Evaluation of hyperactivity produced by pyrethroid treatment on third instar nymphs of Triatoma infestans (Hemiptera: Reduviidae)

The hyperactivity produced in third instar nymphs of Triatoma infestans by their exposure to films of deltamethrin or cis-permethrin was evaluated. Both pyrethroids produced a significant increase in locomotor activity at 26 and 36°C but not at 16°C. At 26°C, only deltamethrin produced hyperactivity when topically applied on the dorsal side of the nymphs abdomen. However, both pyrethroids produced hyperactivity when topically applied on the head of the nymphs. Hyperactivity was not observed when nymphs were treated with N-ethylmaleimide (20 μg/insect) before exposure to the pyrethroids. The effect of both insecticides on locomotor activity reversed the inhibitory influence on locomotion elicited by contact with the walls of the experimental arena (thigmotaxis). Arch. Insect Biochem. Physiol. 35:323–333, 1997.© 1997 Wiley-Liss, Inc.     

Interaction and reactivity of urocanic acid towards peroxyl radicals

Abstract

The capacity of urocanic acid to interact with peroxyl radicals has been evaluated in several systems: oxidation in the presence of a free radical source (2,2′-azobis(2-amidinopropane; AAPH), protection of phycocyanin bleaching elicited by peroxyl radicals, and Cu(II)- and AAPH-promoted LDL oxidation. The results indicate that both isomers (cis and trans) are mild peroxyl radical scavengers. For example, trans-urocanic acid is nearly 400 times less efficient than Trolox in the protection of the peroxyl radical promoted bleaching of phycocyanin. Regarding the removal of urocanic acid by peroxyl radicals, nearly 100 μM trans-urocanic acid is required to trap half of the produced radicals under the employed conditions (10 mM AAPH, 37°C). Competitive experiments show that the cis-isomer traps peroxyl radicals ~30% less efficiently than the trans-isomer. Given the high concentrations that trans-urocanic acid reaches in skin, its capacity to trap peroxyl radicals could contribute to the protection of the tissue towards ROS-mediated processes. Furthermore, both isomers, and particularly the cis-isomer, protect LDL from Cu(II)-induced oxidation.     

The effect of temperature on the behaviour and development of Triatoma brasiliensis

Abstract. The effects of temperature on the development of early stages and the thermopreference of nymphs and adults were analysed in the haematophagous bug Triatoma brasiliensis Neiva, 1911 (Hemiptera, Reduviidae). Egg hatching, mortality of nymphs, feeding and moulting success of the early stages of T. brasiliensis were all affected by temperature. While high rates of egg hatching were observed between 25 and 27 °C, no hatching occurred at 12, 19 and 38 °C. The mortality of first‐instar nymphs was highest at 38 °C, at which no insects survived after 10 days of exposure. Feeding success was only affected at the lowest temperature (12 °C). No ecdysis was observed in the groups exposed to 12, 19 and 21 °C. Recently fed fourth‐instar nymphs preferred to stay at a temperature of approximately 30 °C. The preferred temperature began to decline gradually to approximately 27 °C during ecdysis, reaching 26 °C at 30 days after ecdysis. After a second blood meal, the insects' preferred temperature was again approximately 30 °C. The thermopreference pattern of females was similar to that of nymphs. Nymphs and females showed a daily fluctuation in their preferred temperature, moving towards higher values at the beginning of the dark phase, and choosing lower ones after this time interval, at which they remained until the end of the light phase. The females laid their eggs in all sectors of the arena, although the largest numbers of eggs were found between 28 and 32 °C.     

High temperature determines the ups and downs of small brown planthopper Laodelphax striatellus population

Small brown planthopper, Laodelphax striatellus (Fallén) numbers usually drop sharply in the summer and revive quickly in the autumn. However, it is unclear whether and how the high temperature plays a role in this process. The effects of durations of heat exposure (33°C) on life‐history traits were examined here. Exposure of adults for 1 day during the oviposition stage led to a very low survival of nymphs. The average longevity of L. striatellus exposed for 1–31 days from oviposition was significantly longer than that of the control (27°C). Short‐term (1–5 days) heat exposure of the third instar nymphs did not significantly influence eclosion, but exposure of the fourth instar nymphs significantly increased eclosion. Lifespan from egg to adult was significantly lengthened when the third instar nymphs were exposed to heat for 2–15 days, or the fourth instar were exposed for 10 days. The preoviposition period was prolonged by heat exposure of the third or fourth instar nymphs. Short‐term heat exposure of less than 3 days of the third or fourth instar nymphs did not restrict fecundity, but when the exposure duration exceeded 5 days the total eggs per female and hatchability decreased. Exposure to high temperature increased the brachypter rate of adults. In summary, low survival and slowing development under heat exposure resulted in population decline in the summer, and the relatively high fecundity and brachypter rate led to quick revival in autumn. Temperature in the summer determines the rise and fall in numbers of L. striatellus.     

Effect of Starvation on the Searching Path of the Predatory Bug <Emphasis Type="Italic">Deraeocoris Lutescens</Emphasis>

As physiological factors may influence the predatory behavior of insects, this study was designed to evaluate the possible impact that hunger has on the search path of a polyphagous predator: Deraeocoris lutescens Schilling (Heteroptera: Miridae). D. lutescens was fed a diet of Rhopalosiphum padi (Linné) (Homoptera: Aphididae) nymphs and Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) eggs at 21±1 °C and a photoperiod of 16:8 (L:D). After moulting, third and fifth instar nymphs and adults were placed in Petri dishes containing an abundant food source for 24 h. After 24 h, the food was removed and groups were randomly assigned to increasing periods of prey deprivation. Each D. lutescens nymph or adult was placed at the center of an empty arena and its searching path was recorded to calculate different path parameters and to determine the type of movement mode adopted. Results show an effect of starvation on both nymphal and adult D. lutescens searching paths. Satiated third instar nymphs manifested an intensive search path whereas fifth instar nymphs and adults adopted an intermediate one. When starved, the extensive search path appeared early in young nymphs, and later in the last instar nymphs and adults, according to the degree of prey deprivation.     

Acclimatisation in the green spruce aphid, Elatobium abietinum

Thin layer chromatography separation of 80% ethanol extracts of adult Elatobium abietinum revealed the presence of the polyhydric alcohol mannitol in aphids overwintering outdoors but not in aphids kept permanently indoors at 15°C. After 3 days at 15°C no traces of mannitol were left in overwintering aphids. Mean freezing temperatures of outdoor, unfed instar I nymphs were about 4°C lower than those of unfed instar I nymphs produced at 15°C. Mean freezing temperatures of overwintering adults were considerably higher than those of unfed instar I nymphs and showed no changes associated with time at 15°C following transference indoors. Similarly, mean freezing temperatures of Sitka spruce needles transferred to 15°C did not change. It was concluded that, although freezing was mainly avoided by supercooling, the presence of mannitol lowered the true freezing temperature of aphid haemolymph and, consequently, the actual freezing temperatures of nymphs produced under cold conditions. However, the considerable increase in freezing point temperatures caused by imbibition of plant sap masked these acclimatisation changes in feeding aphids.     

Laboratory evaluation of predation rates of two native natural enemies on the exotic pest Bagrada hilaris

Laboratory studies were conducted to determine the consumption rates of two native predators found attacking the exotic invasive stink bug Bagrada hilaris (Burmeister) (Hempitera: Pentatomidae) in field plots in New Mexico, USA. Individual field‐collected adults of the spined soldier bug, Podisus maculiventris (Say) (Hempitera: Pentatomidae) and the soft‐winged flower beetle, Collops vittatus (Say) (Coleoptera: Melyridae), were provided daily with fixed numbers of different life stages of B. hilaris under controlled conditions. Consumption rates were recorded daily for ten consecutive days for a total of 20 adult P. maculiventris and 20 adult C. vittatus per prey life stage. For P. maculiventris, predation rates were obtained in relation to adult, third and fifth instar prey, and for C. vittatus for first, second and third instar prey. On average, predation on third and fifth instar B. hilaris nymphs by P. maculiventris was 0.6 ± 0.1 and 0.9 ± 0.1 per day respectively. Predation rates on adults were slightly higher (1.3 ± 0.1 per day), with female prey being consumed at a significantly higher rate than male prey when three mating pairs of B. hilaris were provided per day (0.8 ± 0.1 females per day vs. 0.5 ± 0.1 males per day). Collops vittatus adults provisioned daily with 20 first instar B. hilaris nymphs killed a mean total of 4.7 ± 0.4 and 9.3 ± 0.6 prey each day (for male and female beetles respectively), with only approximately half that number of prey being fully consumed. Partial consumption of prey by this species was also observed with second and third instar nymphs, but to a lesser degree. Female beetles consumed significantly more prey than did male beetles when fed first and third instar B. hilaris, but not when given second instar prey.     

Differential Toxicity of Cis and Trans Isomers of Dichlorodiammineplatinum

Nephrotoxicity is the dose-limiting toxic effect of cis-dichlorodiammineplatinum (cis-platin) in humans. Its stereoisomer transplatin does not have any toxicity at equimolar concentrations, and it also possesses little antitumor activity. In this study, subcellular localization of both the platinum isomers was examined in the liver and kidney of the mouse 24 hours following the drug administration. Levels of the platinum isomers were measured using flame-less atomic absorption. The results showed that higher concentrations of the cis isomer were localized in the liver and kidney, while the concentration of the trans isomer was higher in blood. This indicates that trans isomer is sequestered in the central compartment, whereas cis isomer is distributed in the organs. We also measured metallothionein mRNA and protein levels in both liver and kidney following cisdichlorodiammineplatinum and transdichlorodiammine-platinum treatment to distinguish if the differential toxicity of the two stereo-isomers could be related to metallothionein induction. We report here that cisplatin was capable of inducing metallothionein expression in mice in vivo and that there is an inverse relationship between metallothionein expression and the pattern of tissue toxicity induced by the drug.     

Effects of brief exposures to low temperature on the development, longevity and fecundity of the grain aphid Sitobion auenae (Hemiptera: Aphididae)

First instar nymphs and adults of the grain aphid Sirobion auenae that had been reared at 10°C and 20°C over a number of generations, were cooled to -5°C and -10°C for 1 h and 6 h and returned to 20°C to assess the effects of brief exposures to low temperatures (cold-pulses) on their survival. rate of development, longevity and fecundity. A strong acclimation response was observed in first instar nymphs, with significantly less mortality in groups reared to 10°C compared to 20°C. Mean development time from first instar to adult was not significantly affected by low temperature exposure at the first nymphal stage. Longevity in all groups cooled as first instars was reduced by the sub-zero cold-pulses, and was also dependent on temperature and exposure time. Acclimated aphids survived longer than non-acclimated individuals. Reproductive rate, in terms of the number of nymphs born per aphid per day, was unaffected by cold stress applied at the first instar stage. Total fecundity was however reduced, being a function of the number and longevity of the survivors. Adult aphids were less cold hardy than nymphs; mortality was higher at -10°C than -5°C increasing with duration of exposure from 1 h to 6 h. Mean fecundity was reduced significantly in aphids cooled at the adult stage, the number of aphids born per day decreasing as the exposure period of the cold-pulse increased, suggesting that low temperature had affected embryogenesis. All the nymphs born to adults surviving exposure to -5°C for 6 h died within 48 h of birth, indicating that low temperature has a pre-natal effect on mortality.     

Quantitative analysis of histidine and cis and trans isomers of urocanic acid by high-performance liquid chromatography: a new assay method and its application

To elucidate the factors involved in dry skin and the skin damage caused by UV light, it is necessary to analyze small amounts of stratum corneum to determine amino acid contents. A new assay method for this purpose is described. Dabsylated amino acids including histidine and the cis and trans isomers of urocanic acid were analyzed quantitatively by high-performance liquid chromatography (HPLC), using a reversed-phase column. Histidine and the isomers of urocanic acid were separated from 36 other amino acids thought to be present in the extract of stratum corneum. In the presence of the 36 amino acids, standard calibration curves were obtained from 0.25 to 2.5 pmol/μl, for histidine and for both isomers of urocanic acid. The coefficients of variation for the reproducibility of the analysis at 1.0 pmol/μl were 3.8%, 2.9% and 2.5% for the cis and trans isomers of urocanic acid and for histidine, respectively. Amounts of 2 to 50 pmol of cis and trans isomers of urocanic acid and histidine in the stratum corneum were detected. The ratio of the cis to the trans isomer of urocanic acid in sunburned stratum corneum was more than three times that in normal stratum corneum. This method appears to be useful for the determination of small amounts of histidine and of the cis and trans isomers of urocanic acid in the stratum corneum.     

Identification of enriched conjugated linoleic acid isomers in cultures of ruminal microorganisms after dosing with 1-<Superscript>13</Superscript>C-linoleic acid

Most studies of linoleic acid biohydrogenation propose that it converts to stearic acid through the production of cis-9 trans-11 CLA and trans-11 C18:1. However, several other CLA have been identified in ruminai contents, suggesting additional pathways may exist. To explore this possibility, this research investigated the linoleic acid biohydrogenation pathway to identify CLA isomers in cultures of ruminai microorganisms after dosing with a ¹³C stable isotope. The ¹³C enrichment was calculated as [(M+1/M)×100] in labeled minus unlabeled cultures. After 48 h incubation, significant ¹³C enrichment was observed in seven CLA isomers, indicating their formation from linoleic acid. All enriched CLA isomers had double bonds in either the 9,11 or 10,12 position except for trans-9 cis-11 CLA. The cis-9 trans-11 CLA exhibited the highest enrichment (30.65%), followed by enrichments from 21.06 to 23.08% for trans-10 cis-12, cis-10 trans-12, trans-9 trans-11, and trans-10 trans-12 CLA. The remaining two CLA (cis-9 cis-11 and cis-10 cis-12 CLA) exhibited enrichments of 18.38 and 19.29%, respectively. The results of this study verified the formation of cis-9 trans-11 and trans-10 cis-12 CLA isomers from linoleic acid biohydrogenation. An additional five CLA isomers also contained carbons originating from linoleic acid, indicating that pathways of linoleic acid biohydrogenation are more complex than previously described.     

Geometrical Isomers of Monohydroperoxides Formed by Autoxidation of Methyl Linoleate

Isomeric monohydroperoxides produced from autoxidized methyl linoleate were separated into two geometrical isomers (cis-trans and trans-trans) by silver nitrate TLC. Purified monohydroperoxides were converted into hydroxy octadecadienoates. Trimethylsilyl (TMS) derivatives of these compounds (four components) were separated into three peaks in the gas chromatogram; the mixture of 9-hydroxy-cis,trans-isomer and 13-hydroxy-cis,trans-isomer, 9-hydroxy-trans,trans-isomer and 13-hydroxy-trans,trans-isomer. The trans-trans isomers became more dominant than the cis-trans isomers in the later stage of autoxidation and with the rise of temperature. At the degradation of monohydroperoxides, the decrease of trans- trans isomers was apparently slower than that of cis-trans isomers. It is proposed that cis,trans isomerization of monohydroperoxides takes place at the process of autoxidation of methyl linoleate.     

Effect of Temperature on Development and Fecundity of the Predaceous Plant Bug <Emphasis Type="Italic">Deraeocoris brevis</Emphasis> Reared on <Emphasis Type="Italic">Ephestia kuehniella</Emphasis> Eggs

Temperature-dependent development and oviposition component models were developed for Deraeocoris brevis (Uhler) (Hemiptera: Miridae). Egg development times decreased with increasing temperature and ranged from 35.8 d at 15 °C to 6.7 d at 32 °C. Total development times of nymphs reared on frozen Ephestia kuehniella (Zeller) (Lepidoptera: Pyralidae) eggs decreased from an estimated 55.6 d at 15 °C to 9.2 d at 32 °C and 10.0 d at 34.6 °C. By fitting linear models to the data the lower developmental threshold temperatures for eggs, small nymphs (1st to 3rd instar), large nymphs (4th to 5th instar), and all nymphs combined were calculated as 10.5, 12.5, 11.8, and 11.9 °C, respectively. The thermal constants were 144.1, 90.3, 95.0, and 190.8 degree-days for each of the above stages. The non-linear model was based on a Gaussian equation, which fit the relationship between development rate and temperature well for all stages. The Weibull function provided a good fit for the distribution of development times of each stage. Adult longevity decreased with increasing temperature and ranged from 52.9 d at 21.7 to 16.8 d at 32.0 °C. D. brevis had a maximum fecundity of 471 eggs per female at 24 °C, which declined to 191 eggs per female at 32 °C. Also, three temperature-dependent components for an oviposition model of D. brevis were developed including models for total fecundity, age-specific cumulative oviposition rate, and age-specific survival rate.     

9-cis-canthaxanthin exhibits higher pro-apoptotic activity than all-trans-canthaxanthin isomer in THP-1 macrophage cells

All-trans-canthaxanthin (4, 4′-diketo β-carotene) but not 9-cis-canthaxanthin has been shown to induce apoptosis in some cell lines. In this study apoptotic activity of 9-cis-canthaxanthin on THP-1 macrophage is reported. Comparison of apoptotic activities of the two canthaxanthin isomers on this cell line by annexin V-cy3 and TUNEL assays indicated the higher pro-apoptotic activity of 9-cis-isomer than the all-trans-isomer. Canthaxanthin-induced apoptosis in this cell line was found to be accompanied by increased caspase-3 and caspase-8 activities, indicating its progression via caspase cascade. Induction of both caspase activities was higher by 9-cis-canthaxanthin than that by trans-canthaxanthin. All these results suggest that canthaxanthin stereoisomers differentially induce apoptosis of THP-1 monocyte/macrophage.     

Impact of heat shock on heat shock proteins expression, biological and commercial traits of Bombyx mori

We report the thermotolerance of new bivoltine silkworm, Bombyx mori strains NB₄D₂, KSO₁, NP₂, CSR₂ and CSR₄and differential expression of heat shock proteins at different instars. Different instars of silkworm larva were subjected to heat shock at 35°C, 40°C and 45°C for 2 hours followed by 2 hours recovery. Heat shock proteins were analyzed by SDS‐PAGE. The impact of heat shock on commercial traits of cocoons was analyzed by following different strategies in terms of acquired thermotolerance over control. Comparatively NP₂ exhibited better survivability than other strains. Resistance to heat shock was increased as larval development proceeds in the order of first instar > second instar > third instar > fourth instar > fifth instar in all silkworm strains. Expression of heat shock proteins varies in different instars. 90 kDa in the first, second and third instars, 84 kDa in the fourth instar and 84, 62, 60, 47 and 33 kDa heat shock proteins in fifth instar was observed in response to heat shock. Relative influence of heat shock on commercial traits that correspond to different stages was significant in all strains. In NB₄D₂, cocoon and shell weight significantly increased to 17.52% and 19.44% over control respectively. Heat shock proteins as molecular markers for evaluation and evolution of thermotolerant silkworm strains for tropics was discussed.     

Life history ofAphis gossypii on cucumber: influence of temperature, host plant and parasitism

Life table data forAphis gossypii Glover (Homoptera: Aphididae), an important pest in glasshouse cucumber crops, were studied at 20, 25 and 30°C on two cucumber cultivars (Cucumis sativus L.) in controlled climate cabinets. The development time on the cucumber cv. ‘Sporu’ ranged from 4.8 days at 20°C to 3.2 days at 30°C. Immature mortality was approximately 20% and did not differ between temperatures. Most mortality occurred during the first instar. Reproduction periods did not differ among temperatures, but at 25 and 30°C more nymphs were produced (65.9 and 69.8 nymphs/♀, respectively) than at 20°C (59,9 nymphs/♀) because of a higher daily reproduction. Intrinsic rate of increase was greatest at 25°C (r _m =0.556 day⁻¹). At 20 and 30°C the intrinsic rate of increase was 0.426 and 0.510, respectively. On cv. ‘Aramon’, the development time ofA. gossypii was approximately 20% longer at all temperatures. Immature mortality did not differ between the two cultivars. The intrinsic rate of increase on cv. ‘Aramon’ was 15% smaller than on cv. ‘Sporu’. The use of cucumber cultivars partially resistant to aphids is discussed in relation to biological control of cotton aphid in glasshouses. Development time and immature mortality on leaves of the middle and upper leaf layer of glasshouse grown cucumber plants (cv. ‘Aramon’) were comparable to development in the controlled climate cabinets. On the lower leaves immature mortality was much higher (approximately 82%) than on leaves of the middle (24.0%) and upper leaf layer (24.5%). Reproduction was less on the lower leaf layer (45.9, 70.5 and 70.1 nymphs/♀ on leaves of the lower, middle and upper leaf layer, respectively). Aphids, successfully parasitized byAphidius colemani Viereck (Hymenoptera: Braconidae) only reproduced when they were parasitized after the third instar. Fecundity was 0.1 to 0.9 and 10.5 to 13.3 nymphs/♀ for aphids parasitized in the fourth instar or as adults, respectively. Reproduction of aphids that were stung but survived the attack was lower than for aphids not stung. Average longevity of these aphids was equal to the longevity of aphids not stung byA. colemani.     

Positional specificity in the incorporation of isomeric cis- and trans-octadecenoic acids into glycerolipids of cultured soya cells

Heterotrophically grown cell suspension cultures of soya (Glycine max L.) were incubated with two different mixed substrates consisting of positional isomers of either cis-[1-¹⁴C]octadecenoic acids (8 to 15) or trans-[1-¹⁴C]octadecenoic acids (8 to 16), each with known composition. With both substrates, about one-fourth of the radioactivity supplied was incorporated into the diacylglycerophosphocholines, while another one-fourth of the radioactivity was almost equally distributed between diacylglycerophos-phoethanolamines and triacylglycerols. All the positional isomers of cis-and trans-octadecenoic acids supplied to the cells were readily incorporated into various classes of glycerolipids. None of the octadecenoic acids was isomerized, elongated or desaturated during incubation. From the cis-octadecenoic acids, only the naturally occurring 9-isomer (oleic acid) was preferentially incorporated into position 2 of diacylglycerophosphocholines, diacylglycerophospho-ethanolamines, and triacyglycerols; all the other isomers exhibited a strong affinity for position 1 of the glycerophospholipids and positions 1 and 3 of the triacylglycerols. From the trans-octadecenoic acids, only the 9-isomer (elaidic acid) was preferentially incorporated into position 2 of diacylglycerophospho-cholines and triacylglycerols; all the other isomers preferred position 1 and positions 1 and 3, respectively, of these lipids. In diacylglycerophospho-ethanolamines, however, each of the trans-octadecenoic acids, including the 9-isomer, exhibited a strong affinity for position 1. Apparently, the enzymes involved in the incorporation of exogenous monounsaturated fatty acids into membrane lipids of plant cells can recognize the preferred substrate in a mixture of closely related isomers.     

Studies on fluorescence polarization of 1-acyl-2-cis- or trans-parinaroyl sn-3-glycerophosphorylcholines in model systems and microsomal membranes

Fluorescent lecithin probes containing cis- or trans-parinaric acid (PnA) at the 2-position cis-parinaroylphosphatidylcholine (cis-PnPC) and trans-parinaroyl phosphatidylcholine (trans-PnPC)) showed similar behavior to that of the free cis- or trans-parinaric acids (cis-PnA or trans-PnA) in bilayer vesicles of synthetic saturated lecithins. Transition temperatures detected by cis-PnPc were about 1°C lower than those observed with trans-PnPc. In mixed lecithin vesicles, the trans-PnPc probe monitored a higher temperature melting component than did the cis-probe. Both probes were readily incorporated into microsomal membranes and into sonicated vesicles prepared from the microsomal phospholipids. With either cis- or trans-PnPc no change in polarization ratio was observed for microsomal membranes between 40°C and 0°C but this ratio increased with decreasing temperature between 0°C and ?5°C. However, vesicles of extracted phospholipids showed a continuous increase in polarization ratio with decreasing temperature between 20°C and ?15°C with trans-PnPc and bewteen 5°C and ?15°C with cis-PnPc. These results suggest that the two lecithin probes monitor different environments in the membranes and phospholipid vesicles prepared from them.     

High-performance liquid chromatography with fluorometric detection for monitoring of etoposide and its cis-isomer in plasma and leukaemic cells

The podophyllotoxin derivative etoposide, extensively used in anticancer therapy, is highly protein-bound (95%) in plasma. It is a chiral drug and only the trans-isomer is pharmacologically active. Isomerisation to the inactive cis-lactone occurs in plasma. The cis-lacrone is often present in ultrafiltrates of plasma from patients treated with etoposide, therefore it is important to separate the isomers when free etoposide concentrations are assayed. There is reason to believe that free and cellular concentrations are more important for the effect of etoposide therapy than total plasma concentrations. A high-performance liquid chromatographic (HPLC) method for quantification of etoposide and its cis-isomer in plasma, total and non-protein-bound concentrations, and in leukaemic cells is described. After addition of teniposide as internal standard the drugs were extracted with chloroform. Etoposide, its cis-isomer, teniposide and endogenous substances were separated isocratically on a Spherisorb phenyl reversed-phase column. Detection was performed fluorometrically, λ_ex/em = 230/330 nm. Non-protein-bound concentrations were determined after ultrafiltration. The detection limit for etoposide was 10 ng/ml plasma, 25 ng/ml ultrafiltrate and 10 ng/50 · 10⁶ cells. The sensitivity of the assay for the cis-lactone was twice as high due to higher fluorescence. The protein binding of the cis-lactone in plasma from ten healthy blood donors was 54.5±4.8% (mean ± S.D.). Thus, the free fraction was about ten-fold higher than that of the mother compound. The assay is convenient and sensitive enough for the determination of free and cellular fractions of etoposide.     

Life history parameters for Nesidiocoris tenuis (Reuter) (Het., Miridae) under different temperature regimes

The development time for eggs and nymphs and female fertility were determined for Nesidiocoris tenuis Reuter (Het., Miridae: Dicyphini) at 15, 20, 25, 30, 35 and 40 ± 1°C, using tomato, Solanum esculentum (Miller), as substrate and eggs of Ephestia kuehniella Zeller as substitute prey. At 40°C, N. tenuis was unable to develop and barely reproduced. Egg development ranged from 30.8 days at 15°C to 6.3 days at 35°C. The cumulative thermal requirements for the eggs were 148.6 degree days (°d) and the lower thermal threshold, 10.3°C. The duration of the nymphal instar decreased from 55.9 days at 15°C to 8.6 days at 35°C. The thermal constant for the nymphs was 182.3 °d and the lower thermal threshold 11.7°C. No nymphs survived at 40°C, and the highest mortalities were at extreme temperatures (15 and 35°C). Female and male weights were influenced significantly by temperature. The fertility of N. tenuis females was reduced greatly at 15 and 40°C. The highest fertility during an observation period of 18 days following female emergence (79.5–60.0 nymphs per female) was within the temperature range of 20 to 35°C. Fertility was related directly to female weight and temperature (r² = 0.932). Based on development, reproduction data and thermal requirements, the optimum temperature range for N. tenuis was established as being between 20 and 30°C. Overall, N. tenuis is the most thermophilous of all dicyphines from vegetable crops in the Mediterranean area studied so far.