Theory of Diffusion in Gels

It has been shown that when concentration of solute is expressed as amount per unit volume of gel—solvent—solute system, Fick's laws for diffusion in a gel take the same form as for diffusion in solvent alone, except that the usual coefficient must be replaced by a new coefficient, D′, equal to D(1 - α)/(1 - ), where is the effective volume fraction of the gel substance and α is a coefficient of obstruction equal to 5/3 if the gel substance can be considered to be made up of randomly oriented rods. An equation was derived for the total amount of solute entering the gel, which is analogous to but not identical with the equation for the total amount of solute crossing the initial boundary in free diffusion. The effect of slice thickness was investigated by a mathematical procedure involving the solutions of approximate differential equations. It was shown that even for slices so thick that 95 per cent of the solute in the gel is contained in the first two, a correction factor equal to the square of the slice thickness divided by 48D't permits one to obtain accurate measurement of D′ from the mean concentration and the position of the midplane of the slice.     

Diffusion of Small Solutes in the Lateral Intercellular Spaces of MDCK Cell Epithelium Grown on Permeable Supports

The diffusion coefficients of four solutes ranging in molecular weight from 238 to 10,000 in the lateral intercellular spaces (LIS) of cultured kidney cells (MDCK) grown on permeable supports were determined from the spread of fluorescence produced after the release of caged compounds by a pulse from a UV laser. Two types of experiments were performed: measurement of the rate of change of fluorescence after releasing a caged fluorophore, and measurement of the change in fluorescence of a relatively static fluorescent dye produced by the diffusion of an uncaged ligand for the dye. Fluorescence intensity was determined by photon-counting the outputs of a multichannel photomultiplier tube. Diffusion coefficients were determined in free solution as well as in the LIS of MDCK cells grown on permeable supports and the hindrance factor, θ, determined from the ratio of the free solution diffusivity to that in the LIS. The hindrance factors for 3000-MW dextran, 8-hydroxypyrene-1,3,6-trisulfonic acid (HPTS, MW 524) and N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid (HEPES, MW 238) were not significantly different from 1. The diffusion of 10,000-MW dextran was substantially reduced in the LIS with a θ of 5.6 ± 0.3. Enzymatic digestion by neuraminidase of the sialic acid residues of the glycosylation groups in the LIS increased the diffusivity of the 10,000-MW dextran 1.8-fold indicating hindrance by the glycocalyx. We conclude that small solutes, such as Na⁺ and Cl⁻, would not be significantly restricted in their diffusion in the LIS and that solute concentration gradients could not develop along the LIS under physiologic conditions. Received: 7 October 1999     

The Tauber Pollen Trap,a Discussion of Its Usefulness in Pollen Deposition Studies

In connection with pollen deposition studies some discrepancies between Tauber traps and open collectors were discovered. Field observations showed that the aerodynamic lid of the Tauber trap itself acted as a pollen trap by redeposition e.g. because of rainsplash. In an experiment coal dust in the size range of pollen was introduced in a turbulent air stream passing over the trap. Great quantities of coaldust were captured on the aerodynamic lid. By exposure to artificial rain it was observed that the splashing effect of raindrops brought coal dust into the sampler. By application of Corylus pollen on the aerodynamic lid and exposure to rain, it was found that great amount of pollen is transferred by raindrop splash to the trap. This contamination of the pollen contents of the trap is so serious that the usefulness of the Tauber trap for quantitative evaluation of the pollen deposition is doubted unless the trap is protected in some way. Further tests of this effect have been planned.     

荞麦水合花粉和花粉管中的被膜小泡

荞麦水合花粉粒和生长中的花粉管中内质网潴泡形成的囊袋状结构较少见,但内质网囊袋中含有丰富的被膜小泡,直径约为100－150nm。刚刚形成的花粉管中,被膜小泡主要来自于花粉粒营养细胞的细胞质。生长中的花粉管的被膜小泡可由高尔基体分泌形成。另外还观察到内质网的碎裂也是荞麦花粉管中产生被膜小泡的一种机制。花粉管的被膜小泡中含有花粉管壁的前体物质,与花粉管的壁融合参与花粉管的生长。被膜小泡可能含有与脂体和造粉质体水解有关的酶,参与此类物质的降解。荞麦花柱和柱头细胞内含物的解体物质参与花粉管的生长。     

花柱和花粉胞外钙调素对花粉萌发和花粉管伸长的影响

以烟草为材料,通过半体内实验,就花柱和花粉胞外钙调素对花粉萌发和花粉管伸长的影响进行了观察。发现用EGTA及钙调素抗血清处理柱头或花粉均可抑制花粉在柱头上的萌发;向花柱引导组织中显微注射纯化钙调素可促进花粉管束伸长,而注射钙调素抗血清可抑制花粉管束伸长;同时证实玉米花柱和花粉细胞壁中均存在钙调素及钙调素结合蛋白,而且花粉和花柱细胞壁中钙调素结合蛋白的种类有差异。结果表明存在于花粉和花柱细胞外的钙调素对花粉萌发和花粉管伸长均有促进作用。     

The Role of Liver in Determining Serum Colon-Derived Uremic Solutes

Evidence has shown that indoxyl sulfate (IS) and p-cresyl sulfate (PCS) may be alternative predictors of clinical outcomes in chronic kidney disease (CKD). Both toxins are derived from the gastrointestinal tract and metabolised in the liver. However, it is unclear whether the liver affects the production of IS and PCS. Here, we explore the association between IS and PCS levels in liver cirrhosis and a CKD-based cohort (N = 115). Liver and kidney function was assessed and classified by a Child-Pugh score (child A–C) and a modified version of the Modification of Diet in Renal Disease (MDRD) equation (Stages 1–4), respectively. An animal model was also used to confirm the two toxin levels in a case of liver fibrosis. In patients with early liver cirrhosis (child A), IS and PCS were significantly associated with CKD stages. In contrast, serum IS and PCS did not significantly change in advanced liver cirrhosis (child C). A stepwise multiple linear regression analysis also showed that T-PCS was significantly associated with stages of liver cirrhosis after adjusting for other confounding factors (B = -2.29, p = 0.012). Moreover, the serum and urine levels of T-PCS and T-IS were significantly lower in rats with liver failure than in those without (p<0.01, p<0.05 and p<0.01, p<0.05, respectively). These results indicated that in addition to the kidneys, the liver was an essential and independent organ in determining serum IS and PCS levels. The production rate of IS and PCS was lower in patients with advanced liver cirrhosis.     

Transport of Compatible Solutes in Extremophiles

Salt-tolerant as well as moderately halophilic and halophilic organisms have to maintain their turgor. One strategy is to accumulate small organic compounds, compatible solutes, by de novo synthesis or uptake. From a bioenergetic point of view, uptake is preferred over biosynthesis. The transport systems catalyzing uptake of compatible solutes are of primary or secondary nature and coupled to ATP hydrolysis or ion (H+, Na+) symport. Expression of the transporter genes as well as the activity of the transporters is regulated by salinity/osmolarity and one of the key questions is how salinity or osmolarity is sensed and the signal transmitted as far as to gene expression and transporter activation. Recent studies shed light on the nature and the activation mechanisms of solute transporters in extremophiles, and this review summarizes current knowledge on the structure, function and osmo- or salt-regulation of transporters for compatible solutes in extremophiles.     

Solutes and succulence in southern African mistletoes

The inorganic ion and organic solute composition of Tapinanthus oleifolius (Loranthaceae) and Viscum rotundifolium (Viscaceae) growing on various host trees in Namibia were investigated. Organic osmolytes accounted for 22.8–45.1% of the total solutes determined in leaves of the mistletoes parasitizing Acacia nebrownii, Catophractes alexandri, Grewia flava and Ziziphus mucronata. On other hosts (Acacia karroo, Euphorbia virosa, Salvadora persica and Tamarix usneoides) T. oleifolius showed distinct succulence with increasing leaf age, with leaves more than 3 mm thick on E. virosa. In the more succulent leaves (> 1.1 kg H₂O m^–2 leaf area) organic solutes were only of minor importance. Water content per leaf area was significantly correlated with chloride content per leaf area, suggesting that succulence served as a means to keep ion concentration at a physiological tolerable level. At whole plant levels 93.7% of the total sodium of the mistletoe bush was located in leaves thicker than 1 mm for T. oleifolius parasitizing Tamarix usneoides. This pronounced sequestration of sodium in older leaves as well as the high variability of the K/Na ratio in various parts of the parasite-host system point to highly selective ion distribution processes in this association.     

Isoenzyme Polymorphism in Flowering Plants I. Diffusion of Enzymes Out of Intact Pollen Grains

Esterases, leucine aminopeptidases, catalases, amylases and acid phosphatases diffuse out of intact and ungerminated pollen grains of Oenothera organensis, whether suspended in 1 % sodium chloride or in pollen medium. A total of 15 esterase isozymes are recorded; 5 of them appear within 5 minutes, 8 within 30 minutes, 9 within 2 hours, and 13 within 19 hours. Pollen grains suspended for 19 hours gave much stronger isozyme bands than macerated pollen grains. However, one esterase hand was consistently missing from the 19 hour suspensions, although present in all others. It is suggested as a working hypothesis that the early growth of pollen tubes and possibly even germination of pollen grains may be influenced by the metabolic products of pistillate tissues caused by the diffused pollen enzymes, and that inactivation of these enzymes by stigmatic or stylar components could lead to incompatibility reactions.     

Composition, Variation, and Dynamics of Major Osmotic Solutes in Methanohalophilus Strain FDF1

Methanohalophilus strain FDF1, a member of the halophilic genus of methanogens, can grow over a range of external NaCl concentrations from 1.2 to 2.9 M and utilize methanol, trimethylamine, and dimethyl sulfide as substrates for methanogenesis. It produces the osmolytes glycine betaine, beta-glutamine, and N-acetyl-beta-lysine with increasing external NaCl, but the relative ratio of these zwitterions depends primarily on the methanogenic substrate and less on the external osmolarity. When the cells are grown on methanol in defined medium, accumulation of glycine betaine predominates over the other zwitterionic solutes. The cells also synthesized a carbohydrate which was not detected in cells grown on trimethylamine. This negatively charged compound, identified as alpha-glucosylglycerate from the C and H chemical shifts, does not act as an osmoregulatory solute in the salt range 1.4 to 2.7 M in this methanogen as evidenced by its invariant intracellular concentration. CH(3)OH-pulse/CH(3)OH-chase experiments were used to determine half-lifes for these organic solute pools in the cells. l-alpha-Glutamate showed a rapid loss of heavy isotope, indicating that l-alpha-glutamate functions as a biosynthetic intermediate in these cells. Measurable turnover rates for both beta-glutamine, which acts as an osmolyte, and alpha-glucosylglycerate suggest that they function as metabolic intermediates as well. Molecules which function solely as osmolytes (glycine betaine and N-acetyl-beta-lysine) showed a slower turnover consistent with their roles as osmotic solutes in Methanohalophilus strain FDF1.     

Disc-Agar Diffusion Microbiological Assay Procedure for Determining Serum and Urine Levels of Sulfacytine and Other Sulfonamides

A reasonably precise, reproducible, and sensitive microbiological procedure for directly assaying sulfacytine and other sulfonamides as antibacterially active drugs has been developed by appropriately modifying the standard disc-agar diffusion technique. Blood and urine levels as low as 3 mug/ml may be determined through the use of a strain of Escherichia coli and a chemically defined agar medium devoid of sulfonamide antagonists. Results indicate that this assay method should be a useful adjunct to the Bratton-Marshall colorimetric procedure, by permitting the direct measurement of antibacterially active drug in clinical specimens.     

Pollen, Mating and Paternity in Agamospermous Angiosperms

Abstract This review article deals with the male function in agamospermous plants, which is still not fully recognized. Of particular interest are mating and paternity problems. Pollen of agamospermous donors may influence female fitness in early mating phases by contributing the recognition substances or inducing spontaneous seed development. Various interactions between the sperm cell(s) and the female gametophyte at later phases may include (1) occasional double fertilization, (2) occasional syngamy, (3) pseudogamy, and (4) hemigamy. In the first two processes the offspring are sired normally. In pseudogamy, the offspring is maternal, the male contribution being restricted to endosperm paternity. In hemigamy, male function may include induction of the egg cell development only (maternal offspring), or the induction of the egg cell development as well as independent participation of the sperm cell in the offspring development leading to the formation of chimeras. It is concluded that the sperm may be a limiting resource in at least some agamospermous plants. Possible directions of future research are briefly outlined.     

Pollen presenters in the flowering plants–form and function

Pollen presenters are specific floral structures, other than anthers, from which pollen is distributed for cross fertilization between flowers. They occur in only five families of monocotyledons and 20 families of the dicotyledons. Presenters in 15 families are described here. In the largest plant family (Asteraceae) all taxa have pollen presenters, while in others (e.g. Myrtaceae) they occur in only some species in a few genera. Most presenters are associated with the gynoecium and there is a wide range of forms involving hairs or outgrowths of the stylar tissue. Despite the placement of self pollen close to the stigma most taxa are outcrossing and avoid self fertilization by protandry, with the stigma being covered at anthesis, or by precise placement of pollen so that is does not contact the stigma. Likely selective advantages in the development of pollen presenters include greater accuracy in pollen transfer than in the normal anther to stigma movement and avoidance of interference between male and female organs in the flower. In some groups there is enhanced ability for female choice because effective pollen delivery enables a choice from among many pollen grains, while in others pollen presenters enhance male reproductive success. Study of the pollination biology of plants requires a knowledge of which plants have pollen presenters and a full understanding of the structures in the flower which are associated with the pollen presentation action.     

Immunological Reactions of Single Pollen Grains, Electrophoresis and Enzymology of Pollen Protein Exudates

Single intact pollen grains of Oenothera organensis, when placedupon a thin layer of agar containing pollen antiserum, producecircular areas of precipitate. Pollen grains from an S₂S₂ plantdo not produce precipitate in S₆ antiserum. Pollen grains froman S₆S₆ plant and an S₂'S₄' self-compatible plant produce precipitatesin S₆ antiserum. Fifty per cent of the pollen grains from anS₂S₆ plant produce precipitate in S₆ antiserum. Protein diffusesinto buffer solutions from intact pollen grains within 212 min.As much as 40 per cent of the total protein diffuses out inan hour. Amylase and invertase were detected in the diffusatefrom pollen grains. Alkaline and acid phosphatases were confinedto the pollen grains and did not diffuse out. The serologicalprecipitates are specific to the incompatibility system.     

A Unique Pool of Compatible Solutes on Rhodopirellula baltica,Member of the Deep-Branching Phylum Planctomycetes

The intracellular accumulation of small organic solutes was described in the marine bacterium Rhodopirellula baltica, which belongs to the globally distributed phylum Planctomycetes whose members exhibit an intriguing lifestyle and cell morphology. Sucrose, α-glutamate, trehalose and mannosylglucosylglycerate (MGG) are the main solutes involved in the osmoadaptation of R. baltica. The ratio and total intracellular organic solutes varied significantly in response to an increase in salinity, temperature and nitrogen content. R. baltica displayed an initial response to both osmotic and thermal stresses that includes α-glutamate accumulation. This trend was followed by a rather unique and complex osmoadaptation mechanism characterized by a dual response to sub-optimal and supra-optimal salinities. A reduction in the salinity to sub-optimal conditions led primarily to the accumulation of trehalose. In contrast, R. baltica responded to salt stress mostly by increasing the intracellular levels of sucrose. The switch between the accumulation of trehalose and sucrose was by far the most significant effect caused by increasing the salt levels of the medium. Additionally, MGG accumulation was found to be salt- as well as nitrogen-dependent. MGG accumulation was regulated by nitrogen levels replacing α-glutamate as a K⁺ counterion in nitrogen-poor environments. This is the first report of the accumulation of compatible solutes in the phylum Planctomycetes and of the MGG accumulation in a mesophilic organism.