The oviduct musculature of the horsefly, Tabanus sulcifrons, and its response to 5-hydroxytryptamine and proctolin

ABSTRACT. A delicate lace-like membrane covers the ovaries of Tabanus sulcifrons. These membranes were found to contain muscle fibres that provide the organs with motile properties. The lateral oviducts consist of a single layer of longitudinal muscles that form a structural syncytium by means of an extensive anastomosis of the fibres comprising it. The common oviduct is composed of two muscle layers, an outer sheath of circular muscle and an inner substratum of longitudinal muscle. Both of these layers showed evidence of a structural syncytium. When isolated in saline, the oviduct was spontaneously active and gave a simple phasic pattern of contraction. Such muscle preparations were sensitive to both 5-hydroxytryptamine (5HT) and the insect myotropic peptide, proctolin. Excitation was generally indicated by a rise in muscle tonus or an increase in the frequency and amplitude of individual phasic contractions, or all three characteristics. The threshold for activation with 5HT was variable, ranging from as low as 4 × 10^-9M to 1 × 10^-7M. Proctolin evoked a noticeable increase in the tonus of most oviducts at 10^-10M. However, several preparations responded to as little as 3 × 10^-11M proctolin.     

Visual contrast thresholds in the cod Gadus morhua L.

The contrast discrimination of Gadus morhua L. was studied by means of a cardiac conditioning technique. Fish were trained to respond to a projected pattern of spots and the contrast of pattern and background then reduced until the minimum contrast required to elicit a response was established. This was repeated at seven different background light levels to cover the range of light conditions naturally encountered by the cod. The minimum detectable contrast decreased with increasing light level to a minimum of around 2.0%. The contrast threshold curve showed a discontinuity at a light level of approximately 8.0 × 10^-6 W sr^-1 m^-2. This was thought to be linked to the change from photopic to scotopic vision. Significant differences were found in cod taken from two different locations. Overall, cod contrast detection compared very favourably with figures available for other species including man. Using the obtained contrast discrimination figures some estimates were made of the cod sighting distance of hypothetical targets under natural conditions.     

Joint action of l-leucine and sodium phosphate buffer solutions as feeding stimulants for protein-deprived females of the house fly Musca domestica

ABSTRACT. Potentiation in joint action was demonstrated between solutions of L-leucine and sodium phosphate buffer (pH 6.3) as feeding stimulants for protein-deprived females of the house fly, Musca domestica L. Both components alone elicited feeding. In two-choice feeding tests, mixtures consisting of equi-stimulating concentrations of the two components were taken in greater quantities than either component alone at twice the concentration in the mixture.
The presence of 1×10^-1 M phosphate buffer markedly lowered the threshold for detection of L-leucine. The presence of phosphate buffer strengthened the preferences shown by flies given choices of concentrations of L-leucine differing by a factor of 2 and enabled them to display preferences at lower concentrations.
The presence of 1×10^-3 M L-leucine increased, somewhat, the ability of flies to detect low concentrations of phosphate buffer. Its presence had relatively little effect on the strength of preference shown between two-fold differences in concentration of phosphate buffer when the higher concentration was 6.3×10^-3 M or less, but markedly strengthened the preferences when the higher concentration was 2.5×10^-2M or greater. Leucine increased the optimal concentration of phosphate buffer by a factor of more than 2 and converted 2×10^-1 M phosphate buffer from a mild feeding deterrent to a powerful feeding stimulant.     

Sequestration of 125I-Labeled p Nerve Growth Factor by Embryonic Sensory Neurons

Abstract: Nerve growth factor (NGF) binds to two specific receptors on sensory nerve cells. These two receptors are characterized by different equilibrium dissociation constants. The higher affinity (type I) receptors have an equilibrium dissociation constant of 3.3 × 10^-11 M. The lower affinity (type II) receptors have an equilibrium dissociation constant of 1.7 × 10^-9M. These two receptors are not a result of negative cooperatively, but apparently are different receptors. At 22°C the rate of association is 1 × 10⁷ M^-1 S^-1 and the rates of dissociation are 6.5 × 10^-4 s^-1 (type I) and 3.2 × 10^-2 s^-1'(type II). After binding, a time-dependent process occurs that makes the NGF inaccessible to the external milieu (sequestered). The sequestration process is energy-dependent, but apparently temperature-independent. The data suggest that only the type I receptors are involved in the sequestration process. This process is similar to that observed on sympathetic neurons and may be the first step in the internalization of NGF by responsive cells.     

Effects of di-n-butyl phthalate on growth and photosynthesis in algae and on isolated organelles from higher plants

Di- n -butyl phthalate (DBF) is widely used as a plasticizer and has been found in all types of ecosystems. It inhibits growth and photosynthesis of green algae ( Chlorella emersonii CCAP strain 211/8 h and Selenastrum capricornutum CCAP strain 278/4) at concentrations higher than 10-⁵ M . The IC₅₀ value for CO₂-dependent oxygen evolution in algae was 3 × 10^-4M. The CO₂-reduction in isolated protoplasts prepared from barley ( Hordeum vulgare L. cv. Simba) was also inhibited by phthalate. The IC₅₀ value was 2 × 10^-4 M . The electron transport in isolated thylakoids prepared from spinach was inhibited with an IC₅₀ value of 3 × 10^-4 M . The IC₅₀ value for uncoupled electron transport extrapolated to zero chlorophyll concentration was 2.5 × 10^-5 M . The effect of di-n-butyl phthalate was localized to reactions in photosystem II. Di-n-butyl phthalate could thus be a pollutant which affects growth and photosynthesis of plants. The reported IC₅₀ values may be underestimated since di- n -butyl phthalate can attach to surfaces. The results are discussed in relation to observed effects of di- n -butyl phthalate on other organisms.     

Behavioural responses of glass eels of Anguilla anguilla to non-protein amino acids

Glass eels reacted chemotactically towards five non-protein constitutional amino acids, D-glutamine, D-asparagine, D-glutamic acid, D-alanine and β -alanine, dissolved in fresh water or salt water, with behavioural thresholds ranging from 10^-9 M for the most effective (D- and β ;-alanine) to 10^-7 M for the other three. With the exception of D-asparagine in fresh water and D-alanine at concentrations ≥10^-7 M, these amino acids were strong attractants. The results are compared with previous findings on the respective L-isomers. The non-protein amino acids may influence behaviour in the search for food and the recognition of conspecifics.     

Detection and quantification of Desulforhabdus amnigenus in anaerobic granular sludge by dot blot hybridization and PCR amplification

A species-specific 16S rRNA oligonucleotide probe (ASRB1) was developed for the detection of Desulforhabdus amnigenus in anaerobic granular sludge. The presence of nucleic acids from cells of D. amnigenus in granular sludge was determined using ASRB1 as a specific primer for polymerase chain reaction (PCR) amplification or as a probe for dot blot hybridizations. The detection threshold and the reproducibility of these two methods were determined with sludge amended with 10⁴–10¹⁰ D. amnigenus cells per gram of volatile suspended solids (VSS). For D. amnigenus cells with a ribosomal RNA content of 15 fg cell⁻¹, the lowest number of target cells detected by hybridization was 1 × 10⁸ cells g⁻¹ VSS. With the PCR amplification method the lowest number of target cells which could be detected was 1 × 10⁷ g⁻¹ VSS. This corresponds to a threshold level for hybridization of 0·1–0·001‰ of the total bacterial sludge population, while the threshold level obtained with the PCR approach amounted to 0·01–0·0001‰. The rRNA content of D. amnigenus was found to be affected by the growth rate and the growth phase, and it ranged from 19 fg cell⁻¹ in slow-growing cultures to 90 fg cell⁻¹ in fast-growing cultures. Therefore, the detection threshold of the dot blot hybridization method for fast-growing cells is lower than for slow-growing cells.     

Olfactory sensitivity of the herbivorous grass carp, Ctenopharyngodon idella, to amino acids

The olfactory sensitivity of a herbivore, the grass carp, Ctenopharyngodon idella , to ten amino acids was investigated using electrophysiological recording techniques. Underwater electroolfactogram recordings increased exponentially with logarithmic increases of stimulus concentration from threshold to 1 × 10⁻³ m. Calculated detection thresholds ranged from 1 × 10^−7.15 m to 1 × 10^−8.7 M and are similar to those previously reported for carnivorous fish species. The relative stimulatory efficacy (RSE) of the ten amino acids determined at 1 × 10⁻⁴ M revealed five groups of stimuli, with cysteine the most stimulatory and proline the least. Measures of amino acid RSE did not differ significantly from those of previously studied carnivores, except for arginine. Grass carp showed strong responses to this compound, whereas it has produced only limited responses in carnivores. Differences in the responsiveness of herbivores and carnivores were not observed, suggesting that feeding niche segregation is probably not facilitated by differential olfactory sensitivities to feeding stimuli.     

Studies on the Production of Extracellular Proteinases by a Non-pigmented Strain of Chromobacterium lividum Isolated from Abattoir Effluent

A highly proteolytic bacterium isolated from abattoir effluent was identified as a non-pigmented strain of Chromobacterium lividum. Ferrous or ferric ions at concentrations between 1·8 × 10^-5 and 9 × 10^-4 g ions/1, which is 2–3 orders of magnitude greater than that required for growth, were essential for extracellular proteinase production in aerated but not in static culture. Co²⁺, Ni²⁺, Mn²⁺, Cu²⁺ or Zn²⁺ ions could not replace iron. Four proteinases (I-IV) were produced in static culture, but only proteinase I was formed in significant quantities in aerated culture. With both forms of culture amino nitrogen was essential for proteinase production; glucose inhibited formation in aerated, but not static, cultures. Growth occurred over the range 1–33 °C, whereas proteinase production ceased at 27 °C, with maximum activity at 13 °C. Proteinase production appeared to be controlled by an interaction between iron, oxygen tension and glucose.     

Fast locomotion of some African ungulates

Ten species of ungulate were filmed, galloping in their natural habitat. They ranged in size from Thomson's gazelle (about 20 kg) to giraffe (about 1000 kg). They were pursued to make them run as fast as possible. The films have been analysed to determine speed, stride frequency, stride and step lengths, and duty factors. The dependence of these quantities on body size is discussed.  

Summary:

Fast locomotion of zebra, giraffe, warthog and seven species of Bovidae has been studied. The animals were filmed from a pursuing vehicle while galloping in their natural habitat.
Stride frequency was more closely correlated with limb length (represented by hip height) than with body mass. Mean stride frequency was proportional to (hip height)^-0·51 and maximum stride frequency to (hip height) ^-0·63.
Maximum speed was between 10 and 14 m s ^-1 for all species except buffalo (7 m s ^-1). It was not significantly correlated with body mass.
Since the small species ran at least as fast as the large ones they attained higher Froude numbers. Relative stride length was approximately 1·8 (Froude number)^0·39 for all species, irrespective of size. Relative step length was approximately 0·65 (Froude number)^0·2, both for the fore feet and for the hind ones. The vertical forces exerted by the feet are proportional to (body weight)×(Froude number)^0·2 so the forces at maximum speed are larger multiples of body weight for small species than for large ones.     

Neuroendocrine stimulation of uterine gland protein synthesis in the tsetse fly, Glossina morsitans

ABSTRACT. The uterine gland of the tsetse fly Glossina morsitans morsitans Westw. synthesizes a secretion which nourishes the developing larva in the uterus. Aqueous extracts of the brain have been shown to stimulate the synthesis of the protein and amino acid components of this secretion from L- [U-¹⁴C]leucine by uterine gland tubules in vivo and in vitro. A linear dose response relationship was demonstrated in vitro with extract concentrations ranging from 1 × 10^-4 to 1 × 10^-2 brains μl^-1. The maximum response, a > 300% increase in the rate of protein and amino acid synthesis, was achieved with as little as 1 × 10^-2 brains μl^-1 The concentration of active factor(s) in the brain declined during a single interlarval period coincident with the period of release of secretion associated with larval growth. The stimulatory activity in brain extracts was destroyed by proteolytic enzymes indicating that it is probably a protein or peptide. Results suggest that the active factor(s) is a hormone responsible for the stimulation of uterine gland protein synthesis essential for larval nutrition.     

EFFECTS OF CYTOCHALASIN B ON THE UPTAKES OF MONOSACCHARIDES BY RAT BRAIN SYNAPTOSOMES

Abstract— Synaptosomal uptakes of a number of simple carbohydrates were strongly inhibited by cytochalasin B (K₁= 7-9 × 10⁻⁸M). Phloretin (K₁= 2-4 × 10⁻⁶M) and phloridzin (K₁= 3-4 × 10⁻⁴M) were less inhibitory. Cytochalasin B competitively inhibited the uptakes of carbohydrates with saturable transport kinetics. Inhibition of sugar uptake was immediate on addition of cytochalasin B but was promptly reversed upon removal of the drug. Cytochalasin B had no effect on the efflux of D-glucos-amine or of the phosphorylated sugar, and did not affect intrasynaptosomal hexokinase(s). The synapto-somal uptakes of L-glucose, D-mannitol, L-fucose and the N -acetylated amino sugars were non-saturable and uninhibited by cytochalasin B. In the case of sugars which enter synaptosomes by both saturable and non-saturable processes, cytochalasin B could be used to selectively inhibit the saturable uptake components. The resultant non-saturable cytochalasin-insensitive uptake rates obtained were found to be widely different among the sugars studied, and must be corrected for in order to estimate accurate kinetic constants of the saturable processes.     

Increased ELISA sensitivity using a modified extraction buffer for detection of Xanthomonas campestris pv. vesicatoria in leaf tissue

In vitro and in planta sensitivity of an indirect enzyme-linked immunoassaytechnique, using a monoclonal antibody specific for the lipopolysaccharide (LPS) of Xanthomonas campestris pv. vesicatoria , was increased 10-foldby using a newextraction buffer (gl of : KH₂PO₄, 2; NaHPO₄, 11·5; EDTAdisodium, 0·14; thimerosal, 0·02; and lysozyme, 0·2). The procedure improvedsensitivity without increasing background levels. In vitro , the limit of detection wasbetween 1×10⁷ and 1×10⁸ cells ml⁻¹ with the conventionalextraction buffer phosphate-buffered saline (PBS) and less than 1×10⁶ cells ml⁻¹ when lysozyme extraction buffer was substituted for PBS. In comparing 22 X. c.vesicatoria strains, absorbance readings were increased close to three-fold with the lysozymeextraction buffer as opposed to PBS. When leaf tissue extract was spiked with the bacterium, thelimit of detection was 1×10⁷ cfu ml⁻¹ and 1×10⁸ cfu ml⁻¹ with the lysozyme solution and PBS, respectively, as the extraction buffers. Whenusing the lysozyme extraction buffer in combination with a commercial amplification system, thelimit of detection was decreased to less than 1×10⁵ cfu ml⁻¹ in leaftissue. The addition of the lysozyme and EDTA to the phosphate buffer resulted in release of asignificant quantity of LPS and concomitant dramatic increase in sensitivity. The new procedure,termed lysozyme ELISA (L-ELISA), should increase sensitivity of ELISA reactions where LPS isthe reacting epitope.     

Stimulatory effectiveness of amino acids on the olfactory response in an algivorous marine teleost, the rabbitfish Siganus fuscescens Houttuyn

The detection of threshold concentrations and relative stimulatory effectiveness (RSE) for 19 amino acids were studied by olfactory bulbar electroencephalogram (EEG) in the algivorous rabbitfish. The threshold concentrations for 19 amino acids ranged from 10⁻¹⁰ to 10⁻⁵ M. l -Alanine was the most effective amino acid, and the threshold concentration was estimated to be from 10⁻¹⁰ to 10⁻⁹M. These results were compared with the olfactory response in herbivorous and carnivorous fishes previously reported. The comparison of RSE for rabbitfish with those for other fishes showed correlation coefficients ranging from r = 0.97 to r = 0.50, but high similarities did not correspond with difference in feeding habits for herbivores and carnivores. The role of amino acids in fish chemosensory behaviour and the olfactory response of the rabbitfish was compared with the gustatory response reported previously. It was found that l -serine was the most potent stimulant in both chemoreceptors, l -proline and l -glutaminic acid were the most potent gustatory stimulants, and l -alanine, l -glutamine, l -arginine and l -lysine were the most potent olfactory stimulants.     

Interaction of triacontanol with gibberellin in control of lettuce seed germination

Triacontanol at concentrations from 2.3 × 10^-9 M to 2.3 × 10^-7 M did not affect the germination of lettuce ( Lactuca sativa L., cv. Grand Rapids) seeds in darkness, stimulated by light at 25°C or by benzyladenine at 31°C. Stimulation of seed germination by gibberellin A₃ (10^-5 M ) was significantly inhibited by triacontanol; the most effective concentration was 4.6 × 10^-8 M. Pulse experiments demonstrated that triacontanol was ineffective when applied later than gibberellin, whereas an inverse sequence of treatment caused an inhibition comparable to that resulting from continuous treatment of seeds with both factors. Possible interaction of triacontanol with gibberellin receptor is discussed.     

Effect of alcohols on the association of photosynthetic fructose-1,6-bisphosphatase to thylakoid membranes

Aliphatic alcohols have a positive effect on the assoociation of pea ( Pisum sativum L. cv. Lincoln) chloroplast fructose- 1,6-bisphosphatase (FBPase; EC 3.1.3.11) with thylakoid membranes. The alcohol concentration needed to obtain a fixed percentage of enzyme association decreased with increased length of the aliphatic chain of the alcohol; maximum binding was obtained when the lysis medium contained, in molar fractions (or v/v percentages): 48×10^-4(T4 (2.4%), 26×10^-3 (10%), 40×10^-3 (15%), 76×10^-3 (21%), and 13×10^-2 (24%), of 1-butanol, 1-propanol, 2-propanol, ethanol, and methanol, respectively. A good correlation of binding with the octanol/water partition coefficient was observed. Since this coefficient constitutes a measure of hydrophobicity, we suggest that the binding of FBPase to the membranes occurs via hydrophobic clusters of both components.     

Cell-cycle dependence of a ganglioside glycosyltransferase activity and its inhibition by enkephalin in a neurotumor cell line

Abstract: Rat glioma mouse neuroblastoma hybrid neurotumor cells (NG108-15), synchronized by amino acid deprivation, showed a cell-cycle-dependent peak of activity of a ganglioside N-acetylgalactosaminyl transferase 14-24 h following release from the cell cycle block (S/G₂ phase). Maximal expression of two typical lysosomal hydrolases, N-acetyl-β-hexosaminidase and β-galactosidase, occurred between 18 and 21 h following release (S phase), declining to G₁ phase levels during the peak of N-acetylgalactosamine (GalNAc) transferase activity. In addition, glycosyltransferase activity in G₂ phase cells showed an increase in apparent V_max (suggesting the presence of more enzyme/mg of cell protein) and apparent binding affinity for uridine diphosphate N-acetylgalactosamine (UDP-GalNAc) (32 versus 14 M) when compared to transferase activity in the G₁ phase. However, the opioid peptide enkephalin [D-Ala², o-Leu⁵], which inhibits ganglioside GalNAc transferase activity in unsynchronized NG108-15 cultures, was much more inhibitory in whole cells 8 h after release from the cell cycle block (G₁ phase) than in cells 20 h after release (G, phase), with 50% inhibition occurring at 2 ± 10^-9M and 2 ± 10^-7M, respectively. These results suggest that the GalNAc transferase activity is regulated in more than one way during the cell cycle, since both V_max and K_m changes are observed, and that the cyclic AMP-dependent mechanism by which opiates reduce transferase activity is receptor mediated and cell cycle dependent.     

ADENOSINE KINASE OF MAMMALIAN BRAIN: PARTIAL PURIFICATION AND ITS ROLE FOR THE UPTAKE OF ADENOSINE

Abstract— Adenosine metabolism in the homogenate of brain mainly undergoes deamination to inosine and hypoxanthine, while uniformly labelled [¹⁴C]adenosine injected into the carotid artery or [8-¹⁴C]adenosine incubated with brain slices was mostly phosphorylated to [¹⁴C]adenine nucleotides in brain cells. Adenosine kinase has now been partially purified from homogenates of guinea pig brain. The kinase preparation was free of adenosine deaminase, almost free of adenosine triphosphatase and had a K_m of the order of 2 × 10^-5M for adenosine.
Kinetic studies with brain slices showed that adenosine reached the cells by diffusion and that the diffusion was facilitated by subsequent phosphorylation to adenine nucleotides. From the following experimental results, it is concluded that the phosphorylation is catalysed by adenosine kinase quantitatively. (1) During the uptake and phosphorylation of adenosine by brain slices, the nucleoside did not split to adenine and ribose moieties. (2) The rate of formation of adenine nucleotides in the slices was a hyperbolic function of the concentration of adenosine in the medium, showing an apparent K_m foradenosine of the order of 2 × 10^-5 M. (3) Some analogues of adenosine inhibited both the facilitated diffusion of adenosine and the kinase activity, but ouabain (0.005 mM) did not inhibit either.     

The effect of calcium concentration on the toxicity of copper, lead and zinc to yolk-sac fry of brown trout, Salmo trutta L., in soft, acid water

Yolk-sac fry of brown trout were exposed to three levels of single trace metals (Cu, 20,40,80 nmol 1^-1; Pb, 12·5,25,50 nmol 1^-1; Zn, 75,150,300 nmol 1^-1) typical of concentrations reported for acid soft waters, in flowing, artificial, soft water media maintained at pH 4·5 and [Ca] of 20 or 200 μmol 1^-1for 30 days.
Mortalities were high in fry subjected to all levels of the three trace metals at [Ca] 20 μmol 1^-1, with 80% of the total deaths occurring between days 11 and 15 of the experiment. 25% mortality occurred at low [Ca] and pH 4·5 in the absence of trace metals, with only one death recorded at [Ca] 200 μmol1^-1'(Cu, 80 nmol 1^-1). At high [Ca] all three levels of Cu and Pb impaired net Na and K uptake; Cu was the only metal to reduce the uptake of Ca. The Zn treatments had no significant effect on mineral uptake. Calcification of centra was reduced by all three Cu treatments at [Ca] 200 μmol 1^-1. The lowest Zn concentration (75 nmol 1^-1) was the only other treatment to impair skeletal development. In the absence of trace metals, low [Ca] significantly reduced Ca, Na and K uptake, skeletal calcification and dry mass at pH 4·5.
The deleterious effects of low Cu, Pb and Zn concentrations at low pH and low [Ca], and the ameliorative effect of higher ambient [Ca], are discussed in relation to fishery status in soft, acid waters.     

Toxicity versus avoidance response of golden shiner, Notemigonus crysoleucas, to five metals

Avoidance thresholds and 96-h LC50 values were determined for golden shiners, Noiemigonus crysoleucas , for five individual elements: chromium, Cr; copper, Cu; cadmium, Cd; arsenic, As; selenium, Se. The avoidance concentrations were 73, 26 and 28 μgl^-1 for Cr-VI, Cu and As-III, respectively. Cadmium and Se were not avoided at experimental concentrations up to 68 and 3489 μg1^-1, respectively. Acute flow-through 96-h LC50 values were Cr-VI 55·0, Cu 84·6, and As-III 12·5 mg 1^-1, which were more than two orders of magnitude above avoidance concentrations. The acute flow-through 96-h LC50 values for Cd and Se were 2·8 and 11·2 mg 1^-1, respectively. These concentrations are 31 and 2·2 times the highest concentration employed in the avoidance tests, neither of which were avoided by the test organisms. Thus, simple toxicity tests do not identify the environmental hazard of some elements, and the most toxic elements may not elicit a behavioural response. When used in concert with tests of organism function, more realistic indicators of environmental hazard or safety may be determined.