Behaviorally mediated contacts between scent organs: Another prerequisite for pheromone production inDanaus chrysippus males (lepidoptera)

Summary Males ofDanaus butterflies possess two binate glandular organs: abdominal hairpencils, which release aphrodisiac pheromones during courtship behavior, and pockets on the hindwings. Between these two types of organs contacts are established behaviorally: the hairpencils are dipped into the alar pockets (Fig. 2). GLC analyses of hairpencil and wing pocket extracts fromDanaus chrysippus show that this contact behavior is a prerequisite for the synthesis of the ketonic pheromone component (for which the term danaidone is suggested here) in physiologically normal amounts. Danaidone occurs on the hairpencils only i) after the males have ingested pyrrolizidine alkaloids as precursors, and ii) after the hairpencils have been dipped into the wing pockets. The appearance of danaidone in the wing pockets also depends on the ingestion of alkaloidal precursor, but is not affected by the above mentioned contact behavior. Mechanisms by which contact behavior might control pheromone biosynthesis are discussed.We appreciate the skillful and reliable greenhouse and laboratory assistance of C. Adrian and H. Mayr, and we thank Drs. R.A. Steinbrecht and P. Lounibos for kindly supplying us with danaids from Kenya. Partial support of this research by Grant No. AI-12020, from the National Institutes of Health (USA), is acknowledged with pleasure.     

Protein-protein interactions: hot spots and structurally conserved residues often locate in complemented pockets that pre-organized in the unbound states: implications for docking

Energetic hot spots account for a significant portion of the total binding free energy and correlate with structurally conserved interface residues. Here, we map experimentally determined hot spots and structurally conserved residues to investigate their geometrical organization. Unfilled pockets are pockets that remain unfilled after protein-protein complexation, while complemented pockets are pockets that disappear upon binding, representing tightly fit regions. We find that structurally conserved residues and energetic hot spots are strongly favored to be located in complemented pockets, and are disfavored in unfilled pockets. For the three available protein-protein complexes with complemented pockets where both members of the complex were alanine-scanned, 62% of all hot spots (DeltaDeltaG>2kcal/mol) are within these pockets, and 60% of the residues in the complemented pockets are hot spots. 93% of all red-hot residues (DeltaDeltaG>/=4kcal/mol) either protrude into or are located in complemented pockets. The occurrence of hot spots and conserved residues in complemented pockets highlights the role of local tight packing in protein associations, and rationalizes their energetic contribution and conservation. Complemented pockets and their corresponding protruding residues emerge among the most important geometric features in protein-protein interactions. By screening the solvent, this organization shields backbone hydrogen bonds and charge-charge interactions. Complemented pockets often pre-exist binding. For 18 protein-protein complexes with complemented pockets whose unbound structures are available, in 16 the pockets are identified to pre-exist in the unbound structures. The root-mean-squared deviations of the atoms lining the pockets between the bound and unbound states is as small as 0.9A, suggesting that such pockets constitute features of the populated native state that may be used in docking.     

Nickel absorption and kinetics in human volunteers

Mathematical modeling of the kinetics of nickel absorption, distribution, and elimination was performed in healthy human volunteers who ingested NiSO4 drinking water (Experiment 1) or added to food (Experiment 2). Nickel was analyzed by electrothermal atomic absorption spectrophotometry in serum, urine, and feces collected during 2 days before and 4 days after a specified NiSO4 dose (12 micrograms of nickel/kg, n = 4; 18 micrograms of nickel/kg, n = 4; or 50 micrograms of nickel/kg, n = 1). In Experiment 1, each of the subjects fasted 12 hr before and 3 hr after drinking one of the specified NiSO4 doses dissolved in water; in Experiment 2, the respective subjects fasted 12 hr before consuming a standard American breakfast that contained the identical dose of NiSO4 added to scrambled eggs. Kinetic analyses, using a compartmental model, provided excellent goodness-of-fit for paired data sets from all subjects. Absorbed nickel averaged 27 +/- 17% (mean +/- SD) of the dose ingested in water vs 0.7 +/- 0.4% of the same dose ingested in food (a 40-fold difference); rate constants for nickel absorption, transfer, and elimination were not significantly influenced by the oral vehicle. The elimination half-time for absorbed nickel averaged 28 +/- 9 hr. Renal clearance of nickel averaged 8.3 +/- 2.0 ml/min/1.73 m2 in Experiment 1 and 5.8 +/- 4.3 ml/min/1.73 m2 in Experiment 2. This study confirms that dietary constituents profoundly reduce the bioavailability of Ni2+ for alimentary absorption; approximately one-quarter of nickel ingested in drinking water after an over-night fast is absorbed from the human intestine and excreted in urine, compared with only 1% of nickel ingested in food. The compartmental model and kinetic parameters provided by this study will reduce the uncertainty of toxicologic risk assessments of human exposures to nickel in drinking water and food.     

Toxins induce ‘malaise’ behaviour in the honeybee (Apis mellifera)

To avoid poisoning and death when toxins are ingested, the body responds with a suite of physiological detoxification mechanisms accompanied by behaviours that in mammals often include vomiting, nausea, and lethargy. Few studies have characterised whether insects exhibit characteristic ‘malaise-like’ behaviours in response to intoxication. Here, we used the honeybee to investigate how intoxication produced by injection or ingestion with three toxins with different pharmacological modes of action quinine, amygdalin, and lithium chloride affected behaviour. We found that toxin-induced changes in behaviour were best characterised by more time spent grooming. Bees also had difficulty performing the righting reflex and exhibited specific toxin-induced behaviours such as abdomen dragging and curling up. The expression of these behaviours also depended on whether a toxin had been injected or ingested. When toxins were ingested, they were least 10 times less concentrated in the haemolymph than in the ingested food, suggesting that their absorption through the gut is strongly regulated. Our data show that bees exhibit changes in behaviour that are characteristic of ‘malaise’ and suggest that physiological signalling of toxicosis is accomplished by multiple post-ingestive pathways in animals.     

Morphology of the intestine of prefeeding and feeding adult lampreys,Petromyzon marinus (L): The mucosa of the posterior intestine and hindgut

The epithelium of the posterior intestine and hindgut of recently metamorphosed adult lampreys (Petromyzon marinus L.) prior to and during spontaneous feeding was examined using light and electron microscopy. These two regions differ slightly in their general morphology but possess the same mucosal cell types. Included are caveolated absorptive and mucous cells, which are not present in more cephalic regions of the intestine, and ciliated and enteroendocrine cells. During feeding, the caveolated cells undergo dramatic transformation in their structure, namely, through the acquisition of numerous heterophagic vacuoles. Due to their morphology and to the fact that there are low amounts of lipid, it is suspected that caveolated cells are primarily involved in the absorption of protein components from the ingested host blood and body fluids. Iron in caveolated cells may result from the degradation of ingested heme or reflect the excretion of bile products at this location in the intestine. Mucous cells are likely responsible for lubrication of the luminal surface and may be important as a stem cell for the mucosal epithelium.     

Evidence for ligandable sites in structured RNA throughout the Protein Data Bank

RNA has attracted considerable attention as a target for small molecules. However, methods to identify, study, and characterize suitable RNA targets have lagged behind strategies for protein targets. One approach that has received considerable attention for protein targets has been to utilize computational analysis to investigate ligandable “pockets” on proteins that are amenable to small molecule binding. These studies have shown that selected physical properties of pockets are important parameters that govern the ability of a structure to bind to small molecules. This work describes a similar analysis to study pockets on all RNAs in the Protein Data Bank (PDB). Using parameters such as buriedness, hydrophobicity, volume, and other properties, the set of all RNAs is analyzed and compared to all proteins. Considerable overlap is observed between the properties of pockets on RNAs and proteins. Thus, many RNAs are capable of populating conformations with pockets that are likely suitable for small molecule binding. Further, principal moment of inertia (PMI) calculations reveal that liganded RNAs exist in diverse structural space, much of which overlaps with protein structural space. Taken together, these results suggest that complex folded RNAs adopt unique structures with pockets that may represent viable opportunities for small molecule targeting.     

Heme-CO as a probe of the conformational state of calmodulin

The interaction of heme-CO with calmodulin, in the presence of calcium, leads to a complex of four heme-CO molecules per protein. No interaction was observed in the absence of calcium. The binding of heme-CO to calmodulin was monitored by the shift in the Soret absorption band from 407 to 420 nm (bound form); the four sites are not spectrally identical. The ligand CO can be photodissociated from the calmodulin-heme-CO complex and the biomolecular recombination kinetics also indicate a heterogeneous mixture. The complex does not bind oxygen reversibly. As calmodulin has only one histidine, the hemes are apparently not bound by the iron atom as in hemoglobin, but are probably loosely associated (Kd = 0.5 microM) in hydrophobic pockets which apparently open when the protein is activated by calcium.     

Sperm storage in the oviduct of the tropical rock lizard,Psammophilus dorsalis

The female rock lizard, Psammophilus dorsalis (Agamidae), lays multiple clutches of eggs over a period of 6 months (June–December). The later clutches of eggs are presumably fertilized by sperm stored from earlier matings, since testes and epididymides are regressed after August. Sperm storage is seen in pockets of the anterior vaginal region of the oviduct. Sperm recovered from the uterovaginal region are intact and motile. Discrete granules resembling the secretory granules present in the vas deferens also occur along with sperm in the vaginal sperm storage structures. The PAS-positive granules and acid phosphatase form important components of the secretions present along with sperm in the vaginal sperm storage pockets. The epithelium of the vaginal sperm storage pockets is PAS-positive and contains lipid. Several enzymes, including hydroxysteroid dehydrogenases and hydrolases, are localized histochemically in the epithelium of the vaginal sperm storage pockets. A possible role is suggested for the secretions from the male reproductive tract during sperm storage in the oviduct, in which physiological “dormancy” of the sperm during their storage may be maintained by the metabolic “milieu” in the vaginal sperm storage pockets by a mechanism similar to that effecting dormancy of the epididymal sperm in the male. © 1995 Wiley-Liss, Inc.     

Exogenous and endogenous contributions to nitrogen fluxes in the digestive tract of pigs fed a casein diet. II. Ileal and faecal digestibilities and absorption of amino acids.

The present work aimed at quantifying nitrogen (N) and amino acid (AA) fluxes in the digestive tract of growing pigs fed a casein diet. In this paper we report on digesta passage at the terminal ileum, on apparent balances at the ileal and faecal levels, and on nutrients appearance in the portal vein. Digesta flow-rate at the terminal ileum was maximum between 6 and 12 h after the meal. About 10% of N and 5% of total AA ingested were recovered within 24 h. AA absorption started 30 min after the meal, and was measurable until 13 to 14 h. The total AA absorbed in 24 h accounted for 128% of the AA ingested. The AA composition of ileal digesta was very different from that of casein, closely resembling that of endogenous proteins. The AA composition of faeces was very close to that of bacterial proteins. The ileal digestibilities of AA, though lower than their faecal values, were very high. This was confirmed by AA absorption balances greater than 100%. These data suggest that casein was almost totally digested by the terminal ileum, and that endogenous AA were substantially reabsorbed. These findings are supported by data on endogenous N recycling (15N), reported in a following paper.     

The gastrointestinal absorption of organically bound forms of plutonium in fed and fasted hamsters

Values of about 0.005-0.01 per cent were obtained for the absorption in fed hamsters of plutonium ingested as Pu4+ citrate, isocitrate, phytate and malate complexes and Pu3+ ascorbate compared with about 0.003-0.004 per cent for Pu4+ nitrate. Replacing drinking water with tea did not affect the result for Pu4+ nitrate. Fasting hamsters for 8 h before the administration of plutonium citrate increased absorption to 0.1-0.2 per cent. An extra period of fasting for 4 h after administration did not lead to a further increase in absorption. Similar values were also obtained when plutonium citrate was administered after a 24 h fast, followed either by immediate access to food or a further 4 h fast. In hamsters fasted for 24 h before administration of either Pu3+ ascorbate or Pu4+ nitrate, about 6-7 per cent of the ingested plutonium was retained in the gastrointestinal tract after one week. At three weeks after ingestion of Pu3+ ascorbate, gastrointestinal retention had fallen 100-fold without an increase in absorption.     

Transient pockets on XIAP-BIR2: toward the characterization of putative binding sites of small-molecule XIAP inhibitors

Protein-protein interactions are abundant in signal transduction pathways and thus of crucial importance in the regulation of apoptosis. However, designing small-molecule inhibitors for these potential drug targets is very challenging as such proteins often lack well-defined binding pockets. An example for such an interaction is the binding of the anti-apoptotic BIR2 domain of XIAP to the pro-apoptotic caspase-3 that results in the survival of damaged cells. Although small-molecule inhibitors of this interaction have been identified, their exact binding sites on XIAP are not known as its crystal structures reveal no suitable pockets. Here, we apply our previously developed protocol for identifying transient binding pockets to XIAP-BIR2. Transient pockets were identified in snapshots taken during four different molecular dynamics simulations that started from the caspase-3:BIR2 complex or from the unbound BIR2 structure and used water or methanol as solvent. Clustering of these pockets revealed that surprisingly many pockets opened in the flexible linker region that is involved in caspase-3 binding. We docked three known inhibitors into these transient pockets and so determined five putative binding sites. In addition, by docking two inactive compounds of the same series, we show that this protocol is also able to distinguish between binders and nonbinders which was not possible when docking to the crystal structures. These findings represent a first step toward the understanding of the binding of small-molecule XIAP-BIR2 inhibitors on a molecular level and further highlight the importance of considering protein flexibility when designing small-molecule protein-protein interaction inhibitors.     

II. Quantitative aspects of phosphorus absorption in ruminants

Phosphorus absorption in ruminants was analysed from a database described in a previous article. For common values of ingested phosphorus (2.5-5.0 g x kg(-1) of DM), 0.73 of dietary phosphorus is absorbed. The remaining variability is probably due to phosphorus quality. Phosphorus absorbed from silage, cereal, cereal by-products and hay differs greatly. The current true absorption coefficient used to calculate daily phosphorus supply is a constant value in the current systems and often it underestimates the true absorption resulting in an excess of phosphorus being supplied in the diets. Adjusting the true absorption coefficient values requires better characterisation of the phosphorus supplied by each feedstuff. Dietary influences (phytate phosphorus, crude fibre, etc.) were investigated but trials assessing the ration effect on phosphorus absorption are lacking. Since rumen microbes have specific phosphorus requirements, particularly for cell-wall degradation, the feedstuffphosphorus availability for the rumen ecosystem is discussed.     

应用于贝类生理生态学研究的生物沉积法

绝大多数软体动物门瓣鳃纲Ｌａｍｅｌｌｉｂｒａｎｃｈｉａ(又名双壳纲Ｂｉｖｉｌｖｉａ)动物 ,如贻贝 (Ｍｙｔｉｌｉｄａｅ)、扇贝 (Ｐｅｃｔｉｎｉｄａｅ)、牡蛎 (Ｏｓｔｒｅｉｄａｅ)等 ,属于滤食性动物 ,这些动物通过纤毛摆动、鳃和唇瓣过滤水体中的悬浮颗粒物 (包括浮游藻类、浮游细菌、中型动物和有机碎屑等 ) ,代表着一种摄食机制。双壳贝类在自然界水域的栖息范围比较广泛 ,包括河流、淡水湖、咸水湖、河口湾以及各种浅海和深海环境 ,但以沿岸海水环境为最重要的栖息地 ,其种类多 ,数量大[3] 。近些年来 ,海水双壳贝类养殖在世界范…     

Absorption efficiencies of Lytechinus variegatus (Lamarck) (Echinodermata: Echinoidea) for selected marine plants

The absorption efficiencies of the sea urchin Lytechinus variegatus (Lamarck) were calculated by an indicator method. Absorption efficiencies for total organic material, protein, lipid, and carbohydrate were highest with Halimeda incrassata Ellis as food (43 %, 65 %, 71 % and 35 %, respectively), and lowest with Eucheuma isiforme C. Agardh as food (?35%, ?42%, ?56%and ?34%, respectively). Moderately positive absorption efficiencies for total organic material were found with Thalassia testudinum König & Sims and Ulva lactuca L. as food, and low positive absorption efficiencies for total organic material are found with Syringodium filiformis Kützing and Sargassum sp. as food. Absorption efficiencies of the sea urchin Strongylocentrotus purpuratus (Stimpson) (calculated from the literature) were much greater than those of Lytechinus for carbohydrate, but less for protein and lipid. Both sea urchins showed negative efficiencies for some nutrient fractions of their foods. The absorption index (mg nutrient fraction absorbed/g dry wt ingested) showed that Thalassia supplied the most protein and total organic material (43 mg/g dry wt ingested and 142 mg/g dry wt ingested, respectively). In general, the absorption indices indicate that Lytechinus obtains more protein and lipid, but less carbohydrate from its food than does Strongylocentrotus. In the field Lytechinus has primarily a detrital diet.     

Elimination of protein-food ingested into the crop, and failure of ovarian development in female mosquitoes, Culex pipiens pallens

ABSTRACT. A protein-food rarely activates oogenesis in female mosquitoes when the protein is mixed with sugar and therefore ingested into the crop. A high occurrence of ovarian development occurs, however, when the protein is stored in the midgut. Examination of female C. pipiens pallens fed on egg albumin containing sugar revealed that the protein, once stored in the crop, was easily eliminated without any absorption and that ovarian development occurred only in those few females which retained some part of the ingested albumin for more than 24 h after feeding. Sugar mixed with the albumin was not detected in any excreta of fed females, suggesting that the sugar was digested irrespective of the protein elimination.     

Comparative surface geometry of the protein kinase family

Identifying conserved pockets on the surfaces of a family of proteins can provide insight into conserved geometric features and sites of protein–protein interaction. Here we describe mapping and comparison of the surfaces of aligned crystallographic structures, using the protein kinase family as a model. Pockets are rapidly computed using two computer programs, FADE and Crevasse. FADE uses gradients of atomic density to locate grooves and pockets on the molecular surface. Crevasse, a new piece of software, splits the FADE output into distinct pockets. The computation was run on 10 kinase catalytic cores aligned on the αF‐helix, and the resulting pockets spatially clustered. The active site cleft appears as a large, contiguous site that can be subdivided into nucleotide and substrate docking sites. Substrate specificity determinants in the active site cleft between serine/threonine and tyrosine kinases are visible and distinct. The active site clefts cluster tightly, showing a conserved spatial relationship between the active site and αF‐helix in the C‐lobe. When the αC‐helix is examined, there are multiple mechanisms for anchoring the helix using spatially conserved docking sites. A novel site at the top of the N‐lobe is present in all the kinases, and there is a large conserved pocket over the hinge and the αC‐β4 loop. Other pockets on the kinase core are strongly conserved but have not yet been mapped to a protein–protein interaction. Sites identified by this algorithm have revealed structural and spatially conserved features of the kinase family and potential conserved intermolecular and intramolecular binding sites.     

Scanning electron microscopy of intraovarian embryos of the viviparous teleost, Micrometrus minimus (Gibbons), (Perciformes: Embiotocidae)

Scanning electron microscopy of the hypertrophied hindgut and spatulate expansions of the fin membrane in embryos of a viviparous surfperch Micrometrus minimus (Gibbons) has revealed evidence as to the role of these secondary embryonic adaptations in foetal-maternal exchange processes. The villified hindgut probably functions primarily in the absorption of dissolved nutrient material from ingested ovarian fluid, while the vascular spatulate expansions facilitate the respiratory exchange of gases between embryonic and maternal circulations.     

Effects of acute hypobaric hypoxia on the appearance of ingested deuterium from a deuterium oxide-labelled carbohydrate beverage in body fluids of humans during prolonged cycling exercise

To determine whether or not acute hypobaric hypoxia alters the rate of water absorption from a carbohydrate beverage ingested during exercise, six men cycled for 80 min on three randomly assigned different occasions. In one trial, exercise was performed in hypoxia (barometric pressure, P(B) = 594 hPa, altitude 4,400 m) at an exercise intensity selected to elicit 75% of the individual's maximal oxygen uptake (VO2max) previously determined in such conditions. In the two other experiments, the subjects cycled in normoxia (P(B) = 992 hPa) at the same absolute and the same relative intensities as in hypoxia, which corresponded to 55% and 75%, respectively, of their VO2max determined in normoxia. The subjects consumed 400 ml of a 12.5% glucose beverage just prior to exercise, and 250 ml of the same drink at 20, 40 and 60 min from the beginning of exercise. The first drink contained 20 ml of deuterium oxide to serve as a tracer for the entry of water into body fluids. The heart rate (HR) during exercise was higher in hypoxia than in normoxia at the same absolute exercise intensity, whereas it was similar to HR measured in normoxia at the same relative exercise intensity. Both in normoxia and hypoxia, plasma noradrenaline concentrations were related to the relative exercise intensity up to 40 min of exercise. Beyond that duration, when exercise was performed at the highest absolute power in normoxia, the noradrenaline response was higher than in hypoxia at the same relative exercise intensity. No significant differences were observed among experimental conditions, either in temporal profiles of plasma D accumulation or in elimination of water ingested in sweat. Conversely, elimination in urine of the water ingested appeared to be related to the severity of exercise, either high absolute power or the same relative power combined with hypoxia. We concluded that water absorption into blood after drinking a 12.5% glucose beverage is not altered during cycling exercise in acute hypobaric hypoxia. It is suggested that the elimination of water ingested in sweat and urine may be dependent on local circulatory adjustments during exercise.     

Histology and histochemistry of the oviductal sperm storage pockets of the agamid lizard Calotes versicolor

Some reptiles are known to possess sperm storage in the female genital tract. Examination of the oviduct of the lizard Calotes versicolor shows the presence of sperm in storage pockets of the uterovaginal region throughout the reproductive cycle. Histochemical studies indicate that the epithelium of these pockets resembles the local oviductal epithelium and that a considerable amount of activity of steroidogenic and metabolic enzymes exists.     

The regulation of growth by locusts through post-ingestive compensation for variation in the levels of dietary protein and carbohydrate

Summary Post-ingestive responses to four different diets containing protein and digestible carbohydrate in balanced (7% protein, 7% digestible carbohydrate or 21% protein, 21% digestible carbohydrate) or unbalanced proportions (7% protein, 21% digestible carbohydrate or 21% protein, 7% digestible carbohydrate) were assessed in fifth stadium Locusta migratoria. Amounts eaten, defaecated and grown were measured across the fifth stadium. Results showed that locusts differentially utilized ingested protein and carbohydrate and consequently achieved similar growth on all four diets despite having ingested very different amounts of protein and carbohydrate. Nutrients in excess of requirements were voided from the animal mainly after digestion and absorption from the gut. In the case of excess nitrogen, a major route for removal was through increased uric acid excretion, while for carbohydrate the major removal mechanism was apparently via respiration.