The impact of stress and anxiety on the pressure pain threshold of myofascial pain patients

The purpose of this study was to evaluate the influence of stress and anxiety on the pressure pain threshold (PPT) of masticatory muscles and on the subjective pain report. Forty-five women, students, with mean age of 19.75 years, were divided into two groups: group 1:29 presenting with masticatory myofascial pain (MFP), according to the Research Diagnostic Criteria for Temporomandibular Disorders and group 2: 16 asymptomatic controls. An electronic algometer registered the pain thresholds on four different occasions throughout the academic year. To measure levels of stress, anxiety and pain, the Beck Anxiety Inventory, Lipp Stress Symptoms Inventory and Visual Analog Scale (VAS) were used. Three-way anova and Tukey`s tests were used to verify differences in PPT between groups, times and sites. Levels of anxiety and VAS were compared using Mann-Whitney test, while Friedman`s test was used for the within-groups comparison at different times (T1 to T4). The chi-squared and Cochran tests were performed to compare groups for the proportion of subjects with stress (alpha = 0.05). Differences in PPT recordings between time (P = 0.001) and sites (P < 0.001) were detected. Higher levels of anxiety and lower PPT figures were detected at T2 (academic examination) (P = 0.001). There was no difference between groups for anxiety and stress at any time (P > 0.05). The MFP group also has shown significant increase of VAS at the time of academic examination (P < 0.001). External stressors such as academic examinations have a potential impact on masticatory muscle tenderness, regardless of the presence of a previous condition such as masticatory myofascial pain.

Tyrosine residue 271 of the norepinephrine transporter is an important determinant of its pharmacology

The aim was to examine the functional importance in the norepinephrine transporter (NET) of (i) the phenylalanine residue at position 531 in transmembrane domain (TMD) 11 by mutating it to tyrosine in the rat (rF531Y) and human (hF531Y) NETs and (ii) the highly conserved tyrosine residues at positions 249 in TMD 4 of human NET (hNET) (mutated to alanine: hY249A) and 271 in TMD 5, by mutating to alanine (hY271A), phenylalanine (hY271F) and histidine (hY271H). The effects of the mutations on NET function were for uptake of the substrates, examined by expressing the mutant and wildtype NETs in COS-7 cells and measuring the K-m and V-max for uptake of the substrates, [H-3]norepinephrine, [H-3]MPP+ and [H-3]dopamine, the K-D and B-max for [H-3]nisoxetine binding and the K-i of the inhibitors, nisoxetine, desipramine and cocaine, for inhibition of [H-3]norepinephrine uptake. The K-m values of the substrates were lower for the mutants at amino acid 271 than hNET and unaffected for the other mutants, and each mutant had a significantly lower than NET for substrate uptake. The mutations at position 271 caused an increase in the K-i or K-D values of nisoxetine, desipramine and cocaine, but there were no effects for the other mutations. Hence, the 271 tyrosine residue in TMD 5 is an important determinant of NET function, with the mutants showing an increase in the apparent affinities of substrates and a decrease in the apparent affinities of inhibitors, but the 249 tyrosine and 531 phenylalanine residues do not have a major role in determining NET function. (C) 2001 Elsevier Science B.V. All rights reserved.

'Neighbourhood' size, dispersal and density estimates in the prickly forest skink (Gnypetoscincus queenslandiae) using individual genetic and demographic methods

Dispersal, or the amount of dispersion between an individual's birthplace and that of its offspring, is of great importance in population biology, behavioural ecology and conservation, however, obtaining direct estimates from field data on natural populations can be problematic. The prickly forest skink, Gnypetoscincus queenslandiae, is a rainforest endemic skink from the wet tropics of Australia. Because of its log-dwelling habits and lack of definite nesting sites, a demographic estimate of dispersal distance is difficult to obtain. Neighbourhood size, defined as 4 piD sigma (2) (where D is the population density and sigma (2) the mean axial squared parent-offspring dispersal rate), dispersal and density were estimated directly and indirectly for this species using mark-recapture and microsatellite data, respectively, on lizards captured at a local geographical scale of 3 ha. Mark-recapture data gave a dispersal rate of 843 m(2)/generation (assuming a generation time of 6.5 years), a time-scaled density of 13 635 individuals * generation/km(2) and, hence, a neighbourhood size of 144 individuals. A genetic method based on the multilocus (10 loci) microsatellite genotypes of individuals and their geographical location indicated that there is a significant isolation by distance pattern, and gave a neighbourhood size of 69 individuals, with a 95% confidence interval between 48 and 184. This translates into a dispersal rate of 404 m(2)/generation when using the mark-recapture density estimation, or an estimate of time-scaled population density of 6520 individuals * generation/km(2) when using the mark-recapture dispersal rate estimate. The relationship between the two categories of neighbourhood size, dispersal and density estimates and reasons for any disparities are discussed.

Familial corticosteroid-binding globulin deficiency due to a noval null mutation: Association with fatigue and relative hypotension

Corticosteroid-binding globulin is a 383-amino acid glycoprotein that serves a hormone transport role and may have functions related to the stress response and inflammation. We describe a 39-member Italian-Australian family with a novel complete loss of function (null) mutation of the corticosteroid-binding globulin gene. A second, previously described, mutation (Lyon) segregated independently in the same kindred. The novel exon 2 mutation led to a premature termination codon corresponding to residue -12 of the procorticosteroid-binding globulin molecule (c.121G->A). Among 32 family members there were 3 null homozygotes, 19 null heterozygotes, 2 compound heterozygotes, 3 Lyon heterozygotes, and 5 individuals without corticosteroid-binding globulin mutations. Plasma immunoreactive corticosteroid-binding globulin was undetectable in null homozygotes, and mean corticosteroid-binding globulin levels were reduced by approximately 50% at 18.7 ± 1.3 µg/ml (reference range, 30–52 µg/ml) in null heterozygotes. Morning total plasma cortisol levels were less than 1.8 µg/dl in homozygotes and were positively correlated to the plasma corticosteroid-binding globulin level in heterozygotes. Homozygotes and heterozygote null mutation subjects had a high prevalence of hypotension and fatigue. Among 19 adults with the null mutation, the systolic blood pressure z-score was 12.1 ± 3.5; 11 of 19 subjects (54%) had a systolic blood pressure below the third percentile. The mean diastolic blood pressure z-score was 18.1 ± 3.4; 8 of 19 subjects (42%) had a diastolic blood pressure z-score below 10. Idiopathic chronic fatigue was present in 12 of 14 adult null heterozygote subjects (86%) and in 2 of 3 null homozygotes. Five cases met the Centers for Disease Control criteria for chronic fatigue syndrome. Fatigue questionnaires revealed scores of 25.1 ± 2.5 in 18 adults with the mutation vs. 4.2 ± 1.5 in 23 healthy controls (P < 0.0001). Compound heterozygosity for both mutations resulted in plasma cortisol levels comparable to those in null homozygotes. Abnormal corticosteroid-binding globulin concentrations or binding affinity may lead to the misdiagnosis of isolated ACTH deficiency. The mechanism of the association between fatigue and relative hypotension is not established by these studies. As idiopathic fatigue disorders are associated with relatively low plasma cortisol, abnormalities of corticosteroid-binding globulin may be pathogenic.

Traffic and residue cover effects on infiltration

Wheel traffic can lead to compaction and degradation of soil physical properties. This study, as part of a study of controlled traffic farming, assessed the impact of compaction from wheel traffic on soil that had not been trafficked for 5 years. A tractor of 40 kN rear axle weight was used to apply traffic at varying wheelslip on a clay soil with varying residue cover to simulate effects of traffic typical of grain production operations in the northern Australian grain belt. A rainfall simulator was used to determine infiltration characteristics. Wheel traffic significantly reduced time to ponding, steady infiltration rate, and total infiltration compared with non-wheeled soil, with or without residue cover. Non-wheeled soil had 4-5 times greater steady infiltration rate than wheeled soil, irrespective of residue cover. Wheelslip greater than 10% further reduced steady infiltration rate and total infiltration compared with that measured for self-propulsion wheeling (3% wheelslip) under residue-protected conditions. Where there was no compaction from wheel traffic, residue cover had a greater effect on infiltration capacity, with steady infiltration rate increasing proportionally with residue cover (R-2 = 0.98). Residue cover, however, had much less effect on infiltration when wheeling was imposed. These results demonstrated that the infiltration rate for the non-wheeled soil under a controlled traffic zero-till system was similar to that of virgin soil. However, when the soil was wheeled by a medium tractor wheel, infiltration rate was reduced to that of long-term cropped soil. These results suggest that wheel traffic, rather than tillage and cropping, might be the major factor governing infiltration. The exclusion of wheel traffic under a controlled traffic farming system, combined with conservation tillage, provides a way to enhance the sustainability of cropping this soil for improved infiltration, increased plant-available water, and reduced runoff-driven soil erosion.

A single beta subunit M2 domain residue controls the picrotoxin sensitivity of alpha beta heteromeric glycine receptor chloride channels

This study investigated the residues responsible for the reduced picrotoxin sensitivity of the alpha beta heteromeric glycine receptor relative to the alpha homomeric receptor. By analogy with structurally related receptors, the beta subunit M2 domain residues P278 and F282 were considered the most likely candidates for mediating this effect. These residues align with G254 and T258 of the alpha subunit. The T258A, T258C and T258F mutations dramatically reduced the picrotoxin sensitivity of the alpha homomeric receptor. Furthermore, the converse F282T mutation in the beta subunit increased the picrotoxin sensitivity of the alpha beta heteromeric receptor. The P278G mutation in the beta subunit did not affect the picrotoxin sensitivity of the alpha beta heteromer. Thus, a ring of five threonines at the M2 domain depth corresponding to alpha subunit T258 is specifically required for picrotoxin sensitivity. Mutations to alpha subunit T258 also profoundly influenced the apparent glycine affinity. A substituted cysteine accessibility analysis revealed that the T258C sidechain increases its pore exposure in the channel open state. This provides further evidence for an allosteric mechanism of picrotoxin inhibition, but renders it unlikely that picrotoxin las an allosterically acting 'competitive' antagonist) binds to this residue.

An aggregate sales model for consumer durables incorporating a time-varying mean replacement age

Forecasting category or industry sales is a vital component of a company's planning and control activities. Sales for most mature durable product categories are dominated by replacement purchases. Previous sales models which explicitly incorporate a component of sales due to replacement assume there is an age distribution for replacements of existing units which remains constant over time. However, there is evidence that changes in factors such as product reliability/durability, price, repair costs, scrapping values, styling and economic conditions will result in changes in the mean replacement age of units. This paper develops a model for such time-varying replacement behaviour and empirically tests it in the Australian automotive industry. Both longitudinal census data and the empirical analysis of the replacement sales model confirm that there has been a substantial increase in the average aggregate replacement age for motor vehicles over the past 20 years. Further, much of this variation could be explained by real price increases and a linear temporal trend. Consequently, the time-varying model significantly outperformed previous models both in terms of fitting and forecasting the sales data. Copyright (C) 2001 John Wiley & Sons, Ltd.

Inferring genome trees by using a filter to eliminate phylogenetically discordant sequences and a distance matrix based on mean normalized BLASTP scores

Darwin's paradigm holds that the diversity of present-day organisms has arisen via a process of genetic descent with modification, as on a bifurcating tree. Evidence is accumulating that genes are sometimes transferred not along lineages but rather across lineages. To the extent that this is so, Darwin's paradigm can apply only imperfectly to genomes, potentially complicating or perhaps undermining attempts to reconstruct historical relationships among genomes (i.e., a genome tree). Whether most genes in a genome have arisen via treelike (vertical) descent or by lateral transfer across lineages can be tested if enough complete genome sequences are used. We define a phylogenetically discordant sequence (PDS) as an open reading frame (ORF) that exhibits patterns of similarity relationships statistically distinguishable from those of most other ORFs in the same genome. PDSs represent between 6.0 and 16.8% (mean, 10.8%) of the analyzable ORFs in the genomes of 28 bacteria, eight archaea, and one eukaryote (Saccharomyces cerevisiae). In this study we developed and assessed a distance-based approach, based on mean pairwise sequence similarity, for generating genome trees. Exclusion of PDSs improved bootstrap support for basal nodes but altered few topological features, indicating that there is little systematic bias among PDSs. Many but not all features of the genome tree from which PDSs were excluded are consistent with the 16S rRNA tree.

A cassette ligation strategy with thioether replacement of three Gly-Gly peptide bonds: Total chemical synthesis of the 101 residue protein early pregnancy factor [psi(CH2S)(28-29,56-57,76-77)]

The 101 residue protein early pregnancy factor (EPF), also known as human chaperonin 10, was synthesized from four functionalized, but unprotected, peptide segments by a sequential thioether ligation strategy. The approach exploits the differential reactivity of a peptide-NHCH2CH2SH thiolate with XCH2CO-peptides, where X = Cl or I/Br. Initial model studies with short functionalized (but unprotected) peptides showed a significantly faster reaction of a peptide-NHCH2CH2SH thiolate with a BrCH2CO-peptide than with a CICH2CO-peptide, where thiolate displacement of the halide leads to chemoselective formation of a thioether surrogate for the Gly-Gly peptide bond. This rate difference was used as the basis of a novel sequential ligation approach to the synthesis of large polypeptide chains. Thus, ligation of a model bifunctional N-alpha-chloroacetyl, C-terminal thiolated peptide with a second N-alpha-bromoacetyl peptide demonstrated chemoselective bromide displacement by the thiol group. Further investigations showed that the relatively unreactive N-alpha-chloroacetyl peptides could be activated by halide exchange using saturated KI solutions to yield the highly reactive No-iodoacetyl peptides. These findings were used to formulate a sequential thioether ligation strategy for the synthesis of EPF, a 101 amino acid protein containing three Gly-Gly sites approximately equidistantly spaced within the peptide chain. Four peptide segments or cassettes comprising the EPF protein sequence (BrAc-[EPF 78-101] 12, ClAc-[EPF 58-75]-[NHCH2CH2SH] 13, ClAc-[EPF 30-55]-[NHCH2CH2SH] 14, and Ac-[EPF 1-27]-[NHCH2CH2SH] 15) of EPF were synthesized in high yield and purity using Boc SPPS chemistry. In the stepwise sequential ligation strategy, reaction of peptides 12 and 13 was followed by conversion of the N-terminal chloroacetyl functional group to an iodoacetyl, thus activating the product peptide for further ligation with peptide 14. The process of ligation followed by iodoacetyl activation was repeated to yield an analogue of EPF (EPF psi(CH2S)(28-29,56-57,76-77)) 19 in 19% overall yield.

Fate of N-15-labelled nitrate in a wet summer under different residue management regimes in young hoop pine plantations

A field study was conducted to investigate the fate of N-15-labelled nitrate applied at 20 kg N ha(-1) in a wet summer to microplots installed in areas under different residue management regimes in second-rotation hoop pine (Araucaria cunninghamii) plantations aged 1-3 years in south-east Queensland, Australia. PVC microplots of 235 mm diameter and 300 mm long were driven into 250 mm soil. There were three replications of each of eight treatments. These were areas just under and between 1-year-old windrows (ca. 2-3 m in width) of harvesting residues spaced 15 m apart, and with and without incorporated foliage residues (20 t DM ha(-1)); the areas just under and between 2- or 3-year-old windrows spaced 10 m apart. Only 7-29% of the added N-15 was recovered from the top 750 mm of the soil profile with the leaching loss estimated to be 70-86% over the 34-day period. The N-15 loss via denitrification was 3.7-6.3% by directly measuring the N-15 gases emitted. The microplots with the incorporated residues at the 1-year-old site had the highest N-15 loss (6.3%) as compared with the other treatments. The N-15 mass balance method together with the use of bromide (Br) tracer applied at 100 kg Br ha(-1) failed to obtain a reliable estimate of the denitrification loss. The microplots at the 1-year-old site had higher N-15 immobilisation rate (7.5-24.7%) compared with those at 2- and 3-year-old sites (2.1-3.6%). Incorporating the residues resulted in an increase in N-15 immobilisation rate (24.5-24.7%) compared with the control without the incorporated residues (8.4-14.3%). These findings suggest that climatic conditions played important roles in controlling the N-15 transformations in the wet summer season and that the residue management regimes could also significantly influence the N-15 transformations. Most of the N-15 loss occurred through leaching, but a considerable amount of the N-15 was lost through denitrification. Bromide proved to be an unsuitable tracer for monitoring the N-15 leaching and movement under the wet summer conditions. (C) 2002 Elsevier Science B.V. All rights reserved.