Effect of prey stage on life-table attributes of a genetically manipulated strain ofMetaseiulus occidentalis (Acari: Phytoseiidae)

Metaseiulus occidentalis females from the carbaryl-organophosphate-sulfur resistant strain (COS) lived longer (25.3 days versus 19.7 days), had a higher total fecundity (43.8 eggs female^–1 versus 33.6 eggs female^–1) and a higher daily fecundity rate (2.4 eggs female^–1 day^–1 versus 2.0 eggs female^–1 day^–1), and exhibited a higher intrinsic rate of increase (0.243 individuals female^–1 day^–1 versus 0.182 individuals female^–1 day^–1) and shorter generation time (13.9 days versus 17.0 days), at 24–28°C, 47–56%rh under continuous fluorescent light, when reared on a diet of 0–48-h-old eggs rather than a diet of mixed actives ofTetranychus pacificus McGregor on bean leaf disks. The sexratio of the progeny was female-biased for both diets, 2.1 females to 1 male forM. occidentalis reared on eggs and 2.0:1 : forM. occidentalis reared on mixed actives, suggesting that diet influences sex-ratio in some unknown way.There was no significant difference in oviposition rates for repeatedly-mated and once-matedM. occidentalis females reared on a diet of younger (0–24-h old) eggs compared to a diet of older (72–96-h old) eggs ofT. pacificus.The COS strain ofM. occidentalis exhibited life-table parameters comparable to the other strains reported in the literature, suggesting that the reproductive attributes of this acarine predator were not reduced as a result of artificial laboratory selection. Diet, a biotic factor, produced substantial differences in life-table parameters, suggesting that this factor can influence conclusions regarding the potential efficacy of biological control agents.     

Adrenergic regulation of chloride secretion across the opercular epithelium: The role of cyclic AMP

Summary Activation of the -adrenergic receptors of the opercular epithelium ofFundulus heteroclitus stimulates Cl^– secretion, while activation of the -adrenergic receptors inhibits Cl^– secretion (Degnan and Zadunaisky, 1979). The possible involvement of adenosine 3, 5-monophosphate (cAMP) in these adrenergic responses was investigated. Isolated opercular epithelia incubated in Ringer, containing 10 mM theophylline, had cAMP levels ranging between 5.3 and 19.3 pmoles·mg protein^–1 (mean=9.5±1.0 pmoles·mg protein^–1). Activation of the -receptors by 10^–5 M isoproterenol increased the mean cAMP level 430% (P<0.001). Blockage of the -receptors with propranolol greatly reduced the increase in cAMP in response to isoproterenol. Activation of the -receptors by 10^–5 M arterenol stimulated the mean cAMP level 270% (P<0.01). However, when the -receptors were blocked with propranolol, arterenol had no effect on the cAMP level. The possible involvement of Ca⁺⁺ in these adrenergic responses was investigated. Neither the stimulatory effect of isoproterenol, nor the inhibitory effect of arterenol on the Cl^– secretion were diminished in the absence of extracellular Ca⁺⁺. The Ca⁺⁺ ionophore, A23187, and the calmodulin inhibitor, trifluoperazine, had no effects on the Cl^– secretion. The Ca⁺⁺-channel blocker, D600, had a significant inhibitory effect (P<0.005). Guanosine 3,5-monophosphate (cGMP) had no effect on the Cl^– secretion.The results indicate that -adrenergic stimulation of Cl^– secretion across the opercular epithelium is accompanied by an elevation in tissue cAMP levels. -adrenergic inhibition of Cl^– secretion does not involve changes in the tissue cAMP. Neither of these responses appear to require Ca⁺⁺.     

Thymus mast cell numbers following perinatal and adult exposures to low intensity 0.5 Hz magnetic fields

In a factorial design, 40 male 200-day-old rats that had been exposed from 2.5 days before to 2.5 days after birth to either 0.5 Hz rotating magnetic fields (RMFs) between 10^–3T to 10^–6T or to sham fields and maintained after weaning in one of two typical caging conditions were exposed as adults to either one of three 0.5 Hz RMF intensities (10^–6T, 10^–7T or 10^–8T) or to sham fields or to colony room control conditions. The numbers of mast cells (MCs/mm²) were determined for thymus tissues stained with thionin and toluidine blue. Thymuses from adult rats that had been perinatally exposed to the RMF displayed a marginally significant 20% to 35% elevation in MC numbers relative to sham-field controls. However the adult exposures did not sïgnificantlÿ affect the MC numbers. The two postweaning caging conditions, a non-magnetic field comparator variable, induced a significant 35% difference in MC numbers. The absence of sïgnificant perinatal by adult RMF exposure interactions indicated that early magnetic field exposure did not alter adult thymus responsivity to weaker but more natural intensity levels.     

Uptake and release of63Ni2+ byXenopus embryos during early cleavage stages

Summary Uptake and release of⁶³Ni was studied in dejelliedXenopus laevis embryos exposed to⁶³Ni²⁺ (0.3–30 mol/1) for 0.5-h intervals during the period 1–4.5 h post-fertilization (i.e. from first cleavage to early blastula stage). At first cleavage, the mean uptake of⁶³Ni by embryos was 12-17 times that by non-fertilized eggs, suggesting that conversion of the vitelline envelope to the fertilization envelope enhanced integumental permeability to⁶³Ni²⁺. 63 Ni uptake by embryos at the 1-2-cell stage averaged 1.8–2.5 times that at the early blastula stage. An average of 5% of total⁶³Ni in washed embryos was recovered in isolated fertilization envelopes, indicating that⁶³Ni²⁺ passed through the envelope into internal compartments. Progressive increases of⁶³Ni uptake were seen with increasing exposure levels; after exposure during 1–1.5 h post-fertilization to the highest concentration of⁶³Ni²⁺ (30 mol/1),⁶³Ni uptake averaged 11.4 (SD±5.1) pmol/embryo. Rapid efflux of⁶³Ni was noted after⁶³Ni²⁺-exposed embryos were transferred to nickel-free medium; mean⁶³Ni contents at 0.25 h and 2 h post-exposure diminished to 50% and 15% of the initial values, regardless of the exposure level. The finding thatXenopus embryos are permeable to⁶³Ni²⁺ during early cleavage stages provides a convenient experimental system to investigate the embryotoxicity and teratogenicity of nickel.     

Combined effects of cadmium,copper and lead on developing herring eggs and larvae

Eggs of Baltic herring were incubated (10°C; 16S) in sea water containing mixtures of Cd, Cu and Pb at concentrations of 0.56–5.0, 0.0167–0.15, 0.56–5.0 mg metal/l; embryonic survival until hatching, viable hatch and uptake of metals by embryos and early larvae were measured. Negative effects of metals on embryonic survival and viable hatch were additive in the case of Cu and Cd. Pb did not exer detrimental effects. Uptake of metals with exposure time was non-linear in eggs and linear in larvae. Total uptake of Cu and Cd by eggs was subjected to antagonistic or synergistic action of the other two metals present. Accumulation of Pb by eggs was enhanced when Cu was also present.     

On the dynamics of exploited populations ofTisbe holothuriae (Copepoda,Harpacticoidae)

A total of 90, weekly exploited populations of the harpacticoid copepodTisbe holothuriae Humes were exposed to 148, 222, 333, 500, 750 or 1125µg Cd⁺⁺ l^–1, combined with exploitation rates of 10, 30, 50, 70 or 90% under conditions of surplus food supply at 22° C and 30 S. All populations exposed to concentrations down to 500µg Cd⁺⁺ l^–1 and 3 populations (out of 15) exposed to 333µg l^–1 became extinct within the experimental period of 30 weeks. Survival time significantly depended on concentration. A recovery phase from an initially high mortality preceded eventual population extinction after adding 500µg Cd⁺⁺ l^–1. In the initial phase, higher nauplii mortality prevailed. During these experiments on acute intoxication, no relationship could be established between survival and exploitation rate. However, experiments on the effects of stepwise increases in Cd concentration (results not yet published) produced evidence of such a relationship. In spite of increased mortality, no significant numbers of dead copepods were detected in weekly samples because of their rapid decomposition and cannibalism, which depends on the amount of food available. Sampling regimes of 5 times per week yielded significant numbers of dead individuals.     

Induction of desiccation tolerance in microspore-derived embryos ofBrassica napus

Summary A method was developed to induce desiccation tolerance in microspore-derived embryos ofBrassica napus. Treatment of 14- to 20-day-old embryos with 1&#x00D7;10^&#x2212;4 M abscisic acid, in the light, induced tolerance to slow desiccation over a 6-day period. Under these conditions 88 to 100% of embryos of the five cultivars tested survived (as measured by moisture uptake, greening, and growth of the shoot and root meristem) after storage for 1 wk at tissue water content levels of less than 20%. The response was found to be dependent on the abscisic acid concentration in the culture medium and time of exposure of the embryos to the abscisic acid-containing medium, with exposure times of as little as 1 day having a beneficial effect. Exposure times to abscisic acid (ABA) of 5&#x2013;7 days resulted in the highest survival rates. Embryo age and size at the time of ABA exposure also affected the subsequent survival and development of embryos, with older and larger embryos exhibiting the best responses.     

Enhanced plant regeneration from microspore-derived embryos of Brassica napus by chilling,partial desiccation and age selection

Germination was readily induced in recalcitrant microspore-derived embryos of Brassica napus Topas when they were exposed to a period of chilling (9–12 days at 4°C) or partial desiccation (rapid or slow air drying) prior to germination. In general, embryos thirty-five days old had the highest germination rates as compared to younger or older ones. Populations of embryos were induced to germinate at a rate of over 90% under specific temperature, desiccation and age conditions. Comparisons to an embryogenic B. napus winter line, F346, are made.     

Response of egg temperature,heart rate and blood pressure in the chick embryo to hypothermal stress

Summary Chicken eggs incubated for 12–18 days were catheterized via the allantoic artery and temperature was monitored simultaneously using a probe positioned in the allantoic fluid adjacent to the embryo. Fluid temperature (referred to as egg temperature), arterial pressure and heart rate were measured following abrupt exposure to a lower environmental temperature (ca., 2628°C). Egg temperature and heart rate diminished exponentially: The rate of decline of egg temperature approximated Newton's law of cooling, the rate coefficient being 0.0220.025°C/min·°C throughout the incubation period from 12 to 18 days. The half time of temperature response of the egg was 2728 min. The response was much slower than that of fertile unincubated eggs (Kaplan et al. 1978), suggesting that the extraembryonic fluids act as a thermal buffer in embryonated eggs. The heart rate response in older embryos (1718 days) changed in the same manner as egg temperature, while in younger embryos (1216 days) the heart rate diminished more quickly than the change in egg temperature. During development the cardiac pacing of the embryo suggests that it becomes resistive to mild cold stress. The systolic pressure remained almost unchanged or even increased during one hour of exposure as the embryo developed, while the diastolic pressure decreased steadily after exposure irrespective of development. The 18-day-old embryos retained the systolic pressure unaltered during 3-hour exposure. In embryos 34 days prior to hatching the functional capacity of the heart apparently allows continued pumping even after prolonged exposure to low environmental temperature. Symbols and Abbreviations: See definitions at the end of the Materials and methods section     

Voltage-gated chloride currents in cultured canine tracheal epithelial cells

Summary Chloride ions (Cl^–) are concentrated in airway epithelial cells and subsequently secreted into the tracheal lumen by downhill flux through apical Cl^– channels. We have studied Cl^– currents in cultured canine tracheal cells using the whole-cell voltage-clamp technique. Ultrastructural techniques demonstrated that the cells used in the electrophysiological experiments possessed apical membrane specializations known to be present in the intact, transporting cell type. Cultured cells 2–6 days old were characterized by an input resistance of 3.4±0.8 G (n=11) and a capacitance of 63.8±10.8 pF (n=26). A comparison of 3 and 4 day-old cells with 5 and 6 day-old cells showed that the input resistance decreased almost 50%, and the cell capacitance and the inward and outward currents increased concomitantly approximately 200%. Cultured cells 3–4 days old held at –40 mV produced currents of 196±22 pA at 50 mV and –246±27 pA at –90 mV (n=212) with pipette and bath solutions containing primarily 140 KCl and 140 NaCl, respectively. The chloride channel blocker diphenylamine-2-carboxylate (DPC, 100 m) suppressed whole-cell currents by 76.8% at 60 mV; however, currents were unaffected by the stilbenes SITS (1mm) and DNDS (1–30 m). Replacement of K⁺ with Cs⁺ in the pipette solution did not affect the outward current, the current reversal potential, or the input resistance of the cells, indicating that the current was not significantly K⁺ dependent when the intrapipette solution was buffered to a Ca²⁺ concentration of 20nm. The Cl^–/Na⁺ permeability ratio was estimated to be greater than 11 as calculated from reversal potential measurements in the presence of an internal to external NaCl concentration ratio of 12. Current equilibrium permeabilities, relative to Cl^– were: I^– (2.9)NO ₃ ^– (1.1)Br^– (1.1)Cl^– (1.0)F^– (0.93)MeSO ₄ ^– (0.19)gluconate (0.18)aspartate (0.14). Depolarizations to potentials greater than 20 mV elicited a time-dependent component in the outward current in 71% of the cells studied. Currents inactivated with a double exponential time course at the most depolarized voltages. Recovery from inactivation was fast, holding potential-dependent, and followed a double exponential time course. Current amplitude was increased via a cAMP-dependent pathway as has been demonstrated for single Cl-selective channels in cell-attached patches from cultured canine and human tracheal epithelial cells. Forskolin, an activator of adenylate cyclase, produced a 260% increase in the outward current at +50 mV. In summary, cultured canine tracheal cells have a single resting conductance that is Cl^– selective, voltage-dependent, and modulated by a cAMP-dependent mechanism. This preparation appears to be appropriate for analysis of cellular modulation of airway Cl^– channels and Cl^– secretion.     

Die Erzeugung von Chemilumineszenzen wÄ\riger TlI-Salzlösungen durch Röntgenstrahlen

Zusammenfassung Die Lumineszenz wÄ\riger Tl⁺-Lösungen unter Einwirkung von 8,5-keV_eff-Röntgenstrahlen wird untersucht, und zwar 1. in neutraler Lösung, 2. in saurer Lösung, 3. in alkalischer Lösung, 4. bei Zusatz von NaCl.Bei Änderung des pH-Wertes nimmt die Lichtausbeute sowohl auf der sauren Scite (beipH=3,4) als auch auf der alkalischen Scite (beipH=12) ein Maximum an; sie erreicht dort den 2- bis 3fachen Wert der Ausbeute in neutraler Lösung. Andererseits ist bei Zusatz von 1 M NaCl in neutraler Lösung die Ausbeute fünfmal höher als bei Abwesenheit von NaCl; die Maxima im sauren und alkalischen Bereich verschwinden jedoch bei 1 M NaCl vollkommen, und man erhÄlt bei gro\en und bei kleinen pH-Werten lediglich eine Löschung der Lumineszenz.Die Lumineszenz kommt durch Bildung von Tl^+* bei der Anlagerung von e_aq an Tl⁺⁺ zustande. Tl⁺⁺ entsteht jedoch nicht unmittelbar gemÄ\ Tl⁺ + OH Tl⁺⁺ + OH^–, sondern zunÄchst bildet sich Tl⁺OH, und dieses dissoziiert erst nach einer Verzögerung von > 10^–7 sec in Tl⁺⁺ und OH^–. Die Sensibilisierung durch H⁺ wird durch die Reaktion Tl⁺OH + H⁺ Tl⁺⁺ + H₂O und die durch OH^– durch Tl⁺OH + OH^– Tl⁺⁺O^– + H₂O erklÄrt. Die Reaktion der Komplexe Tl⁺Cl^– und Tl⁺Cl₂ ^– mit OH erfolgt offenbar ohne zeitliche Verzögerung, d. h. Tl⁺⁺Cl^–– bzw. Tl⁺⁺Cl₂ ^–– wird dabei innerhalb einer Zeit 10^–7 sec gebildet.

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Chemiluminescences of aqueous Tl^I solutions produced by irradiation with 8.5-keV-X-rays

Summary The luminescence of aqueous solutions of Tl⁺ during irradiation with 8.5-keV-X-rays has been investigated. In particular we have studied the Tl⁺-luminescence in 1. neutral solutions, 2. acid solutions, 3. alkaline solutions, 4. solutions containing various concentrations of NaCl.Varying thepH the luminescence yield passes a maximum on the acid side as well as on the alkaline side (atpH=3.4 respectivelypH=12). Compared with neutral solutions the luminescence yield is increased by a factor 2 to 3 at these pH-values.By adding 1 M NaCl to neutral Tl⁺-solutions the luminescence yield is enhanced by a factor five, however in dependence ofpH no further increase has been observed, but only quenching of the luminescence at low and highpH.The luminescence origins from the formation of Tl^+* by the reaction of e_aq with Tl⁺⁺ formed by oxidation of Tl⁺ by OH. However, Tl⁺⁺ is not formed directly by Tl⁺+OH Tl⁺⁺ + OH^– but via dissociation of the intermediate Tl+OH after a delay > 10^–7 sec.We explain the increase of the luminescence yield in acid solutions by the reaction Tl⁺OH + H⁺ Tl⁺⁺ + H₂O and in alkaline solutions by the reaction Tl⁺OH + OH^– Tl⁺⁺O^– + H₂O. In alkaline solutions the luminescence spectrum shifts to longer wavelengths; we conclude, that this spectrum is attributed to Tl^+*O^–. Evidently the reaction of Tl+Cl^– and Tl+Cl₂ ^–– with OH leads to formation of Tl⁺⁺Cl^– and Tl⁺⁺Cl₂ ^–– without any efficient delay.

     

Glucan-induced modification of the increased susceptibility of cyclophosphamide-treated mice to Staphylococcus aureus infection

Summary Glucan, a 1–3-polyglucosidic component of the cell wall of Saccharomyces cerevisiae, was evaluated for its ability to modify experimentally induced S. aureus septicemia in an immunosuppressed mouse model. AKR/J mice were injected with glucan (0.45 mg), cyclophosphamide (0.6 mg), isovolumetric saline (0.5 ml), or glucan (0.45 mg) and cyclophosphamide (0.6 mg) on days –10, –7, –4 and –1 prior to intravenous challenge with 1.0×10⁹ S. aureus on day 0.In contrast to the significant (P<0.05) decrease in leukocytes observed in the cyclophosphamide-treated mice, the administration of glucan to cyclophosphamide-treated mice resulted in maintenance of the peripheral leukocyte counts. Furthermore, glucan, as a single pretreatment regimen, resulted in a median survival time of 12.5 days, as against only 7.5 days in the saline control group. A 1.4-day median survival was observed in mice pretreated with cyclophosphamide and subsequently challenged with S. aureus. However, when glucan and cyclophosphamide were administered together, a median survival time of 9.0 days was observed.Histopathologic examination revealed that glucan administration inhibited the renal necrosis observed in both normal and cyclophosphamide-treated mice following staphylococcal challenge. Glucan also produced a marked reduction of hepatic pathology in cyclophosphamide-treated mice following S. aureus challenge. These data denote that glucan administration is effective in altering morbidity and mortality due to systemic S. aureus disease in cyclophosphamide-treated mice.     

Acid-base and electrolyte regulation,and haemolymph gas transport in crayfish,Astacus astacus,exposed to soft,acid water with and without aluminium

Summary Intermoult crayfish (Astacus astacus) were exposed to acid (pH 4), soft water ([Ca⁺⁺]=100 mol·l^–1) in the absence and presence of aluminium (25 mol·l^–1) for variable time periods (up to 21 days) in order to assess the consequences for acid-base and electrolyte balance and haemolymph gas transport. Haemolymph osmolality and concentration of major ions decreased drastically and to a similar extent in acid and acid-aluminium water. Muscle tissue ion concentrations were, however, regulated at an almost constant level. A severe metabolic acidosis was gradually developed, attaining a haemolymph metabolic acid load of 6–7 mequiv·l^–1 after 12–21 days. The acidosis was partially compensated by ventilatory means, with the postbranchial haemolymph PCO₂ decreasing earlier in acidaluminium-exposed than in acid-exposed specimens. Hyperventilation seemed to be a direct acid-base regulatory response, since the rise in postbranchial PCO₂ had only minimal influence on haemolymph O₂ transport. The Bohr effect of Astacus astacus haemocyanin was low (log P₅₀/GdpH=-0.24), and the mean P₅₀ only increased from 15 to 19 mmHg after 21 days of acid exposure. The decrease in O₂ affinity with decreasing pH was accompanied by a decrease in the cooperativity of O₂ binding. The haemolymph haemocyanin concentration was not affected by acid and acid-aluminium exposure, but decreased after 21 days due to starvation. Muscle tissue aluminium concentrations were unaffected, whereas gill tissue concentrations increased in acid-aluminium exposed crayfish, most likely due to accumulation of aluminium on the gill surface. Mortality was low, and an internal hypoxia and lactacidosis was not developed in either of the experimental groups. This suggests that the gas transfer qualities of the chitincovered gills of crayfish are much less sensitive to acid and acid-aluminium stress than the gills of teleost fish.Abbreviations Hc haemocyanin - SO₂ saturation of Hc with O₂ - P ₅₀ oxygen tension of haemolymph at 50% SO₂ - n ₅₀ Hills coelficient around 50% SO₂     

Morphological changes of integumental chloride cells to ambient cadmium during the early development of the teleost, Oreochromis mossambicus

Synopsis Newly-hatched embryos of Oreochromis mossambicus were reared in freshwater and treated with 0 (control), 50 (low level) or 200 (high level) ppb cadmium for 4 days. Changes in the numbers and dimensions of chloride cell apical crypts on the skin of the free embryos were examined daily using scanning electron microscopy. The apical crypts of the chloride cells were rarely observed on the skin of the embryo trunk, and unevenly distributed on the surface of the yolksac. Two days after hatching, the chloride cells of the free embryos exposed to 50 ppb Cd were more active than those of the other two groups. Compared with the control group, the maximum dimensions of the developing apical crypts were stimulated by 50 ppb Cd and inhibited by 200 ppb Cd. The results indicated that the development of chloride cells in tilapia free embryos was provoked by low level Cd exposure and stunted by high level Cd exposure, suggesting the existence of structure/function relationships in which the activation of chloride cells may be related to the ionoregulatory mechanism in adaptation to Cd exposure.     

Technical considerations for assessing alterations in skeletal muscle sarcoplasmic reticulum Ca++-sequestration functionin vitro

A multiple measurement system for assessing sarcoplasmic reticulum (SR) Ca⁺⁺-ATPase activity and Ca⁺⁺-uptake was used to examine the effects of SR fractionation and quick freezing on rat white (WG) and red (RG) gastrocnemius muscle.In vitro measurements were performed on whole muscle homogenates (HOM) and crude microsomal fractions (CM) enriched in SR vesicles before and after quick freezing in liquid nitrogen. Isolation of the CM fraction resulted in protein yields of 0.96±0.1 and 0.99±0.1 mg/g in WG and RG, respectively. The percent Ca⁺⁺-ATPase recovery for CM compared to HOM was 14.5% (WG) and 10.1% (RG). SR Ca⁺⁺-activated Ca⁺⁺-ATPase activity was not affected by quick freezing of HOM or CM, but basal ATPase was reduced (P<0.05) in frozen HOM (5.12±0.18–3.98±0.20 mole/g tissue/min in WG and from 5.39±0.20–4.48±0.24 mole/g tissue/min in RG). Ca⁺⁺-uptake was measured at a range of physiological free [Ca⁺⁺] using the Ca⁺⁺ fluorescent dye Indo-1. Maximum Ca⁺⁺-uptake rates when corrected for initial [Ca⁺⁺]_f were not altered in HOM or CM by quick freezing but uptake between 300 and 400nM free Ca⁺⁺ was reduced (P<0.05) in quick frozen HOM (1.30±0.1–0.66±0.1 mole/g tissue/min in WG and 1.04±0.2–0.60±0.1 mole/g tissue/min in RG). Linear correlations between Ca⁺⁺-uptake and Ca⁺⁺-ATPase activity measured in the presence of the Ca⁺⁺ ionophore A23187 were r=+0.25, (P<0.05) and r=+0.74 (P<0.05) in HOM and CM preparations, respectively, and were not altered by freezing. The linear relationships between HOM and CM maximum Ca⁺⁺-uptake (r=+0.44, P<0.05) and between HOM and CM Ca⁺⁺-ATPase activity (r=+0.34, P<0.05) were also not altered by tissue freezing. These data suggest that alterations in maximal SR Ca⁺⁺-uptake function and maximal Ca⁺⁺-ATPase activity may be measured in both HOM and CM fractions following freezing and short term storage. (Mol Cell Biochem139, 41–52, 1994)     

Acetylcholine activates a chloride channel as well as glutamate and GABA

Summary In conventional two microelectrode experiments, acetylcholine had qualitatively the same effect as GABA and glutamate on membrane potential and input resistance of muscle fibres of the opener and intrinsic stomach muscles of crayfish (Austropotamobius torrentium). In patch-clamp experiments, acetylcholine occasionally elicited single channel openings in cell-attached patches on these muscles. If outside-out patches were excised and the Cl^– concentration was high on both sides of the membrane, acetylcholine at concentrations of 1 nM regularly elicited single channel currents. The amplitude of single channel currents depended strongly on the intracellular concentration of Cl^–. The reversal potential of the channel, determined after replacing intracellular K⁺ with Cs⁺, corresponded to the Nernst potential for Cl^–. The voltage dependence and the reversal potential of single channel current amplitudes elicited by ACh, glutamate and GABA were identical. The distribution of life times of openings (>1 ms) elicited by ACh and glutamate could be fitted by a single exponential with a time constant of about 2.5 ms, corresponding to the mean open time. ACh and glutamate applied to the same outside-out patch showed cross-desensitization, and thus ACh and glutamate activate the same channels. An excitatory, cationic ACh-activated channel could not be identified. Permeabilities of the chloride channel were calculated according to the Goldman-Hodgkin-Katz equation at different membrane potentials. Negative single channel current amplitudes (inward currents) could be fitted with a permeability of ₂= 3.9×10^–14 cm³s^–1. For positive currents (outward) the channel had a permeability of ₁= 1.4× 10^–14 cm³s^–1. The permeability of the channel declined from 16×10^–14 cm³s^–1 to 2.3×10^–14 cm³s^–1 if the intracellular Cl^–-concentration was raised from 6 to 257 mM. The activation elicited by acetylcholine was inhibited by extracellular Ca⁺⁺. The mean current activated by ACh was reduced by a factor of 50 if the extracellular concentration of Ca⁺⁺ was raised from 0.1 mM to the physiological concentration of 13.5 mM.     

The effects of UV-B radiation on the reproduction and mortality of Tigriopus californicus (Copepoda: Harpacticoida)

Survival of adult female copepods and survival of eggs to 1-week-old larvae were assessed following exposure times of 0, 15, 30, 45, 60 and 90 min of UV-B radiation (450 W cm^–2). An increase in both adult and larval mortality occurred between the 45 min (12,150 J m^–2 unweighted) and 60 min (16200 J m^–2 unweighted) treatment levels of radiation. At 90 min (24300 J m^–2 unweighted), all individuals died.     

Effects of intraguild predation on aphid parasitoid survival

To assess the potential selection pressure caused by intraguild predation between predators and parasitoids of aphids an estimate was made of the predation risk to Aphis fabaeScop. mummified by Lysiphlebus fabarum(Marshall, 1896) on sugar beet. Mummified aphids were exposed to a natural community of predators. Their survival time was estimated during a 10-day field survey. Additionallythe role of alternative prey on parasitoid survival was investigated by adding unparasitised aphids to half of the mummy aggregations.The field data were evaluated by survival analysis. Two covariates were tested within a Cox proportional hazard model: (i) the presence of the alternative prey and (ii) the patch structure (number of proximal mummies attacked). Within 4–5 days after exposure predators destroyed approx. 50% of the mummies. The model with both covariates revealed a significant difference concerning survival of the mummies in the two treatments (Likelihood ratio test, ²=78.03, P=0.0001). Alternative prey reduced the predation risk on mummies by 29%, while a high level of predation on proximal mummies increased the individual predation risk by 4%. The results are discussed in the context of prey location by predators and the evolution of anti-predator mechanisms.     

Pilot study of treatment of biochemotherapy-refractory stage IV melanoma patients with autologous dendritic cells pulsed with a heterologous melanoma cell line lysate

Eleven AJCC stage IV melanoma patients with progressive disease after treatment with biochemotherapy were treated with autologous dendritic cells pulsed with heterologous tumor cell lysates. The vaccine used mature DCs (CD1a⁺⁺⁺, CD40⁺⁺, CD80⁺⁺, CD83⁺, and CD86⁺⁺⁺) generated from peripheral blood monocytes in the presence of GM-CSF and IL-4. After 7 days, DCs were matured with a defined cocktail of cytokines (IL-1+IL-6+TNF-+PGE₂) and simultaneously pulsed with lysates of heterologous melanoma cell lines, for 2 days. A total of 4×10⁶ DCs was injected monthly under ultrasound control in an inguinal lymph node of normal appearance. The study was closed when all patients died as a consequence of tumor progression. No sign of toxicity was observed during the study. One patient experienced a partial response lasting 5 months, and two patients showed a mixed response which lasted 3 months. The median survival of the whole group was 7.3 months (range 3–14 months). This vaccination program had specific antitumoral activity in highly pretreated and large tumor burden stage IV melanoma patients and was well tolerated. The clinical responses and the median survival of the group of patients, together with the low toxicity of our DC vaccine, suggest that this approach could be applied to earlier AJCC stage IV melanoma patients.     

Electrophoretic analysis of newly synthesized and stored maternal RNA during oogenesis ofCalliphora erythrocephala (Dipt.)

Summary Ovaries ofC. erythrocephala synthesize large amounts of poly(A)⁺ and poly(A)^– RNA during early and middle stages of oogenesis as shown by labelling with³H-uridine in vivo. After incubation for 1 h, a striking difference in the electrophoretic pattern of newly synthesized labelled poly(A)⁺ RNA and the poly(A)⁺ RNA present in sufficient amounts for optical density measurements (steady state poly(A)⁺ RNA) was observed. During early and mid-oogenesis, in the poly(A)^– RNA fraction, 4S predominantly mature rRNA, 5S RNA and tRNA were labelled. These fractions were no longer synthesized during late oogenesis, whereas poly(A)⁺ RNA was labelled continously During oogenesis stage specific differences in the size distribution of newly synthesized and steady state poly(A)⁺ RNA were not obvious. However, different sizes of labelled poly(A)⁺ RNA species were detected in 0–2h old preblastoderm embryos, after injection of³H-uridine into females either 3–4 days (stage 3–4 of oogenesis) or 24 h before oviposition (stage 5–6 of oogenesis). This difference in RNA synthesis was related to the presence of active nurse cell nuclei. The poly(A)⁺ RNA fraction represents about 2–3% of the total RNA in both ovaries and freshly laid eggs as judged by measurements of optical density and radioactivity bound to oligo(dT). The length of poly(A)-segments in ovarian poly(A)⁺ RNA varied from about 30 to 200 nucleotides.