Some unusual staining properties of tannic acid in plants

Summary Maize root tips were fixed in glutaraldehyde fixatives containing tannic acid and then processed for electron microscopy. Under these conditions, tannic acid selectively stained the contents of the Golgi apparatus secretory vesicles of some outer root cap cells, the cell walls of all cells, and substances in, and adjacent to, intercellular connections of mature primary walls and of secondary walls. Intercellular connections of the young primary walls were not stained. Plasma membranes, and substances associated with the outer leaflets of the plasma membranes, were also stained. Tannic acid-positive material was associated with the cell plate vesicles of forming walls but very little, or none, was associated with the Golgi apparatus vesicles of dividing cells.     

Tropical marine herbivore assimilation of phenolic-rich plants

Phenolics in marine brown algae have been thought to follow a latitudinal gradient with high phenolic species in high latitudes and low phenolic species in low latitudes. However, tropical brown algae from the western Caribbean have been shown to be high in phlorotannin concentration, indicating that latitude alone is not a reasonable predictor of marine plant phenolic concentrations. This study shows that the range of high phenolic phaeophytes is not limited to the western Caribbean but encompasses the western tropical Atlantic, including Bermuda and the Caribbean, where algal phlorotannin concentrations can be as high as 25% dry weight (DW). Assimilation efficiencies (AEs) of phenolic-rich and phenolic-poor plants were examined in three tropical marine herbivores (the parrotfish, Sparisoma radians, and the brachyuran crab, Mithrax sculptus, from Belize and the parrotfish, Sparisoma chrysopterum, from Bermuda). AEs of phenolic-rich food by each of the three herbivore species were uniformly high, suggesting that high plant phenolic concentrations did not affect AEs in these species. This is in contrast to some temperate marine herbivores where phenolic concentrations of 10% DW have been shown to drastically reduce AE. The apparent contradiction is discussed in light of the effects of specific herbivore gut characteristics on successful herbivory of high phenolic brown algae.     

Alpha-L-fucosidases: exoglycosidases with unusual transglycosylation properties

Apha-LFucosidases (EC 3.2.1.51), the only members of the CAZy family GH-29, are widespread glycosidases involved in many biological processes including inflammation, metastasis, and the lysosomal storage disease fucosidosis. Despite their biological significance, information concerning the mechanism of alpha-Lfucosidases has only recently become available. In an attempt to obtain further data concerning their mechanism, we have investigated the hydrolytic and transglycosylation properties of a canine and a mollusk (Pecten maximus) alpha-Lfucosidase. Our results show that, despite the evolutionary distance between these two species, both enzymes have similar hydrolysis and transglycosylation properties. Surprisingly, we found that, starting from monosaccharides, these exoglycosidases were able to catalyze efficiently the synthesis of highly branched fuco-oligosaccharides as large as tetrasaccharides, a unique feature for a wild-type exoglycosidase. The structural analysis of the compounds formed revealed that the regioselectivity of alpha-Lfucosidases is strongly influenced by the structure of the acceptor. This leads us to propose an enzymatic approach for the preparative synthesis of fuco-oligosaccharides. This will not only allow the synthesis of biological determinants containing fucose but also of new fucose-containing oligosaccharides as alpha-glycosynthases appear to be difficult to obtain.     

Cadmium for all meals – plants with an unusual appetite

Since the beginning of the Bronze Age the acquisition of metals and the mastering of metal processing has been a driving force in human civilisation. It is difficult to imagine, however, how a plant could possibly profit from accumulating excessive amounts of a metal for which there is no metabolic need. Leaving behind an environmentally degraded wasteland, human mining and smeltering activities contributed to providing the niches for such plants, sometimes even accomplishing seed dispersal – through seeds attached to the boots of migrating miners. A stunning example of this is found in the healthy populations of Thlaspi caerulescens in the Cevennes (southern France), the leaves of which have been found to contain the toxic and non-essential metal cadmium (Cd) at concentrations between 1000 and 3000 mg kg⁻¹ dry biomass (R. D. Reeves, unpublished). In this issue, experiments under controlled experimental and glasshouse conditions are reported, confirming an extraordinary level of Cd accumulation and tolerance in these populations (Lombi et al ., pp. 11–20).     

A bacterium yielding a polysaccharide with unusual properties

A yellow-pigmented bacterium, isolated in Czapek-Dox medium, yielded copious quantities of extracellular polysaccharide. Liquid cultures became bright yellow and increasingly viscous after incubation for 4–6 d. Characterization tests indicate that it is a starch-negative (amylase-negative), gelatinase-positive strain of Pseudomonas paucimobilis , closely resembling an organism previously described as Pseudomonas elodea. The polysaccharide contained L-rhamnose and D-glucose in a molar ratio of 0.66 : 1.0, together with 16% D-glucuronic acid and 10.4% acetate. Solutions of the polysaccharide were highly viscous: certain ions tended to cause gelation of aqueous solutions. After deacetylation with mild alkali the polymer at a concentration of 0.8% yielded a strong gel.     

Membrane properties of isolated mudpuppy taste cells

The voltage-dependent currents of isolated Necturus lingual cells were studied using the whole-cell configuration of the patch-clamp technique. Nongustatory surface epithelial cells had only passive membrane properties. Small, spherical cells resembling basal cells responded to depolarizing voltage steps with predominantly outward K+ currents. Taste receptor cells generated both outward and inward currents in response to depolarizing voltage steps. Outward K+ currents activated at approximately 0 mV and increased almost linearly with increasing depolarization. The K+ current did not inactivate and was partially Ca++ dependent. One inward current activated at -40 mV, reached a peak at -20 mV, and rapidly inactivated. This transient inward current was blocked by tetrodotoxin (TTX), which indicates that it is an Na+ current. The other inward current activated at 0 mV, peaked at 30 mV, and slowly inactivated. This more sustained inward current had the kinetic and pharmacological properties of a slow Ca++ current. In addition, most taste cells had inwardly rectifying K+ currents. Sour taste stimuli (weak acids) decreased outward K+ currents and slightly reduced inward currents; bitter taste stimuli (quinine) reduced inward currents to a greater extent than outward currents. It is concluded that sour and bitter taste stimuli produce depolarizing receptor potentials, at least in part, by reducing the voltage-dependent K+ conductance.     

Prostaglandin phospholipid conjugates with unusual biophysical and cytotoxic properties

The synthesis of two secretory phospholipase A₂ IIA sensitive 15-deoxy-Δ^12,14-prostaglandin J₂ phospholipid conjugates is described and their biophysical and biological properties are reported. The conjugates spontaneously form particles in the liposome size region upon dispersion in an aqueous buffer and both phospholipids are hydrolyzed by phospholipase A₂, but with different conversion rates and extent of hydrolysis. The cytotoxicity was evaluated in HT-29 and Colo205 cells and the conjugates induced cell death in the presence of phospholipase A₂ and surprisingly also in the absence of the enzyme.     

Immunomodulating properties of Argentine plants with ethnomedicinal use

Five Argentine medicinal plants selected according to folk traditional or ethnomedical use, references and primary pharmacological screening; were chosen to elucidate their immunomodulating properties. Dichloromethane, methanolic and aqueous extracts of the aerial parts of Achyrocline flaccida (A. flaccida), Eupatorium arnottianum (E. arnottianum) and Eupatorioum buniifolium (E. buniifolium), leaves of Lithraea molleoides (L. molleoides) and leaves and stems of Phyllanthus sellowianus (P. sellowianus) were analyzed to disclose their effects on murine normal and tumor cell growth as well as on complement hemolytic activity. Modulation of cell growth was evaluated by tritiated thymidine incorporation while inhibition of complement activity was measured on both classical and alternative complement pathways (CP and AP respectively). The results obtained show that most of the extracts exerted inhibitory effect on tumor as well as on mitogen activated normal spleen cell growth. On tumor cells, IC50 ranged between 1-75 microg/ml for most of the extracts with the exception of dichloromethane of L. molleoides and P. sellowianus which required concentrations higher than 100 microg/ml to produce the effect. On mitogenic activated splenocytes, IC50 ranged between < 1 to 85 microg/ml with the exception of methanolic extract of E. buniifolium or P. sellowianus which were not effective on ConA or LPS stimulated splenocytes respectively. Only E. buniifolium was active on murine normal splenocytes proliferation (IC50 0.5-1.5 microg/ml). Finally, one (7%) of 15 extracts showed inhibition of complement activity on CP and 6 extracts (40%) presented moderate activity on CP. The dichloromethane extract of E. arnottianum was the most active (IC50 5 microg/ml), although remarkable effect was also obtained with dichloromethane and methanolic extracts of P. sellowianus (IC50 11.2 and 17.3 microg/ml respectively). Besides, 2 extracts (13%), dichloromethane extract of E. arnottianum and aqueous extract of P. sellowianus, showed moderate inhibition on AP.     

Some taste molecules and their solution properties.

The solution properties of a variety of different sapid substances from all four basic taste modalities, namely, sweet (n = 24), salty (n = 7), sour (n = 11) and bitter (n = 2), have been investigated. Some multisapophoric molecules, i.e. molecules exhibiting more than one taste, have also been included in the study in an attempt to define their properties in relation to the tastes they exhibit; eight sweet-bitter and three salty-bitter molecules were used. The density and sound velocity of their solutions in water have been measured and their apparent volumes, apparent compressibilities and compressibility hydration numbers calculated and compared. Apparent molar volumes (phi(v)) and apparent specific volumes (ASV) reflect the state of hydration of the molecules, and thus their extent of interaction with water structure. The range of ASVs reported are 0.13-0.49 cm3/g for salty molecules, 0.55-0.68 cm3/g for sweet molecules, 0.53-0.88 cm3/g for sweet-bitter molecules and a much wider range (0.16-0.85 cm3/g) for sour molecules. Isentropic apparent specific compressibilities range from -2.33 x 10(-5) to -8.06 x 10(-5) cm3/g x bar for salty molecules, -3.38 x 10(-7) to -2.34 x 10(-5) cm3/g x bar for sweet molecules, +6.35 x 10(-6) to -2.22 x 10(-5) cm3/g x bar for sweet-bitter molecules and +6.131 x 10(-6) to -2.99 x 10(-5) cm3/g x bar for sour molecules. Compressibility hydration numbers are also determinable from the measurements of isentropic compressibilities and these reflect the number of water molecules that are disturbed by the presence of the solutes in solution. This study also shows that it is possible to group isentropic apparent molar compressibility values by the taste quality exhibited by the molecules in the same order as for ASV.     

Temporal properties of the human taste system.

An attempt to apply linear systems methods to the temporal properties of the human taste system is described. The percentage modulation in concentration required to detect a fluctuation in intensity was measured as a function of frequency of modulation. Both a sine-wave and a square-wave input were used. The sensitivity of the taste system at its maximum, and also at the low frequencies, was greater than expected from previous work based on classical methodology. The sensitivity of the taste system at its maximum, and also at the low frequencies, was greater then expected from previous work base on classical methodology. The sensitivity of the taste system to the various qualities was, in decreasing order, salty, sweet, sour, and bitter. The taste system in insensitive to frequencies above about 5 Hz. Except for bitter, representatives of the various taste qualities yielded similar functions within qualities.     

Physical characterization of a kinetoplast DNA fragment with unusual properties

A 414-base pair fragment from a Leishmania tarentolae kinetoplast DNA minicircle has unusual physical properties. We reported previously that in comparison to phi X174 and pBR322 control fragments, the kinetoplast fragment behaves in gel electrophoresis, gel filtration, and electric dichroism experiments as if it has an unusually compact conformation. We accounted for these unusual properties by proposing that the fragment is a systematically bent helix (Marini, J.C., Levene, S.D., Crothers, D.M., and Englund, P.T. (1982) Proc. Natl. Acad. Sci. U.S.A. 79, 7664-7668). In this paper, we further explore the properties of the kinetoplast fragment. Because of its compact conformation, the kinetoplast fragment has difficulty in snaking through polyacrylamide gels and therefore migrates unusually slowly in electrophoresis experiments. Warming (53 degrees C) and ethanol (5-20%) partially normalize gel migration; glyoxal treatment results in denatured strands with electrophoretic mobility close to that expected for their size. In vivo modification does not appear to be responsible for the fragment's properties; its anomalous electrophoretic behavior persists after proteinase K treatment, phenol extraction, or after cloning into pBR322 and reisolation. Velocity sedimentation experiments rule out fragment aggregation. Secondary structure, such as a cruciform, is not detectable by S1 or mung bean nuclease digestion. The kinetoplast fragment has circular dichroism spectra characteristic of a B-type helix. With increasing temperature, there is an increase in the 270/280 ellipticity ratio. Circular dichroism spectra taken in the presence of ethanol show a B to A helix transition at unusually low ethanol concentrations (between 44 and 54% (w/w]. Thermal denaturation reveals a triphasic melting curve.     

The capability of several toxic plants to condition taste aversions in sheep

Grazing livestock frequently ingest toxic plants, occasionally with fatal results. Behavioral adjustments by livestock may reduce toxin intake; for example they can develop food aversions which may protect animals from over-ingestion of toxic plants. The purpose of this study was to evaluate three plants with different mechanisms of toxicity for their efficacy in conditioning a taste aversion: (1) a seleniferous plant, Xylorhiza glabriuscula, (2) an indolizidine alkaloid-containing plant, Astragalus lentiginosus, and (3) a diterpene acid-containing plant, Gutierrezia sarothrae. For each plant species, 15 sheep were divided into 3 treatment groups and periodically tested for consumption of a novel food, whole corn: (1) controls – given 200 g of ground alfalfa hay by oral gavage, (2) averted – given lithium chloride (LiCl) at 175 mg/kg BW via oral gavage, and (3) given the specific target plant by oral gavage. X. glabriuscula was given at a dose equivalent to 3 mg Se/kg BW; A. lentiginosus was given at a dose equivalent to 3 mg/kg of the toxic alkaloid, swainsonine; freshly thawed G. sarothrae was dosed at 5 g/kg body weight (BW). Both LiCl and Xylorhiza conditioned an aversion to corn, with sheep eating 1.6 and 0.6% of offered corn during the final test; controls were not averted, eating 93% of the corn (P < 0.01). Sheep were partially averted by Xylorhiza after a single dose, and the aversion was complete after the second dose. Sheep were not averted by A. lentiginosus or G. sarothrae. Of the three toxic plants used in this study, only Xylorhiza conditioned a taste aversion. These results likely reflect differing mechanisms of action of the plant toxin(s) on brain and gut structures important for forming conditioned taste aversions. These results suggest that conditioned aversions to Se-containing plants may help to deter consumption of such plants by grazing ruminants on rangelands.     

Greater effort boosts the affective taste properties of food

Actions can create preferences, increasing the value ascribed to commodities acquired at greater cost. This behavioural finding has been observed in a variety of species; however, the causal factors underlying the phenomenon are relatively unknown. We sought to develop a behavioural platform to examine the relationship between effort and reinforcer value in mice trained under demanding or lenient schedules of reinforcement to obtain food. In the initial experiment, expenditure of effort enhanced the value of the associated food via relatively lasting changes in its hedonic attributes, promoting an acquired preference for these reinforcers when tested outside of the training environment. Moreover, otherwise neutral cues associated with those reinforcers during training similarly acquired greater reinforcing value, as assessed under conditioned reinforcement. In a separate experiment, expenditure of effort was also capable of enhancing the value of less-preferred low-caloric reinforcers. Analysis of licking microstructure revealed the basis for this increased valuation was, in part, due to increased palatability of the associated reinforcer. This change in the hedonic taste properties of the food can not only serve as a basis for preference, but also guide decision-making and foraging behaviour by coordinating a potentially adaptive repertoire of incentive motivation, goal-directed action and consumption.     

A novel intracellular compartment with unusual secretory properties in human neutrophils

Human neutrophils contain a novel intracellular compartment that is distinct from the previously characterized azurophil and specific granules. This compartment is distinguished by the presence of cytochemically detectable alkaline phosphatase activity. The alkaline phosphatase-containing compartments are short rod-shaped organelles that rapidly undergo a dramatic reorganization upon cell stimulation with either a chemoattractant or an active phorbol ester. Biochemical analysis shows that in unstimulated neutrophils the majority of the alkaline phosphatase activity is intracellular, but after stimulation essentially all of this activity becomes associated with the cell surface. The exocytotic pathway is unusual in that these small organelles fuse to form elongated tubular structures before their association with the plasmalemma.     

A cyanobacterial hemoglobin with unusual ligand binding kinetics and stability properties

The glbN gene of the cyanobacterium Nostoc commune UTEX 584 encodes a hemoprotein, named cyanoglobin, that has high oxygen affinity. The basis for the high oxygen affinity of cyanoglobin was investigated through kinetic studies that utilized stopped-flow spectrophotometry and flash photolysis. Association and dissociation rate constants were measured at 20 degrees C for oxygen, carbon monoxide, nitric oxide, and methyl and ethyl isocyanides. The association rate constants for the binding of these five ligands to cyanoglobin are the highest reported for any naturally occurring hemoglobin, suggesting an unhindered and apolar ligand binding pocket. Cyanoglobin also shows high rates of autoxidation and hemin loss, indicating that the prosthetic group is readily accessible to solvent. The ligand binding behavior of cyanoglobin was more similar to that of leghemoglobin a than to that of sperm whale myoglobin. Collectively, the data support the model of cyanoglobin function described by Hill et al. [(1996) J. Bacteriol. 178, 6587-6598], in which cyanoglobin sequesters oxygen, and presents it to, or is a part of, a terminal cytochrome oxidase complex in Nostoc commune UTEX 584 under microaerobic conditions, when nitrogen fixation, and thus ATP demand, is maximal.     

A DNA polymerase with unusual properties from the slime mold Physarum polycephalum

Two forms of a DNA polymerase have been purified from microplasmodia of Physarum polycephalum by poly(ethyleneimine) precipitation and chromatography on DEAE-Sephacel, phosphocellulose, heparin Sepharose, hydroxyapatite, DNA-agarose, blue-Sepharose. They were separated from DNA polymerase alpha on phosphocellulose and from each other on heparin-Sepharose. Form HS1 enzyme was 30-40% pure and form HS2 enzyme 60% with regard to protein contents of the preparations. Form HS2 enzyme was generated from form HS1 enzyme on prolonged standing of enzyme preparations. The DNA polymerases were obtained as complexes of a 60-kDa protein associated with either a 135-kDa (HS1) or a 110-kDa (HS2) DNA-polymerizing polypeptide in a 1:1 molar stoichiometry. The biochemical function of the 60-kDa protein remained unknown. The complexes tended to dissociate during gradient centrifugation and during partition chromatography as well as during polyacrylamide gradient gel electrophoresis under nondenaturing conditions at high dilutions of samples. Both forms existed in plasmodia extracts, their proportions depending on several factors including those which promoted proteolysis. The DNA polymerases resembled eucaryotic DNA polymerase beta by several criteria and were functionally indistinguishable from each other. It is suggested that lower eucaryotes contain repair DNA polymerases, which are similar to those of eubacteria on a molecular mass basis.     

Superhelical architecture of the myosin filament-linking protein myomesin with unusual elastic properties

Active muscles generate substantial mechanical forces by the contraction/relaxation cycle, and, to maintain an ordered state, they require molecular structures of extraordinary stability. These forces are sensed and buffered by unusually long and elastic filament proteins with highly repetitive domain arrays. Members of the myomesin protein family function as molecular bridges that connect major filament systems in the central M-band of muscle sarcomeres, which is a central locus of passive stress sensing. To unravel the mechanism of molecular elasticity in such filament-connecting proteins, we have determined the overall architecture of the complete C-terminal immunoglobulin domain array of myomesin by X-ray crystallography, electron microscopy, solution X-ray scattering, and atomic force microscopy. Our data reveal a dimeric tail-to-tail filament structure of about 360 Å in length, which is folded into an irregular superhelical coil arrangement of almost identical α-helix/domain modules. The myomesin filament can be stretched to about 2.5-fold its original length by reversible unfolding of these linkers, a mechanism that to our knowledge has not been observed previously. Our data explain how myomesin could act as a highly elastic ribbon to maintain the overall structural organization of the sarcomeric M-band. In general terms, our data demonstrate how repetitive domain modules such as those found in myomesin could generate highly elastic protein structures in highly organized cell systems such as muscle sarcomeres.