The body temperature of the desert locust Schistocerca gregaria

Summary The internal equilibrium body temperatures of hoppers (nymphs) of Schistocerca gregaria were studied in the field. The data obtained are here compared with theoretical estimates of the thermal balance resulting from the factors assumed to be concerned in heat loss and heat gain.In the field, the equilibrium body temperature of quiescent hoppers shaded from direct sunshine was up to 3.2° higher than the air temperature when the latter was about 25° or less, but was lower than the air temperature when this was above about 31°. In sunlight, body-temperature excesses increased linearly with total radiation intensity between 0.15 and 1.25 cal/cm²/min. Differences in orientation to the sun gave rise to differences in equilibrium body temperature of as much as 6°. At relatively low radiation intensities (about 0.5 cal/cm²/min) the equilibrium body temperatures were found to vary with a power of the wind speed of about 0.35. There were no demonstrable significant differences in equilibrium body temperature or rate of change of body temperature between hoppers having opposite extremes of possible coloration.Except in hoppers shaded from sunlight and those exposed to low radiation intensities there was good agreement between the observed equilibrium temperatures and those expected on theoretical grounds. This provides evidence of the relative importance of the factors concerned in thermal balance.The relation between body temperature and behaviour is discussed.

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Zusammenfassung Das innere Gleichgewicht der Körpertemperatur von vorzugsweise fünften Larvenstadien der Wüstenheuschrecke wurde unter Freilandbedingungen in Beziehung zu Lufttemperatur, relativer Luftfeuchtigkeit, Strahlungsintensität, Windgeschwindigkeit und Aktivität gemessen.Die verschiedenen Faktoren, die das Temperaturgleichgewicht der Heuschrecken beeinflussen, werden theoretisch behandelt. Die unter bestimmten Kombinationen der Umweltbedingungen zu erwartenden Gleichgewichts-Körpertemperaturen werden aus den theoretischen Werten für die verschiedenen beteiligten Faktoren errechnet und mit den unter entsprechenden Freilandbedingungen beobachteten Werten verglichen.Ohne Sonneneinstrahlung und unter relativ windstillen Bedingungen wurde bei ruhenden Larven des fünften Stadiums eine Gleichgewichts-Temperatur des Körpers gefunden, die bis zu 3,2° höher lag als die Lufttemperatur, wenn diese ca. 25° oder weniger betrug. Dieser Überschuß hatte eine abnehmende Tendenz bei Lufttemperaturen bis zu etwa 31° und über dieser Schwelle waren die Gleichgewichts-Temperaturen des Körpers im allgemeinen niedriger als die entsprechenden Lufttemperaturen. Theoretisch würde zu erwarten sein, daß unter diesen Umweltbedingungen das Temperaturgleichgewicht des Körpers in allen Niveaus annähernd der Lufttemperatur entspräche. Folglich scheint hier ein physiologischer Mechanismus zu bestehen, mit dessen Hilfe die Insekten in Abwesenheit von Sonnenschein und unter den Bedingungen schwacher Konvektion ihre Körperausgleichstemperaturen bei niedrigen Lufttemperaturen erhöhen, bei hohen senken können.Es war nicht möglich, im Freiland nachzuweisen, daß Unterschiede in der relativen Luftfeuchtigkeit irgendeinen signifikanten Einfluß auf die ausgeglichenen Körpertemperaturen in dem untersuchten Bereich (Sättigungsdefizit von 2 bis 27 mm) ausübten. Die erwarteten maximalen Differenzen der Gleichgewichts-Körpertemperaturen lagen in diesem Bereich des Sättigungsdefizits in einer Größenordnung von nur 0,5°.Unter relativ ruhigen Freilandbedingungen zeigten die Heuschrecken im Sonnenschein Körpertemperaturüberschüsse, die bei einer totalen Strahlungsintensität zwischen 0,15 und 1,25 cal/cm²/min annähernd linear anstiegen (von einem Mittel von 2–10° bei fünften Larvenstadien). Die in diesem Strahlungsintensitätsbereich gemessenen Übertemperaturen des Körpers stimmten einigermaßen gut mit Werten überein, die von anderen Untersuchern unter Laboratoriumsverhältnissen festgestellt wurden. Unter 0,40 cal/cm²/min waren die beobachteten Körpertemperaturüberschüsse im allgemeinen höher als theoretisch zu erwarten gewesen wäre. Die Körpertemperatur von Larven des ersten Stadiums nahm rascher zu als die von Larven des fünften, jedoch nur bis zu einem niedrigeren Niveau; auch das stimmte gut mit den theoretischen Vorstellungen und mit der Laborerfahrung überein. Die Einstellreaktion der Heuschrecken zur Sonne ergab beobachtete Unterschiede in der Gleichgewichts-Körpertemperatur bis zu 6° zwischen Larven des fünften Stadiums, die sich in Richtung der Sonnenstrahlen eingestellt hatten, und solchen, die im rechten Winkel zur Sonnenrichtung auf dem Boden standen; die erwarteten Unterschiede waren von der gleichen Größenordnung.Im Sonnenschein variierten die erwarteten Intensitätsproportionen zwischen der Ausgleichstemperatur und der Windgeschwindigkeit zwischen 0,5 und 0,6 mit dem Sättigungsdefizit. Bei Strahlungsintensitäten von etwa 1,10 cal/cm²/min stimmten die beobachteten Körpergleichgewichtstemperaturen engstens mit den erwarteten überein, aber bei geringeren Strahlungsintensitäten von etwa 0,50 cal/cm²/min variierten die Meßwerte mit einer Intensitätsproportion zur Windgeschwindigkeit von etwa 0,35. Darin zeigt sich möglicherweise, daß bei niederen Strahlungswerten die Körperausgleichstemperaturen zu höheren Werten tendieren als zu erwarten war.Das beobachtete Ansteigen der Körperausgleichstemperatur von ungefähr 1° bei dritten Larvenstadien und von 2–3° bei fünften Larvenstadien befindet sich in Übereinstimmung mit dem für entsprechende Umweltbedingungen errechneten.Zwischen Larven extrem entgegengesetzter Färbungstypen konnten keine Unterschiede in den Körperausgleichstemperaturen oder dem Ausmaß der Körpertemperaturänderung nachgewiesen werden.Mit Ausnahme der vor direkter Sonnenbestrahlung geschützten Heuschrecken unter relativ ruhigen Bedingungen und der Heuschrecken, die niederen Strahlungsintensitäten ausgesetzt waren, bestand eine gute allgemeine Übereinstimmung zwischen den beobachteten und den erwarteten Gleichgewichts-Körpertemperaturen. Es wurde deshalb geschlossen, daß die durchgeführten Bestimmungen Beweise für die relative Bedeutung der für den Temperaturhaushalt verantwortlichen Faktoren darstellen. Zur Erzeugung beträchtlicher Überschüsse der Körpertemperatur über die Lufttemperatur hat die Strahlung die größte Bedeutung. Für den Wärmeabfluß nach außen werden die durch Konvektion entstehenden Wärmeverluste, die insgesamt 94% oder mehr betragen, als bei weitem am wichtigsten angesehen.Einige mögliche Beziehungen zwischen Körpertemperatur und Verhalten werden besprochen.

     

Responses to mitogenic stimulation of lymphocytes taken during and after pregnancy

Serial blood samples were collected during pregnancy, after delivery and several months postnatally from 28 women. The blastogenic responses of lymphocytes to varying concentrations of phytohaemagglutinin (PHA) were tested using autologous plasma or fetal calf serum (FCS) to support the lymphocyte cultures. Using FCS, the blastogenic response decreased as pregnancy progressed and remained depressed months after delivery. In contrast, when autologous plasma was used a 10-fold higher concentration of PHA was required to give optimal stimulation. Blastogenic responses were still suppressed during pregnancy but had returned to initial values by the time of delivery and were greater still in the post-partum and postnatal periods. We conclude that the inherent ability of lymphocytes to undergo blastogenesis is suppressed during pregnancy but that this is over-shadowed by a humoral effect of pregnancy plasma. The significance of these results is discussed.     

Short-term changes in carbon-isotope discrimination identify transitions between C3 and C4 carboxylation during Crassulacean acid metabolism

Short-term measurements of instantaneous carbon-isotope discrimination have been determined from mass-spectrometric analyses of CO₂ collected online during gas exchange for the epiphytic bromeliad Tillandsia utriculata L. Using this technique, the isotopic signature of CO₂ exchange for each phase of Crassulacean acid metabolism (CAM) has been characterised. During night-time fixation of CO₂ (Phase I), discrimination () ranged from 4.4 to 6.6, equivalent to an effective carbon-isotope ratio (¹³C) of –12.3 to –14.5 versus Pee Dee Belemnite (PDB). These values reflected the gross photosynthetic balance between net CO₂ uptake and refixation of respiratory CO₂, characteristic of CAM in the Bromeliaceae. When for the relative proportion of external (p _a ) and internal (p _i) CO₂ is taken into account, calculated p _i/p _a decreased during the later part of the dark period from 0.68 to 0.48. Measurements of during Phase II, early in the light period, showed the transition between C₄ and C₃ pathways, with carboxylation being increasingly dominated by ribulose bisphosphate carboxylase (Rubisco) as increased from 10.5 to 21.2 During decarboxylation in the light period (Phase III), CO₂ leaked out of the leaf and the inherent discrimination of Rubisco was expressed. The value of calculated from on-line measurements (64.4) showed that the CO₂ lost was considerably enriched in ¹³C, and this was confirmed by direct analysis of the CO₂ diffusing out into a CO₂-free atmosphere ( ¹³C = + 51.6 versus PDB). Instantaneous discrimination was characteristic of the C₃ pathway during Phase IV (late in the light period), but a reduction in showed an increasing contribution from phosphoenolpyruvate carboxylase. The results from this non-invasive technique confirm the observations that double carboxylation involving both phosphoenolpyruvate carboxylase and Rubisco occurs during the transient phases of CAM (II and IV) in the light period.Abbreviations and Symbols CAM Crassulacean acid metabolism - H⁺ (dawn-dusk) variation in titratable acidity - ¹³C carbonisotope ratio of plant organic material, relative to Pee Dee Belemnite (vs. PDB) - discrimination against ¹³CO₂, - p _i, p _a internal, external partial pressures of CO₂ - Rubisco ribulose1,5-bisphosphate carboxylase - PAR photosynthetically active radiation - PEPCase phosphoenolpyruvate carboxylase We are grateful for financial support in respect of research grants (GR3/5360, GR3/6419) and a studentship awarded by the Natural Environment Research Council, UK.     

Different requirements for productive interaction between the active site of HIV-1 proteinase and substrates containing -hydrophobic*hydrophobic- or -aromatic*pro- cleavage sites.

The sequence requirements for HIV-1 proteinase catalyzed cleavage of oligopeptides containing two distinct types of junctions (-hydrophobic*hydrophobic- or -aromatic*Pro-) has been investigated. For the first type of junction (-hydrophobic*hydrophobic-) the optimal residues in the P2 and P2' positions were found to be Val and Glu, respectively, in accord with recent statistical analysis of natural cleavage sites [Poorman, R. A., Tomasselli, A. G., Heinrikson, R. L., & Kézdy, F. J. (1991) J. Biol. Chem. 266, 14554-14561]. For the -aromatic*Pro- type of junction, in the specific sequence context studied here, the value of Glu in the P2' position was again observed. An explanation for the inefficient cleavage observed for peptides with the sequence -Val-Tyr*Pro- has been provided from molecular modeling of the putative enzyme-substrate complex. A significant effect upon cleavage rates due to the amino acid in the P5 position has also been documented. While lysine in the P5 position in one sequence of the -hydrophobic*hydrophobic- type produces a peptide cleaved very efficiently (kcat greater than 15 s-1 for Lys-Ala-Arg-Val-Nle*p-nitrophenylalanine-P2'-Ala-Nle-NH2, for P2' = Glu, Gln, Ile, Val, or Ala), for substrates of the -aromatic*Pro- type, the P5 residue can exert either a positive or negative effect on cleavage rates. These results have again been interpreted in light of molecular modeling. We suggest that interaction of the substrate sequence on the periphery of the active site cleft may influence the match of the enzyme-substrate pair and, hence, control the efficiency of catalysis.(ABSTRACT TRUNCATED AT 250 WORDS)     

The use of amphipathic maleimides to study membrane-associated proteins

A series of amphiphilic polymethylenecarboxymaleimides has been synthesized for use as sulfhydryl reagents applicable to membrane proteins. Physical properties of the compounds which are relevant to their proposed mode of action have been determined. By comparing rates of reaction in aqueous and aprotic solvents, the compounds have been shown to react exclusively with the thiolate ion. The effects of the reagents on three membrane-associated proteins are reported, and in two cases a comparative study has been made of the effects on the proteins in the absence of membranes. A mechanism is proposed whereby the reagents are anchored at the lipid/water interface by the negatively charged carboxyl group, thus siting the reactive maleimide in a plane whose depth is defined by the length of the reagent. Supporting evidence for this model is provided by the inability of the reagents to traverse membranes, and variation of their inhibitory potency with chain length when the proteins are embedded in the membrane, but not when extracted into solution. As examples of general use of the reagents to probe sulfhydryl groups in membrane proteins, the reagents have been used to (a) determine the depths in the membrane at which two populations of sulfhydryl groups occur in the mitochondrial phosphate transporter; (b) locate a single sulfhydryl associated with the active site ofD--hydroxybutyrate dehydrogenase in the inner mitochondrial membrane; (c) examine sulfhydryl groups in theD-3-glyceraldehyde phosphate dehydrogenase associated with the human red blood cell membrane.     

Protein import into yeast mitochondria is accelerated by the outer membrane protein MAS70.

The yeast mitochondrial outer membrane contains a major 70 kd protein with an amino-terminal hydrophobic membrane anchor and a hydrophilic 60 kd domain exposed to the cytosol. We now show that this protein (which we term MAS70) accelerates the mitochondrial import of many (but not all) precursor proteins. Anti-MAS70 IgGs or removal of MAS70 from the mitochondria by either mild trypsin treatment or by disrupting the nuclear MAS70 gene inhibits import of the F1-ATPase beta-subunit, the ADP/ATP translocator, and of several other precursors into isolated mitochondria by up to 75%, but has little effect on the import of porin. Intact cells of a mas70 null mutant import the F1-ATPase alpha-subunit and beta-subunits, cytochrome c1 and other precursors at least several fold more slowly than wild-type cells. Removal of MAS70 from wild-type mitochondria inhibits binding of the ADP/ATP translocator to the mitochondrial surface, indicating that MAS70 mediates one of the earliest import steps. Several precursors are thus imported by a pathway in which MAS70 functions as a receptor-like component. MAS70 is not essential for import of these precursors, but only accelerates this process.     

Hormonal stimulation of tyrosinase activity in human foreskin organ cultures

Summary A human foreskin organ culture system has been developed to study the response of human skin to hormonal stimulation. Foreskins are maintained in culture on floating plastic supports which allows the epidermal surface to be exposed to air while the dermis is bathed in nutrient medium. Both black and white human foreskins can be maintained in organ culture for at least 1 wk with no change in the tissue structure or cell viability as determined by histochemical staining and by dopa reaction staining. Tyrosinase activity in both black and white human foreskin cultures decays markedly during the first 2 d of culture to a new steady state level which remains stable throughout the culture period. Both black and white foreskin cultures consistently demonstrate 2- to 10-fold increases in tyrosinase activity when treated with theophylline (1 mM). Approximately 90% of all skin cultures examined showed an increase in enzyme activity when treated with this phosphodiesterase inhibitor. Dibutyryl cAMP (0.1 mM) and [Nle⁴, D-phe⁷]-alpha MSH (10⁻⁸ M), were also found to markedly stimulate tyrosinase activity in some skin cultures, whereas alpha-MSH and prostaglandin E₁ produced only an inconsistent and small increase in the activity of the enzyme. Histamine (1 μM), vitamin D₃ (1 μM), and retinoic acid (1μM) failed to stimulate tyrosinase activity in either white or black foreskin cultures. This hormone-responsive organ culture system can be utilized to characterize the molecular processes responsible for the regulation of tyrosinase and pigmentation in human skin. This work was supported by a research contract from the Oklahoma Center for the Advancement of Science and Technology (OCAST) and by a research grant from the Presbyterian Health Foundation.