Cardenolide‐defended milkweed bugs do not evoke learning in Nephila senegalensis spiders

Antipredator defense of herbivorous insects often relies on the potential toxicity of defensive chemicals sequestered from their host plants. The colorful Lygaeinae (Heteroptera: Lygaeidae) store a concentrated mixture of toxic cardenolides (cardiac glycosides) in specialized storage compartments of the bugs' integument, from which they are released upon attack. Larvae and adults of the large milkweed bug Oncopeltus fasciatus (Heteroptera: Lygaeinae) are specialized to feed on cardenolide‐containing milkweeds in the plant genus Asclepias and display a conspicuous red and black colorations. To investigate whether O. fasciatus gained improved protection by feeding on a toxic host plant (Asclepias syriaca), compared to a nontoxic alternative (sunflower seeds), we fed nymphs and adults of O. fasciatus to the golden orb‐weaver Nephila senegalensis. While visually oriented vertebrates, such as avian predators, have been intensively investigated for their reaction to defensive compounds and aposematic coloration, less attention has been paid to invertebrate predators. Their different perceptual abilities can provide important opportunities for testing hypotheses on warning coloration and chemical defenses. The predation trials showed that the bugs fed on Asclepias were significantly less likely to be killed than the bugs reared on a cardenolide‐free diet. This suggests that sequestered cardenolides in O. fasciatus nymphs and adults represent a significant fitness advantage on an individual level against this invertebrate predator. Yet, when testing for avoidance learning in the spiders, negative experience did not change the way how similar prey was attacked at the next encounter. In this case, visual or chemical aposematism thus does not seem to matter for predator learning.     

Root damage to apple plants by cockchafer larvae induces a change in volatile signals below‐ and above‐ground

Volatile organic compounds (VOCs) mediate communication between plants and insects. Plants under insect herbivore attack release VOCs either at the site of attack or systemically, indicating within‐plant communication. Some of these VOCs, which may be induced only upon herbivore attack, recruit parasitoids and predatory insects to feed on the attacking insects. Moreover, some plants are able to ‘eavesdrop’ on herbivore‐induced plant volatiles (HIPVs) to prime themselves against impending attack; such eavesdropping exemplifies plant–plant communication. In apple orchards, the beetle Melolontha melolontha L. (Coleoptera: Scarabaeidae) is an important insect pest whose larvae live and feed on roots for about 4 years. In this study, we investigated whether the feeding activity of M. melolontha larvae (1) alters the volatile profile of apple roots, (2) induces the release of HIPVs systemically in the leaves, and (3) whether infested plants communicate to neighbouring non‐infested conspecifics through HIPVs. To answer these questions, we collected constitutive VOCs from intact M9 roots as well as M. melolontha larvae‐damaged roots using a newly designed ‘rhizobox’, to collect root‐released volatiles in situ, without damaging the plant root system. We also collected VOCs from the leaf‐bearing shoots of M9 whose roots were under attack by M. melolontha larvae and from shoots of neighbouring non‐infested conspecifics. Gas chromatography‐mass spectrometry analysis showed that feeding activity of M. melolontha larvae induces the release of specific HIPVs; for instance, camphor was found in the roots only after larvae caused root damage. Melolontha melolontha also induced the systemic release of methyl salicylate and (E,E)‐α‐farnesene from the leaf‐bearing shoots. Methyl salicylate and (E,E)‐α‐farnesene were also released by the shoots of non‐infested neighbouring conspecifics. These phenomena indicate the induction of specific VOCs below‐ and above‐ground upon M. melolontha larvae feeding on apple roots as well as plant–plant communication in apple plants.     

Dynamic changes in plant secondary metabolites during UV acclimation in Arabidopsis thaliana

Plants respond to environmental stress by synthesizing a range of secondary metabolites for defense purposes. Here we report on the effect of chronic ultraviolet (UV) radiation on the accumulation of plant secondary metabolites in Arabidopsis thaliana leaves. In the natural environment, UV is a highly dynamic environmental parameter and therefore we hypothesized that plants are continuously readjusting levels of secondary metabolites. Our data show distinct kinetic profiles for accumulation of tocopherols, polyamines and flavonoids upon UV acclimation. The lipid‐soluble antioxidant α‐tocopherol accumulated fast and remained elevated. Polyamines accumulated fast and transiently. This fast response implies a role for α‐tocopherol and polyamines in short‐term UV response. In contrast, an additional sustained accumulation of flavonols took place. The distinct accumulation patterns of these secondary metabolites confirm that the UV acclimation process is a dynamic process, and indicates that commonly used single time‐point analyses do not reveal the full extent of UV acclimation. We demonstrate that UV stimulates the accumulation of specific flavonol glycosides, i.e. kaempferol and (to a lesser extent) quercetin di‐ and triglycosides, all specifically rhamnosylated at position seven. All metabolites were identified by Ultra Performance Liquid Chromatography (UPLC)‐coupled tandem mass spectrometry. Some of these flavonol glycosides reached steady‐state levels in 3–4 days, while concentrations of others are still increasing after 12 days of UV exposure. A biochemical pathway for these glycosides is postulated involving 7‐O‐rhamnosylation for the synthesis of all eight metabolites identified. We postulate that this 7‐O‐rhamnosylation has an important function in UV acclimation.     

Vanadate‐Based Materials for Li‐Ion Batteries: The Search for Anodes for Practical Applications

While the practical application of electrode materials depends intensively on the Li⁺ ion storage mechanisms correlating ultimately with the coulombic efficiency, reversible capacity, and morphology variation of electrode material upon cycling, only intercalation‐type electrode materials have proven viable for commercialization up to now. This paper reviews the promising anode materials of metal vanadates (M_xV_yO_z, M = Co, Cu, Mn, Fe, Zn, Ni, Li) that have high capacity, low cost, and abundant resource, and also discusses the related Li⁺ ion storage mechanism. It is concluded that most of these (M_xV_yO_z, M = Co, Cu, Mn, Fe, Zn, Ni) exhibit irreversible redox reactions upon lithiation/delithiation accompanied by large volume expansion, which is not favorable for industrial applications. In particular, Li₃VO₄ with specific intercalation Li⁺ ion storage mechanism and compatible merits of safety and energy density exhibits great potential for practical application. This review systematically summarizes the latest progress in Li₃VO₄ research, including the representative fabrication approaches for advanced morphology and state‐of‐the‐art technologies to boost performance and the morphology variation associated with Li⁺ ion storage mechanisms. Furthermore, an outlook on where breakthroughs for Li₃VO₄ may be most likely achieved will be provided.     

Circulation of hemocoelic fluid during slow and fast swimming in the pteropod mollusc Clione limacina

In the pteropod mollusc Clione limacina Phipps 1774, individuals possess an open circulatory system that fills their body cavities and functions as a hydrostatic skeleton. Individuals of C. limacina demonstrate two distinct swimming behaviors, slow and fast swimming, and their wings are supported by their hydrostatic skeleton. We investigated the circulation of fluid within the body cavities of individuals of C. limacina by injecting dye into the hemocoelic compartments to visualize flow during both slow swimming and serotonin‐induced fast swimming. Hemocoelic fluid was observed to have a defined pattern of flow: rostrally from the heart into the wings and head, then following a dorsal pathway caudally into the body and tail before being taken up by the heart again. During patterned attack behavior, the neck constricted in width as the head's buccal cones were hydraulically inflated with hemocoelic fluid.     

Morphological basis of cardiac glycoside sequestration by Oncopeltus fasciatus (Dallas) (Hemiptera: Lygaeidae)

Summary The integument of Oncopeltus fasciatus is made up of a vacuolated and a pigmented epidermal cell layer. This double layered integument is present from late embryo to adult in male and female animals reared on milkweed or sunflower seeds. Experiments with a labelled glycoside as well as retrograde ink injections suggest that O. fasciatus concentrates cardiac glycosides, normally derived from the host plants, within the vacuolated epidermal cell layer throughout its life cycle. In the adult, droplets of glycoside-rich fluid appear at precise points along the dorsolateral margins when external pressure is applied to the thorax and abdomen. This pressure causes separation of cuticular flanges in the metathoracic epimeral lobe and rupture of the cuticle in restricted areas in the mesothorax and abdomen. In addition the pigmented epidermal cell layer and the distal membranes of the vacuolated epidermal cell layer rupture with the result that the contents of the vacuolated cell layer are eliminated onto the surface of the animal where they are retained as discrete droplets by the cuticular morphology. Release of cardiac glycosides into the haemolymph is prevented by a thick basal lamina on the haemolymph side of the vacuolated epidermal cells. No specialized muscles involved with fluid release were observed. The vacuolated epidermal cells do not show ultrastructural features characteristic of actively transporting tissues, i.e., abundant mitochondria and elaborate membrane infoldings. This suggests that glycoside sequestration is essentially a passive process and should not be associated with any physiological cost. Large concentration gradients of cardiac glycosides are maintained across the basal lamina, basal plasma and vacuolar membranes of the vacuolated epidermal cell layer. Possible mechanisms by which O. fasciatus is able to concentrate cardiac glycosides as well as the possible function of this phenomenon are discussed.Abbreviations A abdominal trabeculum - Ap mesofurcal apodeme - C metathoracic supracoxal lobe - D metathoracic stink gland duct - E metathoracic epimeral lobe - Ep pigmented epidermal cell layer - Ev vacuolated epidermal cell layer - G last thoracic ganglion - H haemocoel - M midgut - N nerve cord - P second phragma - R reproductive organ - T trachea - V dorsal vessel - W wing - bl basal lamina - c cuticle - cf cytoplasmic fragments - cv coated vesicle - d hemidesmosome - ep epidermal cell - en endocuticle - ex exocuticle - f flange - fp foot processes - gc glycoside compartment - h hair - is intersegmental region - id ink deposits - l lumen of metathoracic stink gland - m mitochondria - mb mushroom bodies - mt microtrichia - n nucleus - p pigment granule - s slit - sp spine - tsm tergosternal muscle - v vacuole     

Vesicular monoamine transporter 2 mediates fear behavior in mice

A subset of people exposed to a traumatic event develops post‐traumatic stress disorder (PTSD), which is associated with dysregulated fear behavior. Genetic variation in SLC18A2, the gene that encodes vesicular monoamine transporter 2 (VMAT2), has been reported to affect risk for the development of PTSD in humans. Here, we use transgenic mice that express either 5% (VMAT2‐LO mice) or 200% (VMAT2‐HI mice) of wild‐type levels of VMAT2 protein. We report that VMAT2‐LO mice have reduced VMAT2 protein in the hippocampus and amygdala, impaired monoaminergic vesicular storage capacity in both the striatum and frontal cortex, decreased monoamine metabolite abundance and a greatly reduced capacity to release dopamine upon stimulation. Furthermore, VMAT2‐LO mice showed exaggerated cued and contextual fear expression, altered fear habituation, inability to discriminate threat from safety cues, altered startle response compared with wild‐type mice and an anxiogenic‐like phenotype, but displayed no deficits in social function. By contrast, VMAT2‐HI mice exhibited increased VMAT2 protein throughout the brain, higher vesicular storage capacity and greater dopamine release upon stimulation compared with wild‐type controls. Behaviorally, VMAT2‐HI mice were similar to wild‐type mice in most assays, with some evidence of a reduced anxiety‐like responses. Together, these data show that presynaptic monoamine function mediates PTSD‐like outcomes in our mouse model, and suggest a causal link between reduced VMAT2 expression and fear behavior, consistent with the correlational relationship between VMAT2 genotype and PTSD risk in humans. Targeting this system is a potential strategy for the development of pharmacotherapies for disorders like PTSD.     

Chrysanthemum expressing a linalool synthase gene ‘smells good’, but ‘tastes bad’ to western flower thrips

Herbivore‐induced plant volatiles are often involved in direct and indirect plant defence against herbivores. Linalool is a common floral scent and found to be released from leaves by many plants after herbivore attack. In this study, a linalool/nerolidol synthase, FaNES1, was overexpressed in the plastids of chrysanthemum plants (Chrysanthemum morifolium). The volatiles of FaNES1 chrysanthemum leaves were strongly dominated by linalool, but they also emitted small amount of the C11‐homoterpene, (3E)‐4,8‐dimethyl‐1,3,7‐nonatriene, a derivative of nerolidol. Four nonvolatile linalool glycosides in methanolic extracts were found to be significantly increased in the leaves of FaNES1 plants compared to wild‐type plants. They were putatively identified by LC‐MS‐MS as two linalool–malonyl–hexoses, a linalool–pentose–hexose and a glycoside of hydroxy–linalool. A leaf‐disc dual‐choice assay with western flower thrips (WFT, Frankliniella occidentalis) showed, initially during the first 15 min of WFT release, that FaNES1 plants were significantly preferred. This gradually reversed into significant preference for the control, however, at 20–28 h after WFT release. The initial preference was shown to be based on the linalool odour of FaNES1 plants by olfactory dual‐choice assays using paper discs emitting pure linalool at similar rates as leaf discs. The reversal of preference into deterrence could be explained by the initial nonvolatile composition of the FaNES1 plants, as methanolic extracts were less preferred by WFT. Considering the common occurrence of linalool and its glycosides in plant tissues, it suggests that plants may balance attractive fragrance with ‘poor taste’ using the same precursor compound.     

A distinct class of vesicles derived from the trans‐Golgi mediates secretion of xylogalacturonan in the root border cell

Root border cells lie on the surface of the root cap and secrete massive amounts of mucilage that contains polysaccharides and proteoglycans. Golgi stacks in the border cells have hypertrophied margins, reflecting elevated biosynthetic activity to produce the polysaccharide components of the mucilage. To investigate the three‐dimensional structures and macromolecular compositions of these Golgi stacks, we examined high‐pressure frozen/freeze‐substituted alfalfa root cap cells with electron microscopy/tomography. Golgi stacks in border cells and peripheral cells, precursor cells of border cells, displayed similar morphological features, such as proliferation of trans cisternae and swelling of the trans cisternae and trans‐Golgi network (TGN) compartments. These swollen margins give rise to two types of vesicles larger than other Golgi‐associated vesicles. Margins of trans‐Golgi cisternae accumulate the LM8 xylogalacturonan (XGA) epitope, and they become darkly stained large vesicles (LVs) after release from the Golgi. Epitopes for xyloglucan (XG), polygalacturonic acid/rhamnogalacturonan‐I (PGA/RG‐I) are detected in the trans‐most cisternae and TGN compartments. LVs produced from TGN compartments (TGN‐LVs) stained lighter than LVs and contained the cell wall polysaccharide epitopes seen in the TGN. LVs carrying the XGA epitope fuse with the plasma membrane only in border cells, whereas TGN‐LVs containing the XG and PGA/RG‐I epitopes fuse with the plasma membrane of both peripheral cells and border cells. Taken together, these results indicate that XGA is secreted by a novel type of secretory vesicles derived from trans‐Golgi cisternae. Furthermore, we simulated the collapse in the central domain of the trans‐cisternae accompanying polysaccharide synthesis with a mathematical model.     

Manipulation of flavour and aroma compound sequestration and release using a glycosyltransferase with specificity for terpene alcohols

Glycosides are an important potential source of aroma and flavour compounds for release as volatiles in flowers and fruit. The production of glycosides is catalysed by UDP‐glycosyltransferases (UGTs) that mediate the transfer of an activated nucleotide sugar to acceptor aglycones. A screen of UGTs expressed in kiwifruit (Actinidia deliciosa) identified the gene AdGT4 which was highly expressed in floral tissues and whose expression increased during fruit ripening. Recombinant AdGT4 enzyme glycosylated a range of terpenes and primary alcohols found as glycosides in ripe kiwifruit. Two of the enzyme's preferred alcohol aglycones, hexanol and (Z)‐hex‐3‐enol, contribute strongly to the ‘grassy‐green’ aroma notes of ripe kiwifruit and other fruit including tomato and olive. Transient over‐expression of AdGT4 in tobacco leaves showed that enzyme was able to glycosylate geraniol and octan‐3‐ol in planta whilst transient expression of an RNAi construct in Actinidia eriantha fruit reduced accumulation of a range of terpene glycosides. Stable over‐expression of AdGT4 in transgenic petunia resulted in increased sequestration of hexanol and other alcohols in the flowers. Transgenic tomato fruit stably over‐expressing AdGT4 showed changes in both the sequestration and release of a range of alcohols including 3‐methylbutanol, hexanol and geraniol. Sequestration occurred at all stages of fruit ripening. Ripe fruit sequestering high levels of glycosides were identified as having a less intense, earthier aroma in a sensory trial. These results demonstrate the importance of UGTs in sequestering key volatile compounds in planta and suggest a future approach to enhancing aromas and flavours in flowers and during fruit ripening.     

Trafficking of the myrosinase‐associated protein GLL23 requires NUC/MVP1/GOLD36/ERMO3 and the p24 protein CYB

Proteins detrimental to endoplasmic reticulum (ER) morphology need to be efficiently exported. Here, we identify two mechanisms that control trafficking of Arabidopsis thalianaGLL23, a 43 kDa GDSL‐like lipase implicated in glucosinolate metabolism through its association with the β‐glucosidase myrosinase. Using immunofluorescence, we identified two mutants that showed aberrant accumulation of GLL23: large perinuclear ER aggregates in the nuclear cage (nuc) mutant; and small compartments contiguous with the peripheral ER in the cytoplasmic bodies (cyb) mutant. Live imaging of fluorescently tagged GLL23 confirmed its presence in the nuc and cyb compartments, but lack of fluorescent signals in the wild‐type plants suggested that GLL23 is normally post‐translationally modified for ER export. NUC encodes the MVP1/GOLD36/ERMO3 myrosinase‐associated protein, previously shown to have vacuolar distribution. CYB is an ER and Golgi‐localized p24 type I membrane protein component of coat protein complex (COP) vesicles, animal and yeast homologues of which are known to be involved in selective cargo sorting for ER–Golgi export. Without NUC, GLL23 accumulates in the ER this situation suggests that NUC is in fact active in the ER. Without CYB, both GLL23 and NUC were found to accumulate in cyb compartments, consistent with a role for NUC in GLL23 processing and indicated that GLL23 is the likely sorting target of the CYB p24 protein.     

Detrimental effects of latex and cardiac glycosides on survival and growth of first-instar monarch butterfly larvae Danaus plexippus feeding on the sandhill milkweed Asclepias humistrata

1. A novel experimental method was developed to study negative physical and chemical effects of latex and cardiac glycosides on first‐instar monarch butterfly larvae in their natural environment in north central Florida. Forceps were used to nibble through the petioles of leaves of the sandhill milkweed Asclepias humistrata, mimicking the behaviour of mature monarch larvae. This notching cut off the supply of latex to the leaves without significantly reducing either their cardiac glycoside concentration or water content. 2. The mean cardiac glycoside concentration in larvae that fed on intact leaves was nearly two and a half times greater than in larvae that fed on notched leaves. This was probably because more latex is present in the gut of the larvae that fed on the intact leaves. Supporting this is the fact that the mean concentration of cardiac glycosides in the latex was 34–47 times that in the leaves. 3. Wet weights, dry weights, and growth rates of first‐instar larvae that fed on intact leaves over a 72‐h period were less than half those of larvae that fed on notched leaves. 4. Mortality due to miring in the latex was 27% on the intact leaves compared with 2% on the notched leaves. 5. Latex, cardiac glycosides, and other as yet undetermined plant factors all have a negative effect on first‐instar larval survival. 6. Video‐analyses indicated that ingestion of latex caused the larvae to become cataleptic and increased their chances of being mired on the leaf by the setting latex glue. Dysfunction resulting from latex ingestion may lead to the larvae falling off the plant and being killed by invertebrate predators. 7. The difficulty of neonate monarch larvae surviving on A. humistrata – one of the principal milkweed species fed on each spring as monarchs remigrate from Mexico into the southern U.S.A. – is evidence that a co‐evolutionary arms race is operating in this plant–herbivore system.     

FLOW PATTERN AROUND THE RIGID CEPHALIC SHIELD OF THE DEVONIAN AGNATHAN ERRIVASPIS WAYNENSIS (PTERASPIDIFORMES: HETEROSTRACI)

Abstract: Palaeozoic armoured agnathans (or ostracoderms) are characterised by having an external, bone shield enclosing the anterior part of their bodies, which demonstrate great diversity of both forms and sizes. The functional significance of these cephalic shields remains unclear (they may have been a functional analogue of the vertebral column, or merely afforded protection). Here we assess the importance of the cephalic shield in terms of locomotion. In order to do this, we have studied flow patterns of the Devonian heterostracan Errivaspis waynensis ( White, 1935 ), using an anatomically correct model of E. waynensis positioned at different pitching angles. The fluid flow was visualised in a wind tunnel, using planar light sheet techniques, adding vaporised propylene glycol to the fluid. The flow pattern over the cephalic shield of Errivaspis is dominated by the formation of leading‐edge vortices (LEVs). When the model was positioned at angles of attack of ‐2 degrees or higher a pair of nearly symmetrical, counter‐rotating primary vortices were produced, which flowed downstream over the upper surface of the cephalic shield. At moderate angles of attack, LEVs remained attached to the dorsal surface, but, as the angle of attack increased above 7 degrees, vortices began to separate from the surface at posterior locations. At a high angles of attack (around 12 degrees or 13 degrees), vortex breakdown (or vortex burst) occured. The body‐induced vortical flow around the cephalic shield is very similar to the that described over delta wing aircraft. This strategy generates lift forces through vortex generation (vortex lift). Based on this analogue and knowing that Errivaspis lacked pectoral fins or any other obvious control surfaces, vortex lift forces added through this mechanism may have played a major role in the locomotion of these primitive fishes, not only to counteract the negative buoyancy of the fish, but also as a means of manoeuvring.     

Neuronal sub‐compartmentalization: a strategy to optimize neuronal function

Neurons are highly polarized cells that consist of three main structural and functional domains: a cell body or soma, an axon, and dendrites. These domains contain smaller compartments with essential roles for proper neuronal function, such as the axonal presynaptic boutons and the dendritic postsynaptic spines. The structure and function of these compartments have now been characterized in great detail. Intriguingly, however, in the last decade additional levels of compartmentalization within the axon and the dendrites have been identified, revealing that these structures are much more complex than previously thought. Herein we examine several types of structural and functional sub‐compartmentalization found in neurons of both vertebrates and invertebrates. For example, in mammalian neurons the axonal initial segment functions as a sub‐compartment to initiate the action potential, to select molecules passing into the axon, and to maintain neuronal polarization. Moreover, work in Drosophila melanogaster has shown that two distinct axonal guidance receptors are precisely clustered in adjacent segments of the commissural axons both in vivo and in vitro, suggesting a cell‐intrinsic mechanism underlying the compartmentalized receptor localization. In Caenorhabditis elegans, a subset of interneurons exhibits calcium dynamics that are localized to specific sections of the axon and control the gait of navigation, demonstrating a regulatory role of compartmentalized neuronal activity in behaviour. These findings have led to a number of new questions, which are important for our understanding of neuronal development and function. How are these sub‐compartments established and maintained? What molecular machinery and cellular events are involved? What is their functional significance for the neuron? Here, we reflect on these and other key questions that remain to be addressed in this expanding field of biology.     

High elevation Plantago lanceolata plants are less resistant to herbivory than their low elevation conspecifics: is it just temperature?

Traits that mediate species interactions are evolutionarily shaped by biotic and abiotic drivers, yet we know relatively little about the relative importance of these factors. Herbivore pressure, along with resource availability and ‘third‐party’ mutualists, are hypothesized to play a major role in the evolution of plant defence traits. Here, we used the model system Plantago lanceolata, which grows along steep elevation gradients in the Swiss Alps, to investigate the effect of elevation, herbivore pressure, mycorrhizal inoculation and temperature on plant resistance. Over a 1200 m elevation gradient, the levels of herbivory and iridoid glycosides (IGs) declined with increasing elevation. By planting seedlings at three different elevations, we further showed that both low‐elevation growing conditions and mycorrhizal inoculation resulted in increased plant resistance to herbivores. Finally, using a temperature‐controlled experiment comparing high‐ and low‐elevation ecotypes, we showed that high‐elevation ecotypes are less resistant to herbivory, and that lower temperatures impair IGs deployment after herbivore attack. We thus propose that both lower herbivore pressure, and colder temperatures relax the defense syndrome of high elevation plants.     

Effect of fermentation, aging and thermal storage on total glycosides, phenol-free glycosides and volatile compounds of White Riesling (Vitis vinifera L.) wines

There is growing recognition of the significance of the products of glycoside hydrolysis to varietal wine aroma. White Riesling wines were produced from four strains of Saccharomyces cerevisiae. Wines underwent conventional aging or anaerobic thermal storage (20 days at 45°C) either 2 or 40 months post-fermentation to quantify influences on total glycosides, phenol-free glycosides and selected volatiles. Glycoside and free volatile concentrations were estimated by analysis of glycosyl-glucose and gas chromatography/mass spectrometry, respectively. Thermal storage of wines 2 months post-fermentation reduced the total glycosides by an average of 33% for all yeasts and increased the concentration of free benzyl alcohol while decreasing the concentration of free linalool and geraniol. Conventional aging for 40 months reduced the total and phenol-free glycosides equally among yeasts by an average of 60%, with phenol-free glycosides averaging 80% of the total. Thermal storage of aged wines reduced the total glycoside concentration by an additional 29%. The effect of thermal storage on selected volatile phenols, higher alcohols, esters, acids, terpenes, carbonyl compounds, C-13 norisoprenoids and six-carbon alcohols was variable depending upon the component. Received 29 September 1998/ Accepted in revised form 16 January 1999     

Neuron‐specific non‐classical release of prothymosin alpha: a novel neuroprotective damage‐associated molecular patterns

Prothymosin alpha (ProTα), a nuclear protein devoid of signal sequence, has been shown to possess a number of cellular functions including cell survival. Most recently, we demonstrated that ProTα is localized in the nuclei of neurons, while it is found in both nuclei and cytoplasm in the astrocytes and microglia of adult brain. However, the cell type‐specific non‐classical release of ProTα under cerebral ischemia is yet unknown. In this study, we report that ProTα is non‐classically released along with S100A13 from neurons in the hippocampus, striatum and somatosensory cortex at 3 h after cerebral ischemia, but amlexanox (an anti‐allergic compound) reversibly blocks this neuronal ProTα release. We found that none of ProTα is released from astrocytes and microglia under ischemic stress. Indeed, ProTα intensity is increased gradually in astrocytes and microglia through 24 h after the cerebral ischemia. Interestingly, Z‐Val‐Ala‐Asp fluoromethyl ketone, a caspase 3 inhibitor, pre‐treatment induces ProTα release from astrocytes in the ischemic brain, but this release is reversibly blocked by amlexanox. However, Z‐Val‐Ala‐Asp fluoromethyl ketone as well as amlexanox has no effect on ProTα distribution in microglia upon cerebral ischemia. Taken together, these results suggest that only neurons have machineries to release ProTα upon cerebral ischemic stress in vivo.     

Angling-induced cardiac disturbance of free-swimming largemouth bass (Micropterus salmoides) monitored with heart rate telemetry

The sub‐lethal effects of catch‐and‐release angling have been poorly studied because of the difficulties in monitoring physiological parameters in free‐swimming fish. Laboratory studies provide the opportunity to examine sub‐lethal effects in controlled environments, but do not incorporate site‐specific characteristics. In this study we angled free‐swimming largemouth bass (Micropterus salmoides) equipped with heart rate transmitters to exhaustion using rod and reel, and exposed fish to air for 30 s. Experiments were repeated at four water temperatures (13, 17, 21, and 25°C). These field data were compared with published findings from largemouth bass collected at the same water temperatures in a controlled laboratory setting using Doppler flow probes. Field collected heart rate data increased with increasing water temperatures (Q₁₀ values 1.30–1.37). Pre‐disturbance heart rates were ～30% higher for free‐swimming fish in the field than previously collected laboratory data at the same water temperatures. Fish angled in the field exhausted ～40% more rapidly than fish chased in the laboratory. Maximal heart rate was ～15% higher for free‐swimming fish in the field than for data collected from laboratory restrained fish, but scope for heart rate was reduced by up to 20% in the field, especially at higher water temperatures. Heart rate in free‐swimming fish was highly variable at all times, obscuring clear recovery patterns. Conversely, laboratory cardiac parameters exhibited less variable patterns, peaking clearly following disturbances and recovering in about 135 min, independent of water temperature. Based upon these findings, we suggest that comprehensive studies incorporating both laboratory and field experiments are needed for truly understanding the effect of catch‐and‐release angling on fish.