Na3 Citrate Research Articles

Interacting effects of urea and water addition on soil mineral‐bound phosphorus dynamics in semi‐arid grasslands with different land‐use history

AbstractNitrogen (N) addition and precipitation increment can greatly influence soil phosphorus (P) dynamics, with much emphasis on total and available P, yet little is known about their interactive effects on soil mineral‐bound inorganic P (Pi) fractions under different historical land uses of grasslands and old fields. Thus, we compared (i) plant readily available P, (ii) less available Pi (sum of NH4F‐extractable Pi, NaOH‐Na2CO3‐extractable Pi and NaHCO3‐extracted Pi), (iii) refractory forms of Pi (NH4Ac‐extractable Pi, H2SO4‐extractable Pi and Na3(citrate)‐dithionite‐extractable Pi) and (iv) organic P under the same urea and water treatments in an old‐field grassland with a semi‐arid steppe. Soil total P remained unchanged with 10‐year urea addition in both sites, with lower organic P but higher Pi concentrations in the old field. Urea addition promoted transformation of refractory Pi into less available Pi in both sites, which potentially was related to higher plant productivity and thus enhanced plant P accumulation. Specifically, urea addition increased less available Pi fractions by as much as 42%, but decreased refractory Pi fractions by as much as 24%, for two sites under ambient precipitation. Water addition decreased less available Pi in the steppe under higher urea addition rates, whereas it increased less available and refractory Pi but only in the control plot of the old field. Irrespective of urea and water treatments, the old field had a higher Pi pool, which could replenish plant readily available P in the longer term and be less prone to P‐limitation than the steppe. We conclude that conversion of semi‐arid steppe to farmland causes long‐term increases in the soil Pi pool, possibly by enhancing organic P mineralization and anthropogenic fertilization during cultivation. Urea addition accelerated soil P cycling via promoting refractory Pi, transforming into less available Pi, with the process being strongly mediated by water availability, whereas the projected precipitation increment could decrease less available Pi via promoting plant P uptake and P leaching out of the plant–soil system.Highlights Urea addition promoted transformation of refractory Pi into less available Pi fractions. Water addition counteracted the positive urea effect on NH4F‐Pi in both steppe and old field. Less responsive organic P suggested that Pi fractions were more essential in soil P cycling. Water addition decreased NH4F‐Pi in both sites, potentially due to higher plant uptake and leaching. Conversion of steppe to farmland increased the soil Pi pool by enhancing organic P mineralization.

Diagnostic ultrasound activates pure prekallikrein

Diagnostic ultrasound activates the contact phase of human coagulation. This has been seen in human blood or plasma or with purified factor 12. The present work aimed to quantify a possibly triggering action of ultrasound on purified prekallikrein, the second of the two main triggers of the intrinsic hemostasis cascade. Either 2.7 μg/ml human prekallikrein or for control 1 μg/ml kallikrein in 26% glycerol - 0.54% NaCl-10.6 mmol/l Na3 citrate pH 7.4, in emptied polypropylene coagulation monovettes (Sarstedt) were exposed to diagnostic ultrasound (Siemens Acouson Antares, 5 MHz, 0.6 TIB, 0.6 TIS) for 0-5 min at room temperature (RT). Fifty microliter samples were withdrawn in duplicate and placed into an U-wells high quality microtiter plate (Brand 781600). Then 10 μl 2 mmol/l chromogenic substrate HD-CHG-Ala-Arg-pNA in 0.45% NaCl were added, and the increase in absorbance with time (ΔA405 nm /t at 37°C) was determined by a microtiterplate photometer with a 1 mA resolution (PHOmo; anthos). Exposure to diagnostic ultrasound biphasically increased the chromogenic activity of a prekallikrein solution in 26% glycerol. About 3-4 min ultrasound at 23 °C generated about 0.02 μg/ml kallikrein, that means that about 1% of pure prekallikrein in glycerol was converted into kallikrein. Thus, diagnostic ultrasound activates purified human prekallikrein to kallikrein. The ultrasound energy seems to fold the latent proenzyme prekallikrein into the active enzyme kallikrein. This contributes to explain the triggering action of ultrasound on the contact system of plasmatic human coagulation. Conversion of only 1% of prekallikrein into kallikrein is absolutely sufficient to start the intrinsic coagulation cascade. The clinical consequence of this action of ultrasound on intrinsic coagulation is that patients at risk for thrombosis, for example, patients with insufficiencies of hepatocytes, AT-3, C1-ina, or fibrinolysis should be protected by low-molecular-weight-heparin prior to the exposure of ultrasound, especially upon its prolonged exposure.

Formation of ZnO thin films consisting of nano-prisms and nano-rods with a high aspect ratio by a hydrothermal technique at 60 °C

ZnO thin films that consist of elongated nano-prisms and nano-rods were successfully grown on 100 nm-thick ZnO seeded glass substrates by hydrothermal synthesis at 60 °C and pH 10.3 in an aqueous solution containing Zn(NO 3)·6H 2O, Al(NO 3) 3·9H 2O, Na 3-citrate and NH 4OH. The effect of Al(NO 3)·6H 2O and Na 3-citrate, as surfactant chemicals, on the structural, morphological, optical and electrical properties of ZnO thin films were investigated. X-ray diffraction results showed that all the deposited films were grown as a polycrystalline wurtzite hexagonal phase with a c-axis preferred, out-of-plane orientation and without unwanted second phase. ZnO thin films deposited without any surfactant chemicals or deposited only with Al(NO 3)·6H 2O consist of elongated needle shaped nano-rods with a very rough surface morphology. On the other hand, ZnO thin films prepared using Na 3-citrate as a surfactant chemical consist of hexagonal nano-prisms with a very smooth surface morphology. The thickness of the ZnO thin films with the very smooth surface morphology was increased remarkably using both Na 3-citrate and Al(NO 3)·6H 2O as surfactant chemicals, in which ZnO thin films consisted of elongated hexagonal nano-prisms. These results show that relatively thick ZnO thin films with a good surface morphology can be grown easily by the appropriate use of surface modifying chemicals, such as Na 3-citrate and Al(NO 3)·6H 2O. The photoluminescence results showed strong defect-related emission peak centered near 545 nm in the rough surfaced ZnO thin film grown without any surfactant chemicals and strong band-edge peak centered near 368 nm in the smooth surfaced ZnO thin film grown using both Na 3-citrate and Al(NO 3)·6H 2O as the surfactant chemicals.

Effect of different annealing conditions on the properties of chemically deposited ZnS thin films on ITO coated glass substrates

The effects of different annealing conditions such as atmospheres, temperatures and times on the structural, morphological and optical properties of ZnS thin films prepared on ITO coated glass substrates by chemical bath deposition were studied. Aqueous solutions of zinc acetate and thiourea were used as precursors along with stable complexing agents, such as Na 2EDTA and Na 3-citrate, in an alkaline medium. X-ray diffraction patterns showed that the as-deposited and as-annealed ZnS films had an amorphous structure or poor crystallinity below the optimized annealing conditions of 500 °C and 60 min with the exception of the films annealed in N 2+H 2S annealing atmosphere. The ZnS thin films annealed in N 2+H 2S atmosphere for 1 h at 500 °C showed three sharp peaks for the (1 1 1), (2 2 0) and (1 1 3) planes of polycrystalline cubic ZnS without any unwanted secondary ZnO phases. X-ray photoelectron spectroscopy revealed Zn–OH and Zn–S bonding in the as-deposited ZnS thin film. However, the ZnS thin films annealed at 500 °C showed Zn–S bonding regardless of the annealing atmosphere. The sharp absorption edge and band gap energy of the as-deposited and as-annealed ZnS thin films varied from 295 to 310 nm and 3.5 to 3.89 eV, respectively.

The effect of salts on molecular mobility in amorphous sucrose monitored by erythrosin B phosphorescence

Salts are present in most amorphous biomaterials such as dried or frozen solid foods, plant seeds, and bacterial spores, and in some pharmaceutical formulations. However, knowledge of how salts modulate the physical properties of amorphous solid sugars, a major component in these systems, is lacking. We have used phosphorescence of the triplet probe erythrosin B (Ery B) to monitor molecular mobility in amorphous sucrose films (dried against P 2O 5) containing the salts NaCl, MgCl 2, CaCl 2, NaAcetate, Na 3Citrate, NaH 2PO 4, or Na 2HPO 4 at a mole ratio of 0.2:1 (salt/sucrose). All the salts examined, except NaH 2PO 4, significantly increased the phosphorescence lifetime of Ery B over the temperature range from 5 to 100 °C. This increase is due to a reduction in the rate of collisional quenching of the triplet state due to interactions with the matrix, indicating that these salts decreased the matrix molecular mobility. NaAcetate, Na 3Citrate, and Na 2HPO 4 decreased mobility more than NaCl, CaCl 2, or MgCl 2, perhaps due to specific hydrogen bonding interactions between the anion and sucrose. Systematic variations in the probe emission lifetime across the excitation and emission bands at 25 °C indicate that there are sites of different mobilities within amorphous solid sucrose; this dynamic site heterogeneity was enhanced in the presence of the divalent cationic salts MgCl 2 and CaCl 2. These results suggest that salts may play a significant role in modulating the mobility, and thus the long-term stability, of amorphous biological matrixes.

Optimization of bacteriorhodopsin production by Halobacterium salinarium PTCC 1685

This paper is concerned with medium optimization for the production of bacteriorhodopsin (BR) by Halobacterium salinarium PTCC 1685 by one-factor-at-a-time and Taguchi's array methods. The one-factor-at-a-time method was used to study the effects of various carbon and nitrogen sources, temperature and initial pH on bacteriorhodopsin production. Among these variables, corn steep powder, universal peptone and yeast extract were the most suitable sources, respectively. The optimal temperature and initial pH for bacteriorhodopsin production were 38 °C and 7.5, respectively. Subsequently, the concentration of corn steep powder and other medium components were optimized using Taguchi's method. The results showed that corn steep powder, glycerol and meat extract had significant effect on bacteriorhodopsin production by H. salinarium PTCC 1685. The optimal concentrations of nutritional components for improved bacteriorhodopsin production were determined as 10 g l −1 meat extract, 3.75 g l −1 casamino acids, 10 g l −1 glycerol, 50 g l −1 corn steep powder, 1 g l −1 Na 3-citrate, 6 g l −1 KCl, 0.2 g l −1 FeSO 4 7H 2O, 0.2 g l −1 MnSO 4 7H 2O and 10 g l −1 MgSO 4 7H 2O. The maximum bacteriorhodopsin production in shake flask culture was 191.7 mg l −1 about 2.85-fold higher than those obtained with the basal medium. Under optimal culture conditions, the maximum bacteriorhodopsin production in a 15-l stirred-tank bioreactor indicated 234.6 mg l −1 about 3.49-fold higher than the basal medium.

Regional Citrate Anticoagulation for Hemodialysis: Calcium-Free Vs. Calcium Containing Dialysate - A Randomized Trial

The majority of citrate protocols for hemodialysis (HD) use calcium (Ca)-free dialysate, a limited number use dialysate with Ca, aiming to simplify the procedure. This randomized clinical study sought to compare the anticoagulant effect of citrate using Ca-free dialysate and dialysate with Ca 1.25 mmol/L. Fifty HD procedures (in 5 chronic HD patients treated by chronic citrate anticoagulation) were randomly assigned to Ca-free dialysate (25 procedures) or Ca-1.25 dialysate (25 procedures), both with Mg 0.5 mmol/L, Na 138 mmol/L, and bicarbonate 28 mmol/L. Ca-free HD: 15% Na3 citrate 80 ml/hour was infused into the arterial line, and 1 M CaCl2, 14 ml/hour into the venous line. Ca-1.25 group: 15% Na3 citrate 100 ml/hour, 1 M CaCl2 2-4 ml/hour. Polyflux H dialyzers were used. Antithrombotic effect was assessed visually after HD at 3 points: dialyzer, arterial, and venous bubble traps, using a score of 5 (no clotting) to 1 (total clotting). Ca-free group: arterial bubble trap score 4.7 +/- 0.5, dialyzer 4.5 +/- 0.6, venous bubble trap 4.8 +/- 0.6. Ionized calcium (iCa) at dialyzer inlet 0.34 +/- 0.17, outlet 0.21 +/- 0.06 mmol/L. All HDs were completed successfully. Ca-1.25 group: arterial bubble trap score 4.7 +/- 0.5 (NS), dialyzer 2.6 +/- 1.04 (p<0.01), venous bubble trap 2.4 +/- 0.9 (p<0.01). Volume of clot in venous bubble trap was 1.9 +/- 1.8 mL (range 0.5-6 mL). iCa at dialyzer inlet 0.24 +/- 0.05 mmol/L (p<0.05), outlet 0.63 +/- 0.11 mmol/L (p<0.01). Four of 25 HD procedures (16%) were prematurely terminated due to threatening dialyzer clotting, in 6/25 HD procedures (24%), the venous line was changed (p<0.01). Citrate anticoagulation with Ca-1.25 dialysate resulted in significantly worse anticoagulation of dialyzer and venous bubble trap compared with Ca-free dialysate, despite higher citrate dose.

Storage of platelets in a novel additive solution (M‐sol), which is prepared by mixing solutions approved for clinical use that are not especially for platelet storage

To reduce adverse reactions due to platelet (PLT) transfusion, medical solutions on the market, such as saline and ACD-A, are used to replace the plasma of PLT concentrates in Japan; however, they are not strongly preservative. Here, an attempt was made to develop a novel additive solution (M-sol) having the ability to preserve PLTs stably, with only approved solutions for clinical use. M-sol is a mixture of solutions for medical use, which consists of 77 mmol per L NaCl, 3 mmol per L KCl, 1 mmol per L CaCl2, 21 mmol per L Na acetate, 15 mmol per L glucose, 9.4 mmol per L Na3 citrate, 4.8 mmol per L citric acid, 44 mmol per L NaHCO3, and 1.6 mmol per L MgSO4. The in vitro variables of PLTs stored in M-sol, Seto-sol, PASIIIM, or 100 percent plasma were compared during 14 days of storage. The in vitro parameters (pH, P-selectin, %hypotonic shock response, %disk, mean PLT volume, aggregability) of PLTs were better maintained in M-sol containing 3 percent plasma than in 100 percent plasma, PASIIIM with 31 percent plasma, and Seto-sol with 3 percent plasma during 14 days of storage. The 2-week storage of PLTs in M-sol is feasible in terms of the in vitro PLT function. Our results here show that the additive solution, with a high ability to preserve PLTs, can be prepared by mixing solutions approved for clinical use that are not specifically for PLT storage.

Identification of Ribosome-binding Protein p34 as an Intracellular Protein That Binds Acidic Fibroblast Growth Factor

With the aim of identifying new intracellular binding partners for acidic fibroblast growth factor (aFGF), proteins from U2OS human osteosarcoma cells were adsorbed to immobilized aFGF. One of the adsorbed proteins is a member of the leucine-rich repeat protein family termed ribosome-binding protein p34 (p34). This protein has previously been localized to endoplasmic reticulum membranes and is thought to span the membrane with the N terminus on the cytosolic side. Confocal microscopy of cells transfected with Myc-p34 confirmed the endoplasmic reticulum localization, and Northern blotting determined p34 mRNA to be present in a multitude of different tissues. Cross-linking experiments indicated that the protein is present in the cell as a dimer. In vitro translated p34 was found to interact with maltose-binding protein-aFGF through its cytosolic coiled-coil domain. The interaction between aFGF and p34 was further characterized by surface plasmon resonance, giving a K(D) of 1.4 +/- 0.3 microm. Even though p34 interacted with mitogenic aFGF, it bound poorly to the non-mitogenic aFGF(K132E) mutant, indicating a possible involvement of p34 in intracellular signaling by aFGF.

Eco‐friendly durable press finishing of cellulose‐containing fabrics

AbstractTo impart easy‐care properties to cellulose‐containing fabrics along with avoiding any harmful effects of formaldehyde on both the health and the environment, attempts have been made to use citric acid (CA) as an ester crosslinking agent along with different catalytic systems in the absence and presence of certain additives. Further, fixation conditions, type of crosslinking agent, as well as type of substrate have been studied. Results revealed that the enhancement in carboxyl content, performance properties, and the decrease in tear strength (TS) as well as in whiteness indices (WI) of the finished fabric samples were increased by increasing CA concentration up to 80 g/L and by raising thermofixation temperature from 140 up to 180°C for 90 s. Inclusion of triethanolamine hydrochloride (TEA. HCl), decreased the carboxyl content, TS, color strength K/S, as well as oily stain release rating (SRR) of the finished fabric samples along with an increase in bound nitrogen (%N), wrinkle recovery angle WRA, and an improvement in WI without affecting the durable press rating (DP). Within the range examined (0–30 g/L), increasing PEG‐600 concentration improved the wet resiliency, TS, as well as WI properties of the finished samples. Increasing DMDHEU ratio in the CA/DMDHEU crosslinking system gave rise to an increase in %N, WRA (dry and wet), DP, as well as in free CH2O of finished fabrics, along with a slight improvement in WI values. On the other hand, the TS, carboxyl content, K/S, SRR values of the finished fabric samples were lower at a higher DMDHEU ratio. Increase in carboxyl content, %N, WRA (dry and wet), DP and SRR, as well as extent of post dyeing (K/S) of the treated fabric samples upon using different ester crosslinking agents followed the descending order: citric acid &gt; pyromellitic dianhydride. The opposite holds true for the TS, and WI values. Among the esterifying catalysts used, and for a given set of finishing conditions, NaH2PO2 · H2O proved to be the most effective one, and the following order of effectiveness may be drawn: NaH2PO2 · H2O &gt; K2HPO4 &gt; Na3–citrate &gt; Na2–tartrate. Inclusion of silicone softener in the finishing formulation brought about an improvement in softeness degree, WRA, %N, DP, TS as well as K/S values along with a decrease in carboxyl content, SRR, and WI values of the treated fabric samples, regardless of the used silicone softener.The performance properties of the finished fabric samples were determined by the type and nature of the substrate. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 84: 2243–2253, 2002

Traditional Extractants in Nontraditional Solvents: Groups 1 and 2 Extraction by Crown Ethers in Room-Temperature Ionic Liquids

The crown ethers 18-crown-6 (18C6), dicyclohexano-18-crown-6 (DCH18C6), and 4,4‘-(5‘)-di-(tert-butylcyclohexano)-18-crown-6 (Dtb18C6) were dissolved in 1-alkyl-3-methylimidazolium hexafluorophosphate ([Cnmim][PF6], n = 4, 6, 8) room-temperature ionic liquids (RTILs) and studied for the extraction of Na+, Cs+, and Sr2+ from aqueous solutions. In the absence of extractant, the distribution ratios for the metal ions indicate a strong preference for the aqueous phase. With the crown ethers as extractants in RTIL-based liquid/liquid separations, the resulting metal ion partitioning depends on the hydrophobicity of the crown ether and also on the composition of the aqueous phase (e.g., concentration of HNO3 vs Al(NO3)3). Aqueous solutions of HCl, Na3 citrate, NaNO3, and HNO3 (the latter at low concentrations) decrease the metal ion distribution ratios and also decrease the water content of the RTIL phase. High concentrations of HNO3 decompose PF6- and increase both the water content and the water solubility of the RTIL phase. Highly hydrated salts such as Al(NO3)3 and LiNO3 salt out both the RTIL ions and the crown ethers; thus, when the aqueous phase contains Al(NO3)3, the trend more closely resembles traditional solvent extraction behavior where DSr > DCs and the most hydrophobic extracting phase produces the highest partitioning. When [C8mim][PF6] is used as the extracting phase, the metal ions can be loaded from Al(NO3)3 and stripped using water. Dtb18C6 forms 1:1 complexes with Cs+ and Sr2+ and also yields the highest distribution ratios out of the three crowns examined. In comparison to traditional solvent extraction behavior, the metal ion partitioning in these systems exhibits exceptional behavior and, in certain instances, suggests a complicated partitioning mechanism, which necessitates a more thorough understanding of RTILs as solvents before interpretation of the results.

SELECTIVE AND QUANTITATIVE PARTITIONING OF PERTECHNETATE IN POLYETHYLENE-GLYCOL BASED AQUEOUS BIPHASIC SYSTEMS

ABSTRACT The partitioning behavior of technetium has been investigated in polyethylene glycol (PEG)-based aqueous biphasic systems using K3P04, K2C03, Na3(citrate), (NH4) 3(citrate), (NH4) 2S04, and NaOH as biphase-forming salts, and PEGs with average molecular weights of 1500, 2000, 3400, and 6000. Pertechnetate quantitatively partitions to the PEG-rich phase in these systems. The distribution ratios increase with increasing salt or PEG concentration used to form the aqueous biphasic systems, consistent with the increasing phase incompatibility. At a given concentration of salt and PEG, DTc values in the various systems increase in the order NaOH < (NH4) 2S04 < (NH4)3(citrate) < K2C03 < K3P04 and PEG-1500 < PEG-2000 < PEG-3400 < PEG-6000. The distribution ratios are lower at very high or very low pH values than they are in the 3-9 pH range. Matrix ions may also influence the DTcvalues by affecting the aqueous biphasic system composition. Technetium can be stripped from the loaded PEG-rich phase by contact with a clean salt solution containing SnCl2, which reduces TcO4 to an oxy-chloro cationic species that partitions to the salt-rich phase.

Effect of anticoagulant and blood storage time on platelet-bound antibody concentrations in clinically normal dogs

Summary A solid-phase elisa to detect antibodies bound to the surface of canine platelets (platelet-bound antibodies) is described. Using this assay, the effect of anticoagulant and storage time of anticoagulant blood on the concentration of antibodies bound to the surface of platelets from clinically normal dogs was investigated. Blood from 3 clinically normal dogs was anticoagulated with acid citrate dextrose, Na3 citrate, and aqueous K3 edta and stored on ice for up to 48 hours. Platelet-bound antibody concentration was measured on platelets isolated from anticoagulated blood immediately after venipuncture and subsequent to storage of blood for 24 and 48 hours. Differences in platelet-bound antibody concentrations were investigated among dogs, anticoagulants, and storage times by anova and Bonferroni pair-wise comparison of means. There was no effect of dog on platelet-bound antibody concentration. The effect of time was significant (P &lt; 0.0001), with higher concentration of platelet-bound antibodies detected with increasing storage time. Effect of anticoagulant on platelet-bound antibody concentration was not statistically significant; however, there was a trend to increasing concentration of antibodies bound to platelets isolated from Na3 citrate- and K3 edta-anticoagulated blood. Moreover, there was significant (P = 0.02) interaction between anticoagulant and time. Platelet-bound antibody concentration increased with storage of anticoagulated blood prior to platelet isolation and with use of Na3 citrate and K3 edta anticoagulants. The preferred anticoagulant for platelet-bound antibody measurement is acid citrate dextrose. Platelet-bound antibody concentration should be determined not longer than 24 hours after blood collection.

Filtration through a polyester white cell-reduction filter of plasma-poor platelet concentrates prepared with an acetate-containing additive solution.

It is of practical importance to known whether the adsorption of platelets and contaminating white cells (WBCs) by the WBC-reduction filter is altered when platelet concentrates (PCs) are prepared in a plasma-poor condition with an acetate-containing additive solution (Seto sol). Plasma-poor PCs with 11-percent residual plasma were prepared from apheresis platelet-rich plasma by using a sterile docking device with steam-sterilized Seto sol. Seto sol contains 115 mM (115 mmol/L) NaCl, 4 mM (4 mmol/L) KCl, 3 mM (3 mmol/L) MgCl2, 10 mM (10 mmol/L) Na3PO4, 15 mM (15 mmol/L) acetate, 3 mM (3 mmol/L) Na3 citrate, and 10 mM (10 mmol/L) glucose (pH 7.1). The solution was steam-sterilized under nitrogen gas. On Days 1 and 5, pooled Seto sol PCs (2.4 x 10(11) platelets) were filtered with a polyester filter at a flow rate of 10 mL per minute. The WBC-removal rate was over 99.9 percent with a platelet recovery of 88 percent following Day 1 filtration. These values were very similar to those of plasma PCs, and 84-percent recovery was achieved following Day 5 filtration. However, when 1 unit of Seto sol PCs with half the number of platelets was filtered with the polyester filter, platelet recovery was about 16 to 17 percent less than that of plasma PCs. Platelet quality was maintained if pooled Seto sol PCs were filtered on Day 1 and stored for over 4 days. Filtration did not alter platelet function in 1-day-old or 5-day-old Seto sol PCs.(ABSTRACT TRUNCATED AT 250 WORDS)

Plasma‐Depleted Platelet Concentrates Prepared with a New Washing Solution

In certain clinical situations, complete removal of the plasma proteins from the platelet concentrates (PCs) is necessary by washing prior to transfusion. A simple electrolyte solution with a pH of 6.5 was developed for washing PCs. The platelet-rich plasma collected with acid-citrate-dextrose solution by apheresis in a 0.6-liter polyolefin bag was centrifuged. After removal of the supernatant plasma from pelleted platelet buttons, 200 ml of a washing solution consisting of 90 mM NaCl, 5 mM KCl, 3 mM MgCl2, 17 mM NaH2PO4, 8 mM Na2HPO4, 23 mM Na acetate, 17 mM Na3 citrate, 23.5 mM glucose, 2 mM adenine, 0.1% dextran, and 28.8 mM maltose (pH 6.5) was added to the pelleted platelet button. Steam sterilization of the solution was carried out under nitrogen to avoid caramelization of glucose. After resuspension of the pelleted platelet button with a washing solution and a second centrifugation, Seto additive solution (Seto sol, pH 7.4) was introduced into the bag to resuspend the platelet buttons for storage for 3 days at 22 degrees C. All of these procedures were completed within 3 h using a sterile docking device. In washed PCs, 99.1% of the plasma was removed and platelet recovery was 96%. The washed PCs were compared for 3 days with plasma-poor PCs consisting of 11% plasma and 89% Seto solution. There were no significant differences in percent hypotonic shock response, aggregation, energy metabolism, and morphology of platelets between the two groups during 3 days, except for significant swelling of 3-day-old platelets in washed PCs.(ABSTRACT TRUNCATED AT 250 WORDS)

Plateletpheresis with the IBM model 2997. I. Effects of ACD, NIH formula B on selected donor indices.

9 healthy subjects dosed with acid-citrate-dextrose, NIH formula B (ACD-B) while undergoing plateletpheresis with the continuous-flow IBM Model 2997 centrifuge, received on average 59.5 +/- (SD) 3.2 mg Na3 citrate.kg-1.h-1. This rate of infusion of Na3 citrate resulted in a maximum 12-fold rise in the concentration of citrate in serum (plasma) following the processing of 10 litres of whole blood, and subsequently reduced the level of total calcium in serum (plasma) by 9%. This dose of Na3 citrate produced no clinical symptoms suggestive of hypocalcaemia in these subjects, even though the use of acid-citrate-dextrose, NIH formula A (ACD-A) under identical conditions has been reported to reduce significantly the level of total calcium in serum, and concomitantly increase the number or reactions occurring in donors. From previous studies employing the intermittent-flow centrifugation system of plateletpheresis, a rate of infusion of Na3 citrate below 65 mg.kg-1.h-1 can insure maximum donor safety and this rate can be achieved with the IBM Model 2997 by the utilisation of ACD-B.

Plateletpheresis with the IBM Model 2997

9 healthy subjects dosed with acid-citrate-dextrose, NIH formula B (ACD-B) while undergoing plateletpheresis with the continuous-flow IBM Model 2997 centrifuge, received on average 59.5 ± (SD) 3.2 mg Na(3) citrate ˖ kg^-1 ˖ h^-1. This rate of infusion of Na(3) citrate resulted in a maximum 12-fold rise in the concentration of citrate in serum (plasma) following the processing of 10 litres of whole blood, and subsequently reduced the level of total calcium in serum (plasma) by 9%. This dose of Na3 citrate produced no clinical symptoms suggestive of hypocalcaemia in these subjects, even though the use of acid-citrate-dextrose, NIH formula A (ACD-A) under identical conditions has been reported to reduce significantly the level of total calcium in serum, and concomitantly increase the number or reactions occurring in donors. From previous studies employing the intermittent-flow centrifugation system of plateletpheresis, a rate of infusion of Na(3) citrate below 65 mg ˖ kg^-1 ˖ h^-1 can insure maximum donor safety and this rate can be achieved with the IBM Model 2997 by the utilisation of ACD-B.

Localisation du gene de l'α-glucosidase acide (α-GLUa) sur le segment q21→qter du chromosome 17 par l'hybridation cellulaire interspecifique

A rapid detection of human α-GLUa in man-mouse hybrid extracts was described. In each of three glass tubes was put a drop (or a droplet, 2 μl) of hybrid extracts. The second and third tubes was inactivated at 60°C during 10 mn and 20 mn respectively. A drop (or droplet) of a solution containing 5 mg 4-methyl-umbelliferyl-α-D-glycopyranoside in 5 ml of Na3 citrate (0.1 M) citric acid, pH 4, was put in different tubes. The homogeneous mixture was obtained with an agitator of tubes. After 1 h at room temperature a drop (or a droplet) of ammonia solution was put in different tubes. A fluorescence appeared under UV for extracts containing α-GLUa. The hybrids positive for human α-GLUa presented a fluorescence in three tubes. The hybrid negative for human α-GLUa presented a fluorescence only in the first tube (non inactivated; presence of mouse α-GLUa). In some cases to ameliorate the lecture under UV, a drop or a droplet from different tubes was put on a “godet plate” or on a cellulose acetate gel.

Control of colony formation in Coelastrum microporum (Chlorococcales, Chlorophyta)

Abstract Colony formation in Coelastrum microporum Naeg. strains 280 and 281 could be controlled by adding 0.75 g l−1 Na3 citrate and 0.1% (v/v) soil extract to modified Bristol's medium with 0.25 g l−1 CaCl2. When growing in this medium, 92% of colonies in strain 280 and 88% in strain 281 could be obtained. Slightly lower percentages of colonies were observed when both strains were grown in modified Bristol's medium with either Na3 citrate or soil extract added. Modified Bristol's medium with 0.025 g l−1 CaCl2 and without both Na3 citrate and soil extract favoured unicell formation in both strains. However, the percentages of colonies increased when the CaCl2 level was raised to 0.25 g l−1 in this medium. Modified Bristol's medium with 1 g l−1 CaCl2 was found to be inhibitory to colony formation.

CONTROL OF COLONY AND UNICELL FORMATION IN A SYNCHRONIZED SCENEDESMUS(1) (2).

Colony formation in a synchronized Scenedesmus could be controlled by the addition of 0.05% Na3 citrate or 85 μM EDTA to modified Bristol's medium. No unicells were formed; only S. quadricauda-like or S. longispina-type colonies were observed in young cultures grown in that medium. A colony population could be made completely unicellular in 2 days if grown in soil-Bristol's medium and transferred daily. The pleomorphic Scenedesmus was examined in synchronized culture. When the organism was grown in a defined medium in a 15-hr light /9-hr dark cycle on a roller tube rotator at 2-6 rpm and transferred daily, synchrony of cell division and release of the products were achieved. In a synchronized culture 2 doublings/day were recorded, with most cytoplasmic cleavages and all releasing of daughter cells taking place in the dark period. In many observations with several synchronized strains of Scenedesmus, no fixed pattern of release of daughter products from mother cells or colonies was detected. Colonies or unicells had their full spine complement at the time of release. As a cell or colony aged the spines sometimes increased in thickness. Other Scenedesmus strains were examined to provide supporting data.