOB represent multiaffinity states of the convulsant sites depending on the presence of GABA or bicuculline methochloride. Apparent association rate constants do not obey the equation k_app=k_off±k_on [TBOB] without assuming interconvertibility of the kinetic states during binding. Avermectin B_1a (AVM B_1a), a chloride channel opening agent, accelerated the association and dissociation of TBOB and resulted in a biphasic effect on TBOB binding, i.e., enhancement at low concentrations (EC₅₀, 7.8 nM) followed by displacement at high concentrations (IC₅₀ 6.3 μM) of AVM B_1a. AVM B_1a resulted in similar biphasic effects on t- [³⁵S]butylbicyclophosphorothionate binding. DIDS, an isothiocyanatostilbene derivative with irreversible anion channel blocking effect, selectively inhibited basal [³H]TBOB binding (IC₅₀ 125 μM DIDS) leaving the enhancement by AVM B_1a unaffected.     

MORPHOLOGY OF DNA CONTAINING STRUCTURES (NUCLEOIDS) AS A PROSPECTIVE CHARACTER IN CYANOPHYTE TAXONOMY1

DNA containing structures (nucleoids) were visualized by 4′, 6-diamidino-2-phenylindole (DAPI) fluorescent staining in two groups of cyanophytes (59 filamentous oscillatorialean species and 12 coccal Synechococcus-like organisms) to test the possibility of using nucleoid morphology in cyanophyte taxonomy. The morphology of nucleoids (size, shape, and structure) in oscillatorialean species is specific for individual families. The morphology of the nucleoid in Synechococcus-like species agrees from the proposed separation of the genus Cyanothece from Synechococcus. A much different nucleoid morphology in three species of Cyanothece suggests that these species should be separated into a new genus. On the basis of other characters, the species could be returned to the genus Cyanobacterium. My results indicate that the morphology of nucleoids is a valuable character in the classification of the cyanophytes examined; thus, it is a prospective feature that could be used in the taxonomy of other groups of cyanophytes. Additionally, DAPI staining is not a complicated procedure. The new character is easy to see in samples taken from nature, both living and preserved.     

A cladistic analysis of Nassauvia Comm. ex Juss. (Asteraceae, Mutisieae) and related genera

FREIRE, S. E., CRISCI, J. V. & KATINAS, L., 1993. A cladistic analysis of Nassauvia Comm. ex Juss. (Asteraceae, Mutisieae) and related genera. Nassauvia and the most closely related genera Calopappus and Triptilion from the southern Andes and Patagonia of South America, form a monophyletic group diagnosed by the following synapomorphies: cypsela trichomes single two-celled, cypsela testa with strengthened cells, pollen grains spheroidal to spheroidal-oblate, colpi membrane with sexine processes, pappus bristles two to six, and pappus deciduous. Furthermore, Nassauvia, Triptilion, and Calopappus form a group with two other Andean genera, Moscharia and Polyachyrus, diagnosed by occurrence of pseudocephalia and a reduction in the number of flowers to five, three or one. A cladistic analysis of the group was undertaken using 35 characters from morphology, anatomy, and palynology. The monophyletic terminal taxa were the 38 species of Nassauvia, the genus Triptilion, the monotypic genus Calopappus, the genus Polyachyrus, and the genus Moscharia. Character polarity was based on outgroup comparison using Cephalopappus. The analysis resulted in 223 equally parsimonious cladograms, each with 70 steps and a consistency index of 0.57. A successive weighting procedure was applied, resulting in 15 cladograms with a consistency index of 0.82. Results of the cladistic analysis support most of the current systematic classification of Nassauvia, with three exceptions: (1) Nassauvia (excluding Calopappus) is paraphyletic; (2) section Masligophorus appears to be a polyphyletic group (N. pygmaea does not cluster with the remaining species of the section); (3) section Panargyrum (without N. lagascae= section Caloptilium) appears to be a paraphyletic group. The capitula arranged in cymose conflorescences in Triptilion are regarded as a primitive condition which gave rise to all stages present in Nassauvia (conflorescence spicate, pseudocephalium, capitula solitary). The capitula arranged in pseudocephalia in Moscharia and Polyachyrus are regarded as a parallel development to the pseudocephalium found in Nassauvia. Nassauvia, subgenus Strongyloma appears as the most primitive taxon, with its spicate conflorescence, whereas section Masligophorus with its solitary capitulum is thought-derived. These results correspond well with cytological data where species of the subgenus Strongyloma have n= 11 and the species of section Masligophorus are tetraploids (n = 22).     

牛背鹭的繁殖、生长和恒温能力发育的研究

1990年4月—1991年5月在河南信阳南湾水库牌坊鸟岛观察了牛背鹭的15巢54只雏鸟的生长,恒温能力和食性;结果是:(1)巢离地平均13.99±0.71(13.2-15.6)米;(2)平均窝卵数3.60±0.74枚,与文献资料比较,窝卵数有显著地随纬度变化,卵平均重25.93±2.13g(3)幼鸟生长模型为Wt=305/1+e~(2.7620-0.2397t),R~2=0.99(4)恒温能力发育分三个时期,即迅速发育期、缓慢发育期和恒温期。(5)雏鸟食性组成主要有两栖类(64.18%)、昆虫(23.71%)、爬行类(8.29%)、植物(4.14%)和泥鳅(3.80%);各周龄不仅相同。     

(蜓)类旋壁超微构造的研究——以麦(蜓)型旋壁为例

光学显微镜下的观察表明(蜒)类的旋壁属于钙质微粒壳类型,对它们的电镜观察揭示了组成旋壁的晶粒的大小、形态和排列方式.对具有致密层和蜂巢层型两层式旋壁的Triticites的电镜研究表明:其致密层由近等轴晶无定向排列而成;蜂巢层由短柱状晶定向排列组成板状复合体,进而组合成管状孔,镶嵌在不定向排列的近等轴品基质中构成.Pseudofusulina的致密层也由近等轴晶不定向排列构成,蜂巢层由粗柱晶定向排列组成纤状复合体,进而形成管状孔,镶嵌在近等轴晶基质中.     

Issues in the culture of the extremely thermophilic methanogen, Methanothermus fervidus

Basic issues in the culture of the extremely thermophilic archaeon, Methanothermus fervidus, have been investigated, including culture medium formulation, substrate yield and product yield coefficient, growth rate and stoichiometry, and H(2) uptake kinetics. The pH optimum for growth of this organism was estimated at 6.9. Growth medium buffered with PIPES instead of bicarbonate supported both increased growth rate and maximum biomass concentration. Substitution of titanium(III) citrate for the reducing agent sodium sulfide improved culture performance as well. However, independent adjustment of iron and nickel concentrations from 11 to 111 muM, respectively, and carbon dioxide partial pressure from 5 to 20 psia did not impact the culture of M. fervidus significantly. An elemental balance approach was utilized to aid in design of a defined medium to support growth to a target maximum biomass concentration of at least 1.0 g dry wt/L. The growth of this organism was limited by H(2) availability in this reformulated culture medium. The maximum growth rate and biomass concentration achieved in anaerobic vials with the defined medium was 0.16 h(-1) and 0.74 g dry wt/L, respectively. This maximum biomass concentration was a 72% improvement over that obtained with a literature-based defined medium. The Monod parameter, K(s), with H(2) as limiting substrate, was estimated at 1.1 +/- 0.4 psia (55 +/- 20 muM in the broth), based on a H(2) consumption study. Representative values for the substrate yield, Y(X/CO(2) ), and product yield coefficient, Y(CH(4)/) (X), were determined experimentally to be 1.78 +/- 0.04 g dry wt/mol CO(2), and 0.52 +/- 0.01 mol CH(4)/g dry wt, respectively. A bench-scale fermentation system suitable for the culture of extremely thermophilic anaerobes was designed and constructed and proved effective for the culture of M. fervidus. (c) 1993 Wiley & Sons, Inc.     

Continuous measurement of microbial heat production in laboratory fermentors

The possibility of continuously measuring the heat produced by microorganisms in an ordinary laboratory fermentor was studies. An inventory of the heat flows influencing the temperature of the culture was made. The magnitude and standard deviation in these heat flows were studied from theoretical and practical viewpoints. Calibration procedures were tested, and a model describing the heat flows in steady state and during dynamic conditions was made. Microbial heat production could be calculated accurately with the help of this model, appropriate temperature measurements, and equipment properties measured during the calibration procedures. It was found that the measurement of heat production could be done with an accuracy similar to that in the O(2) uptake measurement. (c) 1993 John Wiley & Sons, Inc.     

A novel membrane bioreactor for detoxifying industrial wastewater: I. Biodegradation of phenol in a synthetically concocted wastewater

A novel process has been used to biodegrade phenol present in an acidic (1 M HCI) and salty (5% w/w NaCl) synthetically bioreactor, in which the phenol present in the wastewater is separated from the inorganic components by means of a silicone rubber membrane. Transfer of the phenol from the wastewater and into a biological growth medium allows biodegradation to proceed under controlled conditions which are unaffected by the hostile inorganic composition of the wastewater. At a wastewater flow rate of 18 mL h(-1) (contact time 6 h), 98.5% of the phenol present in the wastewater at an inlet concentration of 1000 mg ( (-1) ) was degraded; at a contact time of 1.9 h, 65% of the phenol was degraded. Phenol degradation was accompanied by growth of a biofilm on the membrane tubes and by conversion of approximately 80% of the carbon entering the system to CO(2) carbon. Analysis of the transport of phenol across the membrane revealed that the major resistance to mass transfer arose in the diffusion of phenol across the silicone rubber membrane. A mathematical model was used to describe the transfer of phenol across the membrane and the subsequent diffusion and reaction of phenol in the biofilm attached to the membrane tube. This analysis showed that (a) the attached biofilm significantly lowers the mass transfer driving force for phenol across the membrane, and (b) oxygen concentration limits the phenol degradation rate in the biofilm. These conclusions from the model are consistent with the experimental results. (c) 1993 Wiley & Sons, Inc.     

Stability of bilirubin oxidase in organic solvent media: a comparative study on two low-water systems

The storage stability of bilirubin oxidase was studied in water-in-oil CTAB microemulsions with a chloroformrich continuous organic phase. The kinetics of the inactivation process were best described by a double exponential equation. Approximately half of enzymatic activity was lost during a "fast" phase with a half life of ca. 50 min, whereas the remaining activity was lost much more slowly (half life ca. 1000 min). Rates of inactivation were not affected significantly by variation of either solvent composition or concentration of water droplets, but inactivation was more rapid when droplet size was very small. Steady-state enzyme kinetics were studied at various stages in the inactivation process, and it was shown that inactivation occurred without change in the K(m) of the enzyme for bilirubin. Stability was also studied in a liquid/solid two-phase system; it was found that the inactivation process in this system; it was found that the inactivation process in this system was best described by a single exponential term. The rate was similar to the "fast" phase rate observed in the water-in-oil microemulsion system. Inactivation of the enzyme slow. Addition of the surfactant CTAB to the aqueous environment increased the rate of inactivation to levels comparable to those of the "slow" phase observed in water-in-oil microemulsions. (c) 1993 Wiley & Sons, Inc.     

Nutrient-split feeding strategy for dialysis cultivation of Escherichia coli

Reduction in nutrient loss during dialysis cultivation of Escherichia coli on a glycerol medium was investigated. A dialysis reactor with an inner fermentation and an outer dialysis chamber was used. Aerobic condition was maintained by limiting the glycerol feed rate to an optimum value which was estimated from the oxygen requirements for glycerol oxidation and oxygen transfer capacity of the reactor. High reduction in nutrient loss was achieved by using water as the dialyzing fluid. However, osmotic movement of water from the dialysis to the fermentation chamber was observed, and the final cell concentration was low. With a nutrient-split feeding strategy (feeding glycerol directly to the fermentation chamber and dialyzing with salt solution), glycerol loss was small, there was no osmotic flux of water to the fermentation chamber, and the cell concentration was high. Both glycerol and salt loss could be avoided, and a cell concentration of 170 g/L was obtained when the dialysis process was substituted by addition of XAD adsorbents to the dialysis chamber. Application of this nutrient-split feeding strategy to cell cultivation in a stirred tank reactor, coupled with dialysis in external dialyzer modules, resulted in low cell concentrations. (c) 1993 Wiley & Sons, Inc.