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PubMed1984

Translation is a non-uniform process. Effect of tRNA availability on the rate of elongation of nascent polypeptide chains.

We reported elsewhere (Varenne et al., 1982) that, during synthesis of a number of colicins in Escherichia coli, intermediate nascent chains of discrete sizes accumulated, suggesting a variable rate of translation. In this paper, a detailed analysis provides arguments that this phenomenon, at least for the proteins under study, is not related to aspects of messenger RNA such as secondary structure. It is linked to the difference in transfer RNA availability for the various codons. Experimental analysis of translation of other proteins in E. coli confirms that the main origin for the discontinuous translation in the polypeptide elongation cycle is the following. For a given codon, the stochastic search of the cognate ternary complex (aminoacyl-tRNA-EF-Tu-GTP) is the rate-limiting step in the elongation cycle: transpeptidation and translocation steps are much faster. The degree of slackening in ribosome movement is almost proportional to the inverse of tRNA concentrations. The verification of this model and its possible physiological significance are discussed.

Bacterial Outer Membrane Proteins↗
PubMed1993

Forestomach motility and behavior of bull calves according to changes in regimen.

Six 23-week-old bull calves were randomly assigned to a crossover design to study the effects of hay and concentrates given separately (A), silage and concentrates given in complete mixed ration (B), or given separately (C) on reticulo-ruminal motility and eating and resting behaviors after an adaptation period of 14 days (day 14). The effects of a change from one treatment to another on the same variables were also investigated (day 1). The differences observed between day 1 and day 14 indicate that a change in regimen, even not drastic, seems to affect reticulo-ruminal motility, rumination pattern, ruminal inactivity, and resting behavior. As expected, the number of triphasic and biphasic reticular spike bursts, as well as eating and resting behaviors, were affected by the type of forage ingested (A vs. C) on day 14 (p < or = 0.05). Moreover, the mode of distribution of concentrates affected also the variables measured (p < or = 0.05). Although the length of particles in complete mixed ration (B) was the same than in silage and concentrates given separately (C), the effect of regimen B on many variables was similar to that of regimen A or intermediate between A and C. The effect of regimens B and C was similar only for the rumination pattern, even though the number of boluses regurgitated for rumination differed (p < or = 0.05).

Age Factors↗
PubMed2004

Spatial patterns of recreation impact on experimental campsites.

Management of camping impacts in protected areas worldwide is limited by inadequate understanding of spatial patterns of impact and attention to spatial management strategies. Spatial patterns of campsite impact were studied in two subalpine plant communities in the Wind River Mountains, Wyoming, USA (a forest and a meadow). Response to chronic disturbance and recovery from acute disturbance were both assessed. Previously undisturbed sites were camped on at intensities of one and four nights/year, for either one or three consecutive years. Recovery was followed for three years on sites camped on for one year. Percent bare ground, assessed in 49 contiguous 1 m2 quadrats, increased with increasing use frequency, particularly on the forest sites. Magnitude of impact varied spatially within campsites, with impact decreasing as distance from the center of the campsite increased. On the more fragile forest sites, this radial impact pattern developed rapidly and remained after three years of recovery. Concentration of camping activities around a centrally located small cooking stove was the apparent cause of this pattern. Maximum variation in magnitude of impact occurred at intermediate levels of campsite use and disturbance. The magnitude, variability and spatial pattern of impact varied with the spatial scale of analysis. Generally, results of these controlled experiments are consistent with earlier studies of campsites and validate the management implications derived from those studies. Even where campers use low-impact techniques, low levels of camping use can cause substantial impact and it is important to concentrate use. On resistant sites, however, it is possible that low levels of use can be sustained with minimal resultant impact.

Conservation of Natural Resources↗
PubMed2002

Thyroid status in juvenile alligators (Alligator mississippiensis) from contaminated and reference sites on Lake Okeechobee, Florida, USA.

Exposure to environmental contaminants has been shown to alter normal thyroid function in various wildlife species, including the American alligator (Alligator mississippiensis). Abnormalities in circulating levels of the thyroid hormone thyroxine (T4) have been reported in juvenile alligators from several contaminated lakes in Florida. To further elucidate these functional thyroid abnormalities, this study examines the structure of thyroids and circulating T4 concentrations from juvenile alligators collected from three sites of varying contamination on Lake Okeechobee, Florida. The following variables were used to characterize thyroid morphology: epithelial cell height, width and area, percent colloid, and follicle area. These variables were compared among study sites and between genders. No difference was detected in epithelial cell height, epithelial cell area, or follicle area among the sites, whereas significant differences in epithelial cell width (p = 0.02) and percent colloid (p = 0.008) were found. Animals from the most contaminated site (Belle Glade) had significantly greater epithelial cell widths and significantly less colloid present in their follicles compared to animals from the reference site (West). Gender did not have a significant interaction with site for any variable measured. Thyroxine (T4) concentrations were elevated in the intermediately contaminated site (Conservation Area 3A) compared to the other sites (p < 0.0001). It is proposed that the disruptions seen in Lake Okeechobee alligators are due to disruptions at both the thyroid and extra-thyroidal tissues.

Alligators and Crocodiles↗
PubMed1997

Cognitive deficits induced by global cerebral ischaemia: prospects for transplant therapy.

Global ischaemia induced by interruption of cerebral blood flow results in damage to vulnerable cells, notably in the CA1 and hilar hippocampal fields, and is frequently associated with memory deficits. This review examines cognitive deficits that occur in animal models of global ischaemia in rats and monkeys, the extent to which these deficits are associated with CA1 cell loss, and the evidence for functional recovery following transplants of foetal CA1 cells and grafts of conditionally immortalised precursor cells. In rats, impairments are seen most consistently in tasks of spatial learning and spatial working memory dependent on use of allocentric environmental cues. In monkeys, ischaemic deficits have been shown to a moderate extent in delayed object recognition tasks, but animals with a selective excitotoxic CA1 lesion show a profound impairment in conditional discrimination tasks, suggesting that these may be a more sensitive measure of ischaemic impairments. Several studies have reported correlational links between the extent of CA1 cell loss following two or four vessel occlusion (2 VO, 4 VO) in rats and behavioural impairments, but recent findings indicate that at intermediate levels of damage these relationships are weak and variable, and emerge clearly only when animals with maximal CA1 cell loss are included, suggesting that the deficits involve more than damage to the CA1 field. Nevertheless, ischaemic rats and CA1-lesioned marmosets with grafts of foetal CA1 cells show substantial improvements; in rats these are not found with grafts from other hippocampal fields. Conditionally immortalised cell lines and trophic grafts are currently being assessed for their functional potential in animal models, because clinical use of foetal cells will not be practicable. Recent findings suggest that an expanded population of neuroepithelial cells derived from the conditionally immortalised H-2Kb-tsA58 transgenic mouse improve spatial learning as effectively as CA1 foetal grafts in rats subjected to 4 VO, and clonal lines from the same source show similar promise. Lines derived from precursor cells have the potential to develop into different types of cell (neuronal or glial) depending on signals from the host brain. These cell lines may therefore have the capacity to repair damaged host circuits more precisely than is possible with foetal grafts, and offer a promising, approach both to functional recovery and to elucidating graft-host interactions.

Animals↗
PubMed2001

Behavioural differences between three breed groups of hunting dogs confronted with domestic sheep.

When running free in open fields, domestic dogs occasionally display predatory behaviour towards domestic sheep. This has not yet been studied scientifically. The aim of the present study was to investigate the inclination to chase sheep in three breed groups of hunting dogs that are most frequently used in areas with grazing sheep. We studied 41 elkhounds, 29 hare hunting dogs and 68 English setters. Behaviours indicative of motivation for chasing or attacking sheep were examined in three different ways. A path test examined functional traits such as hunting ability, contact willingness, reactivity to sudden noise, and response towards a lone sheep. In a sheep confrontation test, loose-leashed dogs were observed in a fenced enclosure with sheep and given electric shocks through an electronic dog collar if within 1-2m from the sheep. A questionnaire to the dog owners supplied information on their dog's previous experience with sheep and behavioural responses to various types of novel stimuli. No significant sex differences were found. The elkhounds showed the highest interest in a lone sheep in the path test, and displayed the highest initial hunting motivation, the highest percentage of dogs starting a sheep attack, the highest attack severity, and were most frequently given el. shocks. The hare hunting dogs were intermediate, while setters showed the lowest values for these variables. Dogs reported as showing low fearfulness more frequently acted as potential sheep chasers in the tests. Dogs up to 3 years of age showed a more pronounced initial hunting motivation and more frequent attacks than older dogs, although there were no age differences in the number of el. shocks given in the test. The latter may be related to the more frequent abruption of attacks among younger dogs. The main factors predicting a high hunting motivation and attack severity were lack of previous opportunity to chase sheep, low fearfulness towards gunshots and unfamiliar people, and general interest in sheep shown when encountering them. Probability of sheep chasing differed between dog breeds and age groups. Previous experience and certain character traits were indicative of a high predatory motivation towards sheep.

Journal Article↗
PubMed2002

Expression of the mouse Macf2 gene during inner ear development.

Plakins, a family of linker proteins that connect cytoskeletal elements to cellular junctions and the extracellular matrix, are primarily responsible for the mechanical properties of cells and tissues. They include desmoplakin, envoplakin, plectin, dystonin/BPAG1, and Kakapo. Mutations in plakins cause several skin, muscular and neurological disorders. Macrophins are a recently discovered subfamily of plakins with binding domains for actin, intermediate filaments and microtubules. Characteristic features of macrophins include variable actin binding domains, a central rod domain containing both plectin and spectrin repeats, and a C-terminus containing EF hands and GAS2/GAR22 domain. We have examined expression of mouse Macf2, encoding macrophin-2, in adult tissues and in the developing, neonatal, and mature inner ear by in situ hybridization. Northern blot analysis identified three large tissue-specific Macf2 transcripts: a 16-kb mRNA in skeletal muscle and heart, a 15-kb mRNA in brain, and a 9-kb mRNA in RNA from ovary plus uterus. In situ hybridization of the developing mouse inner ear indicated that Macf2 is expressed in the otocyst at day 12.5, in the sensory epithelium by embryonic day 16.5, and in both inner and outer hair cells by day 16.5. Macf2 is expressed in the bodies of both sensory and motor neurons in the central and peripheral nervous system, including the auditory pathway. The Macf2 protein could be involved in the regulation of cytoskeletal connections to cellular junctions and play an important structural role in organs, such as the inner ear, that are subjected to strong mechanical forces.

Animals↗
PubMed2006

Syntheses, structures, and magnetic properties of mononuclear CuII and tetranuclear CuII3MII (M = Cu, Co, or Mn) compounds derived from N,N'-ethylenebis(3-ethoxysalicylaldimine): cocrystallization due to potential encapsulation of water.

Syntheses, structures, and magnetic properties of one mononuclear inclusion compound [CuIIL1 within (H2O)] (1) and three tetrametal systems of the composition [{CuIIL1}2{CuIIL1MII(H2O)3}]ClO4)2 (M = Cu (2), M = Co (3), M = Mn (4)) derived from the hexadentate Schiff base compartmental ligand N,N'-ethylenebis(3-ethoxysalicylaldimine) (H2L1) have been described. Compounds 1 and 2 crystallize in orthorhombic Pbcn and monoclinic P21/c systems, respectively, and the space group of the isomorphous compounds 3 and 4 is monoclinic C2/c. The water molecule in 1 is encapsulated in the vacant O4 compartment because of the hydrogen bonding interactions with the ether and phenolate oxygens, resulting in the formation of an inclusion product. The structures of 2-4 consist of the [CuIIL1MII(H2O)3]2+ cation and two mononuclear [CuIIL1] moieties. In the dinuclear [CuIIL1MII(H2O)3]2+ cation, the metal centers are doubly bridged by the two phenolate oxygens. The second metal center, MII (Cu in 2, Co in 3, and Mn in 4), in the [CuIIL1MII(H2O)3]2+ cation is pentacoordinated by the two phenoxo oxygens and three water molecules. Two of these three coordinated water molecules interact, similar to that in 1, with two mononuclear [CuIIL1] moieties, resulting in the formation of the tetrametal [{CuIIL1}2{CuIIL1MII(H2O)3}]2+ system that consists of the cocrystallized dinuclear (one) and mononuclear (two) moieties. Evidently, the cocrystallization observed in 2-4 is related to the tendency of a water molecule to be encapsulated in the vacant O4 compartment of the mononuclear [CuIIL1] species. In the case of 2, there are two independent [CuIIL1CuII(H2O)3]2+ units. The tau ((beta-alpha)/60, where beta and alpha are the largest and second largest bond angles, respectively) values in the pentacoordinated environment of the two copper(II) centers in 2 are 0.04 and 0.37, indicating almost ideal and appreciably distorted square pyramidal geometry, respectively. In contrast, the tau values (0.54 for 3 and 0.49 for 4) indicate that the coordination geometry around the cobalt(II) and manganese(II) centers in 3 and 4 is intermediate between square pyramidal and trigonal bipyramidal. The variable-temperature (2-300 K) magnetic susceptibilities of compounds 2-4 have been measured. The magnetic data have been analyzed in the model of one exchange-coupled dinuclear CuIIMII moiety and two noninteracting CuII centers. In all three cases, the metal ions in the dinuclear core are coupled by a weak antiferromagnetic interaction (J = -17.4 cm-1, -8 cm-1, and -14 cm-1 for 2, 3, and 4, respectively). The observation of a weak interaction has been explained in terms of the structural parameters and symmetry of the magnetic orbitals.

Journal Article↗
PubMed2003

Dynamics of heart rate and sleep stages in normals and patients with sleep apnea.

Sleep is not just the absence of wakefulness but a regulated process with an important restorative function. Based on electroencephalographic recordings and characteristic patterns and waveforms we can distinguish wakefulness and five sleep stages grouped into light sleep, deep sleep, and rapid-eye-movement (REM) sleep. In order to explore the functions of sleep and sleep stages, we investigated the dynamics of sleep stages over the night and of heart-rate variability during the different sleep stages. Sleep stages and intermediate wake states have different distributions of their duration and this allows us to create a model for the temporal sequence of sleep stages and wake states. Heart rate is easily accessed with a high precision by the recording and analysis of the electrocardiogram (ECG). Heart-rate regulation is part of the autonomous nervous system and sympathetic tone is strongly influenced by the sleep stages.

Heart Rate↗
PubMed2001

Estimating disease severity of congenital diaphragmatic hernia in the first 5 minutes of life.

BACKGROUND/PURPOSE: Congenital diaphragmatic hernia (CDH), occurring approximately once in every 2,400 live births, remains a significant cause of perinatal death and morbidity. Risk assessment tools for congenital diaphragmatic hernia derived at single institutions fail to predict outcome at other institutions. Without a generally applicable risk assessment tool it is impossible to determine whether the current variation in outcomes is caused by differences in treatment or to variations in the types of patients treated. The authors report a broadly applicable risk assessment tool for newborns with CDH derived from multiinstitutional data. METHODS: Survival data on 322 consecutive liveborn infants with CDH were collected using data from 71 institutions. Demographic and early treatment results were evaluated by univariate analysis. Items useful in an early stratification system were examined using a multivariate logistic regression analysis. The predictive equation developed was applied to the next series of evaluable patients. RESULTS: A total of 1,054 patients with CDH were evaluated from 1995 to 1999 with an overall survival rate of 64%. For the first 322 patients, factors associated with outcome included birth weight, Apgar scores, gestational age, race, immediate distress, presence of a cardiac anomaly, and prenatal diagnosis. Multivariate analysis showed that birth weight and 5-minute Apgar scores were most useful in a predictive equation. A logistic equation using these 2 variables could separate the next 673 patients into high, intermediate, and low risk of death, and this correlated closely with the actual outcome. CONCLUSION: Stratifying neonates with CDH into broad risk groups should allow better comparison of outcomes data from different centers, reserving novel and high-risk strategies for patients with a high likelihood of dying.

Chi-Square Distribution↗
PubMed2002

Sensitivity of HIV-1 to entry inhibitors correlates with envelope/coreceptor affinity, receptor density, and fusion kinetics.

HIV entry inhibitors include coreceptor antagonists and the fusion inhibitor T-20. T-20 binds the first helical region (HR1) in the gp41 subunit of the viral envelope (Env) protein and prevents conformational changes required for membrane fusion. HR1 appears to become accessible to T-20 after Env binds CD4, whereas coreceptor binding is thought to induce the final conformational changes that lead to membrane fusion. Thus, T-20 binds to a structural intermediate of the fusion process. Primary viruses exhibit considerable variability in T-20 sensitivity, and determinants outside of HR1 can affect sensitivity by unknown mechanisms. We studied chimeric Env proteins containing different V3 loop sequences and found that gp120coreceptor affinity correlated with T-20 and coreceptor antagonist sensitivity, with greater affinity resulting in increased resistance to both classes of entry inhibitors. Enhanced affinity resulted in more rapid fusion kinetics, reducing the time during which Env is sensitive to T-20. Reduced coreceptor expression levels also delayed fusion kinetics and enhanced virus sensitivity to T-20, whereas increased coreceptor levels had the opposite effect. A single amino acid change (K421D) in the bridging sheet region of the primary virus strain YU2 reduced affinity for CCR5 and increased T-20 sensitivity by about 30-fold. Thus, mutations in Env that affect receptor engagement and membrane fusion rates can alter entry inhibitor sensitivity. Because coreceptor expression levels are typically limiting in vivo, individuals who express lower coreceptor levels may respond more favorably to entry inhibitors such as T-20, whose effectiveness we show depends in part on fusion kinetics.

3T3 Cells↗
PubMed1977

Three-dimensional structure of an intact human immunoglobulin.

We have examined the low-resolution structure of a complete human IgG1 using known domain coordinates from crystallographic investigations of immunoglobulin fragment structures. Our results indicate that the Fc portion of this molecule has a structure similar to that of an isolated Fc fragment, with the carbohydrate moiety playing a central role as the principal contact between the CH2 domains. Carbohydrate also forms a large part of the interface between the Fc and Fab regions. The relative orientations of the variable and constant portions of the Fab regions are intermediate between those reported previously, emphasizing the flexibility of the switch region. These data do not support a two-state allosteric model such as has been proposed for antibody effector functions.

Carbohydrates↗
PubMed1997

Incipient species formation in salamanders of the Ensatina complex.

The Ensatina eschscholtzii complex of plethodontid salamanders, a well-known "ring species," is thought to illustrate stages in the speciation process. Early research, based on morphology and coloration, has been extended by the incorporation of studies of protein variation and mitochondrial DNA sequences. The new data show that the complex includes a number of geographically and genetically distinct components that are at or near the species level. The complex is old and apparently has undergone instances of range contraction, isolation, differentiation, and then expansion and secondary contact. While the hypothesis that speciation is retarded by gene flow around the ring is not supported by molecular data, the general biogeographical hypothesis is supported. There is evidence of a north to south range expansion along two axes, with secondary contact and completion of the ring in southern California. Current research targets regions once thought to show primary intergradation, but which molecular markers reveal to be zones of secondary contact. Here emphasis is on the subspecies E. e. xanthoptica, which is involved in four distinct secondary contacts in central California. There is evidence of renewed genetic interactions upon recontact, with greater genetic differentiation within xanthoptica than between it and some of the interacting populations. The complex presents a full array of intermediate conditions between well-marked species and geographically variable populations. Geographically differentiated segments represent a diversity of depths of time of isolation and admixture, reflecting the complicated geomorphological history of California. Ensatina illustrates the continuing difficulty in making taxonomic assignments in complexes studied during species formation.

Animals↗
PubMed1977

Evaluation of control populations: body weight and rate of gain in Japanese quail.

1. Two control populations of quail of different origin were maintained by cyclical mating, one for 4 and the other for 6 generations. Means, heritabilities, genetic and phenotypic correlations of body weights at 1, 14 and 35 d and rates of gain from 1 to 14 d and from 14 to 35 d did not show any trend of change with generation. 2. The variability between generations was greater in the characters having intermediate and lower values of heritability, genetic and phenotypic correlations than in those with higher values.

Animals↗
PubMed1991

The radiosensitivity of human keratinocytes: influence of activated c-H-ras oncogene expression and tumorigenicity.

We have investigated the gamma-ray sensitivity of several activated c-H-ras (EJ) containing clones that have been established after transfection of the spontaneously immortalized non-tumorigenic human keratinocyte cell line HaCaT. The clones were grouped according to their tumorigenic potential after subcutaneous injection into nude mice, and fell into three classes: Class I clones A-4 and I-6 are non-tumorigenic and express very low levels of c-H-ras mRNA and no mutated ras protein (p21); Class II clones I-5 and I-7 grow to large (benign) epidermal cysts, express intermediate to high c-H-ras mRNA and variable levels of mutated ras p21 protein with clone I-5 expressing little and clone I-7 expressing high levels of p21; Class III clones II-3 and II-4 grow to solid squamous cell carcinomas, express high c-H-ras mRNA and high level of mutated p21 ras protein similar to clone I-7. Comparison of the single-hit multitarget or linear-quadratic survival curve parameters, and survival at 2 Gy (S2) indicate that there appears to be no general correlation with either activated c-H-ras expression level or tumorigenic potential, and increased radioresistance.

Animals↗
PubMed2002

The degree of difficulty of embryo transfer is an independent factor for predicting pregnancy.

BACKGROUND: The role of embryo transfer as regards the success of IVF/ICSI treatments is recognized but has not been comprehensively evaluated. In order to determine its importance, the degree of difficulty of 4807 embryo transfers after IVF/ICSI was analysed retrospectively. METHODS: Logistic regression analysis identified the age of the subject, type of treatment (IVF versus ICSI), number of embryos transferred and degree of difficulty of embryo transfer as independent factors predicting pregnancy. The main focus of the study was to evaluate the importance of the difficulty of embryo transfer after taking into account the other confounding variables. RESULTS: Embryo transfer was classified as easy (2821), intermediate (1644) or difficult (342). The transfer was considered difficult if it was time consuming, the catheter met great resistance, there was a need to change the catheter, if sounding or cervical dilatation was needed or if blood was found in any part of the catheter. Easy or intermediate transfers resulted in a 1.7-fold higher pregnancy rate than difficult transfers (P < 0.0001; 95% confidence interval: 1.3-2.2). CONCLUSIONS: This study demonstrates that the degree of difficulty of embryo transfer is an independent factor as regards achieving pregnancy after IVF/ICSI. All efforts should be made to avoid difficult embryo transfers. Physicians should be alert to the factors associated with embryo transfer and should be instructed to use a stepwise approach in difficult transfers.

Catheterization↗
PubMed1977

Disk diffusion susceptibility testing of Nocardia species.

The effectiveness of 13 antimicrobial agents against 51 clinical isolates of Nocardia was determined with use of agar dilutions and disk diffusion method. Amikacin inhibited less than 90% of isolates and, like the other aminoglycosides, showed good correlation between minimal inhibitory concentrations and sizes of zones of inhibition around the disks. Both sulfisoxazole and trimethoprim-sulfamethoxazole were very active, although they required a 2- to 3-log lower inoculum for demonstration of susceptibility. Results with the two sulfa disks were variable, but they did allow distinction between sensitive and intermediate strains. All of the isolates of Nocardia were inhibited by 6.3 microng of minocycline; however, the degree of susceptibility could not be determined by zone diameters. Only two-thirds of these clinical isolates of Nocardia grew rapidly enough to be assayed by either susceptibility method.

Anti-Bacterial Agents↗
PubMed1999

A study of leukaemia in Glasgow in connection with chromium-contaminated land.

BACKGROUND: In 1991, soil pollution was found around the site of a former chromium-processing factory in Glasgow, Scotland. Levels of chromium in soil were above limits considered as safe, although a risk assessment concluded that population exposure was likely to be below occupational levels. As an excess incidence of leukaemia has been suspected in the area, it was decided to investigate a possible relationship between the pollutant and the illness. METHODS: The ensuing study was descriptive-geographical. In the absence of better data, levels of exposure were assumed to decrease with distance from the centre of the polluted area. Leukaemia and population figures were obtained for each of nine concentric rings by aggregation of data available at the Enumeration District level. The null study hypothesis was that relative risk (as measured by Poisson regression) would not follow a definite trend with distance from the centre. Sex, age and levels of deprivation were taken into account. RESULTS: Relative risks by variables other than distance followed previously known patterns for leukaemia. No evident pattern by distance was found. After regroupings inside the variables, a significant excess of leukaemia was found for intermediate distances from the pollutant. CONCLUSIONS: No evidence was found of a possible relationship between soil pollution by chromium and leukaemia in the general population. Nonetheless, the excess noticed by the study warrants further research.

Adolescent↗