Production rates of prostaglandin F, 6-keto-PGF1α and thromboxane B2 by perifused human endometrium

Human endometrium obtained from fresh hysterectomy specimens was perifused for 7 hr in 95% O₂/5% CO₂ at 37°C. The phase of the menstrual cycle was determined by histological examination. The concentrations of PGF, 6-keto-PGF_1α and TxB₂ in 20 min fractions of the perifusion medium were measured by radioimmunoassay and production rates were calculated in terms of dry weight of tissue. Biphasic patterns of production were observed; high initial values fell to about 20% at 2 hr and then increased to relatively stable values at about 4 hr which were maintained for the next 2 hr. During this latter period, production rates in endometria taken at different phases of the cycle differed markedly from each other; the production rates of PGF in secretory and early proliferative endometria were low (15.8 ± 2.6, mean ± SEM and 67.2 ± 8.3 ng/min/g respectively) whereas they were high in late proliferative and premenstrual endometria (188.0 ± 16.7 and 196.4 ± 16.9 ng/min/g respectively). The patterns of production of 6-keto-PGF_1α and TxB₂ were similar to those of PGF but the absolute values were much lower (<10%). We conclude that the observed rates of production of prostaglandins by perifused human endometrium are consistent with synthesis being stimulated either by estrogen or withdrawal of hormonal support and being inhibited by progesterone.     

Alterations in premating behavior and pheromone biology of gamma-irradiated Trichoplusia ni (Lepidoptera: Noctuidae)

Exposure of female cabbage looper pupae, Trichoplusia ni, to gamma irradiation at 0 to 72 hr before eclosion resulted in significantly reduced moving and calling activity of the adults. The effects were dose-related from 10 to 40 kR. Generally, irradiation in a nitrogen atmosphere slightly reduced deleterious effects, but fractionation of the radiation dose did not. Four-day-old irradiated females contained significantly more sex pheromone than untreated females, indicating that loss of pheromone is positively correlated to the amount of calling activity.Males exposed to 20 kR or 40 kR in nitrogen were not affected in their flight response in an olfactometer to a level of synthetic pheromone ca. equal to that released by a calling female or to 0.1 x this level. The response of males exposed to 40 kR in air, however, was significantly decreased at the lower pheromone concentration.

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Veränderungen im vorkopulationsverhalten und in der pheromobiologie bei gammabestrahltentrichoplusia ni (Lepitoptera: Noctuidae)

Zusammenfassung Puppen von Trichoplusia ni wurden 0–72 Stunden vor dem Schlüpfen der Falter einer Gamma-bestrahlung ausgesetzt. Die Bestrahlung betrug 10–40 kR und erfolgte entweder in Luft (ununterbrochen oder fraktioniert) oder in einer Stickstoffatmosphäre. Die Effekte der verschiedenen Behandlungen auf das Rufverhalten der Weibchen (CB), die Bewegungsaktivität (MA) und den Pheromontiter wurden beurteilt. Die Reaktion bestrahlter Männchen auf das synthetische Sexual-pheromon (Z-7-Dodecen-1-o1-azetat) wurde gemessen bei einer Konzentration, wie sie von einem Weibchen erzeugt wird. und bei einem Zehntel dieser Menge.Die Bestrahlung reduzierte bei sitzenden Weibchen gesichert CB und MA (Tabelle II). Die Reduktion dieser Aktivitäten war im allgemeinen dosisabhängig. Die Fraktionierung der Bestrahlung und die Bestrahlung in Stickstoffatmosphäre konnten diese Effekte nicht gesichert vermindern.Bestrahlte Weibchen riefen gesichert weniger als nicht bestrahlte Tiere und gaben weniger Pheromon ab. Als Folge davon enthielten ihre Pheromondrüsen im Alter von 4 Tagen gesichert mehr Pheromon als Drüsen unbehandelter Weibchen (Tabelle IV).In Olfaktometertests reagierten Männchen, die mit bis zu 40 kR bestrahlt worden waren, ungefähr gleich wie unbestrahlte Männchen, wenn sie einer Verdampfungsrate von einem Weibchenaequivalent des Pheromons ausgesetzt wurden (Tabelle V). Männchen, die mit 40 kR in Stickstoffatmosphäre behandelt worden waren, reagierten auf eine niedrigere Pheromonkonzentration gleich gut wie Männchen, die mit 20 kR in Luft behandelt worden waren. Sie waren gesichert reaktionsfähiger als solche, die mit 40 kR in Luft bestrahlt worden waren.Die mögliche Bedeutung dieser Effekte für das Autozidverfahren wird diskutiert.

     

Studies on preservation of schistosomula of Schistosoma mansoni and S. mattheei

Schistosomula were not damaged by exposure for 1 hr at room temperature to the cryoprotectant dimethylsulphoxide (DMSO) providing that concentrations greater than 10% were not used. Rapid dilution to remove the DMSO was less harmful to the organisms than was slow dilution. Schistosomula were not damaged by thermal shock (cooling in the absence of freezing) but were damaged by conditions produced by freezing. Although the freezing damage rendered schistosomula noninfective they retained flame cell activity and certain contractile properties in the oral sucker, gut, and musculature. The least damage was produced by slow cooling (at approximately 0.3 °C/min) and fast warming (approximately 300 °C/min). Schistosomula remained infective following freezing and slow cooling to −20 °C in DMSO (10%) and storage for 2 hr at this temperature but were damaged at temperatures below −26 °C and at −20 °C for longer time periods.     

CYTOKINETIC ANALYSIS OF THE JB-1 ASCITES TUMOUR AT DIFFERENT STAGES OF GROWTH

The cell kinetics of the murine JB-1 ascites tumour have been investigated on days 4, 7 and 10 after transplantation of 2·5 × 10⁶ cells. The experimental data, growth curve, percentage of labelled mitoses curves, continuous labelling curves and cytophotometric determination of single-cell DNA content have been analysed by means of a mathematical model for the cell kinetics. The important result was the existence of 8% non-cycling cells with G₂ DNA content in the 10-day tumour, while only 0·2 and 0% were observed in the 7- and 4-day tumours, respectively. The doubling times determined from the growth curve were 22·8, 70 and 240 hr, respectively, in the 4-, 7- and 10-day tumours. Growth fractions of 76, 67 and 44% were calculated for the same tumour ages. The mean cell cycle time increased from 14 to 44 hr from day 4 to 7 due to a proportional increase in the mean transit time of all phases in the cell cycle. In the 10-day tumour, the mean cell cycle changed to 41 hr and T _G1 decreased to 0·5 hr. The cell production rate was 4·3%/hr in the 4-day tumour, 1·2%/hr in the 7-day tumour and 1·0%/hr in the 10-day tumour. The cell loss rates in the same tumours were 1·3, 0·2 and 0·7%/hr, respectively. The analysis made it probable that the mode of cell loss was an age-specific elimination of non-cycling cells with postmitotic DNA content.     

Formulation of a Streptomyces Biocontrol Agent for the Suppression of Rhizoctonia Damping-off in Tomato Transplants

Formulations of a Streptomyces biological control agent for Rhizoctonia damping-off in tomato seedlings were developed for the first time from vegetative propagules obtained from actively growing, nonsporulating liquid cultures. Alginate beads, durum flour (starch) granules, and talcum powder formulation of this new actinomycetous antagonist (Streptomyces sp. Di-944) isolated from the rhizosphere of field-grown tomato (Lycopersicon esculentum) suppressed damping-off caused by Rhizoctonia solani in tomato plug transplants (cv. Bonny Best) in a peat-based, soilless potting mix under greenhouse conditions. For formulations, vegetative biomass of Streptomyces sp. Di-944 from 3-day-old liquid fermentation in yeast extract–malt extract–glucose broth was lyophilized and pulverized to obtain fragments of viable vegetative filaments. The pulverized biomass had an initial viable count of 2 × 10⁷colony forming units/g and retained 100% viability for 2 weeks when stored at 4°C. Formulations stored at 4°C had a longer shelf life than those stored at 24°C based on viability at 2-week intervals over a 6-month storage period. In addition, dual culture tests showed declining efficacy for surviving Streptomyces propagules in formulations during this storage period. At 4°C, the powder and granular formulations were found to be the most stable and were shown to be 100% viable after 14 and 10 weeks of storage, respectively. However, at the end of 24 weeks, the number of viable propagules in the powder and granular formulations declined to 1.2 × 10⁵ and 7 × 10³ colony forming units/g, respectively. Alginate beads were the least stable in storage. Even at 4°C, 6.9 × 10⁴ and 7.3 × 10² viable propagules/g formulation were detected at the end of 12 and 24 weeks, respectively. The talcum powder formulation delivered to tomato seeds as a seed-coating was the most effective biocontrol treatment. It suppressed damping-off in 10-day-old tomato transplants by almost 90% compared to 30 and 22% damping-off reduction when alginate beads or starch granules were delivered concomitantly with tomato seeds. Seed-coating with powder formulation of the biocontrol agent was as effective as drench application of the fungicide, oxine benzoate (No-Damp), in controlling Rhizoctonia damping-off and superior to the commercial biocontrol agent, Streptomyces griseoviridis (Mycostop), applied to tomato seeds as seed-coating.     

Effects of indomethacin on cardiac output distribution in normal and asphyxiated piglets

We determined the effect of breathing 9% CO₂/10% O₂/81% N₂ (asphyxia) on cardiac output distribution (microspheres) in 4–5 day old unanesthetized, chronically instrumented piglets prior to and following intravenous indomethacin administration. Thirty minutes of asphyxia caused Pa_CO2 to increase from 35 ± 2 mmHg to 66 ± 2 mmHg, Pa_O2 to decrease form 73 ± 4 mmHg to 41 ± 1 mmHg, and pH to decrease from 7.52 ± 0.05 to 7.21 ± 0.07. Arterial pressure was increased slightly but cardiac output was not changed significantly. Asphyxia caused blood flow to the brain, diaphragm, liver, heart, and adrenal glands to increase while causing decreases in blood flow to the skin, small intestine, and colon. Blood flows to the stomach and kidneys tended to decrease, but the changes were not significant. Treatment with indomethacin during asphyxia did not alter arterial pressure or cardiac output but decreased cerebral blood flow to the preasphyxiated level and decreased adrenal blood flow about 20%. Indomethacin did not alter blood flow to any other systemic organ. At this time the piglet was allowed to breathe air for 2.5 hr undisturbed. Two and a half hours after indomethacin administration, blood flows to all organs returned to the preasphyxia control levels with the exception of cerebral blood flow which was reduced (93 ± 13 to 65 ± 5 ml/100 g·min. Three hours after indomethacin administration, the cerebral hyperemia caused by asphyxia was less (134 ± 17b ml/100 g·min) than prior to indomethacin (221 ± 15 ml/100 g·min. Indomethacin did not alter the asphyxia-induced changes to any other systemic organ. We conclude that in newborn pigs, systemic treatment with indomethacin decreases cerebral blood flow and cerebral hyperemia in response to asphyxia, without affecting blood flow to any other systemic organ.     

Optimum conditions for the activation of the alternative complement pathway of a cyprinid fish (Tinca tinca L.). Seasonal variations in the titres

The present work describes the existence of a haemolytic activity in the serum of tench, Tinca tinca, against rabbit red blood cells (RRBC) which was identified as belonging to the alternative complement pathway from the following findings: haemolytic activity disappeared when the serum was heated to 45°C for 20 min; 10 mM EDTA, which chelates Ca²⁺ and Mg²⁺, induced a complete loss of haemolysis; Mg²⁺, but not Ca²⁺, was required for the activity, and the use of sheep red blood cells (SRBC), which have a high content of sialic acid, resulted in the serum activity falling to a very low degree of haemolysis. The ACH50 value (units ml^-1 serum) was defined as the reciprocal of the serum dilution necessary to lyse 50% of 4 × 10⁷ RRBC in a buffered medium of normal ionic strength (μ=0·15) containing 10 mM EGTA and optimum concentrations of Mg²⁺. The optimum conditions for the ACH50 assay were: pH 7·2-7-7; reaction temperature, 15°C; concentration of Mg²⁺, 5 mM; and reaction time, 90 min. Under these conditions, the values of ACH50 in spring, summer, autumn and winter for male tench were 69±13, 91±22, 90±36 and 137±41, and for female tench 100±11, 108±13, 82±12 and 145±17. The highest serum activity was found in the winter, suggesting the importance of this pathway during cold periods when the specific immune response is depressed in ectothermic vertebrates.     

Effect of gamma-irradiation on oocysts of Eimeria necatrix.

Effect of gamma radiation on oocysts of Eimeria necatrix was investigated. It was observed that oocysts exposed to 200 kR or above did not sporulate. Irratiation at 10-150 kR caused a progressive decrease in sporulation. Irradiation affected normal development of unsporulated oocysts as the zygote protoplasm divided into unequal masses or was shattered into granules. Increase in the intensity of irradiation of sporulated oocysts resulted in the progressive decrease in severity of the resultant infections in chicks and their effects - mortality, type of lesions developed, total oocyst production and immunity produced - were comparable with infections induced by decreasing the number of unirradiated oocysts. Infection produced by 1000 unirradiated oocysts was comparable with that resulting from 50 000 oocysts irradiated at 25 kR. Infection obtained with 20 000 unexposed oocysts approximated to that produced by 50 000 oocysts irradiated at 2-5 kR. It was concluded that irradiation abolished infectivity of the oocysts/sporozoites rather than bringing about attenuation of the parasite.     

Dose-dependent effect of radiation on angiogenic and angiostatic CXC chemokine expression in human endothelial cells

Blood vessel growth is regulated by angiogenic and angiostatic CXC chemokines, and radiation is a vasculogenic stimulus. We investigated the effect of radiation on endothelial cell chemokine signaling, receptor expression, and migration and apoptosis. Human umbilical vein endothelial cells were exposed to a single fraction of 0, 5, or 20 Gy of ionizing radiation (IR). All vasculogenic chemokines (CXCL1–3/5–8) increased 3–13-fold after 5 or 20 Gy IR. 20 Gy induced a marked increase (1.6–4-fold) in angiostatic CXC chemokines. CXCR4 expression increased 3.5 and 7-fold at 48 h after 5 and 20 Gy, respectively. Bone marrow progenitor cell chemotaxis was augmented by conditioned media from cells treated with 5 Gy IR. Whereas 5 Gy markedly decreased intrinsic cell apoptosis (0 Gy = 16% ± 3.6 vs. 5 Gy = 4.5% ± 0.3), 20 Gy increased it (21.4% ± 1.2); a reflection of pro-survival angiogenic chemokine expression. Radiation induces a dose-dependent increase in pro-angiogenic CXC chemokines and CXCR4. In contrast, angiostatic chemokines and apoptosis were induced at higher (20 Gy) radiation doses. Cell migration improved significantly following 5 Gy, but not 20 Gy IR. Collectively, these data suggest that lower doses of IR induce an angiogenic cascade while higher doses produce an angiostatic profile.     

A quantitative electron microscope autoradiographic study on I-human choriogonadotropin internalization in bovine luteal slices

Very few silver grains were seen on the cell surface and none intracellularly after incubation for 2 h at 4 °C. However, numerous grains were seen in various subcellular organelles when the tissues were incubated for 2 h at 22 ° or 38 °C. The grain distribution was qualitatively similar, but quantitatively, there were fewer grains at 22 ° than at 38 °C. Co-incubation of ¹²⁵I-hCG with excess unlabelled hCG resulted in the virtual disappearance of silver grains from all the subcellular organelles. Excess unlabelled human luteinizing hormone (but not follicle-stimulating hormone or prolactin) inhibited the appearance of silver grains in luteal tissue. There were no silver grains in bovine liver slices incubated with ¹²⁵I-hCG.The plasma membrane-associated grains progressively decreased, while intracellular organelle-associated grains increased with time at 38 °C. There were no grains in nuclei at 5 min, but they appeared at 10 min and increased until 120 min. After correction for radiation spread by three-step mask analysis, smooth endoplasmic reticulum and mitochondria did not contain any grains. The grain density was the highest in Golgi, followed by lysosomes, rough endoplasmic reticulum, nuclei, and plasma membranes after incubation for 2 h at 38 °C. Thus, the electron microscope autoradiography approach confirmed our biochemical data in the preceding paper (Chegini et al., Exp cell res 151 (1984) 466 [5]) on time, temperature dependency and specificity of ¹²⁵I-hCG internalization, association of internalized hormone with a variety of intracellular organelles, and the highest uptake in Golgi.     

The Determination of Phosphorus in Haemophilus influenzae Type b Conjugate Vaccines by Inductively Coupled Plasma-Atomic Emission Spectrometry

This study describes a method for the determination of phosphorus in lyophilized Haemophilus influenzae type b conjugate vaccines by inductively coupled plasma-atomic emission spectroscopy (ICP-AES). The concentration of polysaccharide is directly related to the concentration of phosphorus as measured in the laboratory. Phosphorus is present in the polyribosyl-ribitol phosphate (PRP) group of the Haemophilus influenzae type b conjugate vaccine. The repeating unit of PRP is 3-B-D ribose[1-1]ribitol-5-phosphate. Phosphorus in the final container is measured in μ g per dose. The amount of PRP is calculated from this and reported in μ g per dose. The Haemophilus influenzae type b conjugate vaccine was analyzed for phosphorus content within the range of 1·34 to 2·02 μg phosphorus per ml. The relative difference of phosphorus concentrations determined by the ICP-AES method from the phosphorus concentrations determined by the traditional colorimetric molybdate method ranged from 2·2 to 10·6%. Phosphorus spike recovery for the vaccine ranged from 93 to 99% (1·93±0·13 μ g P/ml). The phosphorus determination of NIST SRM 3139 phosphorus spectrometric solution differed by 3·0% from the certified phosphorus value (10·00 mg P/ml).     

Membrane Permeability Characteristics of Metaphase II Mouse Oocytes at Various Temperatures in the Presence of Me2SO

In this study, the hydraulic conductivity (L_p), Me₂SO permeability ( _Me2SO), and the reflection coefficients (ς) and their activation energies were determined for Metaphase II (MII) mouse oocytes by exposing them to 1.5 M Me₂SO at temperatures of 30, 20, 10, 3, 0, and −3°C. These data were then used to calculate the intracellular concentration of Me₂SO at given temperatures. Individual oocytes were immobilized using a holding pipette in 5 μl of an isosmotic PBS solution and perfused with precooled or prewarmed 1.5 M Me₂SO solutions. Oocyte images were video recorded. The cell volume changes were calculated from the measurement of the diameter of the oocytes, assuming a spherical shape. The initial volume of the oocytes in the isoosmotic solution was considered 100%, and relative changes in the volume of the oocytes after exposure to the Me₂SO were plotted against time. Mean (means ± SEM) L_pvalues in the presence of Me₂SO ( ^Me₂SO_p) at 30, 20, 10, 3, 0, and −3°C were determined to be 1.07 ± 0.03, 0.40 ± 0.02, 0.18 ± 0.01, 7.60 × 10⁻²± 0.60 × 10⁻², 5.29 × 10⁻²± 0.40 × 10⁻², and 3.69 × 10⁻²± 0.30 × 10⁻²μm/min/atm, respectively. The _Me2SOvalues were 3.69 × 10⁻³± 0.3 × 10⁻³, 1.07 × 10⁻³± 0.1 × 10⁻³, 2.75 × 10⁻⁴± 0.15 × 10⁻⁴, 7.83 × 10⁻⁵± 0.50 × 10⁻⁵, 5.24 × 10⁻⁵± 0.50 × 10⁻⁵, and 3.69 × 10⁻⁵± 0.40 × 10⁻⁵cm/min, respectively. The ς values were 0.70 ± 0.03, 0.77 ± 0.04, 0.81 ± 0.06, 0.91 ± 0.05, 0.97 ± 0.03, and 1 ± 0.04, respectively. The estimated activation energies (E_a) for ^Me₂SO_p, _Me2SO, and ς were 16.39, 23.24, and −1.75 Kcal/mol, respectively. These data may provide the fundamental basis for the development of more optimal cryopreservation protocols for MII mouse oocytes.     

Effect of furosemide on urinary excretion of prostaglandin E in normal volunteers and pateints with essential hypertension

Urinary excretion of prostaglandin E was measured radioimmunologically in 19 healthy persons ( 15 men and 4 women ) and in 16 patients ( 10 men and 6 women ) with essential hypertension before and after the administration of furosemide. The excretion rates were increased from 26.3±3.0 to 64.5±11.3 ng/hr in the former and from 11.9±2.7 to 26.9±85 ng/hr in the latter. There was a significant difference between them, healthy subjects showing a greater increase than patients with essential hypertension.There was an obvious sexual difference in urinary excretion of prostaglandin. In men, greater increase in the excretion rates was found than in the women. Greater increases were also obtained in healthy men than in hypertensive men and in healthy women than in hypertensive women. The present results suggest that furosemide enhances urinary excretion of prostaglandin E by mechanisms which entails either an increase in prostaglandin synthesis or a decrease in renal metabolism.     

Effect of variable dew temperatures on infection of green foxtail by Pyricularia setariae, Drechslera gigantea, and Exserohilum rostratum

A thermogradient apparatus was used to investigate the effect of variable dew temperatures on infection of green foxtail by the indigenous pathogen Pyricularia setariae (Ps) and the exotic pathogens Drechslera gigantea (Dg), and Exserohilum rostratum (Er) from the southern USA that showed bioherbicide potential against several grassy weeds. This device is capable of creating multiple diurnal temperature cycles, mimicking daily temperature fluctuations that occur under field conditions. Seven temperature regimes, i.e., 15/10 °C, 20/5 °C, 20/15 °C, 25/10 °C, 25/20 °C, 30/15 °C, and 30/25 °C (maximum/minimum), were used with temperature cycling from maximum to minimum and then back up to maximum in a 24 h period. Ps and Dg were much more virulent than Er on green foxtail, resulting in higher levels of disease and weed control. Dg was little affected by the dew temperatures in terms of plant infection and was more efficacious than Ps under cooler dew temperatures (15/10 °C and 20/5 °C), causing twice as much disease. This greater amount of disease coincided with higher conidial germination, appressorial formation and infection-hypha frequency by Dg at the lower temperatures. The efficacy of Ps improved as dew temperature increased, accompanied by a higher percentage of germination and more frequent appressorial production. Dg caused severe disease 2 d after inoculation whereas Ps required 4 d to initiate disease symptoms. These observations suggest that Dg is a superior candidate than Ps for green foxtail control on the Canadian prairies.     

Effect of extrusion under controlled temperature and moisture conditions on ileal apparent amino acid and starch digestibility in peas determined with young broilers

The objective of the experimental programme was to examine the influence of extrusion of peas under controlled temperature and moisture conditions on coefficient of ileal apparent digestibility (CIAD) of amino acids and to examine the changes to starch structure (crystallisation) and subsequent effects on CIAD of starch in young broilers. Experiment 1 used one pea cultivar (2.94 mg trypsin inhibitor units (TIU)/g DM) processed under two extrusion temperatures (exit temperatures of 70 and 140 °C) and two moisture levels (0 and 1.96l water added/h) giving a 2 × 2 factorial design (with a 5th treatment being the raw pea sample). All pea samples were added at a rate of 500 g/kg into a purified diet where peas were the only source of protein/amino acids. Processing generally led to a reduction in CIAD of amino acids but CIAD of starch for Raw, Treatments 1 and 3 (added water) was lower than that for Treatments 2 and 4 (no water) with data of 0.730, 0.808 and 0.759 vs. 0.852 and 0.834, respectively. Experiment 2 evaluated two near-isogenic lines of peas (HA5 and HB5) processed under the same conditions as for Experiment 1. Extrusion at 70 °C/addition of water had no significant effect on trypsin inhibitor activity but the inhibitor was eliminated at 140 °C with or without addition of water (P<0.001). Pea line HA5 generally had higher CIAD than HB5 (P<0.05 for cystine, lysine, leucine, histidine, alanine; P<0.01 for threonine). There were significant temperature × water addition interactions for CIAD (P<0.001 for all amino acids except methionine; P=0.006). For example, data for lysine were 0.760, 0.828, 0.860 and 0.759, respectively for 140 °C/no water, 140 °C/1.96 l/h, 70 °C/no water and 70 °C/1.96 l/h. CIAD for starch improved following processing, attributed to the loss of crsytallinity. The data indicate that peas are sensitive to high processing temperatures which are associated with a reduction in nutritive value, in terms of amino acid digestibility, for young poultry. In contrast, the results reinforce the adverse effect of crystallinity on starch digestibility and show that extrusion in the absence of water improves the digestibility of raw starches due to granular disorganisation (as evidenced by X-ray diffraction patterns) and changes in crystallinity.     

A comparison between the concentration of prostaglandin F in human plasma and serum

The concentration of prostaglandin F (PGF) has been measured in serum and plasma samples prepared under different conditions from the antecubital vein blood of 4 non-pregnant and 7 pregnant women. Prostaglandin F concentrations were less than 41 pg/ml in 19 samples of serum or plasma prepared by centrifugation within 30 minutes of collection. When the blood was allowed to clot at room temperature for 24 hours, highly variable, but usually markedly increased concentrations of PGF (<30 - 3020 pg/ml) were found in the serum. Plasma obtained from blood which stood at 23°C for 24 hours contained undetectable amounts of PGF in 4 out of 6 samples and less than 75 pg/ml in the 2 remaining samples. Plasma and serum obtained from blood which stood at 4°C for 24 hours contained less than 45 pg PGF/ml. These results show that (i) incubation of blood at room temperature may markedly elevate concentrations of PGF in serum, (ii) plasma samples rather than serum should be used for measurements of PGF concentrations.     

Single crystals of dextran: High temperature polymorph

Lamellar single crystals of a high temperature polymorph of synthetic dextran were prepared at temperatures ranging from 120 to 200°C in a mixture of water and polyethylene glycol. Individual crystals with lath-like shapes gave well resolved electron diffraction diagrams from which the reciprocal unit cell parameters a^*, b^* and γ^* could be measured. The direct cell parameters were then determined from a series of electron diffraction diagrams obtained by sequential tilting of the crystal about the b^* axis. This gave a = 0·922 ± 0·001 nm, b = 0·922 ± 0·001 nm, c (chain axis) = 0·78 ± 0·01 nm, α = γ = 90° and β = 91·3° ± 0·5°. The crystal symmetry was P2₁ with b as the unique monoclinic axis. These data coupled with the observed density of the crystals, indicated that the unit cell contained two antiparallel dextran chains of two residues each. When the crystals were grown at temperatures between 90 and 120°C, a percentage of crystals containing both low and high temperature polymorphs were obtained. These mixed crystals had most likely grown in syntaxy.     

In vitro studies of the effect of Environmental Conditions on the anthacnose pathogen of bananas, Colletotrichum musae

In vitro studies were undertaken to determine the effect of pH, temperature, water availability and carbon dioxide (CO₂) concentration on germination and growth of Colletotrichum musae, the causal pathogen of anthracnose of bananas. The optimum pH for germination and growth varied between 4·0 and 5·0 depending on temperature. At low pH (< 3·0) and 15°C, both germination and growth were significantly reduced, with a marked increase in the lag time, in days, prior to growth. C. musae germinated and grew over a wide range of water activities (a_w; 0·995−0·94 and 0·995−0·92, respectively) at 20, 25 and 30°C. In all cases where germination occurred appresoria were subsequently produced. Optimum growth occurred at 30°C and 0·995 a_w, although this changed to 0·98 a_w at 35°C. Increasing CO₂ concentration to 15% or reducing oxygen concentration to 1% resulted in a significant (P < 0·05) reduction in growth, but did not inhibit growth completely.     

Crosslinked potato starch as an affinity adsorbent for bacterial α-amylase

Crosslinked potato starch was prepared as an affinity adsorbent for bacterial α-amylase. To this end, reaction parameters for crosslinking in an ethanol/water solvent were investigated. The degree of crosslinking, and consequently the suitability of crosslinked starch as an adsorbent for α-amylase, changed by altering these parameters. An increase in the degree of crosslinking of the adsorbent caused lower affinity for bacterial α-amylase which resulted in an unfavourable decrease in adsorption capacity and a favourable decrease in the degradation of the adsorbent by the enzyme. 1 g of a suitable adsorbent for bacterial α-amylase, prepared with an epichlorohydrin/glucose monomer ratio of 0·65 (starch concentration 150 mg/ml, ethanol/water ratio 2·0, sodium hydroxide/epichlorohydrin ratio 1·0), can adsorb 9·8 mg of an α-amylase from B. licheniformis at 4°C in 20 h.The equilibrium constant between bound and unbound α-amylase is dependent on the temperature. An effective desorption was possible by a shift to higher temperatures. Degradation values smaller than 0·1% were measured after an incubation of 1 h at 70°C in a desorption buffer with 20% glycerol.It was concluded that coulombic interactions and hydrogen bonds are of no or little importance in enzyme adsorption. Van der Waals forces, which are responsible for the large temperature effect, are the main forces in the interaction between α-amylase and crosslinked starch.