IMMPACT-recommended outcome measures and tools of assessment in burning mouth syndrome RCTs: an international Delphi survey protocol.

BACKGROUND: A core outcome set (COS) represents the agreed minimum set of domains and measurement instruments that should be measured and reported in any clinical trial for a given condition. In BMS randomized controlled trials (RCTs), the outcomes identified in the existing literature regarding the efficacy of therapeutic interventions are numerous and diverse. Although the standardized IMMPACT core outcome domains has been developed for measurement of outcomes in chronic pain RCTs, no BMS-specific COS have been adopted and validated. With the evolving landscape of BMS management end points and the development of new therapies, a consensus on a COS for use in future BMS trials is paramount to reduce heterogeneity in outcome reporting. The aim of this study was to reach a consensus for adopting the standardized Initiative on Methods, Measurement, and Pain Assessment in Clinical Trials (IMMPACT) outcome domains, and their tools of assessment, for burning mouth syndrome (BMS) clinical trials and clinical practice. METHODS: A BMS-specific COS will be developed using the method recommended by the Core Outcome Measures in Effective Trials (COMET) initiative (Registration: http://www.comet-initiative.org/studies/details/1357 ). Selection of questionnaire outcome measures was informed by the IMMPACT consensus and previous systematic review of RCTs in BMS conducted by the consortium. An international group of clinicians and researchers will be invited to participate in several rounds of a Delphi survey. A consensus meeting will be held with the objective of ratifying the outcomes for inclusion in the COS. A finalized COS explanatory document will be drafted, including all outcomes and measurements as determined by the Delphi rounds and consensus meeting. DISCUSSION: A COS for the management of BMS will improve the quality of future RCTs, reduce outcome reporting heterogeneity, and facilitate more vigorous data synthesis of management interventions for systematic reviews and meta-analysis. This would ensure enhanced quality evidence for clinical management of the condition.

Rapid Communication: 16S ribosomal ribonucleic acid characterization of liver abscesses in feedlot cattle from three states in the United States.

Liver abscesses are a major economic burden to beef producers. Although a few causative organisms have been cultured from purulent material, the full polymicrobial diversity of liver abscesses has not been reported. The objective of this study was to characterize purulent material collected from liver abscess in beef cattle produced in different production systems in 3 cattle producing states in the United States using 16S rRNA gene sequencing. Differences between purulent material microbial communities among geographic region of feeding and application of a common antimicrobial were also investigated. Cattle included in the study were fed in California (dairy type) and Colorado and Texas (both beef type). Liver abscesses from a cross section of feedlots, geographic areas, and tylosin phosphate-administered groups were collected at harvest; DNA from 34 liver abscess samples was extracted; and the V4 region of the 16S rRNA gene was amplified and sequenced. Sequences were classified into 5 phyla, 13 classes, and 17 orders in the domain Bacteria. The phyla identified included Bacteroidetes (35.2% of reads), Proteobacteria (28.6%), Fusobacteria (18.2%), Firmicutes (12.4%), and Actinobacteria (5.5%). Sequences matching the genera and , which have previously been identified as causative agents in liver abscesses, were both present in the abscess bacterial communities at a relative abundance of 15.1 and 3.2%, respectively, of the overall relative abundance. Furthermore, 3 of the most common phyla were Gram-negative bacteria. An analysis-of-similarities test was conducted on Euclidean distances to assess differences between cattle treated and not treated with tylosin as well as to assess differences between geographic regions. Geographical region and treatment with tylosin did affect the microbiome ( = 0.002 and = 0.026 respectively); however, a more robust sample scheme is needed to explore these differences. To our knowledge, this is the first publication describing the complex community of liver purulent material using next generation sequencing in cattle. These data provide a framework for research on a more targeted approach to liver abscess prevention and treatment.

Smoking and Posttraumatic Stress Disorder Symptomatology in Orofacial Pain.

To explore the impact of interactions between smoking and symptoms of posttraumatic stress disorder (PTSD) on pain intensity, psychological distress, and pain-related functioning in patients with orofacial pain, a retrospective review was conducted of data obtained during evaluations of 610 new patients with a temporomandibular disorder who also reported a history of a traumatic event. Pain-related outcomes included measures of pain intensity, psychological distress, and pain-related functioning. Main effects of smoking status and PTSD symptom severity on pain-related outcomes were evaluated with linear regression analyses. Further analyses tested interactions between smoking status and PTSD symptom severity on pain-related outcomes. PTSD symptom severity and smoking predicted worse pain-related outcomes. Interaction analyses between PTSD symptom severity and smoking status revealed that smoking attenuated the impact of PTSD symptom severity on affective distress, although this effect was not found at high levels of PTSD symptom severity. No other significant interactions were found, but the present results identifying smoking as an ineffective coping mechanism and the likely role of inaccurate outcome expectancies support the importance of smoking cessation efforts in patients with orofacial pain. Smoking is a maladaptive mechanism for coping with pain that carries significant health- and pain-related risks while failing to fulfill smokers' expectations of affect regulation, particularly among persons with orofacial pain who also have high levels of PTSD symptom severity. Addressing smoking cessation is a critical component of comprehensive treatment. Further research is needed to develop more effective ways to help patients with pain and/or PTSD to replace smoking with more effective coping strategies.

Trigeminal Inflammatory Compression (TIC) injury induces chronic facial pain and susceptibility to anxiety-related behaviors.

Our laboratory previously developed a novel neuropathic and inflammatory facial pain model for mice referred to as the Trigeminal Inflammatory Compression (TIC) model. Rather than inducing whole nerve ischemia and neuronal loss, this injury induces only slight peripheral nerve demyelination triggering long-term mechanical allodynia and cold hypersensitivity on the ipsilateral whisker pad. The aim of the present study is to further characterize the phenotype of the TIC injury model using specific behavioral assays (i.e. light-dark box, open field exploratory activity, and elevated plus maze) to explore pain- and anxiety-like behaviors associated with this model. Our findings determined that the TIC injury produces hypersensitivity 100% of the time after surgery that persists at least 21 weeks post injury (until the animals are euthanized). Three receptive field sensitivity pattern variations in mice with TIC injury are specified. Animals with TIC injury begin displaying anxiety-like behavior in the light-dark box preference and open field exploratory tests at week eight post injury as compared to sham and naïve animals. Panic anxiety-like behavior was shown in the elevated plus maze in mice with TIC injury if the test was preceded with acoustic startle. Thus, in addition to mechanical and cold hypersensitivity, the present study identified significant anxiety-like behaviors in mice with TIC injury resembling the clinical symptomatology and psychosocial impairments of patients with chronic facial pain. Overall, the TIC injury model's chronicity, reproducibility, and reliability in producing pain- and anxiety-like behaviors demonstrate its usefulness as a chronic neuropathic facial pain model.

Soil carbon sequestration resulting from long-term application of biosolids for land reclamation.

Investigations on the impact of application of biosolids for land reclamation on C sequestration in soil were conducted at Fulton County, Illinois, where 41 fields (3.6-66 ha) received biosolids at a cumulative loading rate from 455 to 1654 dry Mg ha(-1) for 8 to 23 yr in rotation from 1972 to 2004. The fields were cropped with corn, wheat, and sorghum and also with soybean and grass or fallowed. Soil organic carbon (SOC) increased rapidly with the application of biosolids, whereas it fluctuated slightly in fertilizer controls. The peak SOC in the 0- to 15-cm depth of biosolids-amended fields ranged from 4 to 7% and was greater at higher rates of biosolids. In fields where biosolids application ceased for 22 yr, SOC was still much higher than the initial levels. Over the 34-yr reclamation, the mean net soil C sequestration was 1.73 (0.54-3.05) Mg C ha(-1) yr(-1) in biosolids-amended fields as compared with -0.07 to 0.17 Mg C ha(-1) yr(-1) in fertilizer controls, demonstrating a high potential of soil C sequestration by the land application of biosolids. Soil C sequestration was significantly correlated with the biosolids application rate, and the equation can be expressed as y = 0.064x - 0.11, in which y is the annual net soil C sequestration (Mg C ha(-1) yr(-1)), and x is annual biosolids application in dry weight (Mg ha(-1) yr(-1)). Our results indicate that biosolids applications can turn Midwest Corn Belt soils from current C-neutral to C-sink. A method for calculating SOC stock under conditions in which surface soil layer depth and mass changes is also described.

Effect of long-term application of biosolids for land reclamation on surface water chemistry.

Biosolids are known to have a potential to restore degraded land, but the long-term impacts of this practice on the environment, including water quality, still need to be evaluated. The surface water chemistry (NO3-, NH4+, and total P, Cd, Cu, and Hg) was monitored for 31 yr from 1972 to 2002 in a 6000-ha watershed at Fulton County, Illinois, where the Metropolitan Water Reclamation District of Greater Chicago was restoring the productivity of strip-mined land using biosolids. The mean cumulative loading rates during the past 31 yr were 875 dry Mg ha(-1) for 1120-ha fields in the biosolids-amended watershed and 4.3 dry Mg ha(-1) for the 670-ha fields in the control watershed. Biosolids were injected into mine spoil fields as liquid fertilizer from 1972 to 1985, and incorporated as dewatered cake from 1980 to 1996 and air-dried solids from 1987 to 2002. The mean annual loadings of nutrients and trace elements from biosolids in 1 ha were 735 kg N, 530 kg P, 4.5 kg Cd, 30.7 kg Cu, and 0.11 kg Hg in the fields of the biosolids-amended watershed, and negligible in the fields of the control watershed. Sampling of surface water was conducted monthly in the 1970s, and three times per year in the 1980s and 1990s. The water samples were collected from 12 reservoirs and 2 creeks receiving drainage from the fields in the control watershed, and 8 reservoirs and 4 creeks associated with the fields in the biosolids-amended watershed for the analysis of NO3- -N (including NO2- N), NH4+-N, and total P, Cd, Cu, and Hg. Compared to the control (0.18 mg L(-1)), surface water NO3- -N in the biosolids-amended watershed (2.23 mg L(-1)) was consistently higher; however, it was still below the Illinois limit of 10 mg L(-1) for public and food-processing water supplies. Biosolids applications had a significant effect on mean concentrations of ammonium N (0.11 mg L(-1) for control and 0.24 mg L(-1) for biosolids) and total P (0.10 mg L(-1) for control and 0.16 mg L(-1) for biosolids) in surface water. Application of biosolids did not increase the concentrations of Cd and Hg in surface water. The elevation of Cu in surface water with biosolids application only occurred in some years of the first decade, when land-applied sludges contained high concentrations of trace metals, including Cu. In fact, following the promulgation of 40 CFR Part 503, the concentrations of all three metals fell below the method detection level (MDL) in surface water for nearly all samplings. Nitrate in the surface water tends to be higher in spring, and ammonium, total P, and total Hg in summer and fall. Mean nitrate, ammonium, and total phosphorus concentrations were found to be greater in creeks than reservoirs. The results indicate that application of biosolids for land reclamation at high loading rates from 1972 to 2002, with adequate runoff and soil erosion control, had only a minor impact on surface water quality.

Psychological and sleep quality differences between chronic daily headache and temporomandibular disorders patients.

The aim of this study was to investigate whether chronic daily headache (CDH) and temporomandibular disorders (TMD) patients present with different psychological and sleep quality characteristics. Sixty-seven patients diagnosed with CDH, according to classification criteria from Silberstein et al., were matched by age and sex with 67 patients who had a primary diagnosis of myofascial pain (MP) and 67 patients with a primary diagnosis of TMJ intracapsular pain (IC) according to the Research Diagnostic Criteria for TMD. The CDH group was comprised of three mutually exclusive diagnostic groups: chronic migraine (n = 35); chronic tension-type headache (n = 26); 'other CDH' (n = 6). All patients completed a battery of psychological and sleep quality questionnaires. All CDH subgroups showed similar psychological and sleep quality profiles. Pain intensity and duration were controlled in the multivariate analyses (Mancova) by treating them as covariates. The CDH and MP groups revealed higher levels of psychological distress than the IC group on most psychological domains. The MP group also revealed numerically higher levels of psychological distress in most psychological domains than the CDH group, although these differences were generally not significant. We did not find significant differences between the three groups on post traumatic stress symptoms either. Sleep quality was significantly worse in the MP group than in the CDH and IC groups. These results are discussed in the context of multimodal patient evaluation and treatments that are often necessary for successful clinical management.

Regulation of anchoring of the RIIalpha regulatory subunit of PKA to AKAP95 by threonine phosphorylation of RIIalpha: implications for chromosome dynamics at mitosis.

CDK1 phosphorylates the A-kinase regulatory subunit RIIalpha on threonine 54 (T54) at mitosis, an event proposed to alter the subcellular localization of RIIalpha. Using an RIIalpha-deficient leukemic cell line (Reh) and stably transfected Reh cell clones expressing wild-type RIIalpha or an RIIalpha(T54E) mutant, we show that RIIalpha associates with chromatin-bound A-kinase anchoring protein AKAP95 at mitosis and that this interaction involves phosphorylation of RIIalpha on T54. During interphase, both RIIalpha and RIIalpha(T54E) exhibit a centrosome-Golgi localization, whereas AKAP95 is intranuclear. At mitosis and in a mitotic extract, most RIIalpha, but not RIIalpha(T54E), co-fractionates with chromatin, onto which it associates with AKAP95. This correlates with T54 phosphorylation of RIIalpha. Disrupting AKAP95-RIIalpha anchoring or depleting RIIalpha from the mitotic extract promotes premature chromatin decondensation. In a nuclear reconstitution assay that mimics mitotic nuclear reformation, RIIalpha is threonine dephosphorylated and dissociates from AKAP95 prior to assembly of nuclear membranes. Lastly, the Reh cell line exhibits premature chromatin decondensation in vitro, which can be rescued by addition of wild-type RIIalpha or an RIIalpha(T54D) mutant, but not RIIalpha(T54E, A, L or V) mutants. Our results suggest that CDK1-mediated T54 phosphorylation of RIIalpha constitutes a molecular switch controlling anchoring of RIIalpha to chromatin-bound AKAP95, where the PKA-AKAP95 complex participates in remodeling chromatin during mitosis.

CDK1-mediated phosphorylation of the RIIalpha regulatory subunit of PKA works as a molecular switch that promotes dissociation of RIIalpha from centrosomes at mitosis.

Protein kinase A regulatory subunit RIIalpha is tightly bound to centrosomal structures during interphase through interaction with the A-kinase anchoring protein AKAP450, but dissociates and redistributes from centrosomes at mitosis. The cyclin B-p34(cdc2) kinase (CDK1) has been shown to phosphorylate RIIalpha on T54 and this has been proposed to alter the subcellular localization of RIIalpha. We have made stable transfectants from an RIIalpha-deficient leukemia cell line (Reh) that expresses either wild-type or mutant RIIalpha (RIIalpha(T54E)). When expressed, RIIalpha detaches from centrosomes at mitosis and dissociates from its centrosomal location in purified nucleus-centrosome complexes by incubation with CDK1 in vitro. By contrast, centrosomal RIIalpha(T54E) is not redistributed at mitosis, remains mostly associated with centrosomes during all phases of the cell cycle and cannot be solubilized by CDK1 in vitro. Furthermore, RIIalpha is solubilized from particular cell fractions and changes affinity for AKAP450 in the presence of CDK1. D and V mutations of T54 also reduce affinity for the N-terminal RII-binding domain of AKAP450, whereas small neutral residues do not change affinity detected by surface plasmon resonance. In addition, only RIIalpha(T54E) interacts with AKAP450 in a RIPA-soluble extract from mitotic cells. Finally, microtubule repolymerization from mitotic centrosomes of the RIIalpha(T54E) transfectant is poorer and occurs at a lower frequency than that of RIIalpha transfectants. Our results suggest that T54 phosphorylation of RIIalpha by CDK1 might serve to regulate the centrosomal association of PKA during the cell cycle.

Social constraints, cognitive processing, and adjustment to breast cancer.

This cross-sectional study of 70 breast cancer survivors examined relationships among social constraints, behavioral and self-report indicators of cognitive processing, depression, and well-being. On the basis of a social-cognitive processing (SCP) model, it was predicted that social constraints would inhibit cognitive processing of the cancer experience, leading to poorer adjustment. Constraints were positively associated with intrusions, avoidance, and linguistic uncertainty in cancer narratives. Greater uncertainty, intrusions, and avoidance, as well as less talking about cancer were associated with greater depression and less well-being. Intrusions partially mediated the positive constraints-depression relationship. Talking about cancer partially mediated the inverse avoidance-well-being relationship. Findings support the SCP model and the importance of using behavioral indicators of cognitive processing to predict positive and negative psychosocial outcomes of cancer.

Posttraumatic growth following breast cancer: a controlled comparison study.

Cancer may be viewed as a psychosocial transition with the potential for positive and negative outcomes. This cross-sectional study (a) compared breast cancer (BC) survivors' (n = 70) self-reports of depression, well-being, and posttraumatic growth with those of age- and education-matched healthy comparison women (n = 70) and (b) identified correlates of posttraumatic growth among BC survivors. Groups did not differ in depression or well-being, but the BC group showed a pattern of greater posttraumatic growth, particularly in relating to others, appreciation of life, and spiritual change. BC participants' posttraumatic growth was unrelated to distress or well-being but was positively associated with perceived life-threat, prior talking about breast cancer, income, and time since diagnosis. Research that has focused solely on detection of distress and its correlates may paint an incomplete and potentially misleading picture of adjustment to cancer.

Binding of PKA-RIIalpha to the Adenovirus E1A12S oncoprotein correlates with its nuclear translocation and an increase in PKA-dependent promoter activity.

The adenovirus type 12 (Ad12) E1A12S oncoprotein utilizes the cAMP/protein kinase A (PKA) signal transduction pathway to activate expression of the viral E2 gene, the products of which are essential for viral replication. A central unsolved question is, however, whether E1A12S interacts directly with PKA in the process of promoter activation. We show here that E1A12S binds to the regulatory subunits (R) of PKA in vitro and in vivo. Interaction depends on the N-terminus and the conserved region 1 (CR1) of E1A12S. Both domains are also essential for the activation of viral E2 gene expression. Infection of cells with Ad12 leads to the cellular redistribution of RIIalpha from the cytoplasm into the nucleus. Furthermore, RIIalpha is also located in the nucleus of cells transformed by E1 of Ad12 and transient expression of E1A12S leads to the redistribution of RIIalpha into the nucleus in a N-terminus- and CR1-dependent manner. Cotransfection of E1A12S with RIIalpha results in strong activation of the E2 promoter. Based on these results we conclude that E1A12S functions as a viral A-kinase anchoring protein redistributing RIIalpha from the cytoplasm into the nucleus where it is involved in E1A12S-mediated activation of the E2 promoter.

The role of anger and depression in recurrent headache.

Individuals with tension-type headache report significant anger, depression, anxiety, and stressors. However, it is not clear to what extent these variables are interrelated. The objective of the present study was to explore the role of anger in headaches, and to examine its relationship to anxiety, depression, and daily life stressors. Participants were 65 young adult women who suffered from recurrent headaches. The sample was obtained in a large-scale screening of young adult women using the Headache Symptoms List to identify those with recurrent headache. Those individuals reporting headaches completed a battery of assessment measures that included the State-Trait Anger Expression Inventory, the Mood and Anxiety Symptoms Questionnaire, and the Hassles Scale. Results revealed a significant relationship between anger suppression and depression (r = 0.40, P<.01), as well as anger expression and anxiety (r = 0.41, P<.01) for those with headache. The use of the Mood and Anxiety Symptoms Questionnaire allowed for the separate analysis of general distress symptoms and symptoms more specific to anxiety and depression. Results indicated that those with headache experience more general, nonspecific distress rather than symptoms indicative of anxiety and depression (P<.01). In addition, the mixed headache group scored high on both general (mean, 28.96) and specific measures of depression (mean, 65.76) and on anger suppression (mean, 20.12), suggesting that they might experience more psychological distress than those with tension-type headache. The present results indicate the need to distinguish the unique dimensions of anxiety and depression that should be assessed in the population with recurrent headache.

Physical self-regulation training for the management of temporomandibular disorders.

AIMS: To evaluate the long-term effectiveness of a brief skills training program for the management of chronic facial muscle pain. This program of physical self-regulation (PSR) involved primarily training in breathing, postural relaxation, and proprioceptive re-education. METHODS: Physical self-regulation training was presented by a dentist during two 50-minute sessions spaced at 3-week intervals and was compared to a standard dental care (SDC) program that included a flat-plane intraoral appliance and self-care instructions provided by a dentist. Participants (n = 44) were initially evaluated by a dentist experienced in the diagnosis and management of orofacial pain and were determined to have myofascial pain (Type 1a and 1b diagnoses per the Research Diagnostic Criteria) prior to random assignment to either the PSR or SDC conditions. Posttreatment evaluations 6 weeks and 26 weeks after treatment had begun were conducted by a dentist who was not aware of which treatment the participants received. RESULTS: Initial results indicated that pain severity and life interference from pain were reduced in both groups (P < 0.001), while perception of control was increased (P < 0.001), as was incisal opening without pain (P < 0.05). At the 26-week follow-up, the PSR group reported less pain (P < 0.04) and greater incisal opening, both with (P < 0.04) and without (P < 0.01) pain, than the SDC group. There were also significant decreases (P < 0.05) in affective distress, somatization, obsessive-compulsive symptoms, tender point sensitivity, awareness of tooth contact, and sleep dysfunction for both groups over time. CONCLUSION: The findings support the use of PSR for the short- and long-term management of muscle pain in the facial region. These results are discussed in terms of the potential mechanisms by which self-regulation treatment strategies are effective for the management of these pain disorders.

A fission yeast general translation factor reveals links between protein synthesis and cell cycle controls.

In two independent screens we isolated fission yeast mutations with phenotypes suggesting defects in B-cyclin function or expression. These mutations define a single gene which we call ded1. We show that ded1 encodes a general translation factor that is related in sequence and function to RNA helicases required for translation in other species. Levels of the B-cyclins Cig2 and Cdc13 are dramatically reduced upon inactivation of Ded1, and this reduction is independent of degradation by the anaphase promoting complex. When a ded1 mutant is grown under semi-restrictive conditions, the translation of Cig2 (and to a lesser extent Cdc13), is impaired relative to other proteins. We show that B-cyclin translation is specifically inhibited upon nitrogen starvation of wild-type cells, when B-cyclin/Cdc2 inactivation is a prerequisite for G(1) arrest and subsequent mating. Our data suggest that translational inhibition of B-cyclin expression represents a third mechanism, in addition to cyclin degradation and Rum1 inhibition, that contributes to Cdc2 inactivation as cells exit from the mitotic cell cycle and prepare for meiosis.

Psychosocial profiles of patients with burning mouth syndrome.

AIMS: Burning mouth syndrome (BMS) is estimated to affect 1 to 5% of the adult population, with women experiencing symptoms more frequently than men. The purpose of this study was to examine the psychosocial profiles of BMS patients to determine whether psychologic factors are related to pain reports. Based on previous literature, it was hypothesized that patients with BMS would be characterized by clinical elevations on standardized psychologic assessment instruments that included the Revised Symptom Checklist (SCL-90R) and the Multidimensional Pain Inventory (MPI). METHODS: Thirty-three BMS patients completed the McGill Pain Questionnaire, MPI, and SCL-90R during their initial clinical evaluation session. The SCL-90R and MPI data were then summarized and presented in standardized format (T-scores) to enable meaningful comparisons with larger population samples that included both a chronic pain population and a normal nonclinical sample. RESULTS: The T-score for the overall pain severity on the MPI was 40.8 (SD 12.8). For the entire BMS sample, there was no evidence for significant clinical elevations on any of the SCL-90R subscales, including depression, anxiety, and somatization. Moreover, patients reported significantly fewer disruptions in normal activities as a result of their oral burning pain than did a large sample of chronic pain patients. CONCLUSION: These findings indicate that, as a group, this sample of BMS patients did not report significant psychologic distress. There were, however, individual cases (7 of 33, or 21%) where psychometric data indicated a likelihood of psychologic distress, and further evaluation by a competent health professional would be warranted for those individuals.

Influence of biased clinician statements on patient report of referred pain.

AIMS: The purpose of this study was to examine the influence of clinician bias on patients' reports of referred pain. Diagnosis of temporomandibular disorders is dependent on subjective reports of pain and referred pain upon manual muscle palpation. The influence of biased clinician statements in such subjective reports has not been previously investigated. METHODS: Forty subjects with pain and who met specific inclusion criteria were randomly assigned to 1 of 2 experimental groups. One group was subjected to a standardized biasing statement, while the other group was not. Tender points in the masseter muscle were then stimulated with a pressure algometer to the pressure-pain threshold. Subjects then recorded the presence or absence, location, intensity, and unpleasantness of any referred pain. State-trait anxiety and social desirability were also assessed to explore the possibility that anxiety levels or subjects' desires to please the experimenter influenced results. RESULTS: The biased group reported increased presence (P < 0.01), intensity (P < 0.001), and unpleasantness (P < 0.003) of referred pain as compared to the non-biased group. There were no differences between groups on state-trait anxiety or social desirability (P > 0.05). CONCLUSION: These data suggest that patient reports of pain referral may be subject to clinician bias, and recommendations to control this bias are offered.

Regulation of the start of DNA replication in Schizosaccharomyces pombe.

Cells of Schizosaccharomyces pombe were grown in minimal medium with different nitrogen sources under steady-state conditions, with doubling times ranging from 2.5 to 14 hours. Flow cytometry and fluorescence microscopy confirmed earlier findings that at rapid growth rates, the G1 phase was short and cell separation occurred at the end of S phase. For some nitrogen sources, the growth rate was greatly decreased, the G1 phase occupied 30-50% of the cell cycle, and cell separation occurred in early G1. In contrast, other nitrogen sources supported low growth rates without any significant increase in G1 duration. The method described allows manipulation of the length of G1 and the relative cell cycle position of S phase in wild-type cells. Cell mass was measured by flow cytometry as scattered light and as protein-associated fluorescence. The extensions of G1 were not related to cell mass at entry into S phase. Our data do not support the hypothesis that the cells must reach a certain fixed, critical mass before entry into S. We suggest that cell mass at the G1/S transition point is variable and determined by a set of molecular parameters. In the present experiments, these parameters were influenced by the different nitrogen sources in a way that was independent of the actual growth rate.

Measurement of nuclear DNA content in fission yeast by flow cytometry.

Cell division cycle (cdc) mutants of Schizosaccharomyces pombe are arrested at specific points in the cell cycle when grown at restrictive temperature. Flow cytometry of such cells reveals an anomalous increase in the DNA fluorescence signal, which represents a problem in experiments designed to determine the cell cycle arrest point. The increased fluorescence signal is due to cytoplasmic constituents and has been attributed to mitochondrial DNA synthesis (S. Sazer and S. W. Sherwood, J. Cell Sci. 97: 509-516, 1990). Here we have studied the cdc10 mutant by flow cytometry using different DNA-binding fluorochromes and found no evidence that the increased fluorescence signal was caused by mitochondrial DNA synthesis. To determine more accurately the nuclear DNA content we have developed a novel method to remove most of the cytoplasmic material by exposing the cells to Triton X-100 and hypotonic conditions after cell wall digestion. The DNA fluorescence from cells treated in this way was more constant with time of incubation at restrictive temperature in spite of a considerable increase in cell size. With this method we could determine that the recently isolated temperature sensitive orp1 mutant is arrested with a 1C DNA content. Premature and abnormal mitosis ('cut') could be observed for the orp1 mutant after only 4 h at restrictive temperature.

Effects of stretch-based progressive relaxation training on the secretion of salivary immunoglobulin A in orofacial pain patients.

There is a growing body of evidence that psychologic stressors can affect physical health and proneness to disease through depletion of the body's immune system. Relatively little research, however, has investigated the potential immunoenhancing effect of stress-relieving strategies such as progressive muscle relaxation. This study explored the relationship between immune functioning and relaxation training with persons experiencing persistent facial pain. In a single experimental session, 21 subjects either received relaxation training or rested for an equivalent time period. Salivary immunoglobulin A, mood, pain, and tension levels were measured before and after relaxation and rest periods. Results indicated that a greater proportion of those receiving relaxation training had increases in secretion of salivary immunoglobulin A. These findings suggest that immunoenhancement may be another potential benefit of progressive relaxation training for persons with chronic pain conditions.