Denitrification Performance of Pseudomonas denitrificans in a Fluidized‐Bed Biofilm Reactor and in a Stirred Tank Reactor

Denitrification of a synthetic wastewater containing nitrates and methanol as carbon source was carried out in two systems – a fluidized‐bed biofilm reactor (FBBR) and a stirred tank reactor (STR) – using Pseudomonas denitrificans over a period of five months. Nitrogen loading was varied during operation of both reactors to assess differences in the response to transient conditions. Experimental data were analyzed to obtain a comparison of denitrification kinetics in biofilm and suspended growth reactors. The comparison showed that the volumetric degradation capacity in the FBBR (5.36 kg _N · m^–3 · d^–1) was higher than in the STR, due to higher biomass concentration (10 kg _BM · m^–3 vs 1.2 kg _BM m^–3).     

Feed-forward and feed-backward amplification model from cochlear cytoarchitecture: an interspecies comparison

The high sensitivity and wide bandwidth of mammalian hearing are thought to derive from an active process involving the somatic and hair-bundle motility of the thousands of outer hair cells uniquely found in mammalian cochleae. To better understand this, a biophysical three-dimensional cochlear fluid model was developed for gerbil, chinchilla, cat, and human, featuring an active “push-pull” cochlear amplifier mechanism based on the cytoarchitecture of the organ of Corti and using the time-averaged Lagrangian method. Cochlear responses are simulated and compared with in vivo physiological measurements for the basilar membrane (BM) velocity, V_BM, frequency tuning of the BM vibration, and Q₁₀ values representing the sharpness of the cochlear tuning curves. The V_BM simulation results for gerbil and chinchilla are consistent with in vivo cochlea measurements. Simulated mechanical tuning curves based on maintaining a constant V_BM value agree with neural-tuning threshold measurements better than those based on a constant displacement value, which implies that the inner hair cells are more sensitive to V_BM than to BM displacement. The Q₁₀ values of the V_BM tuning curve agree well with those of cochlear neurons across species, and appear to be related in part to the width of the basilar membrane.     

Robust One-Way ANOVA under Possibly Non-Regular Conditions

Consider the one-way ANOVA problem of comparing the means m₁, m₂, …, m_c of c distributions F₁(x) = F(x — m₁), …, F_c(x) = F(x — m_c). Solutions are available based on (i) normal-theory procedures, (ii) linear rank statistics and (iii) M-estimators. The above model presupposes that F₁, F₂, …, F_c have equal variances (= homoscedasticity). However practising statisticans content that homoscedasticity is often violated in practice. Hence a more realistic problem to consider is F₁(x) = F((x — m₁)/σ₁), …, F_c(x) = F((x — m_c)/σ_c), where F is symmetric about the origin and σ₁, …, σ_c are unknown and possibly unequal (= heteroscedasticity). Now we have to compare m₁, m₂, …, m_c. At present, nonparametric tests of the equality of m₁, m₂, …, m_c are available. However, simultaneous tests for paired comparisons and contrasts and do not seem to be available. This paper begins by proposing a solution applicable to both the homoscedastic and the heteroscedastic situations, assuming F to be symmetric. Then the assumptions of symmetry and the identical shapes of F₁, …, F_c are progressively relaxed and solutions are proposed for these cases as well. The procedures are all based on either the 15% trimmed means or the sample medians, whose quantiles are estimated by means of the bootstrap. Monte Carlo studies show that these procedures tend to be superior to the Wilcoxon procedure and Dunnett's normal theory procedure. A rigorous justification of the bootstrap is also presented. The methodology is illustrated by a comparison of mean effects of cocaine administration in pregnant female Sprague-Dawley rats, where skewness and heteroscedascity are known to be present.     

Mesophyll CO2 conductance and leakiness are not responsive to short‐ and long‐term soil water limitations in the C4 plant Sorghum bicolor

Breeding economically important C₄ crops for enhanced whole‐plant water‐use efficiency (WUE_plant) is needed for sustainable agriculture. WUE_plant is a complex trait and an efficient phenotyping method that reports on components of WUE_plant, such as intrinsic water‐use efficiency (WUE_i, the rate of leaf CO₂ assimilation relative to water loss via stomatal conductance), is needed. In C₄ plants, theoretical models suggest that leaf carbon isotope composition (δ¹³C), when the efficiency of the CO₂‐concentrating mechanism (leakiness, ?) remains constant, can be used to screen for WUE_i. The limited information about how ? responds to water limitations confines the application of δ¹³C for WUE_i screening of C₄ crops. The current research aimed to test the response of ? to short‐ or long‐term moderate water limitations, and the relationship of δ¹³C with WUE_i and WUE_plant, by addressing potential mesophyll CO₂ conductance (g_m) and biochemical limitations in the C₄ plant Sorghum bicolor. We demonstrate that g_m and ? are not responsive to short‐ or long‐term water limitations. Additionally, δ¹³C was not correlated with gas‐exchange estimates of WUE_i under short‐ and long‐term water limitations, but showed a significant negative relationship with WUE_plant. The observed association between the δ¹³C and WUE_plant suggests an intrinsic link of δ¹³C with WUE_i in this C₄ plant, and can potentially be used as a screening tool for WUE_plant in sorghum.     

Chemically reacting systems of the type A + B ⇄ AB. II. Osmometry

The osmotic pressure equation for nonideal, associating systems of the type nA +mB ? A_nB_m, has been derived, by using the assumption yA _nB _m/yA ⁿyB ^m = 1. This treatment can also be applied to related associations such as nA + mB ? AB + AB₂ + A₂B + …. From osmotic pressure experiments on the pure reactants it is possible to obtain the molecular weights (M_A and M_B) of the reactants and also the virial coefficients (B_AA and B_BB) of the reactants. The osmotic pressure of a nonreacting mixture of A and B can be calculated from these measurements. It can be used along with osmotic pressure measurements on equilibrium mixtures of A and B to obtain expressions containing the equilibrium constant (or constants) and the cross-virial coefficients (B_AB and B_BA). Several procedures are described for the evaluation of the equilibrium constant (or constants) and the B_AB or B_BA terms. It appears that this procedure is a general one which is applicable to associations of the type nA + mB ? AB + A₂B + AB₂ + …. By correcting for nonideal behavior, one should then be able to apply it to any method available for analyzing ideal associations of the types considered here. In addition it is possible, subject to certain restrictions, to analyze associations of the type 3A + B ? A₂ + AB.     

In vitro asymbiotic germination and seedling development of Limodorum abortivum (Orchidaceae)

Abstract

Germination tests were carried out using immature seeds of Limodorum abortivum and applying in vitro techniques. The results proved that BM₁ culture medium is suitable to promote both germination and further growth stages. Details of the developmental pattern, and some micromorphological features, are described.     

Sulphate transport in the cyanobacterium Synechococcus R-2 (Anacystis nidulans, S. leopoliensis) PCC 7942

Synechococcus R-2 (PCC 1942) actively accumulates sulphate in the light and dark. Intracellular sulphate was 1.35 ± 0.23 mol m^?3 (light) and 0.894 ± 0.152 mol m^?3 (dark) under control conditions (BG-11 media: pH_o, 7.5; [SO₄^2?]_o, 0.304 mol m^?3). The sulphate transporter is different from that found in higher plants: it appears to be an ATP-driven pump transporting one SO₄^2?/ATP [ΔμSO₄^2?_i,o=+ 27.7 ± 0.24 kJ mol^?1 (light) and + 24 ± 0.34 kj mol^?1 (dark)]. The rate of metabolism of SO₄^2?at pH_o, 7.5 was 150 ± 28 pmol m^?2 s^?1 (n = 185) in the light but only 12.8 ± 3.6 pmol m^?2 s^?1 (n = 61) in the dark. Light-driven sulphate uptake is partially inhibited by DCMU and chloramphenicol. Sulphate uptake is not linked to potassium, proton, sodium or chloride transport. The alga has a constitutive over-capacity for sulphate uptake [light (n= 105): K_m= 0.3 ± 0.1 mmol m^?3, V_max, = 1.8 ± 0.6 nmol m^?2 s^?1; dark (n= 56): K_m= 1.4 ± 0.4 mmol m^?3, V_max= 41 ± 22 pmol m^?2 s^?1]. Sulphite (SO₃^2?) was a competitive inhibitor of sulphate uptake. Selenate (SeO₄^2?) was an uncompetitive inhibitor.     

Canopy scale measurements of CO2 and water vapor exchange along a precipitation gradient in southern Africa

Short‐term measurements of carbon dioxide, water, and energy fluxes were collected at four locations along a mean annual precipitation gradient in southern Africa during the wet (growing) season with the purpose of determining how the observed vegetation–atmosphere exchange properties are functionally related to the long‐term climatic conditions. This research was conducted along the Kalahari Transect (KT), one in the global set of International Geosphere‐Biosphere Program transects, which covers a north–south aridity gradient, all on a homogenous sand formation. Eddy covariance instruments were deployed on a permanent tower in Mongu, Zambia (879 mm of rainfall per year), as well as on a portable tower in Maun (460 mm yr^?1), Okwa River Crossing (407 mm yr^?1), and Tshane (365 mm yr^?1), Botswana for several days at each site. The relationships between CO₂ flux, F_c, and photosynthetically active radiation were described well by a hyperbolic fit to the data at all locations except for Mongu, the wettest site. Here, there appeared to be an air temperature effect on F_c. While daytime values of F_c routinely approached or exceeded ?20 μmol m^?2 s^?1 at Mongu, the magnitude of F_c remained less than ?10 μmol m^?2 s^?1 when the air temperature was above 27°C. Canopy resistances to water vapor transfer, r_c, displayed an overall decline from the wetter sites to the more arid sites, but the differences in r_c could be almost exclusively accounted for by the decrease in leaf area index (LAI) from north to south along the KT. Ecosystem water use efficiency (WUE), defined as the ratio of net carbon flux to evapotranspiration, showed a general decrease with increasing vapor pressure deficit, D, for all of the sites. The magnitudes of WUE at a given D, however, were dissimilar for the individual sites and were found to be stratified according to the position of the sites along the long‐term aridity gradient. For example, Mongu, which has the wettest climate, has a much lower WUE for like levels of D than Tshane, which historically has the most arid climate. Given the similar inferred stomatal resistances between the sites, the disparate carbon uptake behavior for the grass vs. woody vegetation is the likely cause for the observed differences in WUE along the aridity gradient. The short‐term flux measurements provide a framework for evaluating the vegetation's functional adaptation to the long‐term climate and provide information that may be useful for predicting the dynamic response of the vegetation to future climate change.     

Effect of basal medium, growth regulators and Phytagel concentration on initiation of embryogenic cultures from immature zygotic embryos of loblolly pine (Pinus taeda L.)

Low initiation frequency is one of the main barriers in applying somatic embryogenesis to the clonal production of Pinus species. Factors affecting initiation, including basal medium, plant growth regulators, and Phytagel concentration, have been investigated in loblolly pine (Pinus taeda L.). BM₁ basal medium proved superior to DCR₁ and LP (LP basal salts plus BM₁ organic nutrients). No extrusion from megagametophytes was exhibited on LP medium. The combination of 3 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg/l 6-benzylaminopurine (BA) resulted in a higher extrusion frequency than that of 11 mg/l 2,4-D, 4.5 mg/l BA and 4.3 mg/l kinetin. Phytagel at 1 g/l resulted in the highest explant browning, but the lowest extrusion frequency, while 4 g/l Phytagel induced some dry embryogenic extrusions. Phytagel at 2 g/l was regarded as the best level for initiation of embryogenic cultures. Received: 23 December 1996 / Revision received: 22 July 1997 / Accepted: 2 September 1997     

Recent climatic drying leads to age‐independent growth reductions of white spruce stands in western Canada

Since 2001, climatic conditions have been notably drier than normal across large areas of the western Canadian interior, leading to widespread impacts on the forests of this region. This poses a major concern for the future, given climate change projections for continued warming and drying. We conducted tree‐ring analysis in 75 pure stands of white spruce (Picea glauca) across Alberta and west‐central Saskatchewan to examine the effects of recent climatic drying on the growth of this important boreal tree species. Allometric equations were used to calculate annual growth in aboveground tree biomass (G_BM) from ring width measurements. Results showed an increasing trend in G_BM from the 1960s to the 1990s, followed by a sharp decline during the severe drought of 2001–2002. Of the 75 stands, only 18 recovered sufficiently to cause an increase in mean G_BM from the predrought decade of 1991–2000 to the subsequent decade of 2001–2010. The remaining 57 stands exhibited a decline in mean G_BM between these decades. Climatic drying was a major cause of the growth decline, as shown by the significant stand‐level relationship between percentage change in decadal mean G_BM and the change in decadal mean values of a climate moisture index from 1991–2000 to 2001–2010. Subsequent analyses of boreal stands sampled across Alberta during 2015 revealed that white spruce growth had declined even further as drought conditions intensified during 2014–2015. Overall, there was a 38% decrease in mean G_BM between 1997 and 2015, but surprisingly, the percentage decrease was not significantly different for young, productive stands compared with older, less productive stands. Thus, stand ageing cannot explain the observed decline in white spruce growth during the past quarter century, suggesting that these forests are at risk if the trend towards more frequent, severe drought continues in the region.     

Mesophyll conductance decreases in the wild type but not in an ABA‐deficient mutant (aba1) of Nicotiana plumbaginifolia under drought conditions

Under drought conditions, leaf photosynthesis is limited by the supply of CO₂. Drought induces production of abscisic acid (ABA), and ABA decreases stomatal conductance (g_s). Previous papers reported that the drought stress also causes the decrease in mesophyll conductance (g_m). However, the relationships between ABA content and g_m are unclear. We investigated the responses of g_m to the leaf ABA content [(ABA)_L] using an ABA‐deficient mutant, aba1, and the wild type (WT) of Nicotiana plumbaginifolia. We also measured leaf water potential (Ψ_L) because leaf hydraulics may be related to g_m. Under drought conditions, g_m decreased with the increase in (ABA)_L in WT, whereas both (ABA)_L and g_m were unchanged by the drought treatment in aba1. Exogenously applied ABA decreased g_m in both WT and aba1 in a dose‐dependent manner. Ψ_L in WT was decreased by the drought treatment to ?0.7 MPa, whereas Ψ_L in aba1 was around ?0.8 MPa even under the well‐watered conditions and unchanged by the drought treatment. From these results, we conclude that the increase in (ABA)_L is crucial for the decrease in g_m under drought conditions. We discuss possible relationships between the decrease in g_m and changes in the leaf hydraulics.     

Competitive substrate inhibition of amyloglucosidase from Rhizopus sp. by vanillin and curcumin

Kinetic studies of two glucosylation reactions catalyzed by an amyloglucosidase from Rhizopus sp. leading to the synthesis of vanillin-α/β-D-glucoside from D-glucose and vanillin and curcumin-bis-α-D-glucoside from D-glucose and curcumin were investigated in detail. Initial reaction rates were determined from kinetic runs involving different concentrations of D-glucose and vanillin (5?mM to 0.1?M) or D-glucose and curcumin (5?mM to 0.1?M). Graphical double reciprocal plots showed that the kinetics of the two enzyme catalyzed reactions exhibited Ping-Pong Bi-Bi mechanism where competitive substrate inhibition by vanillin/curcumin led to dead-end amyloglucosidase–vanillin/curcumin complexes at higher concentrations of vanillin/curcumin. An attempt to obtain the best fit of this kinetic model through computer simulation yielded in good approximation, the values of four important kinetic parameters, vanillin-α/β-D-glucoside: k_cat=35.0±3.2 10^?5M?h^?1·mg, K_i=10.5±1.1?mM, K_mD-glucose=60.0±6.2?mM, K_mvanillin=50.0±4.8?mM; curcumin-bis-α-D-glucoside: k_cat=6.07±0.58 10^?5M?h^?1·mg, K_i=3.0±0.28?mM, K_mD-glucose=10.0±0.9?mM, K_mcurcumin=4.6±0.5?mM.     

Attribution of seasonal leaf area index trends in the northern latitudes with “optimally” integrated ecosystem models

Significant increases in remotely sensed vegetation indices in the northern latitudes since the 1980s have been detected and attributed at annual and growing season scales. However, we presently lack a systematic understanding of how vegetation responds to asymmetric seasonal environmental changes. In this study, we first investigated trends in the seasonal mean leaf area index (LAI) at northern latitudes (north of 30°N) between 1982 and 2009 using three remotely sensed long‐term LAI data sets. The most significant LAI increases occurred in summer (0.009 m² m^?2 year^?1, p < .01), followed by autumn (0.005 m² m^?2 year^?1, p < .01) and spring (0.003 m² m^?2 year^?1, p < .01). We then quantified the contribution of elevating atmospheric CO₂ concentration (eCO₂), climate change, nitrogen deposition, and land cover change to seasonal LAI increases based on factorial simulations from 10 state‐of‐the‐art ecosystem models. Unlike previous studies that used multimodel ensemble mean (MME), we used the Bayesian model averaging (BMA) to optimize the integration of model ensemble. The optimally integrated ensemble LAI changes are significantly closer to the observed seasonal LAI changes than the traditional MME results. The BMA factorial simulations suggest that eCO₂ provides the greatest contribution to increasing LAI trends in all seasons (0.003–0.007 m² m^?2 year^?1), and is the main factor driving asymmetric seasonal LAI trends. Climate change controls the spatial pattern of seasonal LAI trends and dominates the increase in seasonal LAI in the northern high latitudes. The effects of nitrogen deposition and land use change are relatively small in all seasons (around 0.0002 m² m^?2 year^?1 and 0.0001–0.001 m² m^?2 year^?1, respectively). Our analysis of the seasonal LAI responses to the interactions between seasonal changes in environmental factors offers a new perspective on the response of global vegetation to environmental changes.     

Carbon sequestration and ecosystem respiration for 4 years in a Scots pine forest

The net exchange of CO₂ (NEE) between a Scots pine (Pinus sylvestris L.) forest ecosystem in eastern Finland and the atmosphere was measured continuously by the eddy covariance (EC) technique over 4 years (1999–2002). The annual temperature coefficient (Q₁₀) of ecosystem respiration (R) for these years, respectively, was 2.32, 2.66, 2.73 and 2.69. The light‐saturated rate of photosynthesis (A_max) was highest in July or August, with an annual average A_max of 10.9, 14.6, 15.3 and 17.1 μmol m^?2 s^?1 in the 4 years, respectively. There was obvious seasonality in NEE, R and gross primary production (GPP), exhibiting a similar pattern to photosynthetically active radiation (PAR) and air temperature. The integrated daily NEE ranged from 2.59 to ?4.97 g C m^?2 day^?1 in 1999, from 2.70 to ?4.72 in 2000, from 2.61 to ?4.71 in 2001 and from 5.27 to ?4.88 in 2002. The maximum net C uptake occurred in July, with the exception of 2000, when it was in June. The interannual variation in ecosystem C flux was pronounced. The length of the growing season, based on net C uptake, was 179, 170, 175 and 176 days in 1999–2002, respectively, and annual net C sequestration was 152, 101, 172 and 205 g C m^?2 yr^?1. It is estimated that ecosystem respiration contributed 615, 591, 752 and 879 g C m^?2 yr^?1 to the NEE in these years, leading to an annual GPP of ?768, ?692, ?924 and ?1084 g C m^?2 yr^?1. It is concluded that temperature and PAR were the main determinants of the ecosystem CO₂ flux. Interannual variations in net C sequestration are predominantly controlled by average air temperature and integrated radiation in spring and summer. Four years of EC data indicate that boreal Scots pine forest ecosystem in eastern Finland acts as a relatively powerful carbon sink. Carbon sequestration may benefit from warmer climatic conditions.     

The role of optimal control in assessing the most cost-effective implementation of a vaccination programme: HPV as a case study

Consider a population that develops over units of time labelled by zero and the negative integers. It is assumed that at any time r ? 0 there are respectively N(m, a) males of age a and N(f, b) of females of age b, where a = 1, … , A_m and b = 1, … , A_f. At time 0 a sample of n copies of a gene are assumed to be observed, where n ? min_ab(N(m, a), N(f, b)). It is assumed that at any particular time r any possible mating is equally probable and that numbers of gametes contributed to offspring of age 1 and sex u by parents of sex v are exchangeable within age groups and independently distributed among age-groups. Coalescent theory is then derived, with time measured in multiples 2N_eC of the effective population size N_eC, which depends on a measure T of the generation interval. Theory is developed for both autosomal and sex-linked loci in two special cases.     

Growth and nitrate uptake properties of plants grown at different relative rates of nitrogen supply. II. Activity and affinity of the nitrate uptake system in Pisum and Lemna in relation to nitrogen availability and nitrogen demand

Abstract Net nitrate uptake rates were measured and the kinetics calculated in non-nodulated Pisum sativum L. cv. Marma and Lemna gibba L. adapted to constant relative rates of nitrate-N additions (R_A), ranging from 0.03 to 0.27 d^?1 for Pisum and from 0.05 to 0.40 d^?1 for Lemna, V_max of net nitrate uptake (measured in the range 10 to 100 mmol m^?3 nitrate, i.e. ‘system I’) increased with R_A in the growth limiting range but decreased when R_A exceeded the relative growth rate (RGR), K_m was not significantly related to changes in R_A. On the basis of previous ¹³N-flux experiments, it is concluded that the differences in V_max at growth limiting R_A are attributable to differences in influx rates. Linear relationships between V_max and tissue nitrogen concentrations were obtained in the growth limiting range for both species, and extrapolated intercepts relate well with the previously defined minimal nitrogen concentrations for plant growth (Oscarson, Ingemarsson & Larsson, 1989). Analysis of V_max for net nitrate uptake on intact plant basis in relation to nitrogen demand during stable, nitrogen limited, growth shows an increased overcapacity at lower R_A values in both species, which is largely explained by the increased relative root size at low R_A. A balancing nitrate concentration, defined as the steady state concentration needed to sustain the relative rate of increase in plant nitrogen (R_N), predicted by R_A, was calculated for both species. In the growth limiting range, this value ranges from 3.5 mmol m^?3 (R_A 0.03 d^?1) to 44 mmol m^?3 (R_A 0.21 d^?1) for Pisum and from 0.2 mmol m^?3 (R_A 0.05 d^?1) to 5.4 mmol m^?3 (R_A 0.03 d^?1) for Lemna. It is suggested that this value can be used as a unifying measure of the affinity for nitrate, integrating the performance of the nitrate uptake system with nitrate flux and long term growth and demand for nitrogen.     

Leaf Wilting Movement Can Protect Water-Stressed Cotton (<Emphasis Type="Italic">Gossypium hirsutum</Emphasis> L.) Plants Against Photoinhibition of Photosynthesis and Maintain Carbon Assimilation in the Field

Under severe water stress, leaf wilting is quite general in higher plants. This passive movement can reduce the energy load on a leaf. This paper reports an experimental test of the hypothesis that leaf wilting movement has a protective function that mitigates against photoinhibition of photosynthesis in the field. The experiments exposed cotton (Gossypium hirsutum L.) to two water regimes: water-stressed and well-watered. Leaf wilting movement occurred in water-stressed plants as the water potential decreased to −4.1 MPa, reducing light interception but maintaining comparable quantum yields of photosystem II (PS II; Yield for short) and the proportion of total PS II centers that were open (qP). Predrawn F _v/F _m (potential quantum yield of PS II) as an indicator of overnight recovery of PS II from photoinhibition was higher than or similar to that in well-watered plants. Compared with water-stressed cotton leaves for which wilting movement was permitted, water-stressed cotton leaves restrained from such movement had significantly increased leaf temperature and instantaneous CO₂ assimilation rates in the short term, but reduced Yield, qP, and F _v/F _m. In the long term, predrawn F _v/F _m and CO₂ assimilation capacity were reduced in water-stressed leaves restrained from wilting movement. These results suggest that, under water stress, leaf wilting movement could reduce the incident light on leaves and their heat load, alleviate damage to the photosynthetic apparatus due to photoinhibition, and maintain considerable carbon assimilation capacity in the long term despite a partial loss of instantaneous carbon assimilation in the short term.     

Long‐term Successful Weight Loss Improves Vascular Endothelial Function in Severely Obese Individuals

Obesity is associated with increased cardiovascular risk. Although short‐term weight loss improves vascular endothelial function, longer term outcomes have not been widely investigated. We examined brachial artery endothelium‐dependent vasodilation and metabolic parameters in 29 severely obese subjects who lost ≥10% body weight (age 45 ± 13 years; BMI 48 ± 9 kg/m²) at baseline and after 12 months of dietary and/or surgical intervention. We compared these parameters to 14 obese individuals (age 49 ± 11 years; BMI 39 ± 7 kg/m²) who failed to lose weight. For the entire group, mean brachial artery flow‐mediated dilation (FMD) was impaired at 6.7 ± 4.1%. Following sustained weight loss, FMD increased significantly from 6.8 ± 4.2 to 10.0 ± 4.7%, but remained blunted in patients without weight decline from 6.5 ± 4.0 to 5.7 ± 4.1%, P = 0.013 by ANOVA. Endothelium‐independent, nitroglycerin‐mediated dilation (NMD) was unaltered. BMI fell by 13 ± 7 kg/m² following successful weight intervention and was associated with reduced total and low‐density lipoprotein cholesterol, glucose, hemoglobin A_1c, and high‐sensitivity C‐reactive protein (CRP). Vascular improvement correlated most strongly with glucose levels (r = ?0.51, P = 0.002) and was independent of weight change. In this cohort of severely obese subjects, sustained weight loss at 1 year improved vascular function and metabolic parameters. The findings suggest that reversal of endothelial dysfunction and restoration of arterial homeostasis could potentially reduce cardiovascular risk. The results also demonstrate that metabolic changes in association with weight loss are stronger determinants of vascular phenotype than degree of weight reduction.     

Interactions of lens proteins. Self-association and mixed-association studies of bovine alpha-crystallin and gamma-crystallin

Concentrated solutions of calf alpha-crystallin (up to 45 g/l) and gamma-crystallin (up to 67 g/l) were subjected to frontal exclusion chromatography at pH 7.3, ionic strength 0.17 and 20 degrees C. The experimental concentration dependence of the weight-average partition coefficient was compared with theoretical expressions, which include considerations of thermodynamic non-ideality effects, for the concentration dependence of a single solute and of a solute undergoing reversible self-association. Two types of association pattern were examined, discrete dimerization and indefinite self-association. The partition chromatography results are consistent with an indefinite self-association of gamma-crystallin, governed by an isodesmic association constant of 6.7 X 10(-3) l/g. alpha-Crystallin appears to self-associate either very weakly, with a maximal association constant of 0.9 X 10(-3) l/g, or not at all; the distinction depends on the assessment of the non-ideality coefficients. The consequences of excluded volume effects on these self-association equilibria at high total protein concentration are discussed. Mixtures of alpha-crystallin and gamma-crystallin were analyzed by frontal exclusion chromatography (up to 14 g/l) and sedimentation velocity (up to 115 g/l): no interaction was observed.     

Ion Secretion and Isotonic Transport in Frog Skin Glands

The aim of this study was to clarify the mechanism of isotonic fluid transport in frog skin glands. Stationary ion secretion by the glands was studied by measuring unidirectional fluxes of ²⁴Na⁺, ⁴²K⁺, and carrier-free ¹³⁴Cs⁺ in paired frog skins bathed on both sides with Ringer's solution, and with 10⁻⁵ m noradrenaline on the inside and 10⁻⁴ m amiloride on the outside. At transepithelial thermodynamic equilibrium conditions, the ¹³⁴Cs⁺ flux ratio, J ^out _Cs/J ⁱⁿ _Cs, varied in seven pairs of preparations from 6 to 36. Since carrier-free ¹³⁴Cs⁺ entering the cells is irreversibly trapped in the cellular compartment (Ussing & Lind, 1996), the transepithelial net flux of ¹³⁴Cs⁺ indicates that a paracellular flow of water is dragging ¹³⁴Cs⁺ in the direction from the serosal- to outside solution. From the measured flux ratios it was calculated that the force driving the secretory flux of Cs⁺ varied from 30 to 61 mV among preparations. In the same experiments unidirectional Na⁺ fluxes were measured as well, and it was found that also Na⁺ was subjected to secretion. The ratio of unidirectional Na⁺ fluxes, however, was significantly smaller than would be predicted if the two ions were both flowing along the paracellular route dragged by the flow of water. This result indicates that Na⁺ and Cs⁺ do not take the same pathway through the glands. The flux ratio of unidirectional K⁺ fluxes indicated active secretion of K⁺. The time it takes for steady-state K⁺ fluxes to be established was significantly longer than that of the simultaneously measured Cs⁺ fluxes. These results allow the conclusion that — in addition to being transported between cells — K⁺ is submitted to active transport along a cellular pathway.Based on the recirculation theory, we propose a new model which accounts for stationary Na⁺, K⁺, Cl⁻ and water secretion under thermodynamic equilibrium conditions. The new features of the model, as compared to the classical Silva-model for the shark-rectal gland, are: (i) the sodium pumps in the activated gland transport Na⁺ into the lateral intercellular space only. (ii) A barrier at the level of the basement membrane prevents the major fraction of Na⁺ entering the lateral space from returning to the serosal bath. Thus, Na⁺ is secreted into the outside bath. It has to be assumed then that the Na⁺ permeability of the basement membrane barrier (P ^BM _Na) is smaller than the Na⁺ permeability of the junctional membrane (P ^JM _Na), i.e., P ^JM _Na/P ^BM _Na > 1. The secretory paracellular flow of water further requires that the Na⁺ reflection coefficients (σ_Na) of the two barriers are governed by the conditions, σ^BM _Na > 0, and σ^BM _Na > σ^JM _Na. (iii) Na⁺ channels are located in the apical membrane of the activated gland cells, so that a fraction of the Na⁺ outflux appearing downstream the lateral intercellular space is recirculated by the gland cells. Based on measured unidirectional fluxes, a set of equations is developed from which we estimate the ion fluxes flowing through major pathways during stationary secretion. It is shown that 80% of the sodium ions flowing downstream the lateral intercellular space is recycled by the gland cells. Our calculations also indicate that under the conditions prevailing in the present experiments 1.8 ATP molecule would be hydrolyzed for every Na⁺ secreted to the outside bath. Received: 30 January 1996/Revised: 12 March 1996