极谱法测定无氧介质中根系氧气输导

介绍了利用极谱法测定凤眼莲 (Eichhorniacras sipesSolm .)、菰 (Zizanialatifolia (Geiseb)Stapf)、石菖蒲(AcorustartarinowiiSchott)、慈菇 (SagitarriasagittifoliaLinn .)、香蒲 (TyphaangustifoliaLinn .)等多种湿生植物根系在无氧介质中氧气的对外扩散速率 (ODR)。实验结果表明 ,对于同种植物而言 ,新生根较老根的氧气扩散速率快。根尖区 (约 0～ 5mm范围内 )氧气扩散速率最大 ,氧化活力最强。愈往上愈小 ,根基部的氧气输出速率接近为零 ,氧化活力几乎可以忽略。对于根长相同的不同种植物而言 ,香蒲的ODR最大 ,凤眼莲的最小     

An Electrical Analogue to Simulate the Oxygen Relations of Roots in Anaerobic Media

The paper describes the construction and the theory underlying a simple electrical analogue which may be used to simulate the internal oxygen relations of roots in the wetland condition. The model represents an unbranched root lying within an oxygen sink and consists of twenty circuit units linked in series. Each unit which can be programmed independently, represents the major oxygen diffusion paths, impedances and sinks recognizable in a centimetre-long segment of root/wet soil system. Simplicity has been achieved by specifying constant radius for the root (0.05 cm), constant potential activity for the external sink, by expressing respiration and radial oxygen loss as a function of root surface area, and by the non-inclusion of oxygen storage capacity.     

Oxygen Diffusion from the Roots of Woody Species

Application of the‘polarographic’ technique for studying oxygen diffusion from roots has provided preliminary results which establish that oxygen passes through the woody species Salix atrocinerea Brot., Salix fragilis L., Salix repens L., and Myrica gale L., in the gaseous phase as it does in other wetland species. Entry into the shoots occurs through the bark directly above the water table and in the willow cuttings the effective length of shoot for gas intake was the basal three centimetres above the water table. The length of shoot involved was longer in Myrica gale and the roots were of the normal (un-nodulated) type. Trials on Alnus glutinosa (L.) Gaertn. provided no conclusive results and it is thought that this was perhaps because measurements were on nodulated roots only.     

The Exudation from Excised Maize Roots Bathed in Sulphate Media

The fluid exudation fluxes of excised maize roots bathed ina variety of K₂SO₄ solutions have been measured, along withthe ionic concentrations and osmolalities of the exudates. Anapparent imbalance of inorganic ionic charge in the exudate,and a discrepancy in the total salt concentrations comparedwith the measured osmolalities, suggest the presence of significantamounts of organic anions in the exuded fluid. Thin-layer chromatographicmethods indicate these to be mainly amino-acids and organicacids of the Kreb's cycle. Linear regression analysis showsthat the exudation flux is satisfactorily described by a simpleosmotic process together with a constant non-osmotic water fluxof 0.43 µ1 cm^-2 h^-1. The osmotic permeability for theroot is found to be 0.18 x 10^-6 cm sec^-1 atm^-1. The exudationfrom sulphate media is contrasted with that from chloride media.     

Measurement of Reactive Oxygen Species in the Culture Media Using Acridan Lumigen PS-3 Assay

Reactive oxygen species (ROS) are generated continuously during aerobic metabolism. ROS are highly reactive molecules and in excessive amounts, can lead to protein and DNA oxidation, protein cross-linking, and cell death. Cell-culture models provide a valuable tool in understanding the mechanisms that lead to cell death. Accumulation of ROS within cells and/or their release into the culture media are highly cell type-specific. The ability to estimate ROS levels in the culture media is an important step in understanding the mechanisms contributing to disease processes. In this paper, we describe the optimization of a simple method to estimate ROS levels in the culture media using the Acridan Lumigen PS-3 reagent provided in the Amersham ECL Plus kit (GE Healthcare, UK). We have shown that the Acridan Lumigen PS-3 assay generates ROS-specific chemiluminescence in fresh as well as media stored at −20°C, in as little as 10–20 μl of samples. The method was able to detect the dose (of stimulants)- and time (acute and chronic)-dependent changes in ROS levels in media collected from various cell types. Our results suggest that the kit reagents, PBS buffer, and various media did not contribute significantly to the overall chemiluminescence generated in the assay; however, we suggest that the unused medium specific for each cell type should be used as blanks and final readings of test samples normalized against these readings. As this method uses commonly available laboratory equipment and commercially available reagents, we believe this assay is convenient, economical, and specific in estimating ROS released extracellularly into the culture media.     

Oxygen Diffusion from the Roots of Rice Grown under Non-waterlogged Conditions

Three rice varieties, cv. Norin 36, cv. Norin 37 and cv. Yubae, were grown in a loam with a 20 cm water-table which gave aerobic conditions to a depth of not less than 15 to 17 cm. Under these conditions Norin 36 grew more vigorously and tillered more frequently than the other two varieties. The rates of oxygen diffusion at 23°C from roots up to 11 cm in length were however appreciably lower for Norin 36 (4.3 × 10^?8g · cm^?2 of root surface · min^?1) than for Norin 37 or Yubae (c. 7.8 × 10^?8g). A considerable increase (up to 200 %) in the oxygen diffusion rate (ODR) from the roots occurred if they were cooled to 3°C, and at this temperature differences in ODR between the varieties were not significant. For a purely physical system, because of the decrease in the diffusion coefficient of oxygen in water, and, the increase in oxygen solubility, a drop of c. 20 % in ODR should accompany the above 20°C drop in temperature. A 16 % drop was recorded for artificial ‘roots’ under these conditions. It was concluded that respiratory activity at the higher temperature must have been responsible for the low readings and intervarietal differences observed at 23°C. By increasing the 3°C values by 25 % a mean value of 14.2 × 10^?8g · cm^?2 of root surface · min^?1 was recorded for the three varieties, being the probable ODR at 23°C in the absence of a respiratory factor. Calculations show that respiratory activity removed enough oxygen to reduce the ODR for Norin 36 by more than 9 × 10^?8g, and for Norin 37 and Yubae by c. 6.7 × 10^?8g · cm^?2 of root surface · min^?1. Anatomical investigations showed that cortical breakdown was always extensive at 4 to 4.5 cm from the apex of the roots. In some cases however breakdown had not occurred in the basal segment of the root. No opinion could be formed as to whether differences in the amount of cortical breakdown between the varieties might have occasioned the respiratory differences observed. An interesting feature of the root anatomy was the failure of breakdown in those regions of the roots through which lateral roots emerged.     

Oxygen Sensitivity of Various Anaerobic Bacteria

Anaerobes differ in their sensitivity to oxygen, as two patterns were recognizable in the organisms included in this study. Strict anaerobes were species incapable of agar surface growth at pO(2) levels greater than 0.5%. Species that were found to be strict anaerobes were Treponema macrodentium, Treponema denticola, Treponema oralis n. sp., Clostridium haemolyticum, Selenomonas ruminatium, Butyrivibrio fibrisolvens, Succinivibrio dextrinosolvens, and Lachnospira multiparus. Moderate anaerobes would include those species capable of growth in the presence of oxygen levels as high as 2 to 8%. The moderate anaerobes could be exposed to room atmosphere for 60 to 90 min without appreciable loss of viability. Species considered as moderate anaerobes were Bacteroides fragilis, B. melaninogenicus, B. oralis, Fusobacteria nucleatum, Clostridium novyi type A, and Peptostreptococcus elsdenii. The recognition of at least two general types of anaerobes would seem to have practical import in regard to the primary isolation of anaerobes from source material.     

Growth Curves of Anaerobic Bacteria in Solid Media

Simple pour plate and spectrophotometric techniques for the evaluation of growth curves of several anaerobic bacteria on solid media are described. Three basic patterns of anaerobic growth were observed. The curves obtained were very reproducible when studied on separate occasions. The curves obtained by spectrophotometric measurement were comparable to those obtained by the pour plate method, especially when a large bacterial inoculum was used. Limitations in the interpretation of the results are discussed. The methods and principles reported could provide the basis for the determination of bacterial growth on solid media using other organisms and different experimental conditions.     

Rhizosphere Oxidation in Rice: An Analysis of Intervarietal Differences in Oxygen Flux from the Roots

The polarographic technique (Armstrong 1967) has been found suitable for the detection of significant intervarietal differences in oxygen diffusion from rice roots. All the evidence obtained supports the thesis that intervarietal differences in oxygen diffusion rate occur and the varieties tested may be arranged in order of increasing oxygen flux as follows N 36 < N 22 < N 29 < N 32 < Yubae. The lowest oxygen flux rates are found in those varieties (N 22 and N 36) susceptible to Akagare disease of rice while the flux rates of varieties resistant to Akagare (N 29 and N 32) are higher. On the basis of these results one would also expect Yubae to be resistant to Akagare. The results are discussed with reference to previous work which has shown the oxidising activity and carbohydrate exudation properties of rice roots to be similarly correlated with resistance or susceptibility to Akagare. Attention is drawn to what it is felt are the shortcomings in existing techniques for measuring the oxidising activity of roots. It is suggested that with certain provisions the polarographic technique may he found useful in any effort to build up high oxidising power into the new plant type proposed for the tropics.     

Factors Related to the Oxygen Tolerance of Anaerobic Bacteria

The effect of atmospheric oxygen on the viability of 13 strains of anaerobic bacteria, two strains of facultative bacteria, and one aerobic organism was examined. There were great variations in oxygen tolerance among the bacteria. All facultative bacteria survived more than 72 h of exposure to atmospheric oxygen. The survival time for anaerobes ranged from less than 45 min for Peptostreptococcus anaerobius to more than 72 h for two Clostridium perfringens strains. An effort was made to relate the degree of oxygen tolerance to the activities of superoxide dismutase, catalase, and peroxidases in cell-free extracts of the bacteria. All facultative bacteria and a number of anaerobic bacteria possessed superoxide dismutase. There was a correlation between superoxide dismutase activity and oxygen tolerance, but there were notable exceptions. Polyacrylamide gel electropherograms stained for superoxide dismutase indicated that many of the anaerobic bacteria contained at least two electrophoretically distinct enzymes with superoxide dismutase activity. All facultative bacteria contained peroxidase, whereas none of the anaerobic bacteria possessed measurable amounts of this enzyme. Catalase activity was variable among the bacteria and showed no relationship to oxygen tolerance. The ability of the bacteria to reduce oxygen was also examined and related to enzyme content and oxygen tolerance. In general, organisms that survived for relatively long periods of time in the presence of oxygen but demonstrated little superoxide dismutase activity reduced little oxygen. The effects of medium composition and conditions of growth were examined for their influence on the level of the three enzymes. Bacteria grown on the surface of an enriched blood agar medium generally had more enzyme activity than bacteria grown in a liquid medium. The data indicate that superoxide dismutase activity and oxygen reduction rates are important determinants related to the tolerance of anaerobic bacteria to oxygen.     

A Detoxifying Oxygen Reductase in the Anaerobic Protozoan Entamoeba histolytica

We report the characterization of a bacterial-type oxygen reductase abundant in the cytoplasm of the anaerobic protozoan parasite Entamoeba histolytica. Upon host infection, E. histolytica is confronted with various oxygen tensions in the host intestine, as well as increased reactive oxygen and nitrogen species at the site of local tissue inflammation. Resistance to oxygen-derived stress thus plays an important role in the pathogenic potential of E. histolytica. The genome of E. histolytica has four genes that encode flavodiiron proteins, which are bacterial-type oxygen or nitric oxide reductases and were likely acquired by lateral gene transfer from prokaryotes. The EhFdp1 gene has higher expression in virulent than in nonvirulent Entamoeba strains and species, hinting that the response to oxidative stress may be one correlate of virulence potential. We demonstrate that EhFdp1 is abundantly expressed in the cytoplasm of E. histolytica and that the protein levels are markedly increased (up to ～5-fold) upon oxygen exposure. Additionally, we produced fully functional recombinant EhFdp1 and demonstrated that this enzyme is a specific and robust oxygen reductase but has poor nitric oxide reductase activity. This observation represents a new mechanism of oxygen resistance in the anaerobic protozoan pathogen E. histolytica.     

Use of the Anaerobic Pouch in Isolating Clostridium botulinum Spores from Fresh Meats

The anaerobic film pouch was demonstrated to be an effective device for the primary isolation of Clostridium botulinum types A and B spores from raw pork, beef, and chicken. Optimal pasteurization of these meats (for reduction of nonspore microflora without affecting indigenous putrefactive anaerobic spore levels) was 50 min at 60 C. C. botulinum spores were recovered with good precision from meat samples inoculated with mixtures of C. botulinum and Putrefactive Anaerobe 3679 at 1:1 and at 1:99 ratios. Verification of C. botulinum isolates was accomplished by protection testing of subcultures in mice.