The mosquito dihydrofolate reductase gene functions as a dominant selectable marker in transfected cells

An Aedes albopictus dihydrofolate reductase gene was used to construct two chimeric DNA vectors that functioned as dominant selectable markers in transfected, wild type mosquito cells. Stably transformed clones were recovered after 10–15 days in the presence of selective medium containing 1 μM methotrexate. The transformed clones contained an estimated 100–500 copies of transfected DNA per nucleus. Combined data from Southern blots and in situ hybridization to metaphase chromosomes indicated that transfected DNA was likely integrated into chromosomes both as repeated structures and as randomly integrated single copy molecules, with minimal rearrangement of coding sequences. Transfected DNA was stably maintained under selective conditions, but in some cases was lost when cells were maintained for prolonged periods in the absence of methotrexate. These observations provide a general framework for further development of stable gene transfer systems for mosquito cells in culture.     

EFFECTS OF FERTILIZATION ON EPICUTICULAR WAX MORPHOLOGY OF NEEDLE LEAVES OF DOUGLAS FIR,PSEUDOTSUGA MENZIESII (PINACEAE)

Fertilized stands of Pseudotsuga menziesii were found to have glaucous needles. We investigated the morphological and quantitative characteristics of the epicuticular waxes of needles of fertilized and control trees. Glaucousness was caused by ornate tubular epicuticular wax. Dipping needles in chloroform, which dissolves waxes, eliminated the glaucous appearance. Based on cryostage scanning electron microscopic observations, the epicuticular waxes in the nonstomatal region were much more ornate on the needles of the fertilized trees (experimental needles) than in unfertilized trees (control needles). The stomatal region in both experimental and control needles showed similarly ornate waxes. Quantities of waxes were similar in experimental and control needles. The glaucousness was not the result of greater quantities of wax; rather, fertilization altered wax morphology in the nonstomatal regions.     

Vigilant Behaviour: Predictability or Randomness? Spectral Analysis of Series of Scan Durations and their Relationship with Inter-scan Intervals

While feeding, many animals frequently look up and visually scan their environment. Using spectral analysis of continuous series of scan durations from a purple sandpiper Calidris maritima and Barbary doves Streptopelia risoria, we show that there are sequential non-random patterns and significant periodicities in all the examined series such that the birds cycled regularly between short and long scans. The cycles are comparable to those for the continuous series of inter-scan intervals of the same behavioural sequences. We suggest a re-examination of the functional costs and benefits of instantaneous randomness versus sequential predictability in alternating between feeding and scanning and a revision of the models of the way animals alternate between these behaviour patterns.     

Twins and non-twin siblings: different estimates of shared environmental influence in early childhood.

Twin studies typically indicate shared environmental influence for cognitive abilities, especially in early childhood. However, across studies, DZ twin correlations tend to be greater than non-twin sibling correlations, suggesting that twin estimates of shared environment are to some extent specific to twins. We tested this hypothesis in a sample of more than 1800 MZ and 1800 same-sex DZ pairs from the Twins Early Development Study (TEDS), a study of twins born in England and Wales in 1994 and 1995. For this analysis, we obtained comparable data from more than 130 same-sex younger siblings of the twins. Twins and their younger siblings were assessed for language, cognitive abilities and behavior problems by their parents at 2 and 3 years of age. For language and cognitive measures at both 2 and 3 years, but not for behavior problems, estimates of shared environment were more than twice as large for twins as compared to non-twin siblings. We conclude that about half of twin study estimates of shared environment for cognitive abilities in early childhood are specific to twins. Although many possibilities exist for explaining the special shared environment effect for twins, we suggest that cognitive-relevant experiences that are not shared by siblings are shared by twins because they are exactly the same age.     

Extracellular matrix components of the placental extravillous trophoblast: immunocytochemistry and ultrastructural distribution

 Invasive extravillous trophoblast cells of the human placenta are embedded in a self-secreted extracellular matrix, the matrix-type fibrinoid. The ultrastructure and molecular composition of the matrix-type fibrinoid of the term human placenta were studied by transmission electron microscopy and immunogold labelling. We used antibodies directed against different matrix proteins such as collagen type IV, laminin, vitronectin, heparan sulfate, various fibronectin isoforms, and against the oncofetal blood group antigen, ”i”. Immunogold labelling patterns of matrix proteins are the basis for the subdivision of the trophoblast-derived matrix-type fibrinoid into mosaic-like patches of structurally and immunocytochemically different compartments. Firstly, fine granular patches with structural similarities to basal lamina material are composed solely of collagen type IV and laminin. Secondly, an ultrastructurally amorphous glossy substance shows reactivity with antibodies against heparan sulfate and vitronectin. A third type of patches, fine fibrillar networks embedded in the above-mentioned glossy matrix, are reactive with antibodies against normal fibronectin isoforms (IST-4, IST-6, IST-9) and oncofetal isoforms (BC-1, FDC-6). The blood group precursor antigen ”i” was not only expressed on the surfaces of the extravillous trophoblast cells but was associated with the fibronectin-positive fibrils. In conclusion, within this extracellular matrix, clear compartments of different composition can be distinguished from each other. Glycosylation with ”i” in this matrix may be involved in immunological masking, thus preventing rejection of placenta and fetus. Accepted: 6 May 1996     

Seasonality ofOrmia depletaand Limits to Its Spread

Ormia depleta(Wiedemann) (Diptera: Tachinidae), a South American fly that homes on the calling songs of its hosts, was brought from Piracicaba, Brazil; colonized; and released in Florida for the biological control ofScapteriscusspp. mole crickets. It became established at some release sites and rapidly spread through most of peninsular Florida. The seasonal distributions of phonotactic females in Florida differed from the single summer peak recorded at Rio Claro and Piracicaba, Brazil (23° S). Near Bradenton (27° N), spring and fall peaks with a summer hiatus were quickly established and remain evident. At two sites near Gainesville (30° N), a strong fall peak and a modest-to-none spring peak developed. Peaks of phonotactic females seem to follow times of peak host availability and sometimes coincide with minima of host availability. Limits to geographical expansion ofO. depletaare poorly understood. At Gainesville, the site of earliest introduction, establishment was rapid and soundtrap catches increased for the first 3 years, exceeding 1000 per year at one site. Catches then declined for 3 years, almost reaching zero in 1994. On the other hand, annual sound-trap catches ofO. depletaat Bradenton exceeded 1000 in <2 years and have remained above that level for 5 years. Winters in Gainesville are more severe than those in Bradenton, but minimum winter temperatures in Gainesville were substantially lower duringO. depleta's increase than during its decline. Perhaps mild winters cause the flies to become active early—only to starve for lack of adequate winter nectar sources.O. depletamay yet spread to the northern limits of itsScapteriscushosts as present populations adapt to new environments. Or farther spread may require that new strains be introduced from more southerly sites in South America.     

Grenzen der Kompensation: Energiebudgets von Ringelg?nsen (Branta b. bernicla) — die Wirkung von St?rreizen

Zusammenfassung Der Einfluß von Störreizen auf das Energiebudget von Ringelgänsen wurde im Frühjahr 1991 in zwei Vorland-Salzwiesen des Nationalparks Schleswig-Holsteinisches Wattenmeer vergleichend untersucht. Die Häufigkeit störreizbedingter Reaktionen der Gänse (Störreizhäufigkeit) wurde als Index für die anthropogene Beeinflussung der Gebiete herangezogen. Die Beeinflussung war in Westerhever mit 1,5±0,7 Reaktionen/h signifikant größer als vor dem Norderheverkoog (1,0±0,6 Reaktionen/h). Mit zunehmender Aktivitätszeit wurde in beiden Gebieten im Verlauf des Frühjahres mehr umsetzbare Energie aufgenommen. In Westerhever haben die Gänse im Vergleich zu denen vor dem Norderheverkoog mit Ausnahme des Mai jeweils mehr umsetzbare Energie aufgenommen. Die energetischen Kosten (DEE) der Gänse wurden anhand der Zeit-Energie-Budget Methode und über die gemessene Körpermasseentwicklung von Fänglingen berechnet. Beide Methoden erzielten vergleichbare Ergebnisse; die Unterschiede betrugen je nach Gebiet 0,4 bis 4,5 %. In Westerhever verzeichneten die Gänse im Vergleich zu denen vor dem Norderheverkoog im Monatsmittel höhere energetische Kosten. Aus dem budgetierten Energieüberschuß (E) wurde die theoretisch mögliche Reservestoffanlagerung (Änderung der Körpermasse) der Gänse berechnet. In Westerhever konnten die Gänse im Verlauf des Frühjahres 357 g Körperreserven anlagern. Bei den Gänsen vor dem Norderheverkoog waren es 399 g. Der Unterschied betrug 11,7 %, war aber nicht signifikant. Anhand von Fangdaten ist bekannt, daß Ringelgänse im langjährigen Mittel Körperreserven von 380 bis 400 Gramm anlagern. Die Störreizhäufigkeit des Tages bewirkte eine signifikante Veränderung der Budgetparameter DME und DEE und E auf stündlicher Basis. In Westerhever stieg die Aufnahme an umsetzbarer Energie signifikant mit steigender Störreizhäufigkeit an. Vor dem Norderheverkoog verringerte sich diese jedoch signifikant. Die Energieaufwendungen stiegen in beiden Gebieten signifikant mit steigender Reizhäufigkeit an. Der Überschuß in der Energiebilanz der Gänse in Westerhever war an Tagen mit einer großen Störreizhäufigkeit um 8,7 % gegenüber den Tagen mit einer geringen Reizhäufigkeit verringert. Aufgrund fehlender kompensatorischer Nahrungsaufnahme war der Überschuß bei den Gänsen vor dem Norderheverkoog um maximal 27,5 % vermindert. Die Gänse in Westerhever haben störreizbedingte Zeitverluste bei der Nahrungsaufnahme und erhöhte energetische Kosten durch Verhaltensänderung und eine erhöhte Nahrungsaufnahme pro Zeit kompensiert. Aufgrund physiologischer Zwänge sind sie jedoch in dem stark vom Menschen beeinflußten Gebiet an die Grenzen ihrer Kompensationsmöglichkeit angelangt. Die errechnete Reservestoffanlagerung und auch der Jungvogelanteil im nachfolgenden Herbst waren geringer als bei den Gänsen vor dem Norderheverkoog.

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Compensatory limits: energy budgets of Brent Geese,Branta b. bernicla, the influence of human disturbance

The impact of human disturbances on the energy budget of Brent Geese,Branta b. bernicla, during spring migration was investigated in two different salt marsh areas within the national park Schleswig-Holsteinisches Wattenmeer, Germany. We used the frequency of disturbance-related reactions of the birds as an indicator of the anthropogenic influencing of the sites. The disturbance frequency was significantly higher in the Westerhever salt marsh (1,5±0,7 reactions/h) than in the Norderheverkoog area (1,0±0,6 reactions/h). Monthly activity and energy budgets of the birds were calculated from March to May, based on several simultaneous day-round field observations per month. The amount of daily metabolizable energy (DME) was calculated via dropping rate and digestability of the food. The daily energy expenditure (DEE) was calculated by means of the time-energy-budget methods and by calculations, using the bodymass changes of caught birds during spring. With both methods very similar results were obtained; they varied between 0,4 to 4,5 % per areas and month. The daily energy budget (Æ E) was calculated from DEE — DME. Due to an increasing daily activity time from March to Mai the DME increased in both areas. Compared to the Norderheverkoog area the DME of the birds in Westerhever was higher in March and April, and similar in Mai. DEE increased in both areas too and was in all months higher in Westerhever than in Norderheverkoog. The calculated bodymass changes of the birds during spring was 357 g for the birds in Westerhever and 399 g for the birds in Norderheverkoog. Bodymass measures from caught birds revealed a mean mass gain of 380 to 400 g during spring. The daily disturbance frequency caused a significant change in the budget parameters DME, DDE and E on a hourly base. In Westerhever DME/h increased, while in Norderheverkoog DME/h decreased significantly with increasing disturbance frequency. At the same time DEE/h increased in both areas significantly. As a consequence E/h was reduced at days with a high disturbance frequency; in Westerhever by 8,7 % and in Norderheverkoog by 27,5%. In the disturbed area the birds compensated the time-loss during feeding and the higher energetic costs during disturbance-related flights by a change in activity pattern and by an increased food consumption per time unit. Based on physiological constraints the birds reached their compensatory abilities. The calculated bodymass gain and the proportion of young birds in the flocks was reduced in autumn.

     

Induction of chitinase and {beta}-1,3-glucanase activity in sunflower suspension cells in response to an elicitor from Phytophthora megasperma f. sp. glycinea (Pmg). Evidence for regulation by ethylene and 1-aminocyclopropane-1-carboxylic acid (ACC)

In heterotrophic cell suspensions of sunflower (Helianthus annuusL. cv. Spanners Allzweck) the effect of Pmg elicitor, a fungalelicitor preparation from Phytophthora megasperma f. sp. glycinea,on the induction of chitinase and ß-1,3-glucanaseactivity was studied in relation to changes in ethylene biosynthesis.Dose-response experiments with Pmg elicitor showed that theonset of the induction of intracellular chitinase and ß-1,3-glucanaseactivity coincided or followed a transient rise in ethyleneand particularly endogenous 1-aminocyclopropane-1-carboxylicacid (ACC) levels within 5 h of application. Treatment with5 µg ml^–1 elicitor stimulated ethylene and ACC levels1.6-fold and 4-fold, relative to control, respectively. Themolar ratio of ACC to ethylene changed from approximately 3:1in controls to 9:1 in treated cells. During further incubation,ethylene formation and, to a lesser degree, ACC levels declinedand the ACC/ethylene ratio increased to 56:1 in elicitor-treatedcells. On a protein basis, the activities of ß-1,3-glucanaseand chitinase increased approximately 5-fold and 8-fold, respectively,48 h after elicitor application. Additional treatment with theACC synthesis inhibitor aminoethoxyvinyiglycine (AVG) decreasedelicitor-induced enzyme activities and the levels of both ethyleneand ACC. Elicitor effects on chitinase and ß-1,3-glucanaseactivities could be fully restored when ACC was additionallyapplied. Concomitantly, the ACC/ ethylene ratio increased. Neithertreatments with ACC alone, which simultaneously increased internalACC and ethylene levels, nor treatments with AVG alone, whichsimultaneously reduced ACC and ethylene levels, could generallystimulate chitinase or ß-1,3-glucanase activitiesin the cells. It is suggested that ACC functions as a promotingfactor in the induction of chitinase and ß-1,3-glucanaseactivity triggered by Pmg elicitor and appears to reverse aninhibiting influence of ethylene. Key words: 1-Aminocyclopropane-1-carboxylic acid, chitinase, ß-1,3-glucanase, ethylene, Helianthus cellsuspension cultures, Phytophthora megasperma-elicitor     

Blood chemistry of the common marmoset (Callithrix jacchus) maintained in an indoor-outdoor environment: Primate comparisons

Blood chemistry values were collected over a three-year period from at least 10 colony-born and 24 wild-born apparently normal common marmosets. BUN, SGOT, creatinine, calcium, phosphorus, alkaline phosphatase, protein, albumin, cholesterol, triglycerides, uric and glucose values were determined. A statistical comparison of baseline values was made between wild-born, colony-born, male and female marmosets. Also the same comparison was made between common marmosets and cotton-top tamarins, white lipped tamarins and human subjects.