Nitrogen sink strength of ectomycorrhizal morphotypes of <Emphasis Type="Italic">Quercus douglasii</Emphasis>, <Emphasis Type="Italic">Q. garryana</Emphasis>, and <Emphasis Type="Italic">Q. agrifolia</Emphasis> seedlings grown in a northern California oak woodland

Little information is known on what the magnitude of nitrogen (N) processed by ectomycorrhizal (ECM) fungal species in the field. In a common garden experiment performed in a northern California oak woodland, we investigated transfer of nitrogen applied as ¹⁵NH₄ or ¹⁵NO₃ from leaves to ectomycorrhizal roots of three oak species, Quercus agrifolia, Q. douglasii, and Q. garryana. Oak seedlings formed five common ectomycorrhizal morphotypes on root tips. Mycorrhizal tips were more enriched in ¹⁵N than fine roots. N transfer was greater to the less common morphotypes than to the more common types. ¹⁵N transfer from leaves to roots was greater when , not , was supplied. ¹⁵N transfer to roots was greater in seedlings of Q. agrifolia than in Q. douglasii and Q. garryana. Differential N transfer to ectomycorrhizal root tips suggests that ectomycorrhizal morphotypes can influence flows of N from leaves to roots and that mycorrhizal diversity may influence the total N requirement of plants.     

Concentration-Dependent Effects on Intracellular and Surface pH of Exposing <Emphasis Type="Italic">Xenopus</Emphasis> oocytes to Solutions Containing NH<Subscript>3</Subscript>/NH<Subscript>4</Subscript><Superscript>+</Superscript>

Others have shown that exposing oocytes to high levels of (10–20 mM) causes a paradoxical fall in intracellular pH (pH_i), whereas low levels (e.g., 0.5 mM) cause little pH_i change. Here we monitored pH_i and extracellular surface pH (pH_S) while exposing oocytes to 5 or 0.5 mM NH₃/NH₄ ⁺. We confirm that 5 mM causes a paradoxical pH_i fall (−ΔpH_i ≅ 0.2), but also observe an abrupt pH_S fall (−ΔpH_S ≅ 0.2)—indicative of NH₃ influx—followed by a slow decay. Reducing [NH₃/NH₄ ⁺] to 0.5 mM minimizes pH_i changes but maintains pH_S changes at a reduced magnitude. Expressing AmtB (bacterial Rh homologue) exaggerates −ΔpH_S at both levels. During removal of 0.5 or 5 mM NH₃/NH₄ ⁺, failure of pH_S to markedly overshoot bulk extracellular pH implies little NH₃ efflux and, thus, little free cytosolic NH₃/NH₄ ⁺. A new analysis of the effects of NH₃ vs. NH₄ ⁺ fluxes on pH_S and pH_i indicates that (a) NH₃ rather than NH₄ ⁺ fluxes dominate pH_i and pH_S changes and (b) oocytes dispose of most incoming NH₃. NMR studies of oocytes exposed to ¹⁵N-labeled show no significant formation of glutamine but substantial accumulation in what is likely an acid intracellular compartment. In conclusion, parallel measurements of pH_i and pH_S demonstrate that NH₃ flows across the plasma membrane and provide new insights into how a protein molecule in the plasma membrane—AmtB—enhances the flux of a gas across a biological membrane.

Anaerobic growth and potential for amino acid production by nitrate respiration in <Emphasis Type="Italic">Corynebacterium glutamicum</Emphasis>

Oxygen limitation is a crucial problem in amino acid fermentation by Corynebacterium glutamicum. Toward this subject, our study was initiated by analysis of the oxygen-requiring properties of C. glutamicum, generally regarded as a strict aerobe. This organism formed colonies on agar plates up to relatively low oxygen concentrations (0.5% O₂), while no visible colonies were formed in the absence of O₂. However, in the presence of nitrate (), the organism exhibited limited growth anaerobically with production of nitrite (), indicating that C. glutamicum can use nitrate as a final electron acceptor. Assays of cell extracts from aerobic and hypoxic cultures yielded comparable nitrate reductase activities, irrespective of nitrate levels. Genome analysis revealed a narK2GHJI cluster potentially relevant to nitrate reductase and transport. Disruptions of narG and narJ abolished the nitrate-dependent anaerobic growth with the loss of nitrate reductase activity. Disruption of the putative nitrate/nitrite antiporter gene narK2 did not affect the enzyme activity but impaired the anaerobic growth. These indicate that this locus is responsible for nitrate respiration. Agar piece assays using l-lysine- and l-arginine-producing strains showed that production of both amino acids occurred anaerobically by nitrate respiration, indicating the potential of C. glutamicum for anaerobic amino acid production.     

Report on non-temperature related variations in CO<Subscript>2</Subscript> efflux rates from young tree stems in the dormant season

Respiration rates are reported to increase exponentially with temperature. Respiration rates of woody tissues are commonly measured as CO₂ efflux rates () from that tissue. However, this paper describes clear variations in stem that were not related to temperature for the case of a young beech (Fagus sylvatica L.) and oak (Quercus robur L.) tree during the dormant season. The CO₂ concentration ([CO₂]) in the xylem of the beech tree showed similar temperature-independent variations. The trees were grown in a growth chamber in which radiation patterns and temperature were kept constant. was measured with an IRGA connected to cuvettes surrounding a stem segment. Xylem [CO₂] was measured in situ using a CO₂ microelectrode. Depressions in and [CO₂] occurred during the light period, despite equal temperatures in the light and dark period. Explanations found in literature for discrepancies in the exponential relationship between temperature and are the influence of (1) sap flow or (2) decreased cell water content. However, (1) the variations were observed in the dormant season, when no sap flow was observed yet, and (2) reduced cell water content was not likely to be apparent as differences in stem transpiration rates between the dark and light period were not significant. Hence, previously formulated theories failed to explain our results. This work therefore provides a new ground for discussion on other possible causes of daytime depressions in . One might be the refixation of respired CO₂ by corticular photosynthesis in the stem parts adjacent to the stem segment enclosed by the cuvette.     

Immobilization of the recombinant invertase INVB from <Emphasis Type="Italic">Zymomonas mobilis</Emphasis> on Nylon-6

The recombinant invertase INVB (re-INVB) from Zymomonas mobilis was immobilized on microbeads of Nylon-6, by means of covalent bonding. The enzyme was strongly and successfully bound to the support. The activity of the free and immobilized enzyme was determined, using 10% (w/v) sucrose, at a temperature ranging between 15 and 60 °C and a pH ranging between 3.5 and 7. The optimal pH and temperature for the immobilized enzyme were 5.5 and 25 °C, respectively. Immobilization of re-INVB on Nylon-6 showed no significant change in the optimal pH, but a difference in the optimal temperature was evident, as that for the free enzyme was shown to be 40 °C. The values for kinetic parameters were determined as: 984 and 98 mM for of immobilized and free re-INVB, respectively. values for immobilized and free enzymes were 6.1 × 10² and 1.2 × 10⁴ s⁻¹, respectively, and immobilized re-INVB showed of 158.73 μmol h min⁻¹ mg⁻¹. Immobilization of re-INVB on Nylon-6 enhanced the thermostability of the enzyme by 50% at 30 °C and 70% at 40 °C, when compared to the free enzyme. The immobilization system reported here may have future biotechnological applications, owing to the simplicity of the immobilization technique, the strong binding of re-INVB to the support and the effective thermostability of the enzyme.     

Alteration of substrate specificity of fructosyl-amino acid oxidase from <Emphasis Type="Italic">Fusarium oxysporum</Emphasis>

Fructosyl-amino acid oxidase (FOD-F) from Fusarium oxysporum f. sp. raphani (NBRC 9972) is the enzyme catalyzing the oxidative deglycation of fructosyl-amino acids such as -fructosyl -benzyloxycarbonyl-lysine (FZK) and fructosyl valine (FV), which are model compounds of the glycated proteins in blood. Wild-type FOD-F has high activities toward both substrates. We obtained a mutant FOD-F, which reacts with FZK but not with FV by random mutagenesis. One amino-acid substitution (K373R) occurred in the mutant FOD-F. In addition to K373R, K373W, K373M, K373T, and K373V, which were selected for optimization of the substitution at position K373, were purified and characterized. Kinetic analysis showed that the catalytic turnover for FV greatly decreased, whereas that for FZK did not. In consequence, the specificities toward FZK were increased in the mutant FOD-Fs. The relation between the substrate specificity of the mutant FOD-Fs and the position of the carboxyl group of the substrates was demonstrated using a series of the substrates having the carboxyl group at the different position. The mutant FOD-Fs are attractive candidates for developing an enzymatic measurement method for glycated proteins such as glycated albumin in serum. This study will be helpful to establish an easier and rapid clinical assay system of glycated albumin.     

Cell-to-cell pathway dominates xylem-epidermis hydraulic connection in<Emphasis Type="Italic"> Tradescantia fluminensis</Emphasis> (Vell. Conc.) leaves

A steady supply of water is indispensable for leaves to fulfil their photosynthetic function. Understanding water movement in leaves, especially factors that regulate the movement of water flux from xylem to epidermis, requires that the nature of the transport pathway be elucidated. To determine the hydraulic linkage between xylem and epidermis, epidermal cell turgor pressure (P _t) in leaves of Tradescantia fluminensis was monitored using a cell pressure probe in response to a 0.2 MPa step change in xylem pressure applied at the leaf petiole. Halftime of P _t changes were 10–30 times greater than that of water exchange across an individual cell membrane suggesting that cell-to-cell water transport constitutes a significant part of the leaf hydraulic path from xylem to epidermis. Furthermore, perfusion of H₂O₂ resulted in increases of both and by a factor of 2.5, indicating that aquaporins may play a role in the xylem to epidermis hydraulic link. The halftime for water exchange did not differ significantly between cells located at the leaf base (2.5 s), middle (2.6 s) and tip (2.5 s), indicating that epidermal cell hydraulic properties are similar along the length of the leaf. Following the pressure application to the xylem (0.2 MPa), P _t changed by 0.12, 0.06 and 0.04 MPa for epidermal cells at the base, middle and the tip of the leaf, respectively. This suggests that pressure dissipation between xylem and epidermis is significant, and that the pressure drop along the vein may be due to its structural similarities to a porous pipe, an idea which was further supported by measurements of xylem hydraulic resistance using a perfusion technique.     

Nitrate and phosphate removal by<Emphasis Type="Italic"> Spirulina platensis</Emphasis>

The cyanobacterium Spirulina platensis was used to verify the possibility of employing microalgal biomass to reduce the contents of nitrate and phosphate in wastewaters. Batch tests were carried out in 0.5 dm³ Erlenmeyer flasks under conditions of light limitation (40 mol quanta m^–2 s^–1) at a starting biomass level of 0.50 g/dm³ and varying temperature in the range 23–40°C. In this way, the best temperature for the growth of this microalga (30°C) was determined and the related thermodynamic parameters were estimated. All removed nitrate was used for biomass growth (biotic removal), whereas phosphate appeared to be removed mainly by chemical precipitation (abiotic removal). The best results in terms of specific and volumetric growth rates ( =0.044 day^–1, Q _x =33.2 mg dm^–3 day^–1) as well as volumetric rate and final yield of nitrogen removal ( =3.26 mg dm^–3 day^–1, =0.739) were obtained at 30°C, whereas phosphorus was more effectively removed at a lower temperature. In order to simulate full-scale studies, batch tests of nitrate and phosphate removal were also performed in 5.0 dm³ vessels (mini-ponds) at the optimum temperature (30°C) but increasing the photon fluence rate to 80 mol quanta m^–2 s^–1 and varying the initial biomass concentration from 0.25 to 0.86 g/dm³. These additional tests demonstrated that an increase in the inoculum level up to 0.75 g/dm³ enhanced both NO₃ ^– and PO₄ ^3– removal, confirming a strict dependence of these processes on biomass activity. In addition, the larger surface area of the ponds and the higher light intensity improved removal yields and kinetics compared to the flasks, particularly concerning phosphorus removal ( =0.032–0.050 day^–1, Q _x =34.7–42.4 mg dm^–3 day^–1, =3.24–4.06 mg dm^–3 day^–1, =0.750–0.879, =0.312–0.623 mg dm^–3 day^–1, and =0.224–0.440).     

Improving heterologous polyketide production in <Emphasis Type="Italic">Escherichia coli</Emphasis> by overexpression of an <Emphasis Type="Italic">S</Emphasis>-adenosylmethionine synthetase gene

An S-adenosylmethionine synthetase gene (metK) from Streptomyces spectabilis was cloned into an expression plasmid under the control of an inducible T7 promoter and introduced into a strain of Escherichia coli (BAP1(pBP130/pBP144)) capable of producing the polyketide product 6-deoxyerythronolide B (6-dEB). The metK coexpression in BAP1(pBP130/pBP144) improved the specific production of 6-dEB from 10.86 to 20.08 mg l⁻¹ . In an effort to probe the reason for this improvement, a series of gene deletion and expression experiments were conducted based on a metK metabolic pathway that branches between propionyl-CoA (a 6-dEB precursor) and autoinducer compounds. The deletion and expression studies suggested that the autoinducer pathway had a larger impact on improved 6-dEB biosynthesis. Supporting these results were experiments demonstrating the positive effect conditioned media (the suspected location of the autoinducer compounds) had on 6-dEB production. Taken together, the results of this study show an increase in heterologous 6-dEB production concomitant with heterologous metK gene expression and suggest that the mechanism for this improvement is linked to native autoinducer compounds.     

Effects of low night temperature on pigments,chl <Emphasis Type="Italic">a</Emphasis> fluorescence and energy allocation in two bitter gourd (<Emphasis Type="Italic">Momordica charantia</Emphasis> L.) genotypes

Using two different inbred lines of Momordica charantia (bitter gourd), Y-106-5 and Z-1-4, the cell membrane stability, leaf water potential, pigment contents and the chlorophyll a fluorescence were investigated with different low night temperature (LNT) treatments over a 7 day time period and the sequent a 7 day recovery. Under LNT treatments, electrolyte leakage increased in both inbred lines and it increased more significantly in Y-106-5 plants than that in Z-1-4. The content of Chl b and total Chl decreased, while the Chl a/b ratio increased in stressed plants of the two lines. Almost all LNT treatments induced little change in Chl a content in Z-1-4 whereas obvious decreases in 5 and 8°C treated Y-106-5 plants were observed. Chilling changed the water status of plants and induced decreases of leaf water potential (LWP) in 5 and 8°C treated plants. LNT treatments also resulted in changes in the chlorophyll fluorescence parameters in bitter gourd leaves. The potential PSII activity (F _v/F _o) was reduced obviously by LNT stress and showed more sensitive to LNT than the maximum quantum efficiency of PSII primary photochemistry (F _v/F _m). The efficiency of open PSII centers exhibited a slight decrease whereas the photochemical quenching efficient (q _P) was affected more seriously by LNT stress in both two inbred lines. The allocation of energy was rearranged by LNT stress. The light fraction used for PSII photochemistry (P) was reduced, while that used for heat dissipation (D) and the third fraction of absorbed light defines excess energy (E) increased due to the chilling stress. The impacts of LNT stress on bitter gourd generally increased with the number of LNT chilling and the severe night chilling. Plants were little affected by 12°C night chilling and the most acute damage was found in 5°C night chilling treatments. A 7 day recovery mitigated the adverse effects of LNT for both lines and almost all LNT treated plants restored to control levels except 5°C night chilling treated Y-106-5 plants. The two lines have a variance in tolerance to LNT stress and display obvious differences of phenotypes under extreme conditions.     

Secondary production,calcification and CO<Subscript>2</Subscript> fluxes in the cirripedes <Emphasis Type="Italic">Chthamalus montagui</Emphasis> and <Emphasis Type="Italic">Elminius modestus</Emphasis>

Calcification, a process common to numerous marine taxa, has traditionally been considered to be a significant source of CO₂ in tropical waters only. A number of relatively recent studies, however, have shown that significant amounts of CO₂ are also produced in temperate waters, although none of these studies was carried out on rocky shores, which are considered to be very productive systems. We compared the CO₂ fluxes due to respiration and calcification in two temperate species, the cirripedes Chthamalus montagui and Elminius modestus. The population dynamics of both species were estimated at two sites during a 1-year experimental period in order to establish mean organic (ash-free dry weight) and CaCO₃ (dry shell weight) production. Based on these parameters, we estimated the CO₂ fluxes due to respiration and calcification. CaCO₃ production was estimated to be 481.0 and at each site, representing 3.4 and respectively, of released CO₂. These fluxes represent each 47% of the CO₂ released as a result of respiration and calcification. The production of CaCO₃ at the high-density site was: (1) among the highest values obtained for temperate organisms, and (2) comparable to the estimated CO₂ fluxes for coral reefs. As calcifying organisms are well represented in temperate ecosystems in terms of both density and biomass, our results provide clear evidence that calcification of temperate organisms should not be underestimated. Additional studies on other rocky shore taxa are needed before the relative importance of calcification in rocky intertidal carbon budgets can be generalized.     

A new medium formulation for <Emphasis Type="Italic">in vitro</Emphasis> rooting of carob tree based on leaf macronutrients concentrations

Experiments were performed to optimize the macronutrients concentrations for in vitro rooting of Ceratonia siliqua micropropagated shoots. Several dilutions of Murashige and Skoog (MS) medium were tested: full-strength MS, half-strength MS ( MS), and MS + full N. The frequency of in vitro rooting was enhanced when the MS was used (50 % rooted shoots). Mature leaves from 20 – 30 year-old carob trees and from 2 year-old micropropagated plants were collected and the concentrations of macronutrients (N, P, K, Ca, Mg) assessed. Based on the mineral composition of the leaves a new medium was formulated and compared with the previous ones showing an increment of the rooting frequency to 80 %. Moreover, shoots rooted in the new medium did not show symptoms of apical necrosis that occurred in the other tested media.     

A method for the destruction and analysis of biogenic silicon in two Antarctic diatom species:<Emphasis Type="Italic"> Thalassiosira</Emphasis> sp. and<Emphasis Type="Italic"> Chaetoceros brevis</Emphasis>

Diatoms in the Southern Ocean are limited by iron and light, and therefore produce little biomass. Sufficient biomass for analysis under these conditions requires large sample volumes, and diatom samples are therefore often pre-concentrated on a filter. A method for the digestion of diatom cells on polycarbonate filters, that is also suitable for trace metal analysis, is described here. Additional analysis by inductively coupled plasma-optical emission spectroscopy (ICP-OES) is used for the determination of biogenic silicon. Although several procedures were tested, the method of Hauptkorn et al., which uses tetramethylammonium hydroxide for the destruction of silicon is adapted here [Hauptkorn et al. (2001) Fres J Anal Chem 370:246–250]. Additional nitric acid is added to destroy the polycarbonate filters. The described method results in clear digests and a good correlation between cell numbers and silicon content. Using this procedure, the cellular silicon content for Chaetoceros brevis was determined as 86 ± 4 fmol cell⁻¹. For Thalassiosira sp. a sensitivity effect was observed, and silicon content was determined as . The obtained cellular silicon contents are in good agreement with values presented in the literature.     

Acclimation to light in seedlings of <Emphasis Type="Italic">Quercus petraea</Emphasis> (Mattuschka) Liebl. and <Emphasis Type="Italic">Quercus pyrenaica</Emphasis> Willd. planted along a forest-edge gradient

Photosynthetic acclimation of two co-occurring deciduous oaks (Quercus petraea and Quercus pyrenaica) to a natural light gradient was studied during one growing season. In the spring of 2003, 90 seedlings per species were planted along a transect resulting from a dense Pinus sylvestris stand, an adjacent thinned area and a 10-m-wide firebreak (16.5–60.9% Global Site Factor (GSF)). In two dates of the following summer, we measured leaf gas exchange, carboxylation efficiency (CE), chlorophyll and nitrogen content, light–response curves of chlorophyll a fluorescence parameters, and leaf mass per area (LMA). Summer was mild, as evidenced by leaf predawn water potential (Ψ_pd), which reduced the interactive effect of water stress on the response of seedlings to light. Q. pyrenaica had higher LMA, CE, stomatal conductance (g _{s max}) and photosynthesis per unit area than Q. petraea at all growth irradiances. , LMA, g _{s max} and electron transport rate (ETR) all increased with light availability (GSF) in a similar fashion in both species. Light had also a clear effect on the organization of Photosystem II (PS II), as deduced by chlorophyll a fluorescence curves. Chlorophyll concentration (Chl_m) decreased with increasing light availability in Q. pyrenaica but it did not in Q. petraea. Seedlings of Q. petraea acclimated to higher irradiances showed a greater non-photochemical quenching (NPQ) than those of Q. pyrenaica. This suggests a higher susceptibility to high light in Q. petraea, which would be consistent with a better adaptation to shade, inferred from the lower LMA or the lower rate of photosynthesis.     

Population genetics of <Emphasis Type="Italic">Collisella subrugosa</Emphasis> (Patellogastropoda: Acmaeidae): evidence of two scales of population structure

Marine invertebrate populations usually show high levels of genetic variability that has frequently been associated with spatial and temporal environmental heterogeneity. One of the most heterogeneous marine environments is the intertidal zone, the habitat of Collisella subrugosa, the most widespread and abundant Brazilian limpet. C. subrugosa has planktonic larvae that can disperse over long distances, what can promote gene flow among shores, working against interpopulational differentiation. In this study we investigated the genetic variability and populational substructure of C. subrugosa through analysis of 24 allozyme loci in 14 samples (590 individuals) collected along 2,700 km of the Brazilian coast. The genetic variability was high ( and ), as expected for intertidal species. Genetic differentiation among samples was low (F _ST = 0.03) what may reflect intensive gene flow associated with larval dispersal. However, we detected an isolation-by-distance pattern of population substructure in one sampled region. High levels of heterozygote deficiency were also observed for many loci in each sample. Alternative hypothesis are discussed, and the “breeding groups” is suggested to explain these pattern, indicating the main cause as environmental heterogeneity.     

Injured and uninjured leaf photosynthetic responses after mechanical injury on <Emphasis Type="Italic">Nerium oleander</Emphasis> leaves,and <Emphasis Type="Italic">Danaus plexippus</Emphasis> herbivory on <Emphasis Type="Italic">Asclepias curassavica</Emphasis> leaves

Insect herbivory has variable effects on plant physiology; so greater understanding is needed about how injury alters photosynthesis on individual injured and uninjured leaves. Gas exchange and light-adapted leaf chlorophyll fluorescence measurements were collected from uninjured and mechanical partial leaf defoliation in two experiments with Nerium oleander (Apocynaceae) leaves, and one experiment with Danaus plexippus herbivory on Asclepias curassavica (Asclepiadaceae) leaves. Gas exchange impairment (lower photosynthetic rate (P _n ), stomatal conductance (g _s)) indicates water stress in a leaf, suggests stomatal limitations causing injury P _n impairment. The same pattern of gas exchange impairment also occurred on uninjured leaves opposite from injured leaves in both N. oleander experiments. This is an interesting result because photosynthetic impairment is rarely reported on injured leaves near injured leaves. No photosynthetic changes occurred in uninjured A. curassavica leaves opposite from D. plexippus-fed leaves. Partially defoliated leaves that had P _n and g _s reductions lacked any significant changes in intercellular leaf [CO₂], C _i. These results neither support, nor are sufficient to reject, stomatal limitations to photosynthesis. Manually imposed midrib vein severance in N. oleander experiment #1 significantly increased leaf C _i, indicating mesophyll limitations to photosynthesis. Maximal light-adapted leaf photochemical efficiency () and also non-photochemical quenching (q _N) were reduced by mechanical or insect herbivory to both study species, suggesting leaf trouble handling excess light energy not used for photochemistry. Midrib injury on N. oleander leaves and D. plexippus herbivory on A. curassavica leaves also reduced effective quantum yield (ΦPSII) and photochemical quenching (q _P); so reduced plastoquinone pools could lead to additional PSII reaction center closure.     

The role of body surfaces and ventilation in gas exchange of the abalone, <Emphasis Type="Italic">Haliotis iris</Emphasis>

The archaeogastropod Haliotis iris possesses paired bipectinate gills and normally four to six shell holes. In still water, endogenous water flow entered the branchial chamber anteriorly to the left of the head and was exhaled primarily from the three most posterior holes. The first or second anterior aperture was occasionally weakly inhalant. Cardiac interaction superimposed an oscillatory component upon ciliary ventilation but did not augment mean flow. At normal endogenous flow rates 49% of oxygen was extracted from the branchial flow, increasing to 71% at lower flows. In still water, normoxic was 0.47 μmol g⁻¹ h⁻¹. Oxyregulation occurred down to with partial oxyregulation down to 45 Torr (P _crit), and oxyconformity below this. The oxyregulatory plateau was absent in artificially ventilated animals but normoxic was higher (0.65 μmol g⁻¹ h⁻¹). Endogenous ventilation was unaffected by hypoxia to 15 Torr. Heart rate decreased by ~20% at 26 Torr before falling more steeply. Oxygen uptake from the branchial ventilation stream fully accounted for normoxic In hypoxia (<30 Torr), no uptake occurred from the head or foot despite extensive eversion of the epipodium. Blood oxygen measurements excluded the right mantle as a significant gas exchange organ. Changes in oxygen uptake caused by changes in the velocity of external water currents support the concept of induced ventilation and suggest that in still water aerobic respiration was ventilation-limited. Although ciliary ventilation appears adequate to support resting aerobic metabolism, induced ventilation may provide increased aerobic scope for activity and repayment of oxygen debt. An erratum to this article can be found at     

Homology Modeling of Hemagglutinin/Protease [HA/P (vibriolysin)] from <Emphasis Type="Italic">Vibrio Cholerae</Emphasis>: Sequence Comparision,Residue Interactions and Molecular Mechanism

Vibrio cholerae produces a zinc-containing and calcium-stabilized soluble hemagglutinin/protease, which has been earlier shown to have the ability to cleave several physiologically important substrates including mucin, fibronectin and lactoferin. This study presents homology modeling of hemagglutinin/protease (vibriolysin) from Vibrio cholerae in the presence of inhibitor HPI [N-(1-carboxy-3-phenylpropyl)-phenylalanyl-alpha-aspargine]. The 3D structure was predicted based on its sequence homology with Pseudomonas aeruginosa elastase (PAE). Comparison of the 3D structures of PAE and HA/P reveals a remarkable similarity having a conserved α + β domain. The inhibitor shows similar binding features as seen in other metalloproteases of M4 peptidase family. The study also highlights the key catalytic residues as well as the residues at the S₁ and binding sub-sites. The similarities between the two proteins provide support for the hypothesis that the two enzymes have similar three-dimensional structures and a common mechanism of action. The fact that both enzymes are secreted as zinc-containing proteases, led us to further hypothesize that they may play similar role in pathogenesis.     

Correlation between photorespiration,CO<Subscript>2</Subscript>-assimilation and spatiotemporal dynamics of photosynthesis in leaves of the C<Subscript>3</Subscript>-photosynthesis/crassulacean acid metabolism-intermediate species <Emphasis Type="Italic">Clusia minor</Emphasis> L. (Clusiaceae)

The tree Clusia minor L. (Clusiaceae) operates with different modes of photosynthesis in response to different combinations of environmental parameters. Here plants were subjected to experimental conditions eliciting performance of C₃-photosynthesis and crassulacean acid metabolism (CAM), respectively. A combination of instruments was used to determine CO₂ and water vapour gas exchange, relative quantum use efficiency of photosynthesis (Φ_PSII) and for the first time in such studies also photorespiration simultaneously with the other parameters. In the C₃-mode photorespiration was constant during the light period, where oxygenase activity of ribulose-bis-phosphate carboxylase/oxygenase (RubisCO) was ranging between 32.1 and 35.7% of total RubisCO activity. In the CAM-mode photorespiration depended on the CAM phases. In phase II in the morning was 15.6%. In phase IV in the afternoon initially it was 37.9% and then declined to 17.6% of total RubisCO activity towards the evening. Anatomically leaves of C. minor are differentiated in palisade and spongy parenchyma with an internal air space of 9.3% of the total volume and therefore could be structurally homobaric. However, heterogeneity of Φ_PSII under both non-photorespiratory and photorespiratory conditions in the C₃- and CAM-mode indicated that lateral diffusion of CO₂ and O₂ were subject to limitations showing that leaves are functionally heterobaric.