Cross pathway regulation: effect of histidine on the synthesis and activity of enzymes of aromatic acid biosynthesis in Bacillus subtilis

l-Histidine and, to a lesser degree, l-phenylalanine at concentrations of 10(-4)m inhibit the growth of leaky mutants (bradytrophs) of Bacillus subtilis that are deficient in the synthesis of p-hydroxyphenylpyruvate, the first intermediate specific to tyrosine synthesis. The inhibition can be overcome by growth factor amounts of l-tyrosine and p-hydroxyphenylpyruvate. Histidine and phenylalanine are capable of inhibiting the synthesis of tyrosine in several ways, and the major physiological effect which results in growth inhibition has not been established. Both l-histidine and l-phenylalanine inhibit the activity of prephenate dehydrogenase at concentrations about 100-fold higher than the inhibitory concentration of l-tyrosine. Histidine also appears to repress the synthesis of prephenate dehydrogenase because a histidine bradytroph growing in histidine-supplemented medium has a twofold lower level of this enzyme than the same cells growing in unsupplemented medium. These same two amino acids also inhibit the growth of a bradytroph deficient in dehydroquinate synthetase, an early enzyme in the pathway of tyrosine, phenylalanine, and tryptophan synthesis. The inhibition is overcome by a combination of tyrosine and phenylalanine. Histidine-resistant derivatives of both the prephenate dehydrogenase and dehydroquinate synthetase-deficient strains, which simultaneously have gained resistance to phenylalanine, have been isolated. Most of these resistant mutants synthesize additional tyrosine compared with the parent strain. One class of resistant mutants excretes tyrosine and has a number of enzymes of aromatic acid synthesis which are no longer repressible by any combination of the aromatic amino acids. Tyrosine inhibits the growth of histidine bradytrophs. Histidine, at growth factor levels, overcomes the inhibition.     

Mutations that alter the covalent modification of glutamine synthetase in Salmonella typhimurium.

glnD and glnE mutant strains of Salmonella typhimurium lack three of the four activities required for reversible covalent modification of glutamine synthetase (GS; EC 6.3.1.2). The glnD strains, which are unable to deadenylylate GS and therefore accumulate the adenylylated or less active form of the enzyme, were isolated as glutamine bradytrophs. They lack the activity of PIIA uridylyl-transferase, one of the proteins required for deadenylylation of GS; in addition, they lack PIID uridylyl-removing activity. Mutations in glnD are suppressed by second-site mutations in glnE that eliminate the activity of GS adenylyltransferase (EC 2.7.7.42) and thus prevent adenylylation of GS. The glnD and glnE strains have one-third to one-half as much total GS as the wild-type strain when they are grown in a medium containing a high concentration of NH4+. The wild-type strain derepresses synthesis of GS fourfold in response to nitrogen limitation; glnD and glnE strains derepress synthesis of the enzyme fourfold and sevenfold, respectively. Thus, mutations that alter covalent modification of GS in Salmonella do not significantly affect derepression of its synthesis. The glnD gene lies at 7 min on the Salmonella chromosome and is 50% linked to pyrH by P22-mediated transduction.     

Structural and regulatory mutations allowing utilization of citrulline or carbamoylaspartate as a source of carbamoylphosphate in Escherichia coli K-12.

Escherichia coli mutants lacking carbamoylphosphate synthase require arginine and uracil for growth. It is, however, possible to obtain mutants in which carbamoylphosphate is obtained by phosphorolysis of citrulline or carbamyolaspartate. Citrulline utilizers are argG bradytrophs or strains in which the synthesis of ornithine carbamoyltransferase (either of the F or I type) is specifically depressed by unstable chromosomal rearrangements or stable mutations that presumably affect the operators of those genes. Carbamoylaspartate utilization as a source of carbamoylphosphate appears to require more than one mutation; the best-understood strains are pyrD pyrH or pyrC pyrH mutants in which aspartate carbamoyltransferase activity is high and the pool of cytidine triphosphate (feedback inhibitor of aspartate carbamoyl-transferase) is presumably low and in which channeling of carbamoylaspartate towards pyrimidine biosynthesis is considerably reduced. Selection of enzyme overproducers based on a metabolic dependency for a reversed enzymatic reaction can be regarded as a means for isolating regulatory mutants.     

L-Methionine SR-sulfoximine-resistant glutamine synthetase from mutants of Salmonella typhimurium

Two mutants of Salmonella typhimurium resistant to growth inhibition by the glutamine synthetase transition state analog, L-methionine SR-sulfoximine, were isolated and characterized. These mutants are glutamine bradytrophs and cannot use growth rate-limiting nitrogen sources. Although this phenotype resembles that of mutants with lesions in the regulatory gene for glutamine synthetase, glnG, these mutations do not lie in the glnG gene. Purification and characterization of the glutamine synthetase from one of the mutants and a control strain demonstrated that the mutant enzyme is defective in the reverse gamma-glutamyltransferase activity but has biosynthetic activity that is resistant to inhibition by L-methionine SR-sulfoximine. The mutant enzyme also has a 4.4-fold higher apparent Km for glutamate (0.2 mM versus 2.1 mM, respectively) and a 13.8-fold higher Km for NH3 (6.4 mM versus 0.46 mM) than the enzyme from the control. These data show that the glutamine synthetase protein has been altered by this mutation, designated as glnA982, and suggest that the L-methionine SR-sulfoximine resistance is conferred by a change in the NH3 binding domain of the enzyme.     

Control of the flux in the arginine pathway of Neurospora crassa. Modulations of enzyme activity and concentration.

The influence of particular enzyme activities on the flux of metabolites in a pathway can be estimated by 'modulating' enzymes (i.e. changing turnover or concentration) and measuring the response in various parts of the system. By controlling the nuclear ration of two genetically different nuclear types in heterokaryons, the enzyme concentrations at four different steps in the arginine pathway were decreased over a range. This range was extended by the use of bradytrophs, mutant strains specifying enzymes with greatly diminished enzyme activities. Strains altered simultaneously at more than one step were also constructed by genetic recombination. By measuring the outputs of the pathway and the steady-state concentrations of intermediate pools, the fluxes in different parts of the pathway were calculated. This allowed the construction of flux/enzyme relationships, the slope of which is a measure of the sensitivity of a flux to the change in enzyme activity at that step. All fluxes were found to be considerably buffered for quite substantial decreases in the activities of all enzymes. Mass action plays an important part in this phenomenon, as do inhibition and repression. Because of the existence of expansion fluxes in growing systems, we find quantitatively different fluxes in different parts of the single pathway. For the same reason some enzyme modulations given decreased fluxes in one part and increased fluxes in another. The understanding of control in the pathway thus involves consideration of many mechanisms operating simultaneously and the estimation of changes in the whole system. The concept of a 'rate-limiting step' is found to be inadequate and is replaced by a quantitative measure, the Sensitivity Coefficient, which takes account of all the interactions. It is shown that control of the flux is shared among all the enzymes of the pathway. The results are discussed in terms of the theory of flux control.     

Enhanced vigilance in monogamous pairs of the lizard, Tiliqua rugosa

The Australian sleepy lizard, Tiliqua rugosa, forms monogamouspairs for up to 8 weeks each spring before mating. We observedthat males had food in their mouths significantly less oftenwhen they were in pain than when they were alone. Females hadfood in the mouth independent of the presence or absence ofmales. Among females that had been feeding, indicated by foodin the mouth, we observed them feeding, as we approached, lessoften when they were in pairs than when they were alone. Amongfemales in pairs with food in their mouths, we observed themfeeding less often when their male partners were not feedingthan when their partners were feeding. This suggests that femalesare alerted to approaching danger earlier when they are in apair, and alerted earlier in a pair when their male partneris not feeding. Enhanced vigilance may be one function of pairingbehavior.     

Evaluation of environmental and plant-associated cues for nymphal settling preference by Trioza eugeniae

Experiments were conducted to determine potential cues used by nymphs when they select the preferred abaxial side of a leaf. Nymphs most frequently settle and develop on the abaxial side of leaves. The degree of preference for the abaxial side was found to be reduced when a high density of nymphs were present, however. The orientation of leaves in a completely darkened chamber did not influence settling preference for the abaxial side, even when leaves were oriented abaxial side up. Coating both sides of a leaf with an antitranspirant compound did not prevent nymphs from preferring the abaxial side for settling. Similar results were obtained when leaves were dipped in pentane. These studies show that nymphs do not use gravity, color, light, or contact chemoreception when selecting suitable settling sites on a leaf.     

Negative frequency-dependent preferences and variation in male facial hair

Negative frequency-dependent sexual selection maintains striking polymorphisms in secondary sexual traits in several animal species. Here, we test whether frequency of beardedness modulates perceived attractiveness of men''s facial hair, a secondary sexual trait subject to considerable cultural variation. We first showed participants a suite of faces, within which we manipulated the frequency of beard thicknesses and then measured preferences for four standard levels of beardedness. Women and men judged heavy stubble and full beards more attractive when presented in treatments where beards were rare than when they were common, with intermediate preferences when intermediate frequencies of beardedness were presented. Likewise, clean-shaven faces were least attractive when clean-shaven faces were most common and more attractive when rare. This pattern in preferences is consistent with negative frequency-dependent selection.     

Kin-biased dispersal behaviour in the mango shield scale, Milviscutulus mangiferae

When fitness decreases with increasing density in a habitat, dispersal behaviour is expected to evolve. To avoid competition between kin, dispersal behaviour based on kin recognition should be more likely to occur when the individuals in a habitat are closely related. I tested this prediction with first-instar larvae (crawlers) of the mango shield scale, Milviscutulus mangiferae. The body size of adult females, a measure of fecundity, was larger when only one female was present on a leaf than when two were present. When I placed two crawlers on a leaf, they emigrated more frequently when they were siblings than when they were not related. I discuss the implication of the results for kin recognition in thelytokous parthenogenetic animals. Copyright 2000 The Association for the Study of Animal Behaviour.     

Inadequate rates are lower when FNAC samples are taken by cytopathologists

Inadequate rates (IR) in FNAC from different sources were compared. The rates were lowest when FNAC was performed by a cytopathologist (12%) and highest when done by a non-cytopathologist (32%). These differences were mirrored in high IRs in breast cancer cases. IR was not significantly improved when non-cytopathologist FNAC was attended by a cytotechnician.     

Visual motion retards alternations between conflicting perceptual interpretations

When the visual system is faced with conflicting or ambiguous stimulus information, visual perception fluctuates over time. We found that perceptual alternations are slowed when inducing stimuli move within the visual field, constantly engaging fresh, unadapted neural tissue. During binocular rivalry, dominance durations were longer when rival figures moved compared to when they were stationary, yielding lower alternation rates. Rate was not reduced, however, when observers tracked the moving targets, keeping the images on approximately the same retinal area. Alternations were reliably triggered when rival targets passed through a local region of the visual field preadapted to one of the rival targets. During viewing of a kinetic globe whose direction of rotation was ambiguous, observers experienced fewer alternations in perceived direction when the globe moved around the visual field or when the globe's axis of rotation changed continuously. Evidently, local neural adaptation is a key ingredient in the instability of perception.     

Evidence of Social Influences on Cache‐Making by Grey Squirrels (Sciurus carolinensis)

Many studies have found that scatter‐hoarding animals change their behaviour when storing food in the presence of conspecifics to minimize the likelihood that their caches will be pilfered; they refrain from caching, move away from conspecifics or choose visually obscured sites. This study reports the first evidence that the presence of conspecifics continues to influence the caching behaviour of a scatter‐hoarding mammal, the grey squirrel, after a suitable cache site has been selected and the hoarder is filling and covering its cache. Wild grey squirrels were filmed when storing preferred and less preferred nuts and when they were alone or with conspecifics present. In line with previous findings, squirrels spent longer travelling from the nut patch and were more vigilant when conspecifics were present. However, squirrels also spent longer disguising their caches and were more likely to stop digging and become vigilant when conspecifics were present than when they were alone. In particular, they were most likely to curtail their digging when storing their preferred nuts in the presence of conspecifics. The results indicate that caching squirrels remain sensitive to the presence of conspecifics until the cache is complete and that they respond flexibly to conspecifics according to the type of food they are storing.     

Studies on reduction of lipolysis in adipose tissue on freezing and thawing.

Adrenaline-induced lipolysis in fat cells was remarkably reduced when the cells were preincubated in a dry ice-aceton bath, but their adenylcyclase and lipase activities were not reduced. In the reconstructed lipid micelles which consisted of lipase-depleted lipid micelles and lipase-containing adipose tissue extract, adrenaline, theophylline and DBcAMP-induced lipolysis was not found when lipase-depleted lipid micelles were preincubated in a dry ice-aceton bath but was found when lipase was preincubated.     

Regulation of cache stores and body mass in Carolina chickadees (Panes carolinensis)

Here I report the results of laboratory experiments on cachingbehavior of Carolina chickadees (Parus carolinensis) designedto test the following predictions from a recent dynamic optimizationmodel: under limited resources, small birds, when fat, shouldcache food instead of eating it, but they should eat insteadof caching when lean; when resources are abundant, birds shouldcache less when fat and more when lean. In addition, when resourcesare abundant the amount of time spent in foraging-related activityshould decrease with an increase in body mass. Chickadees weretested for 2 weeks in either a poor-quality environment (wherethey were given four 5-min periods per day of access to a feederfilled with sunflower seeds) or a richer environment (wherethey were given four 10-min access periods); they were thenswitched to the alternative environment for an additional 2weeks. The entire experiment lasted from October through June.Within-individual comparisons showed that birds in both thepoor and rich environments exhibited the predicted correlationsbetween the probability that a seed was cached and body massmeasured at dawn. The number of seeds already stored had a weakand variable effect on caching decisions. Time budgets changedas predicted; birds spent less time with food-related behaviorswhen their energy stores were high compared to when they werelow, and overall spent a higher fraction of their time foragingin the poor-quality environment compared to the rich environment.Two other variables also affected caching behavior: experienceand season. Individuals were more likely to cache in the poorenvironment when they had been tested in the rich environmentthe previous 2 weeks, as compared with birds started in thepoor environment. However, this was true only for tests donein the winter. In late spring, all birds stopped caching orreduced caching rates when faced with limited food availability,irrespective of previous experience. The birds harvested seedsfaster when they cached instead of eating seeds; as a result,long-term weight regulation in the poor environment was affectedby caching decisions. Birds tested in late spring lost weightin the poor environment, whereas those tested in the wintermaintained a stable weight when switched from the rich to thepoor environment. Thus, annual endogenous cues directly affectcaching decisions and indirectly influence the long-term regulationof body mass     

Dispersal of yearling yellow-bellied marmots (Marmota flaviventris)

Field studies of the social organization of yellow-bellied marmots (Marmota flaviventris) in the vicinity of Gothic, Colorado, were initiated in 1962. Data collected during the subsequent 14 years are sufficient to analyse the behaviours that correlate with the time yearling male and yearling female marmots leave their natal home sites. Dispersal of yearling male yellow-bellied marmots was delayed when there were many yearling males in a harem, when yearling males were underweight, and when the rates of amicable interactions between yearling males and adults were high. Dispersal of yearling females was delayed when adults behaved amicably toward them and when the rates of adult aggression were low. Dispersal of yearling females was independent of the number of females in a harem. These observations are in accord with a model of dispersal that relates sexual differences in dispersal to maternal reproductive tactics, and to responses of yearlings that would maximize their own fitness.     

Resistance to Leptosphaeria coniothyrium in the red raspberry and some related species

Useful resistance to Leptosphaeria coniothyrium was found in the red raspberry cv. Latham, but its effectiveness was affected by the stage of growth reached by the canes when they were inoculated and by the kind of inoculum used: it was effective when field-grown canes were inoculated in mid-August but less so when they were inoculated in early July, and more effective against a spore inoculum than against a mycelial one. Resistance was inherited as a dominant multigenic character, but there were interactions between parents and two very favourable combinations were found.
The species Rubus pileatus, R. mesogaeus, R. coreanus and R. lasiocarpus showed resistance when inoculated in pot experiments, but F₁ hybrids obtained by crossing them with the raspberry were largely susceptible. The problems of using these resistance sources in breeding are discussed.     

Working for a dustbath: are hens increasing pleasure rather than reducing suffering?

Dustbathing is one of the major behavioural systems of domestic fowl that is constrained by commercial caging systems. Although research conducted over the last 20 years has revealed a great deal about causation of dustbathing, attempts at measuring the importance to hens of having access to dustbathing substrate have proven difficult. Various economic techniques, operant conditioning and learning trials have been used to determine whether hens have a 'need' for dustbathing substrate or 'think' about dustbathing in the absence of substrate. In this study, we measured hens' willingness to work to obtain substrate for dustbathing using a vertically swinging door to which weights could be added. Hens were trained to push through the door to enter a goal box containing peat moss. The hens were subjected to two series of trials to compare the maximum weight that they would push to open the door when living in a cage and deprived of dustbathing substrate, with the maximum weight that they would push when living in a pen furnished with litter and a dustbath (non-deprived). Of the 10 hens that opened the door for access to peat moss, six hens pushed more weight when deprived, three hens pushed more weight when non-deprived and one hen pushed an equal amount of weight. Overall, the hens tended to push more weight (860+/-95.6 vs. 682.5+/-83.3 g; P<0.10, one-tailed paired t-test) and tended to make more attempts to open the door (P<0.10) when they were deprived than when they were non-deprived. Significantly more trials resulted in dustbathing when hens were deprived (P<0.01) suggesting that following deprivation, the hens were, in fact, more motivated to dustbathe. The results of these trials indicate that although deprived hens may be more motivated to dustbathe, and that most hens may be willing to work to obtain a dusty substrate when they can see it, they are not necessarily willing to work harder when they are in a state of deprivation than when they have recently dustbathed. These results are very difficult to explain using a 'needs' model of motivation in which deprivation leads to a state of suffering. They are much more consistent with an 'opportunity' model of motivation in which performance of the behaviour, when the opportunity presents itself, leads to a state of pleasure.     

Visual search for orientation of faces by a chimpanzee (<Emphasis Type="Italic">Pan troglodytes</Emphasis>): face-specific upright superiority and the role of facial configural properties

A previous experiment showed that a chimpanzee performed better in searching for a target human face that differed in orientation from distractors when the target had an upright orientation than when targets had inverted or horizontal orientation [Tomonaga (1999a) Primate Res 15:215–229]. This upright superiority effect was also seen when using chimpanzee faces as targets but not when using photographs of a house. The present study sought to extend these results and explore factors affecting the face-specific upright superiority effect. Upright superiority was shown in a visual search for orientation when caricaturized human faces and dog faces were used as stimuli for the chimpanzee but not when shapes of a hand and chairs were presented. Thus, the configural properties of facial features, which cause an inversion effect in face recognition in humans and chimpanzees, were thought to be a source of the upright superiority effect in the visual search process. To examine this possibility, various stimuli manipulations were introduced in subsequent experiments. The results clearly show that the configuration of facial features plays a critical role in the upright superiority effect, and strongly suggest similarity in face processing in humans and chimpanzees.     

Son preference and child replacement in Bangladesh: a new look at the child survival hypothesis

Birth history data from women in the 1975-76 Bangladesh Fertility Survey were used to search for intentions to replace dead children. The median intervals between successive births of orders (i) and (i + 1) were not shorter when some siblings of orders below (i) had died. Nor was the median duration between the death of a child and the first posthumous birth shorter when the dead child was a boy or when it was survived by fewer than two brothers. The median intervals were generally shorter when the mother lived in an urban rather than a rural area but this difference was attributable only to the shorter duration of breast-feeding by urban women. These results disputed the notions that the timing of births was deliberately quicker to replace a dead child, that attempts at replacement were sex-selective, or that child replacement intentions were stronger in urban than in rural populations.     

Differential effects of empty and filled intervals on duration estimation by pigeons: tests of an attention-sharing explanation

Pigeons were trained in a within-subjects design to discriminate durations of an empty interval and a filled interval. Even when different stimuli were used to mark empty intervals and to signal filled intervals, pigeons judged empty intervals to be longer than equal-length filled intervals. This timing difference was not a result of pigeons timing marker duration on empty interval trials. Increasing marker duration did not produce an overestimation of the empty time intervals. It was suggested that this timing difference could be due to a reduction in attention to temporal processing on filled interval trials when visual stimuli are used. Consistent with this hypothesis, it was found that empty intervals were judged longer than filled intervals when testing occurred in a darkened test room, but not when the test room was illuminated. In addition, no timing difference was observed when different auditory stimuli were used as markers for empty intervals and as signals for filled intervals.