Conformational Transitions and Modifications in Ionic Exchange Properties Induced by Alkaline Ions in the Nitella flexilis Cell Wall

The decrease in the cationic exchange capacity (CEC) concomitantto the replacement in the Nitella wall of adsorbed Mn²⁺ ionswas measured in different mixtures of alkaline ions. At lowexternal concentrations, the loss of CEC is enhanced in presenceof Li⁺ ions but is weaker when Na⁺ ions are present in the exchangemixtures. The relative affinity of the wall exchange sites foralkaline ions was Na⁺>K⁺>Rb⁺Cs⁺>Li⁺. As the CEC isprogressively reduced, the wall discrimination between the differentalkaline ions tends to cancel out except for the Na⁺-K⁺ pair.The wall preference for K⁺ is then increased. A diminution ofthe effective pK of the polygalacturonic acids constitutiveof the wall is also observed, while increasing the CEC loss.The simple disruption of divalent cation crosslinks cannot fullyexplain the CEC leakage at low monovalent concentrations. Itis suggested that the alkaline ions also cleave H bonds or solvatation-likebonds between the cell wall polyuronides and then cause a concomitantunfolding of the short pectic chains which involves their solubilization. (Received May 6, 1993; Accepted November 8, 1993)     

Proton-Metal Cation Exchange in the Cell Wall of Nitella flexilis

When protons are exchanging for bivalent cations (Cu²⁺, Zn²⁺, or Ca²⁺) on the carboxylic groups of Nitella flexilis cell wall, the values of the respective global equilibrium constants do not change up to a protonation degree of 80%. These values drastically increase at higher proton concentrations and tend to 3.4, which is the intrinsic pK value of the constitutive α-d-galacturonic acid monomer. These data suggest that the electric field in the matricial polymer and the cation bridges between pairs of negative sites have disappeared.     

Estimation of the Anion-Exchange Capacity of the Cell Wall of Nitella flexilis

Diffusion potentials in KCI and triphenyl phosphonium chloride(TPP) solutions have been measured across an isolated cell wallof Nitella flexilis either after pretreatment by difluoronitrobenzene(DFNB), a compound which reacts with the protein amino groups,or after alkalization of the external solutions. In both cases,it appears that the cationic transport numbers are enhancedwith respect to the controls. These data suggest that the maineffect of the two treatments was to deprive the wall of positivecharges carried by protonized amino groups. The changes in t_Kwere used to roughly estimate the anion-exchange capacity ofthe wall.     

Ionic Diffusion Through the Nitella Cell Wall in the Presence of Calcium

Diffusion potentials (concentration and bi- or multi-ionic potentials)in KC1, NaCl, or LiCl solutions have been measured across anisolated cell wall of Nitella, with or without the same concentrationof CaCl² on either side of the cell wall. The absolute valueof the potentials decreases as the external Ca²⁺concentrationincreases and it may happen that an inversion of the sign ofthe concentration potentials results when the external Ca²⁺solution reaches 1 mM. Dosages of K⁺ and Ca²⁺ in the cell wallhave shown that Ca²⁺ easily displaces the monovalent ion fromthe exchange sites and tends to neutralize the cationic exchanger.However, in most cases, the measured potentials are still morenegative than the theoretical potentials which would be setup by a neutral-site membrane in the same conditions. Theseresults suggest that Ca²⁺ largely reduces the discriminationproperties of the cell wall between cations and anions.     

Infrared Spectra of Nitella Cell Walls and Orientation of Carboxylate Ions in the Walls

Infrared spectra of film specimens of the cell wall of Nitella were recorded in the untreated state, after acid treatment, and after treatment for removal of pectic substances and hemicellulose. Assignment of the bands in the spectrum of the wall was made. Polarization measurements on the wall indicate that in addition to cellulose, carboxylate ions, which are attributable to pectic substances, are oriented in the wall. The nature of the bonds holding the oriented carboxylate ions is described.     

Cell Wall and Morphological Changes Induced by Temperature Shift in Bacillus subtilis Cell Wall Mutants

Bacillus subtilis RUB1012 and RUB1013 have the following phenotype when grown at 45 degrees C: no growth on tryptose blood agar base, growth as clumps of spheres in broth culture, a slow autolysis rate, and a low proportion of teichoic acid to peptidoglycan. Revertants of strain RUB1012 (RUB2032, RUB2012, and RUB2042) that could grow on tryptose blood agar base were isolated. Each revertant had a different proportion of teichoic acid to peptidoglycan. The nanomoles of phosphorus per milligram of cell wall at the nonpermissive temperature were 141, 160, 236, and 541 for strain RUB1012 and revertants RUB2032, 2012, and 2042, respectively, as compared with 1,100 for the parent strain. With most bacteriophage tested, plating efficiency was related to the amount of glucosylated teichoic acid. Scanning electron microscopy was used to study strain RUB2032 during a shift from growth at 30 degrees C to growth at 45 degrees C. The change from rod to sphere began with the thickening of the cylindrical portion of the cell. Caps of the cells appeared to be immune to the thickening process. During growth, the cells became progressively shorter and thicker, and cell separation was inhibited. When cells of strain RUB2032 were shifted from growth at 45 degrees C to growth at 30 degrees C, accumulation of an amorphous material on the outer surfaces of the cells preceded the change from sphere to rod morphology. Cells remained clumped, with rods appearing at the periphery of the clumps. Analysis by DNA-mediated transformation and PBS1-mediated transduction indicated that strains RUB1012 and RUB1013 have multiple mutations mapping in the same region as other cell wall mutations.     

Control of Cell Elongation in Nitella by Endogenous Cell Wall pH Gradients: MULTIAXIAL EXTENSIBILITY AND GROWTH STUDIES

The multiaxial stress of turgor pressure was stimulated in vitro by inflating isolated Nitella cell walls with mercury. The initial in vitro extension at pH 6.5, 5 atmospheres pressure, returned the wall approximately to the in vivo stressed length, and did not induce any additional extension during a 15-minute period. Upon release of pressure, a plastic deformation was observed which did not correlate with cell growth rates until the final stages of cell maturation. Since wall plasticity does not correlate with growth rate, a metabolic factor(s) is implicated. Walls at all stages of development exhibited a primary yield stress between 0 and 2 atmospheres, while rapidly growing cells (1-3% per hour) exhibited a secondary yield stress of 4 to 5 atmospheres. The creep rate and plastic deformation of young walls were markedly enhanced by acid buffers (10 millimolar, pH ≤ 5.3).     

Simultaneous Measurement of the Bioelectric Potential of the Cell Wall and the Vacuoles during the Oscillatory Response to the Nitella Cell

Simultaneous measurements of bioelectric potentials of the vacuole and cell wall in cells of Nitella mucronata were made by inserting glass microelectrodes into the vacuole and cell wall respeclively. During the oscillation of the bioelectric potential of the vacuole. induced by sudden changes of the external bathing solution or by the impalement of the cell with a microelectrode. the cell wall potential also exhibited fluctuations of variable intensities in phase and concomitant with spikes of the vacuolar potential oscillation. However, the polarity of the pulses of the cell wall potential was reverse to that of the spikes of the vacuolar potential. These results suggest that the same event is registered at both sides of the plasmalemma membrane across which these phenomena are occurring. The results also support the voltage clamp and tracer flux measurements on these cells which indicate that during the generation of single action potentials, induced by current, the plasma lemma transiently increases its permeability to Cl^? and K⁺ ions expelling them from the cell. The variable intensity of the transient hyperpolarizations of the cell wall potential is explained by the distance of the microelectrode in the cell wall from the plasmalemma.     

Relations Between Indoleacetic Acid, Calcium Ions and Ethylene in the Regulation of Growth and Cell Wall Composition in Picea abies

Effects of indoleacetic acid, calcium ions and ethylene on thegrowth of and deposition of different cell wall fractions inthe hypocotyl of Norway spruce (Picea abies (L.) Karst.) seedlingswere investigated. Indoleacetic acid progressively stimulated cellulose depositionas the amount of added Ca²⁺ increased. In contrast, indoleaceticacid promoted lignification and the deposition of non-cellulosicpolysaccharides only in the absence of added Ca²⁺ . When Ca²⁺was added, the indoleacetic acid effect disappeared. Similarly,indoleacetic acid promoted non-cellulosic polysaccharide depositiononly in the absence of ethylene. At increasing ethylene levelsthe effect of indoleacetic acid on non-cellulosic polysaccharidedeposition disappeared and indoleacetic acid instead promotedcellulose deposition. The response to indoleacetic acid depended on the Ca²⁺ concentrationand on the rate of ethylene production. The relationship betweenindoleacetic acid and Ca²⁺ seemed complex, but clearly indoleaceticacid could partially overcome a Ca²⁺ deficiency. The resultssuggest that ethylene may be a factor of particular importancefor the type of polysaccharide deposition during cell wall formation. Key words: Calcium, cell wall, conifers, ethylene, indoleacetic acid     

Biochemical Localization of Alkaline Phosphatase in the Cell Wall of a Marine Pseudomonad

The various layers of the cell envelope of marine pseudomonad B-16 (ATCC 19855) have been separated from the cells and assayed directly for alkaline phosphatase activity under conditions established previously to be optimum for maintenance of the activity of the enzyme. Under conditions known to lead to the release of the contents of the periplasmic space from the cells, over 90% of the alkaline phosphatase was released into the medium. Neither the loosely bound outer layer nor the outer double-track layer (cell wall membrane) showed significant activity. A small amount of the alkaline phosphatase activity of the cells remained associated with the mureinoplasts when the outer layers of the cell wall were removed. Upon treatment of the mureinoplasts with lysozyme, some alkaline phosphatase was released into the medium and some remained with the protoplasts formed. Cells washed and suspended in 0.5 M NaCl were lysed by treatment with 2% toluene, and 95% of the alkaline phosphatase in the cells was released into the medium. Cells washed and suspended in complete salts solution (0.3 M NaCl, 0.05 M MgSO(4), and 0.01 M KCl) or 0.05 M MgSO(4) appeared intact after treatment with toluene but lost 50 and 10%, respectively, of their alkaline phosphatase. The results suggest that the presence of Mg(2+) in the cell wall is necessary to prevent disruption of the cells by toluene and may also be required to prevent the release of alkaline phosphatase by toluene when disruption of the cells by toluene does not take place.     

Influence of Magnesium Ions on Porosity of the Bacillus megaterium Cell Wall and Membrane

The presence of magnesium chloride (0.01 M) caused a relatively small but significant reduction in the equivalent porosity of the protoplast membrane, but not in that of the cell wall, in intact Bacillus megaterium cells.     

Measuring Plant Cell Wall Extension (Creep) Induced by Acidic pH and by Alpha-Expansin

Growing plant cell walls characteristically exhibit a property known as ''acid growth'', by which we mean they are more extensible at low pH (< 5) ¹. The plant hormone auxin rapidly stimulates cell elongation in young stems and similar tissues at least in part by an acid-growth mechanism ^{2, 3}. Auxin activates a H⁺ pump in the plasma membrane, causing acidification of the cell wall solution. Wall acidification activates expansins, which are endogenous cell wall-loosening proteins ⁴, causing the cell wall to yield to the wall tensions created by cell turgor pressure. As a result, the cell begins to enlarge rapidly. This ''acid growth'' phenomenon is readily measured in isolated (nonliving) cell wall specimens. The ability of cell walls to undergo acid-induced extension is not simply the result of the structural arrangement of the cell wall polysaccharides (e.g. pectins), but depends on the activity of expansins ⁵. Expansins do not have any known enzymatic activity and the only way to assay for expansin activity is to measure their induction of cell wall extension. This video report details the sources and preparation techniques for obtaining suitable wall materials for expansin assays and goes on to show acid-induced extension and expansin-induced extension of wall samples prepared from growing cucumber hypocotyls.To obtain suitable cell wall samples, cucumber seedlings are grown in the dark, the hypocotyls are cut and frozen at -80 °C. Frozen hypocotyls are abraded, flattened, and then clamped at constant tension in a special cuvette for extensometer measurements. To measure acid-induced extension, the walls are initially buffered at neutral pH, resulting in low activity of expansins that are components of the native cell walls. Upon buffer exchange to acidic pH, expansins are activated and the cell walls extend rapidly. We also demonstrate expansin activity in a reconstitution assay. For this part, we use a brief heat treatment to denature the native expansins in the cell wall samples. These inactivated cell walls do not extend even in acidic buffer, but addition of expansins to the cell walls rapidly restores their ability to extend.Open in a separate windowClick here to view.^{(58M, flv)}     

Surgically Induced Interpregnancy Weight Loss and Prevalence of Overweight and Obesity in Offspring

Introduction

According to the fetal overnutrition hypothesis, obesity in pregnancy predisposes the offspring to obesity. Previous studies have suggested that after biliopancreatic surgery for obesity, the offspring is less likely to be obese. This study aims to further compare the BMI development of children born before and after maternal surgical weight loss.

Method

Women with at least one child born before and one child born after bariatric surgery were identified by record-linkage. Information about maternal BMI was extracted from medical records, as was information about the children''s BMI from birth to 10 years of age. We retrieved BMI data at four years of age for 340 children, born to 223 women (164 children born before surgery (BS), 176 children born after surgery (AS)). We evaluated prevalence of overweight/obesity and mean BMI in children born BS and AS at the ages of four, six and ten using GEE regression models. For 71 families, where we had complete data on mother and both children, we used a fixed-effects regression model to explore the association between differences in maternal BMI in w10 of the pre- and post-operative pregnancies with siblings'' BMI differences at age four.

Results

In no age group did we see a significantly reduced prevalence of overweight/obesity AS. For 10-year-old girls, the AS group had significantly higher rates of obesity. There was no association between differences in maternal BMI in early pregnancy and differences in siblings'' BMI at four years of age (β = −0.01, CI 95% = −0.11; 0.09).

Conclusions

We have been unable to demonstrate any effect of bariatric surgery on weight development in offspring. It seems unlikely that restrictive bariatric surgery conveys a protective effect in offspring with regards to obesity.     

茉莉酸甲酯诱导烟草幼苗抗炭疽病与PAL活性及细胞壁物质的关系

探讨了茉莉酸甲酯(methyl jasmonate, Me-Ja)诱导烟草幼苗抗炭疽病与苯丙氨酸解氨酶(PAL)活性、木质素和富含羟脯氨酸蛋白(HRGP)含量的关系。 Me-Ja处理烟草幼苗不仅可以提高幼苗抗炭疽病的能力,而且明显提高幼苗的PAL活性、木质素和HRGP含量; 3个不同抗性品种的烟草幼苗的PAL酶活性、木质素和HRGP含量三者与感病程度之间的负相关都达到显著水平,表明PAL、木质素和富含羟脯氨酸蛋白(HRGP)在茉莉酸甲酯诱导抗病中起关键作用。     

拟康氏木霉胞壁多糖对黄瓜抗病性的诱导作用

为了更全面地认识拟康氏木霉菌（Trichoderma pseudokoningii SDTP1）的抗病机理,为该菌的开发和科学的施用提供依据,通过灌根处理和人工接病的方式研究了拟康氏木霉菌胞壁多糖（TPWS）对黄瓜幼苗抗病性的诱导作用。结果表明用浓度为200mg／L的TPWS处理后对人工接种尖镰孢分生孢子悬液的黄瓜幼苗的保护作用达到53．8％,浓度进一步提高保护作用升高不明显;用200mg／L的TPWS处理后,黄瓜幼苗下胚轴内苯丙氨酸解氨酶（PAL）和多酚氧化酶（POD）的活性及木质素的含量分别于6、30、30h后开始迅速升高,最高值时分别是对照的2．2、3．1、4．4倍;用200mg／L的TPWS处理后需要4d黄瓜幼苗才能表现出抗病性,不仅降低了伤根接种尖孢镰刀菌分生孢子悬液幼苗的枯萎病发病率,也能降低在茎部和叶面穿刺接种灰葡萄孢分生孢子悬液幼苗的灰霉病发病率。而TPWS在体外对两种病原菌无抑制作用,我们认为经处理的幼苗枯萎病和灰霉病发病率的降低与TPWS诱导了幼苗的抗病性有关,且这种抗性是系统的、非特异性的。抗病性可能与木质素含量的升高有关。     

D-Alanylation of Lipoteichoic Acids Confers Resistance to Cationic Peptides in Group B Streptococcus by Increasing the Cell Wall Density

Cationic antimicrobial peptides (CAMPs) serve as the first line of defense of the innate immune system against invading microbial pathogens. Gram-positive bacteria can resist CAMPs by modifying their anionic teichoic acids (TAs) with D-alanine, but the exact mechanism of resistance is not fully understood. Here, we utilized various functional and biophysical approaches to investigate the interactions of the human pathogen Group B Streptococcus (GBS) with a series of CAMPs having different properties. The data reveal that: (i) D-alanylation of lipoteichoic acids (LTAs) enhance GBS resistance only to a subset of CAMPs and there is a direct correlation between resistance and CAMPs length and charge density; (ii) resistance due to reduced anionic charge of LTAs is not attributed to decreased amounts of bound peptides to the bacteria; and (iii) D-alanylation most probably alters the conformation of LTAs which results in increasing the cell wall density, as seen by Transmission Electron Microscopy, and reduces the penetration of CAMPs through the cell wall. Furthermore, Atomic Force Microscopy reveals increased surface rigidity of the cell wall of the wild-type GBS strain to more than 20-fold that of the dltA mutant. We propose that D-alanylation of LTAs confers protection against linear CAMPs mainly by decreasing the flexibility and permeability of the cell wall, rather than by reducing the electrostatic interactions of the peptide with the cell surface. Overall, our findings uncover an important protective role of the cell wall against CAMPs and extend our understanding of mechanisms of bacterial resistance.     

Derivation of Plant Cell Wall Water Content by Examination of the Water-Holding Capacity of Membrane-Disrupted Tissues10

Plant cell lumina are several orders of magnitude larger thancell wall pores. If the membranes of the plant sample are disruptedand the tissue dried out, a value should be reached at whichthe cell lumina have drained but the cell walls remain relativelyhydrated. The water content of membrane-disrupted tissues atlow may, therefore, serve as a good approximation of plantcell wall water content (CW). The relationship between and water content of membrane-disruptedtissues was measured for four grass spaces over the range of0 to –40 MPa. It was found that most water loss occurredabove a of – 10 MPa. CW was estimated from the watercontent of membrane-disrupted tissues at lower and it was foundthat there was very little intra-or intraspecific variabilityin CW estimates when water content was expressed as a percentagedry weight of the tissue. This is in contrast to the high variabilityin CW estimated from the analysis of inverse water potential—watercontent (IP—WC) curves measured with the thermocouplepsychrometer. Applications of the membrane-disruption methodof estimating CW of plant tissue are discussed. Key words: Cell wall water, thermocouple psychrometer, membrane-disruption