Planktonic primary production in the German Wadden Sea

By combining weekly data of irradiance, attenuation and chlorophylla concentrations with photosynthesis (P) versus light intensity(E) curve characteristics, the annual cycle of planktonic primaryproduction in the estuarine part of the Northfrisian WaddenSea was computed for a 2 year period. Daily water column particulategross production ranged from 5 to 2200 mg C m^–2 day^–1and showed a seasonal pattern similar to chlorophyll a. Budgetcalculation yielded annual gross particulate primary productionsof 124 and 176 g C m^–2 year^–1 in 1995 and 1996,respectively. Annual amounts of phytoplankton respiration, calculatedaccording to a two-compartment model of Langdon [in Li,W.K.W.and Maestrini,S.Y. (eds), Measurement of Primary Productionfrom the Molecular to the Global Scale. International Councilfor the Exploration of the Sea, Copenhagen, 1993, pp. 20–36],and dissolved production in 1996, were both in the range of24–39 g C m^–2 year^–1. Annual total net productionwas thus very similar to particulate gross production (127 and177 g C m^–2 year^–1 in 1995 and 1996, respectively).Phytoplankton growth was low or even negative in winter. Inspring and summer, production/biomass (Pr/B) ratios varied from0.2 up to 1.7. Phytoplankton growth during the growth seasonalways surpassed average flushing time in the area, thus underliningthe potential of local phytoplankton bloom development in thispart of the Wadden Sea. The chlorophyll-specific maximum photosyntheticrate (P^B_max) ranged from 0.8 to 9.9 mg C mg^–1 Chl h^–1and was strongly correlated with water temperature (r² = 0.67).By contrast, there was no clear seasonal cycle in ^B, which rangedfrom 0.007 to 0.039 mg C mg^–1 Chl h^–1 (µmolphotons m^–2 s^–1)^–1. Its variability was muchless than P^B_max and independent of temperature. The magnitudeand part of the variability of P^B_max and ^B are presumably causedby changes in species composition, as evidenced from the rangeof these parameters found among 10 predominant diatom speciesisolated from the Wadden Sea. The ratio of average light conditionsin the water column (E_av) to the light saturation parameterE_k indicates that primary production in the Wadden Sea regionunder study is predominantly controlled by light limitationand that nutrient limitation was likely to occur for a few hoursper day only during 5 (dissolved inorganic nitrogen) to 10 (PO₄,Si) weeks in the 2 year period investigated.     

Exploring the role of galectin 3 in kidney function: a genetic approach

Galectin 3 belongs to a family of glycoconjugate-binding proteinsthat participate in cellular homeostasis by modulating cellgrowth, adhesion, and signaling. We studied adult galectin 3null mutant (Gal 3–/–) and wild-type (WT) mice togain insights into the role of galectin 3 in the kidney. Byimmunofluorescence, galectin 3 was found in collecting duct(CD) principal and intercalated cells in some regions of thekidney, as well as in the thick ascending limbs at lower levels.Compared to WT mice, Gal 3–/– mice had ~11% fewerglomeruli (p < 0.04), associated with kidney hypertrophy(p < 0.006). In clearance experiments, urinary chloride excretionwas found to be higher in Gal 3–/– than in WT mice(p < 0.04), but there was no difference in urinary bicarbonateexcretion, in glomerular filtration, or urinary flow rates.Under chronic low sodium diet, Gal 3–/– mice hadlower extracellular fluid (ECF) volume than WT mice (p <0.05). Plasma aldosterone concentration was higher in Gal 3–/–than in WT mice (p < 0.04), which probably caused the observedincrease in -epithelial sodium channel (-ENaC) protein abundancein the mutant mice (p < 0.001). Chronic high sodium dietresulted paradoxically in lower blood pressure (p < 0.01)in Gal 3–/– than in WT. We conclude that Gal 3–/–mice have mild renal chloride loss, which causes chronic ECFvolume contraction and reduced blood pressure levels.     

On the causes of interspecific differences in the growth-irradiance relationship for phytoplankton. II. A general review

The causes of interspecific differences in the µ-l relationshipare examined in the context of a mechanistic model which relatesµ to irradiance in terms of six factors:, k_c photosyntheticquotient (PQ), Chl a:C, respiration and excretion. The effectof cell size on the light saturated growth rate is also considered.It is shown that photosynthetic efficiency and PQ exhibit remarkablylittle interspecific variability, and average 0.024 ±0.005 µg C(µg Chl a)^–1 h^–1 (µEm^–2 s^–1)^–1 and 1.5 ± 0.2 mol 02 molC^–1 (when NO₃^– is the nitrogen source) respectively.Two useful relationships were derived: (i) between growth efficiency,_g and Chl a:C at µ. = 0; (ii) between the compensationintensity, I_c and the Chl a-specific maintenance respirationrate. Both relationships were independent of temperature anddaylength. Species best adapted to growth at low light werefound to exhibit high Chl a:C ratios and low maintenance respirationrates. As a group, diatoms were consistently the best adaptedfor growth at low irradiance. Chiorophytes, haptophytes, chrysophytesand cryptophytes were intermediate in their performance at lowirradiance. Dinoflagellates exhibited extreme diversity, withspecies spanning the spectrum from very good performance atlow irradiance to very poor. A new µ_max-cell carbon relationshipis given based on growth rates normalized to 15°C. Evidenceis presented to show that noise in this relationship can besignificantly reduced by using only carbon-specific growth ratesand using only data for species grown at the same daylength.     

MYPT1 mutants demonstrate the importance of aa 888-928 for the interaction with PKGIalpha

During nitric oxide signaling, type I cGMP-dependent protein kinase (PKGI) activates myosin light chain (MLC) phosphatase through an interaction with the 130-kDa myosin targeting subunit (MYPT1), leading to dephosphorylation of 20-kDa MLC and vasodilatation. It has been suggested that the MYPT1-PKGI interaction is mediated by the COOH-terminal leucine zipper (LZ) of MYPT1 and the NH₂-terminal LZ of PKGI (HK Surks and ME Mendelsohn. Cell Signal 15: 937–944, 2003; HK Surks et al. Science 286: 1583–1587, 1999), but we previously showed that PKGI interacts with LZ-positive (LZ+) and LZ-negative (LZ–) MYPT1 isoforms (13). Interestingly, PKGI is known to preferentially bind to RR and RK motifs (WR Dostmann et al. Proc Natl Acad Sci USA 97: 14772–14777, 2000), and there is an RK motif within the aa 888–928 sequence of MYPT1 in LZ+ and LZ– isoforms. Thus, to localize the domain of MYPT1 important for the MYPT1-PKGI interaction, we designed four MYPT1 fragments that contained both the aa 888–928 sequence and the downstream LZ domain (MYPT1FL), lacked both the aa 888–928 sequence and the LZ domain (MYPT1TR), lacked only the aa 888–928 sequence (MYPT1SO), or lacked only the LZ domain (MYPT1TR2). Using coimmunoprecipitation, we found that only the fragments containing the aa 888–928 sequence (MYPT1FL and MYPT1TR2) were able to form a complex with PKGI in avian smooth muscle tissue lysates. Furthermore, mutations of the RK motif at aa 916–917 (R⁹¹⁶K⁹¹⁷) to AA decreased binding of MYPT1 to PKGI in chicken gizzard lysates; these mutations had no effect on binding in chicken aorta lysates. However, mutation of R⁹¹⁶K⁹¹⁷ to E⁹¹⁶E⁹¹⁷ eliminated binding, suggesting that one factor important for the PKGI-MYPT1 interaction is the charge at aa 916–917. These results suggest that, during cGMP-mediated signaling, aa 888–928 of MYPT1 mediate the PKGI-MYPT1 interaction. myosin light chain phosphatase; nitric oxide; smooth muscle; calcium desensitization; cGMP-dependent protein kinase; cGMP     

Photoadaptation in Antarctic phytopfankton: variations in growth rate, chemical composition and P versus I curves

The response of phytoplankton to variations in the light regimewas studied during the VULCAN and ACDA cruises in the Antarctic.Unenriched batch cultures of 12–19 days' duration reachedchl concentrations of 10–50 µg^–1 and exhibitedexponential growth rates, with the maximal rate being 0.41 doubl,day^–1. Ice edge algae exhibited maximum growth rates atphoton flux densities (PFD) of 30–100 µE m^–2S^–1and the growth rate was reduced by about 30% at 500–1000µE m^–2S^–1 The chl/C ratio ranged between 0.004and 0.018, with the lowest ratios at PFDs above 500 µEm^–2S^–1 chl/C ratios were also below maximum at PFDsbelow 40–50 µE m^–2S^–1 The C:N:P ratioswere close to the Redfield ratios; the Si/C ratio averaged 0.16(atoms), and the ATP/C ratio averaged from 0.0024 to 0.0050in different culture senes. When thawed after having been frozenfor 10 days, shade-adapted cultures were in a much better conditionthan sun-adapted ones. P versus I data showed that the maximumassimilation number varied from 0.75 to 4.4 µg C (µgchl)^–1h^–1. It varied inversely with the chl/C ratio;therefore the maximum carbon turnover rate varied little betweensamples (0.024/0.035 h^–1). Low biomass communities exhibitedrelatively high values for (the initial slope of P versus Icurves), low values for 1_sat (160–330 µE m^–2S^–1),and they were susceptible to photoinhibition. In contrast, communitiesdominated by Odontella weissflogii exhibited low values for, a high value for I_sat (560 µE m^–2S^–1 andthey tolerated high PFDs. The photo-adaptational status of thephytoplankton in natural water samples is discussed relativeto the profile of water column stability and mixing processes.     

The influence of temperature and light on the upper limit of Microcystis aeruginosa production in a hypertrophic reservoir

Primary production was measured for 7 years, using the in situ¹⁴C-method in hypertrophic Hartbeespoort Dam, South Africa,to examine the influence of light and water temperature on theupper limit of Microcystis aeruginosa production. Water temperaturesvaried from 11 to >25°C and chlorophyll concentrationsreached 6500 mg m^–3. The maximum volumetric rate of production(A_max) was 12->8800 mg C m^–3 h^–1 with areal productions(A) of 69->3300 mg C m^–2 h^–1 for euphotic zonedepths of <0.5–8.4 m. The intrinsic parameters of phytoplanktonproduction (, A_max/B, I_k) indicated that the phytoplankton populationwas adapted to high light levels. Both A_max/B and I_k were correlatedwith temperature. Under optimal conditions, , the theoreticalupper limit of A, was calculated to be 2.8 g Cm^–2 h^–1,while the measured rate was 2.5 g Cm^–2 h^–1. Measuredareal rates exceeding were overestimated due to methodologicalproblems when working with Microcystis scums. Light and watertemperature interacted to yield high production rates: watertemperature through its direct effect on photosynthetic ratesand indirectly in the formation of diurnal mixed layers; lightindirectly through water temperature and directly through itsattenuation and induction of light-adapted physiology in Microcystis.     

A Transient Stimulation of Net Photosynthesis of Rye Leaves by {alpha}-Hydroxy-2-pyridinemethanesulphonic Acid ({alpha}-HPMS) due to Inhibition of Photorespiratory CO2 Release

The effects of -hydroxy-2-pyridinemethanesulphonic acid (-HPMS)upon net photosynthesis (P_n, the CO₂ compensation point (),post-lower illumination burst of CO₂ (PLIB) and post-lower temperatureburst of CO₂ (PLTB) in detached rye (Secale cereale L.) leaveswere investigated. At low concentrations ( 0.5 mol m^–3),-HPMS initially stimulated P_n and decreased the magnitude ofboth PLIB and PLTB. The decreased at all concentrations of-HPMS (0.05–5.0 mol m^–3. The effects of -HPMS onP_n and were time-dependent and, after a few minutes, the P_nwas inhibited while values increased considerably. At a higherconcentration (5.0 mol m ^–3), the transient effects of-HPMS were shorter () or not observed at all (P_n. Both PLIBand PLTB, when expressed in relation to P_n, increased at higherlevels of this compound. Similar data with respect to the effectsof -HPMS on PLIB and PLTB were found for leaves of dandelion(Taraxacum officinale L.). The results suggest that -HPMS may stimulate P_n by inhibitingphotorespiration, as originally suggested by Zelitch (1966),but only at low concentrations and over a short time span. Thedecrease of PLIB and PLTB values at low -HPMS levels is consistentwith these processes being a residual activity of the glycolatepathway. Key words: CO₂ compensation point, -hydroxy-2-pyridinemethanesulphonic acid, photorespiration, photosynthesis     

Sialylation of n-alkyllactosides, galactosides and glucosides by sialyltransferases from rat liver Golgi vesicles

nAlkyl - and -lactosides, galactosides and glucosides with differentalkyl chain lengths (C₂, C₈, C₁₄, and C₂₀) were synthesizedand used as acceptors for sialyltransferases from rat liverGolgi vesicles. The -galactosides, -glucosides, and both - and-lactosides, were sialylated. Keeping the acceptor concentrationconstant, sialylation rates reached a maximum for the n-octyl- and -lactosides, n-Octyl -galactoside and noctyl -glucoside,respectively. noctyl -glucoside, respectivwly. n-Octyl -galactosideand n-octyl -glucoside were not sialylated. The reaction productswere characterized by TLC. With n-octyl lactoside and galactosideas acceptors, two major sialylation products were formed. Thjeycould be separated by preparative TLC, and their structureswere identified as 2–3 and 2–6 sialylated acceptors,respectively, by a combination of periodated oxidation, NaBD₄reduction,permethylation and subsequent analysis by fast atombombardment mass spectrometry (FAB-MS). The structure of thesingle product obtained from n-ictyl -glucoside was determinedto be the 2–6 sialylated glucoside. Competition experimentswith n-octyl lactoside and lactosylceramide and gangliosideGal1-3GalNAc1-4(NeuAc2–3)Gal1–4Glcbeeta1–1Cer(GM1) as acceptors for sialyltransferases suggested that SAT-I[NeuAc2–3Gal1–4Glc1-1Cer (GM3) synthase] was atleast in least in part responsible for the 2–3 sialylationof n-octyl lactoside. alkylgalactosides alkylglucosides alkyllactosides neoglycolipids sialytransferases     

Measurement of Yield Threshold and Cell Wal Extensibility of Intact Wheat Roots under Different Ionic, Osmotic and Temperature Treatments

Yield stress threshold (Y) and volumetric extensibility () arethe rheological properties that appear to control root growth.In this study they were measured in wheat roots by means ofparallel measurement of the growth rate (r) of intact wheatroots and of the turgor pressures (P) of individual cells withinthe expansion zone. Growth and turgor pressure were manipulatedby immersion in graded osmoticum (mannitol) solutions. Turgorwas measured with a pressure probe and growth rate by visualobservation. The influence of various growth conditions on Yand was investigated; (a) At 27 °C.In 0.5 mol m^–3 CaCl₂ r, P, Y and were20.7±4.6 µm min^–1, 0.77±0.05 MPa,0.07±0.03 MPa and 26±1.9 µm min^–1MPa^–1 (expressed as increase in length), respectively.Following 24 h growth in 10 mol m^–3 KC1 these parametersbecame 12.3±3.5 µm min^–1, 0.72±0.04MPa, 0.13±0.01 MPa and 21±0.7 µm min^–1MPa^–1. After 24 h osmotic adjustment in 150 mol m^–3mannitol/0.5 mol m^–3 CaCl₂ r= 19.6±4.2 µmmin^–1, P = 0.68±0.05 MPa and Y and were 0.07±0.04MPa and 30±0.2 µm min^–1 MPa^–01, respectively.After 24 h growth in 350 mol m^–3 mannitol/0.5 mol m^–3CaCl₂ r= 13.3±4.1 µm min^–1, P= 0.58±0.07MPa, Y=0.12±0.01 MPa and ø 32±0.2 tim min^–1MPa^–1. During osmotic adjustment in 200 mol m^–3mannitol/0.5 mol m^–3 CaCl₂, with or without KCl, the recoveryof growth rate corresponded to turgor pressure recovery (t1/2approximately 3 h). (b) At 15 °C. Lowered temperature dramatically influencedthe growth parameters which became r= 8.3±2.8 um min^–1,P=0.78 MPa, r=<0.2 MPa and =15±0.1 µm min^–1MPa^–1. Therefore, Y and are influenced by 10 mol m^–3 K⁺ ionsand low temperature. In each case the effective pressure forgrowth (P-Y) was large indicating that small fluctuations ofsoil water potential will not stop root elongation. Key words: Yield threshold, cell wall extensibility, wheat root growth, temperature, turgor pressur     

Photoinhibition and the diurnal variation of phytoplankton photosynthesis--I. Development of a photosynthesis--irradiance model from studies of in situ responses

Diurnal series of fluorescence and photosynthesis assays wereconducted in high altitude (3803 m), tropical (16°), LakeTiticaca (Peru/Bolivia). Near-surface diurnal thermoclines formedon typical days of high photon flux density (PFD, {small tilde}2000 µE m^–2 s^–1). In the depth range of diurnalstratification profiles of in vivo fluorescence, both without(F_a and with (F_b DCMU, exhibited a mean decrease of 64% frommorning to mid-day, but little change (mean increase of 1.5%)through the afternoon. Three times during the day surface, mid-depth(3–5 m) and deep (15–20 m) phytoplankton sampleswere incubated with H¹⁴CO₃^– under short (<2 h) exposuresto a range of in situ PFDs. Comparison of phytoplankton in differentsamples (ANOVA) showed identical photosynthetic response insunrise (isothermal) samples but a significant drop in surfaceand mid-depth photosynthesis at all PFDs during times of diurnalstratification. Similarly, both low-light () and light-saturated(P² _max photosynthetic parameters were lower in mid-day surfacesamples compared to deep samples. In addition, previously photoinhibitedsamples had a higher threshold intensity for photoinhibition,I_T. These results, together with diurnal time series of fluorescencefrom in situ incubations, demonstrate that recovery from extendedepisodes of photoinhibition during diurnal stratification isslower than suggested by previous observations in vitro. Photosynthesisby near-surface phytoplankton is different in light increasingup to I_T than light decreasing from I_T. This effect can be modeledby reducing and P_max as a function of the maximum photoinhibitingPFD in the diurnal light history. ¹Present address: Division of Molecular Plant Biology, Universityof California, Berkeley, Berkeley, CA 94720, USA     

Ecological studies on the phytoplankton of Boknafjorden, western Norway. II. Environmental control of photosynthesis

Both predicted (incubator) and measured (in situ) ¹⁴C-assimilationrates were studied from February to November 1981 at three stationsin Boknafjorden, a deep silled fjord of western Norway. Sampleswere taken from different light depths within the euphotic zone.A high degree of conformity was found between the two approaches.Daily values of carbon assimilation integrated over the euphoticzone varied between 0.05 and 1.4 g C m^–2. Yearly primaryproduction varied between stations from 82 to 112 g C m^–2(120–148 g C m^–2 when based on average light conditions).The light-saturation curve parameters ^B and P^B_max ranged from0.0056 to 0.0537 mg C mg Chla^–1 h^–1 µE^–1m² and from 0.7 to 8.5 mg C mg Chla^–1 h^–2 (in situassimilation numbers ranged from 0.9 to 9.3 mg C mg Chla^–1h^–1) respectively, which compare well with those publishedfrom the northwestern side of the Atlantic. The overall importanceof light in controlling photosynthesis throughout the year wasrevealed by the light utilization index , estimated to be 0.43mg C mg Chla^–1 E^–1 m². The maximum quantum yieldwas encountered on August 17, with 0.089 mol CE^–1. Chla/Cratios above and below 0.010 were found to be typical for shade-and light-adapted cells respectively. Assimilation numbers andgrowth rates were linearly related only when considering light-adaptedcells. Consistent with the findings of this study, the applicabilityof I_K, ^B and P^B_max as indicators of light-shade adaptation propertiesshould be questioned. Maximum growth rates were encounteredduring an autumn bloom of the dinoflagellate Gyrodinium aureolum(1.0 doublings day^–1), while 0.7–0.8 doublings day^–1were found for a winter bloom (water temperature of 2°C)of the diatom Skeletonema costatum. No unambiguous temperatureeffect on assimilation number and growth of phytoplankton couldbe recognized in Boknafjorden. A tendency towards increasedassimilation numbers coinciding with increased water columnstability was revealed. The highest P^B_max values were oftenencountered at almost undetectable nutrient concentrations.At least during summer this could be attributed to recyclingof nutrients by macro- and/or microzooplankton, responsiblefor a greater part of the primary production now being grazeddown. This study supports the convention that the depth of theeuphotic zone may extend considerably below the 1% light depth.     

Cell-Cell Recognition during Fertilization in a Red Alga, Antithamnion sparsum (Ceramiaceae, Rhodophyta)

A gamete recognition mechanism in Antithamnion sparsum Tokidais proposed based on experiments using various lectins and carbohydrates.Spermatial binding to trichogynes is inhibited by pre-incubationof spermatia with concanavalin A (ConA) and/or L-fucose, whiletrichogyne receptors are blocked by the complementary carbohydrate-methyl D-mannose and/or the lectin Ulex europaeus agglutinin(UeA1). Binding inhibition (40–50%) was observed with10–50 mM carbohydrates and 25–50 µg ml^-1 lectins.The inhibitory effects of ConA and UeA1 is partially reversed(to 80–90% of controls) by addition of -methyl D-mannoseand L-fucose, respectively. Lectin binding to spermatial surfaceswas visualized by Fluorescein isothiocyanate (FITC) conjugatedConA, whereas carbohydrate receptors along the trichogyne andspermatium were localized with -mannosylated-FITC-albumin andL-fucosylated-FITC-albumin, respectively. These results suggestthat gamete recognition in Antithamnion sparsum is mediatedby a double-docking recognition system consisting of spermatiapossessing surface L-fucose receptors and -methyl D-mannosemoiety, and trichogynes possessing the complementary receptors. (Received December 5, 1995; Accepted April 22, 1996)     

RNA Polymerase Activity During Breakage of Seed Dormancy by Low Temperature Treatment of Fruits of Acer platanoides (Norway Maple)

Slater, R. J. and Bryant, J. A. 1987. RNA polymerase activityduring breakage of seed dormancy by low temperature treatmentof fruits of Acer platanoides (Norway maple).—J. exp.Bot. 38:1026–1032. Endogenous RNA polymerase activity has been characterized innuclei isolated from embryo axes of Acer platanoides. Optimalactivity was recorded at 4·0 mol m^–3 MgCl₂ and50 mol m^–3 (NH₄)₂SO₄ and total activity could be inhibitedby up to 30% by -amanitin. Stratification of fruits leads toa stimulation of RNA polymerase activity. A minimum of 3 d coldtreatment is required with at least 3-fold stimulation recordedafter 10 d at 4°C. The increased enzyme activity is resistantto -amanitin suggesting an effect on RNA polymerase I. Key words: Acer platanoides, RNA polymerase, seed dormancy     

UPTAKE OF ACETATE BY NEOTRICULA APERTA (GASTROPODA: POMATIOPSIDAE), THE SNAIL HOST OF SCHISTOSOMA MEKONGI IN THE LOWER MEKONG BASIN

All three races of Neotricula aperta, an epilithic, schistosometransmitting, snail of the Mekong and Mul rivers of NortheastThailand and southern Laos, were found to take up acetate froma dilute solution. After 48 h incubation the mean specific netuptake rates (µmol^–1 g^–1 h^–1), from750 µM acetate, were: 1.86, -race; 1.39, ß-raceand 3.25, y-race. Over 48 h the snails were able to achievereductions in the ambient acetate concentration of up to 60%.Incubations under bacteriostatis suggested that bacteria arenot involved in the uptake of acetate by N. aperta. The uptakecharacteristics conform to the Michaelis-Menten model. The transportconstants, J_max (µmol^–1 g^–1 h^–1) andK_t (µM) were 1.10 and 102 respectively (-race). Racialdifferences in uptake characteristics are discussed in relationto micro-habitat differences. HPLC indicated concentrations of acetate in y-N. aperta microhabitatsof around 325 µM. This suggests a pool size sufficientto satisfy only 6% of the snail's basal metabolic rate (BMR).Levels within the epilithic aufwuchs, however, are probablyhigh enough to provide for more than 50% of the BMR. The possible role of acetate in the energy metabolism of N.aperta is discussed. Short-chain carboxylic acids (such as acetate),arising from the decomposition of the aufwuchs, could representsources of fermentable organics that may be taken up by N. apertasnails and used to supplement their nutrition during times offood shortage. However, further investigations involving ¹⁴C-labellingtechniques are required. The findings of this investigationhave implications for the chemical ecology and life-cycle ofN. aperta. (Received 16 June 1994; accepted 28 July 1994)     

Lipopolysaccharide- and gram-positive bacteria-induced cellular inflammatory responses: role of heterotrimeric Galpha(i) proteins

Heterotrimeric G_i proteins may play a role in lipopolysaccharide (LPS)-activated signaling through Toll-like receptor 4 (TLR4), leading to inflammatory mediator production. Although LPS is a TLR4 ligand, the gram-positive bacterium Staphylococcus aureus (SA) is a TLR2 ligand, and group B streptococci (GBS) are neither TLR2 nor TLR4 ligands but are MyD88 dependent. We hypothesized that genetic deletion of G_i proteins would alter mediator production induced by LPS and gram-positive bacterial stimulation. We examined genetic deletion of G_i2 or G_i1/3 protein in G_i2-knockout (G_i2–/–) or G_i1/3-knockout (G_i1/3–/–) mice. LPS-, heat-killed SA-, or GBS-induced mediator production in splenocytes or peritoneal macrophages (M) was investigated. There were significant increases in LPS-, SA-, and GBS-induced production of TNF- and IFN- in splenocytes from G_i2–/– mice compared with wild-type (WT) mice. Also, LPS-induced TNF- was increased in splenocytes from G_i1/3–/– mice. In contrast to splenocytes, LPS-, SA-, and GBS-induced TNF-, IL-10, and thromboxane B₂ (TxB₂) production was decreased in M harvested from G_i2–/– mice. Also, LPS-induced production of IL-10 and TxB₂ was decreased in M from G_i1/3–/– mice. In subsequent in vivo studies, TNF- levels after LPS challenge were significantly greater in G_i2–/– mice than in WT mice. Also, myeloperoxidase activity, a marker of tissue neutrophil infiltration, was significantly increased in the gut and lung of LPS-treated G_i2–/– mice compared with WT mice. These data suggest that G_i proteins differentially regulate murine TLR-mediated inflammatory cytokine production in a cell-specific manner in response to both LPS and gram-positive microbial stimuli. G_i protein-deficient mice; endotoxin; group B streptococci; Staphylococcus aureus; Toll-like receptors     

Glycosidases from Cotyledons of Pisum sativum L.

-Mannosidase and ß-N-acetylglucosaminidase were purifiedfrom extracts of cotyledons of germinating Pisum sativum L.A 13-fold purification of a-mannosidase free from ß-N-acetylglucosaminidaseactivity was achieved by precipitation in ammonium sulphate,column chromatography on DEAE-cellulose, and treatment with2 M pyridine. ß-N-Acetylglucosaminidase was purified200-fold by the use of (NH₄)₂SO₄, and chromatography on ConcanavalinA¹-Sepharose and Sephacryl-200. This preparation showed no measurablecontamination by -mannosidase activity. Both glycosidases appearto be glycoproteins and demonstrate optimal activity at pH valuesof 4.0–4.5. Both glycosidases appear to have very similarmolecular weights, with -mannosidase being slightly larger thanß-N-acetylglucosaminidase. An extensive search forthe activity of aspartylglycosylamine amido hydrolase in peacotyledons proved unsuccessful.     

In vivo Studies on the Occurrence of Stored mRNA in Embryonic Axes of Vigna unguiculata Seeds

-Amanitin and cordycepin at various concentrations were testedfor their inhibitory effect on the fresh weight increase ofVigna unguiculata embryonic axes after the onset of imbibitionand on the incorporation rate of ³H-labeled leucine into proteinin axes of the 36–38 h stage. -Amanitin at 0.5–5µ/Kg/ml clearly exerted an inhibitory effect on both thefresh weight increase and the protein synthesis. This drug at1 µg/ml, however, showed no significant effect on theprotein synthesis at an early stage of imbibition (4 h), whereascycloheximide was a very potent inhibitor. By experiments inwhich ‘dry’ axes were allowed to imbibe 3H-lebeledadenosine solution for 4 and 12 h in the presence of -amanitin,it was found that poly A⁺RNA was newly synthesized to some extentin axes as early as 4 h after the onset of imbibition and thatthe drug effectively inhibited the poly A⁺RNA synthesis. Theresults may indicate the occurrence of stored mRNA in embryonicaxes of V. unguiculata seeds. (Received June 11, 1983; Accepted August 16, 1983)     

Purification and properties of soluble cytochrome b-560 from spinach leaves

A b-type cytochrome having an -band at 560 nm was isolated fromspinach leaves (Spinacia oleracea). A method is described forpreparing this cytochrome, cytochrome b-560 (spinach), in apurified state. The cytochrome has, in its reduced state, absorption bands at560 nm (), 530 nm (ß) and 427 nm (); and in the oxidizedstate at 562 nm (), 529 nm (ß) and 417 nm (). Thepyridine ferro-haemochrome prepared from cytochrome b-560 hadan -band at 556.5 nm, indicating the protohaem-nature of theprosthetic group. The cytochrome has an oxidation-reduction potential (E'⁰) of+0.13V at pH 7.0, as measured using the ferri-ferro oxalate system. The cytochrome is rapidly reduced on illumination with red orfar-red light in the presence of spinach chloroplasts and isoxidized at a slower rate in the dark. This photoreduction isinhibited by 1x10^–6 M 3-(3,4-dichlorophenyl)-1,1-dimethylurea(DCMU). The molecular weight of the cytochrome is 30,000 asestimated by the dextran gel filtration method. (Received December 3, 1971; )     

Effects of algal concentration, bacterial size and water chemistry on the ingestion of natural bacteria by cladocerans

Journal of Plankton Research, 11, 1273–1295, 1989. The values of P/U⁰ (Table I) and fluid velocity used to calculatethe energy required for sieving (pp. 1289–1290) and severalequations (footnote b of Table I; p. 1290, lines 3–4)are incorrect. The corrected table appears below: Table I. Filter setule measurements (mean and within specimenstandard deviation) of the gnathobases for the cladocerans studied^aGnathobaseof trunklimb number. ^bP = 8µU₀/(b(1 – 21nt + 1/6(t²) - 1/144(t⁴))), whereP = pressure drop in dyn cm^–2, =3.1416, U₀ = fluid velocityin cm s^–1, b = distance between setule centres in cm,t = ( x setule diameter)/b and µ = 0.0101 dyn s^–1cm^–2. Formula from Jørgensen (1983). The text (p. 1289, line 19 to p. 1290, line 10) should read: organism. Using a similar argument, a 0.5 mm Ceriodaphnia witha filter area of 0.025 mm² (Ganf and Shiel, 1985) and pressuredrop P = 2757 dyn cm^–2 (with fluid velocity of 0.07 cms^–1) allocates only 2171 ergs h^–1 to filtrationof a total energy expenditure of 10⁴ ergs h^–1 [filtrationenergy (ergs h^–1) = area (cm²) x pressure drop (dyn cm^–2)x 3600 (s h^–1) x 1/0.2 (efficiency of conversion of biochemicalinto mechanical work); total energy (ergs h^–1) = respiration(0.05 µl O₂ ind^–1 h^–1 consumed; Gophen, 1976)x conversion factor (2 x 10⁵ ergs µl^–1 O₂). Withan estimated 0.034 mm² in filter area, fluid velocity of 0.041cm s^–1 and respiration of 1.8 x 10⁴ ergs h^–1 (calculatedfrom Porter and McDonough, 1984), a 0.5 mm Bosmina uses <4%of its metabolism to overcome filter resistance. The velocities used in the original examples (0.4 cm s^–1for Ceriodaphnia, 0.2 cm s^–1 for Bosmina) were derivedfrom literature values of appendage beat rate and estimatesof the distance travelled by the appendages during each beatcycle. This approach unnecessarily assumes that all water movedpasses through the filter. In the new calculations, the flowacross the filter needed for food to be collected by sieving(0.07 cm s^–1 for Ceriodaphnia and 0.041 cm s^–1 forBosmina) was determined from the maximum clearance rate/filterarea. The amended energy expenditures, although higher, do notrefute the sieve model of particle collection.     

Electrophysiological studies of chemoreceptors sensitive to pyridine on the crayfish walking leg. II. Characteristics of inhibitory molecules

The effect of substituted pyridines on the response of singlepyridine-sensitive cells from crayfish walking legs was investigatedelectrophysiologically. Seven p-substituted pyridines, actingreversibly, were identified as specific antagonists at the pyridinereceptor. The maximum saturation frequency of the response toagonists was reached in the presence of antagonists but thedose–response curves of the agonists were shifted in parallelto the right along the concentration axis. Schild plots of threehighly effective antagonists with six agonists were linear witha slope close to one, indicating competitive antagonism. Theinhibition constants yielded a K₁ value of {small tilde}4–8µmol for the most effective antagonist, 4-(4-nitrobenzyl)pyridine,which had only one order of magnitude lower affinity than themost effective agonist 2,3'-bipyridyl. The antagonists 2,4'-bipyridyland 4-benzylpyridine had K¹ values of 6–10 µmol,followed by 4-acetylpyridine with a K¹ value of 30–70µmol. The rank order of inhibitory potency of the differentantagonists was found to be the same for all units tested. Comparingelectronic effects (Hammett values and pKa values) of the substitutentsin p- and m-position showed that inhibitory effectiveness decreasedwith a decrease in pKa and an increase in Partition coefficientswere determined for 10 agonists and the antagonists which weregenerally more lipophilic than the agonists. A hypotheticalreceptor site is proposed.