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Two different approaches to computer-aided teaching of microbial genetics.

Two computer packages have been developed to teach bacterial genetics on an introductory genetics course for undergraduate students in biology. The first package, 'CONJUGACION', is designed to teach bacterial conjugation and its genetic outcomes. It includes four main parts. Firstly, a tutorial part presents a theoretical framework using screens of text and animated graphics. Secondly, an interactive concept application section requires students to carry out experiments for the determination of the correct sex and genotype of 10 bacterial strains. The third part uses the previously obtained data for simulating interrupted mating experiments and mapping the bacterial genome. Finally, an evaluation section allows the students to test their understanding through a series of multiple choice questions. The second package, 'LURIDEL', is intended for teaching the preadaptative character of mutation in bacterial populations on the basis of the fluctuation test of Luria and Delbrück. It simulates, graphically, the appearance of mutations in microbial cultures and gives results of simulated fluctuation experiments. Programs were written under the PCOS operating system in MBASIC extended to graphics for running on Olivetti M20 microcomputers.

Computer-Assisted Instruction↗

Plasmodium falciparum CyRPA Glycan Binding Does Not Explain Adaptation to Humans.

The human malaria parasite Plasmodium falciparum evolved from a parasite that infects gorillas, termed Plasmodium praefalciparum. The sialic acids on glycans on the surface of erythrocytes differ between humans and other apes. It has recently been shown that the P. falciparum cysteine-rich protective antigen (PfCyRPA) binds human sialoglycans as an essential step in the erythrocyte invasion pathway, while that of the chimpanzee parasite, Plasmodium reichenowi has affinities matching ape glycans. Two amino acid changes, at sites 154 and 209, were shown to be sufficient to switch glycan binding preferences and inferred to reflect adaptation of P. falciparum to humans. However, we show that sites 154 and 209 are identical in P. falciparum and P. praefalciparum, with no other differences located in or near the CyRPA glycan binding sites. Thus, the gorilla precursor appears to have already been preadapted to bind human sialoglycans.

Plasmodium falciparum↗

Sleep and mammalian hibernation: homologous adaptations and homologous processes?

Evidence from electroencephalographic, thermoregulatory and cellular neurophysiological studies suggests that sleep and hibernation may be homologous adaptations for energy conservation. However, despite the similarities between non-rapid eye movement (NREM) sleep and hibernation, the restorative function normally associated with slow wave sleep appears not to occur during hibernation, perhaps because of the low body temperature (Tb). Cellular neurophysiological studies also suggest that a bout of hibernation is not exclusively NREM sleep but is punctuated by periods of wakefulness. The entrance to hibernation involves both an inhibition of cortical activity and activation of hypothalamic regions, whereas the arousal from hibernation is primarily a hypothalamic function. Multiple neurochemical systems are affected by the arousal state change that occurs in hibernation, and a serotonergic-opiatergic interaction, in particular, may be important in regulating these events. Among regulated physiological systems affected by arousal state changes, the episodic respiration evident in hibernation shows striking similarities to the apneas observed during sleep in both humans and other mammals. Although the slight down-regulation of Tb and metabolism that accompanies the transition from wakefulness to NREM sleep may have served as a preadaptation for the evolution of hibernation among the mammals, increasing consideration must be given to the possibility that hibernation represents an arousal state distinct from any known normothermic arousal state.

Animals↗

Adaptive modification of oculomotor pursuit influences manual tracking responses.

We have addressed the question of whether adaptively modifying the oculomotor response to a visual pursuit stimulus has an influence on a related manual tracking response. Subjects used their unseen right hand to track targets moving at constant velocities while visually fixating a stationary LED. Manual tracking performance was compared before and after a 20 min period during which smooth pursuit eye movements alone were adaptively enhanced by adding 50% of the instantaneous eye position signal to target position. Compared with the preadaptation trials, hand gain was markedly increased during the postadaptation period. These results imply that the adaptation occurred at a level common to both motor systems, probably in CNS structures concerned with visual motion processing.

Adaptation, Physiological↗

Effect of accommodative adaptation on static and dynamic accommodation in emmetropia and late-onset myopia.

PURPOSE: Prolonged near work is considered to be an environmental factor leading to the development of late-onset myopia. Accommodation may be the specific mechanism underlying the association between near work and late-onset myopia. To determine whether late-onset myopes have abnormal accommodation, we compared accommodative static responses and dynamic facilities before and after a near task in two groups of subjects, emmetropes and late-onset myopes. METHODS: In experiment 1, the accommodative stimulus/response function with monocular viewing and the dark focus (the accommodative response in the dark) were objectively measured with a Canon R-1 infrared optometer before (preadaptation) and after (postadaptation) a 20-min near task. In experiment 2, monocular accommodative facility (AF) and dark focus were measured before and after the near task. Facility was measured as the subjective time needed to clear an accommodative target (20/40 letters) at 40 cm through +/-2.00 D lenses. The time between when the subject flipped the lenses from viewing through the +2.00 D to the -2.00 D lenses was recorded by a computer. RESULTS: In both experiments, inward shifts of the dark focus were observed after the near task. In experiment 1, after the near task, static accommodative responses also showed a small but statistically significant inward shift. Neither postadaptation effect differed between refractive groups. The only difference between groups was that late-onset myopes had a lower slope of the accommodative stimulus/response function, both pre- and postadaptation. In experiment 2, both refractive groups showed the same results. After the near task, the duration for accommodation from near to far (relaxation) increased but the duration for accommodation from far to near (stimulation) did not change. CONCLUSIONS: Late-onset myopes have shallower accommodative stimulus/response functions. As suggested in a previous study, this may be due to their reduced sensitivity to defocus. In both emmetropes and late-onset myopes, the near task causes an increase in static accommodative responses. Although our results show it to be a small increase, it is consistent with predictions of Hung and Semmlow's model of accommodation. In both emmetropes and late-onset myopes, the near task also increases the duration for relaxing accommodation, but not for stimulating accommodation. This suggests there are two subsystems which may adapt to prolonged accommodation differently.

Accommodation, Ocular↗

Clinical evaluation of toric soft lens reproducibility.

PURPOSE: The manufacturing variability of frequent replacement toric soft lenses is such that some resulting variation in clinical performance can be expected. The purpose of this study was to evaluate the effect of manufacturing variability on the clinical performance of three frequent replacement toric soft contact lenses. METHODS: This was a three-part study in which 12 to 14 subjects were enrolled for each part. In each part, subjects were preadapted to the lens type 1 month prior to the assessment visits. Subjects wore a total of 12 test lenses in one eye and, in each case, the clinical performance was compared with a control lens of the same type worn in the other eye that was removed between assessments but not replaced. The study lenses were Acuvue Toric (Vistakon), Focus Toric (Ciba Vision) and FreshLook Toric (Wesley lessen). RESULTS: Two lens types (Focus Toric and FreshLook Toric) showed significantly greater variability with the test lenses compared with the nonreplaced control lenses with several variables: lens fit, visual acuity, and subjective vision quality. One lens (Focus Toric) also showed greater variability of orientation position, and another (Acuvue Toric) showed greater variability in subjective vision quality. CONCLUSIONS: The study demonstrated variability in aspects of clinical performance, which appear to be linked to variability of manufacturing output. With toric soft lenses used on a frequent replacement basis, better reproducibility is needed if consistent clinical performance is to be maintained.

Contact Lenses, Hydrophilic↗

Palaeoecology and palaeophytogeography of the rhynie chert plants: evidence from integrated analysis of in situ and dispersed spores.

The Rhynie cherts yield exceptionally preserved early land plants, and provide a unique insight into the nature of Lower Devonian vegetation. Hitherto they have been poorly age constrained, and the palaeoecology and palaeophytogeography of the flora are poorly understood. Well-preserved dispersed-spore assemblages have been recovered from a number of borehole cores through the stratigraphical sequence of the Rhynie outlier. They are all similar and belong with the polygonalis-emsiensis (PE) spore zone, indicating an Early Devonian age (Early (but not earliest) Pragian to earliest Emsian). Comparisons with PE spore-zone assemblages from elsewhere suggest that the flora of the Rhynie drainage basin was slightly impoverished, with certain plant taxa that occurred at other locations not represented. This probably reflects differences between the flora of an inland intermontaine basin (Rhynie) and that of the lowland flood-plains. In situ spores have been characterized for all seven Rhynie chert plants. Analysis of the distribution of these spore types in the Rhynie sequence, in addition to those of coeval deposits from elsewhere, enables interpretation of the palaeoecology and palaeophytogeography of the Rhynie chert plants. It is concluded that at least some of the plants were not highly specialized or adapted to the peculiar hot-springs environment in which they are preserved. Rather, they were components of a diverse and widespread flora, but were the only elements able to tolerate the inhospitable hot-springs environment (i.e. they were preadapted).

Age Factors↗

Light dependence of catalase synthesis and degradation in leaves and the influence of interfering stress conditions.

The enzyme catalase (EC 1.11.1.6) is light sensitive and subject to a rapid turnover in light, similar to the D1 reaction center protein of photosystem II. After 3 h of preadaptation to darkness or to different light intensities (90 and 520 mumol m(-2) s(-1) photosynthetic photon flux density), sections of rye leaves (Secale cereale L.) were labeled for 4 h with l-[(35)S]methionine. From leaf extracts, catalase was immunoprecipitated with an antiserum prepared against the purified enzyme from rye leaves. Both incorporation into catalase and degradation of the enzyme polypeptide during a subsequent 16-h chase period increased with light intensity. At a photon flux density of 520 mumol m(-2) s(-1), the apparent half-time of catalase in rye leaves was 3 to 4 h, whereas that of the D1 protein was much shorter, about 1.5 h. Exposure to stress conditions, such as 0.6 m NaCl or a heat-shock temperature of 40 degrees C, greatly suppressed both total protein synthesis and incorporation of the label into catalase and into the D1 protein. Immunoblotting assays indicated that in light, but not in darkness, steady-state levels of catalase and of the D1 protein strongly declined during treatments with salt, heat shock, or translation inhibitors that block repair synthesis. Because of the common property of rapid photodegradation and the resulting dependence on continuous repair, declines in catalase as well as of the D1 protein represent specific and sensitive indicators for stress conditions that suppress the translational activities of leaves.

Journal Article↗

Phenotypical temperature adaptation of protein synthesis in wheat seedlings: time curves for readaptation.

Optimum temperature and temperature coefficient of protein synthesis in young wheat plants exhibit phenotypical temperature adaptation. In plants grown for 2 days at either chilling (4 C), medium (20 C), or high (36 C) temperature the respective values are: 27 C and 14.2 kilocalories per mole, 31 C and 18.2 kilocalories per mole, 35 C and 23.6 kilocalories per mole, based on in vivo [(14)C]leucine incorporation into total protein. The validity of the [(14)C]leucine incubation method has been confirmed by double-labeling experiments. Readaptation time curves are complex: the optimum temperature parameter readjusts within approximately 4 hours to an altered temperature regime, whereas the temperature coefficient needs between 4 and 96 hours for complete readaptation-depending on the temperature conditions prior to the temperature shift. Heat-preadapted plants need a recovery period at medium temperature to regain their cold adaptability with respect to optimum temperature. Cycloheximide (30 micrograms per milliliter) reduces [(14)C]leucine incorporation into protein by 85%, thus indicating that predominantly the cytoplasmic 80S system of protein synthesis is involved in temperature adaptation.

Journal Article↗

Multiple origin of viviparity in Southeast Asian gastropods (Cerithioidea: Pachychilidae) and its evolutionary implications.

This study aims at a better understanding of the evolutionary significance of viviparity in some freshwater gastropods. We use a phylogeny based on partial sequences of the mitochondrial 16S gene of representatives of the limnetic and pantropical Pachychilidae to infer the relationships within this particular group of cerithioideans and the evolution of reproductive strategies. The phylogeny presented herein implies a new systematization and suggests that viviparity has appeared three times among the Pachychilidae. This is supported by the finding of very distinct reproductive morphologies in different lineages of viviparous taxa that are exclusively found in Southeast Asia. Based on the observation that oviparity is the ancestral character state in this freshwater family, we conclude that viviparity has evolved subsequent to the exploration of freshwater. We present data showing that all Pachychilidae produce considerably larger but fewer egg capsules compared to most marine snails. In other studies on freshwater gastropods, this has been discussed as an adaptation to freshwater environments. In this context we hypothesize that the increased parental investment involved in the enlargement of eggs in concert with the reduction of clutch sizes was the driving factor that ultimately lead to the evolution of viviparity in the Asian taxa. Consequently, although not directly correlated with the colonization of the new adaptive zone, viviparity is strongly favored by other consequences of this step. Hence, we hypothesize that the production of large eggs, which is necessitated by the exploration of freshwater, represents a preadaptation existing in those ancestors from which viviparous pachychilid lineages eventually evolved in Southeast Asia.

Animals↗

Heterochrony and the evolution of poecilogony: generating larval diversity.

Poecilogony is the production of more than one type of young within a single species of marine invertebrate. We chose a poecilogonous polychaete to investigate potential differences in morphogenesis among offspring that are polymorphic in dispersal potentials (planktonic, benthic) and trophic modes (planktotrophy, adelphophagy). Differences in morphogenesis occur and are strongly influenced by maternal type. Females that provide extra-embryonic nutrition (as nurse eggs; type III females) also produce offspring with an accelerated onset of juvenile traits, relative to planktotrophic offspring of females that do not provide extra-embryonic nutrition (type I females). Thus, progeny of some females appear morphologically preadapted for a benthic lifestyle. Surprisingly, differences in phenotype among offspring do not parallel offspring ecotype, as offspring with early onset of juvenile traits (III) are ecologically bimodal. Some Type III offspring eat the nurse eggs (adelphophagy), have accelerated development, and hatch as benthic juveniles. In contrast, their siblings hatch as small, planktotrophic, dispersive larvae that are morphologically similar to their type III siblings, but ecologically similar to Type I planktotrophic larvae. We propose that poecilogony evolved through sequence heterochrony in morphogenesis with accelerated onset of juvenile traits in type III offspring. In addition, we suggest that heterochrony in life-history events (hatching, metamorphosis) also occurs, thereby generating offspring that are dimorphic in both phenotype and ecotype. Over time, selection acting on different levels of ontogeny (morphogenesis vs. dispersal) may balance this polymorphism and allow poecilogony to persist.

Animal Nutritional Physiological Phenomena↗

Loss of GLUR2 alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid receptor subunit differentially affects remaining synaptic glutamate receptors in cerebellum and cochlear nuclei.

The alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid (AMPA) type of ionotropic glutamate receptor is the major mediator of fast neurotransmission in the brain and spinal cord. Most AMPA receptors are impermeable to calcium because they contain the GluR2 subunit. However, some AMPA receptors lack GluR2 and pass calcium which can mediate synaptic plasticity and, in excess, neurotoxicity. Previously, we showed a decrease in the density of synaptic AMPA receptors in the hippocampus of mice lacking GluR2. In this study, using these GluR2-lacking mice, we examined other areas of the brain that differ in the amount of GluR2 normally present. Like hippocampal spines, cerebellar Purkinje spines normally express AMPA receptors with high GluR2 and showed a decrease in synaptic AMPA receptors in mutant mice. In contrast, neurons that normally express AMPA receptors with little or no GluR2, such as in the anteroventral cochlear nucleus, showed no decrease in AMPA receptors and even showed an increase in one AMPA receptor subunit. These two different patterns may relate to preadaptations to prevent calcium neurotoxicity; such mechanisms might be absent in Purkinje and hippocampal spines so that these neurons must decrease their total expression of synaptic AMPA receptors (calcium permeable in mutant mice) to prevent calcium neurotoxicity. In addition, we found that another glutamate receptor, GluRdelta2, which is abundant only in parallel fibre synapses on Purkinje cells and in the dorsal cochlear nucleus, is up-regulated at these synapses in mutant mice; this probably reflects some change in GluRdelta2 targeting to these synapses.

Animals↗

Sensitivity of cells to apoptosis induced by iron deprivation can be reversibly changed by iron availability.

We tested the effect of iron deprivation on cell death induction in human Raji cells pre-adapted to differing availability of extracellular iron. Iron deprivation was achieved by incubation in a defined iron-free medium. Original Raji cells have previously been adapted to long-term culture in a defined medium with 5 microg/ml of iron-saturated human transferrin as a source of iron. Raji/lowFe cells were derived from original Raji cells by subsequent adaptation to culture in the medium with 50 microm ferric citrate as a source of iron. Raji/lowFe-re cells were derived from Raji/lowFe cells by re-adaptation to the transferrin-containing (5 microg/ml) medium. Iron deprivation induced cell death in both Raji cells and Raji/lowFe-re cells; that is, cells pre-adapted to a near optimum source of extracellular iron (5 microg/ml of transferrin). However, Raji/lowFe cells preadapted to a limited source of extracellular iron (50 microm ferric citrate) became resistant to the induction of cell death by iron deprivation. We demonstrated that cell death induction by iron deprivation in Raji cells correlates with the activation of executioner caspase-3 and the cleavage of caspase-3 substrate, poly-ADP ribose polymerase. Two other executioner caspases, caspase-7 and caspase-6, were not activated. Taken together, we suggest that in human Raji cells, iron deprivation induces apoptotic cell death related to caspase-3 activation. However, the sensitivity of the cells to death induction by iron deprivation can be reversibly changed by extracellular iron availability. The cells pre-adapted to a limited source of extracellular iron became resistant.

Apoptosis↗

Can maternally transmitted endosymbionts facilitate the evolution of haplodiploidy?

Whilst many invertebrate taxa are haplodiploid, the factors underlying the evolution of haplodiploidy remain unresolved. We investigate theoretically whether haplodiploidy might evolve as an outcome of the co-evolution between maternally inherited endosymbionts and their hosts. First, we substantially extend a recently developed model that involves maternally inherited endosymbionts that kill male offspring by eliminating the paternal genome. We also put forward a new hypothesis and develop a model that involves bacteria that induce cytoplasmic incompatibility (CI). Based on these models, we explore the co-evolutionary events that might occur between hosts and symbionts. We find that both with male-killers and CI-inducing endosymbionts, the hosts are likely to develop increased viability of haploid males, which can be considered a preadaptation to haplodiploidy. In addition, populations with haploidizing male-killers can in some cases evolve directly towards a genetic system of paternal genome elimination, a special form of haplodiploidy. These results are combined with consideration of mechanism and ecology to appraise the likelihood of male-killers and CI inducing bacteria being involved in the evolution of haplodiploidy.

Animals↗

Trypanosoma brucei: differentiation of in vitro-grown bloodstream trypomastigotes into procyclic forms.

Trypanosoma brucei strain 366D trypomastigotes grown at 37 degrees C in the presence of a human fibroblast cell line formed foci underneath the feeder cells whereas trypanosomes grown in the presence of a human epithelial cell line grew only in the culture supernatant. A culture system was developed to study the differentiation of bloodstream trypomastigotes grown in the epithelial cell system into procyclic trypomastigotes at 27 degrees C. The morphological differentiation into the procyclic form was complete by 48 h. Cell division did not occur until 30-40 h after transfer to 27 degrees C. Various characteristics of this system were examined, including the effect of the feeder layer, the type of medium, the presence of the metabolites cis-aconitate and citrate, the preadaptation period, and the trypanosome cell concentration. The respiration of the recently differentiated procyclic cells was less sensitive to inhibition by CN- than that of established procyclic forms, implying a delayed appearance of complete mitochondrial oxidative pathways. This trypanosome differentiation system has the advantage that the animal host is not needed and the entire process is carried out in in vitro culture.

Aconitic Acid↗

Survival of enteric bacteria in seawater.

Enteric bacteria exposed to the marine environment simultaneously encounter a variety of abiotic and biotic challenges. Among the former, light appears to be critical in affecting seawater survival; previous growth history plays a major part in preadaptation of the cells, and stationary phase cells are generally more resistant than exponentially growing ones. Predation, mostly by protozoa, is probably the most significant biotic factor. Using Escherichia coli as a model, a surprisingly small number of genes was found that, when mutated, significantly affect seawater sensitivity of this bacterium. Most prominent among those is rpoS, which was also dominant among genes induced upon transfer to seawater.

Enterobacteriaceae↗

Evolutionary origins of the vertebrate dentition: phylogenetic patterns and developmental evolution.

The theory that teeth evolved from dermal denticles linked with the origin of jaws no longer accounts for the diversity of new data emerging from the fossil record. We have reviewed oropharyngeal dental patterns in all fossil groups of early vertebrates to establish the primitive condition, in order to understand the polarity of change. The evolutionary precedence of dermal denticles before teeth now seems less likely; both may be alternative manifestations of a common morphogenetic system. This developmental system involves regulatory changes affecting the odontode, a fundamental exoskeletal unit, and can explain skeletal diversity. However, tooth and denticle differences may have diverged at loci deep within vertebrate phylogeny, as real differences exist between them. Teeth were conceived as evolving from non-growing odontodes with regulation of precise increase in size, position, sequence of time of development, and polarity of shape. A characteristic feature of teeth is the ability to replace from a developing sequence, programmed with these parameters, prior to demand. Tooth whorls, a feature of denticles in the oropharyngeal region, may be regarded as a preadaptation of this tooth replacement mechanism. The new fossil evidence suggests that teeth may have evolved from these more specialised oropharyngeal denticles in agnathan vertebrates.

Animals↗