The contributions of sex, genotype and age to transcriptional variance in Drosophila melanogaster.

Here we present a statistically rigorous approach to quantifying microarray expression data that allows the relative effects of multiple classes of treatment to be compared and incorporates analytical methods that are common to quantitative genetics. From the magnitude of gene effects and contributions of variance components, we find that gene expression in adult flies is affected most strongly by sex, less so by genotype and only weakly by age (for 1- and 6-wk flies); in addition, sex x genotype interactions may be present for as much as 10% of the Drosophila transcriptome. This interpretation is compromised to some extent by statistical issues relating to power and experimental design. Nevertheless, we show that changes in expression as small as 1.2-fold can be highly significant. Genotypic contributions to transcriptional variance may be of a similar magnitude to those relating to some quantitative phenotypes and should be considered when assessing the significance of experimental treatments.

A vision for a biomedical cloud.

We present a vision for a Biomedical Cloud that draws on progress in the fields of Genomics, Systems Biology and biomedical data mining. The successful fusion of these areas will combine the use of biomarkers, genetic variants, and environmental variables to build predictive models that will drastically increase the specificity and timeliness of diagnosis for a wide range of common diseases, whilst delivering accurate predictions about the efficacy of treatment options. However, the amount of data being generated by each of these fields is staggering, as is the task of managing and analysing it. Adequate computing infrastructure needs to be developed to assemble, manage and mine the enormous and rapidly growing corpus of 'omics' data along with clinical information. We have now arrived at an intersection point between genome technology, cloud computing and biological data mining. This intersection point provides a launch pad for developing a globally applicable cloud computing platform capable of supporting a new paradigm of data intensive, cloud-enabled predictive medicine.

The diagnosis and management of Piriformis Syndrome: myths and facts.

Piriformis Syndrome (PS) is an uncommon, controversial neuromuscular disorder that is presumed to be a compression neuropathy of the sciatic nerve at the level of the piriformis muscle (PM). The diagnosis is hampered by a lack of agreed upon clinical criteria and a lack of definitive investigations such as imaging or electrodiagnostic testing. Treatment has focused on stretching, physical therapies, local injections, including botulinum toxin, and surgical management. This article explores the various sources of controversy surrounding piriformis syndrome including diagnosis, investigation and management. We conclude with a proposal for diagnostic criteria which include signs and symptoms, imaging, and response to therapeutic injections.

Coordination of Drosophila metamorphosis by two ecdysone-induced nuclear receptors.

The functions of the ecdysone-induced DHR3 and E75B orphan nuclear receptors in the early stages of Drosophila metamorphosis were investigated. DHR3 represses the ecdysone induction of early genes turned on by the pulse of ecdysone that triggers metamorphosis. It also induces betaFTZF1, an orphan nuclear receptor that is essential for the appropriate response to the subsequent prepupal pulse of ecdysone. The E75B receptor, which lacks a complete DNA binding domain, inhibits this inductive function by forming a complex with DHR3 on the betaFTZF1 promoter, thereby providing a timing mechanism for betaFTZF1 induction that is dependent on the disappearance of E75B.

Patterns of gene expression during Drosophila mesoderm development.

The transcription factor Twist initiates Drosophila mesoderm development, resulting in the formation of heart, somatic muscle, and other cell types. Using a Drosophila embryo sorter, we isolated enough homozygous twist mutant embryos to perform DNA microarray experiments. Transcription profiles of twist loss-of-function embryos, embryos with ubiquitous twist expression, and wild-type embryos were compared at different developmental stages. The results implicate hundreds of genes, many with vertebrate homologs, in stage-specific processes in mesoderm development. One such gene, gleeful, related to the vertebrate Gli genes, is essential for somatic muscle development and sufficient to cause neural cells to express a muscle marker.

Microarray analysis of Drosophila development during metamorphosis.

Metamorphosis is an integrated set of developmental processes controlled by a transcriptional hierarchy that coordinates the action of hundreds of genes. In order to identify and analyze the expression of these genes, high-density DNA microarrays containing several thousand Drosophila melanogaster gene sequences were constructed. Many differentially expressed genes can be assigned to developmental pathways known to be active during metamorphosis, whereas others can be assigned to pathways not previously associated with metamorphosis. Additionally, many genes of unknown function were identified that may be involved in the control and execution of metamorphosis. The utility of this genome-based approach is demonstrated for studying a set of complex biological processes in a multicellular organism.

Nfkb1 is a haploinsufficient DNA damage-specific tumor suppressor.

NF-κB proteins play a central and subunit-specific role in the response to DNA damage. Previous work identified p50/NF-κB1 as being necessary for cytotoxicity in response to DNA alkylation damage. Given the importance of damage-induced cell death for the maintenance of genomic stability, we examined whether Nfkb1 acts as a tumor suppressor in the setting of alkylation damage. Hprt mutation analysis demonstrates that Nfkb1(-/-) cells accumulate more alkylator-induced, but not ionizing radiation (IR)-induced, mutations than similarly treated wild-type cells. Subsequent in vivo tumor induction studies reveal that following alkylator treatment, but not IR, Nfkb1(-/-) mice develop more lymphomas than similarly treated Nfkb1(+/+) animals. Heterozygous mice develop lymphomas at an intermediate rate and retain functional p50 in their tumors, indicating that Nfkb1 acts in a haploinsufficient manner. Analysis of human cancers, including therapy-related myeloid neoplasms, demonstrates that NFKB1 mRNA expression is downregulated compared with control samples in multiple hematological malignancies. These data indicate that Nfkb1 is a haploinsufficient, pathway-specific tumor suppressor that prevents the development of hematologic malignancy in the setting of alkylation damage.

Work disability evaluation and the fibromyalgia syndrome.

Fibromyalgia syndrome (FMS) is a common and costly cause of work disability. Patients with FMS, nevertheless, encounter considerable difficulties in their assessment of claims for disability payments. Factors that contribute to FMS as an important cause of disability are its high prevalence, the patients' perception of severe discomfort, and poor function. Disability evaluation in FMS is controversial for several reasons including lack of acceptance of the diagnosis, concurrent psychological abnormalities, difficulties in objectifying disability, deficiencies in instruments of evaluation, the uncertain efficacy of treatment, and physician attitudes. Third parties appear to have inappropriate expectations of the physician's role in determining disability. We suggest that the process of disability evaluation be improved by more objective assessments and by the inclusion of other health professionals in assessing disability and necessary retraining. Further research is needed to develop better instruments for measuring disability, to assess the long-term effects of various treatments, and to clarify the contributions of the work place and of compensation in causing or aggravating FMS.

Trauma and fibromyalgia: is there an association and what does it mean?

OBJECTIVES: The primary objective is to review current research with respect to the role of trauma in fibromyalgia (FM). A secondary objective is to hypothesize which steps need to be taken, first to determine whether such an association truly exists, and second to clarify what such an association might mean. METHODS: An extensive literature review was undertaken, including Medline from 1979 to the present. RESULTS: The strongest evidence supporting an association between trauma and FM is a recently published Israeli study in which adults with neck injuries had greater than a 10-fold increased risk of developing FM within 1 year of their injury, compared with adults with lower extremity fractures (P= .001). Several other studies provide a hypothetical construct for such an association. These include studies on (1) postinjury sleep abnormalities; (2) local injury sites as a source of chronic distant regional pain; and (3) the concept of neuroplasticity. There are, however, several primary arguments against such an association: (1) FM may not be a distinct clinical entity; (2) FM may be a psychological, rather than physical, disease; (3) the evidence supporting any association is limited and not definitive; (4) the Israeli study, itself, has some methodological limitations; and (5) other factors may be more important than the injurious event in determining chronic symptoms after an acute injury. CONCLUSIONS: Although there is some evidence supporting an association between trauma and FM, the evidence is not definitive. Further prospective studies are needed to confirm this association and to identify whether trauma has a causal role.

Reliability of the six-minute walk test in people with fibromyalgia.

OBJECTIVE: To determine the test-retest reliability of the 6-minute walk test in people with fibromyalgia. METHODS: Twenty-six subjects (27-59 years of age) performed 3 walk tests over consecutive days before and after a 4-week treatment program. Reliability was determined using a one-way repeated measures analysis of variance and the intraclass correlation coefficient (ICC2,1). RESULTS: Reliability of the 6-minute walk test was excellent both at program intake (ICC2,1 = 0.91) and program completion (ICC2,1 = 0.98). On program intake, significant differences (P < 0.01) were found between test 1 (478 +/- 61 m) and test 2 (492 +/- 57 m), and between test 1 and test 3 (495 +/- 60 m). On program completion, there were no significant differences across the 3 replicate tests (507 m, 505 m, and 509 m). CONCLUSIONS: The 6-minute walk test is a reliable measure in people with fibromyalgia. In this study, two trials were required to achieve a stable walk performance before a treatment program. This learning effect was not present following the intervention.

A comparison of rheumatoid arthritis and fibromyalgia patients and healthy controls exposed to a pulsed (200 microT) magnetic field: effects on normal standing balance.

Specific weak time varying pulsed magnetic fields (MF) have been shown to alter animal and human behaviors, including pain perception and postural sway. Here we demonstrate an objective assessment of exposure to pulsed MF's on Rheumatoid Arthritis (RA) and Fibromyalgia (FM) patients and healthy controls using standing balance. 15 RA and 15 FM patients were recruited from a university hospital outpatient Rheumatology Clinic and 15 healthy controls from university students and personnel. Each subject stood on the center of a 3-D forceplate to record postural sway within three square orthogonal coil pairs (2 m, 1.75 m, 1.5 m) which generated a spatially uniform MF centered at head level. Four 2-min exposure conditions (eyes open/eyes closed, sham/MF) were applied in a random order. With eyes open and during sham exposure, FM patients and controls appeared to have similar standing balance, with RA patients worse. With eyes closed, postural sway worsened for all three groups, but more for RA and FM patients than controls. The Romberg Quotient (eyes closed/eyes open) was highest among FM patients. Mixed design analysis of variance on the center of pressure (COP) movements showed a significant interaction of eyes open/closed and sham/MF conditions [F=8.78(1,42), P<0.006]. Romberg Quotients of COP movements improved significantly with MF exposure [F=9.5(1,42), P<0.005] and COP path length showed an interaction approaching significance with clinical diagnosis [F=3.2(1,28), P<0.09]. Therefore RA and FM patients, and healthy controls, have significantly different postural sway in response to a specific pulsed MF.

Oral cavity tumors in younger patients show a poor prognosis and do not contain viral RNA.

BACKGROUND: Oral cavity and in particular oral tongue cancers occur with a rising incidence in younger patients often lacking the typical risk factors of tobacco use, alcohol use, and human papilloma virus (HPV) infection. Their prognosis when treated with chemoradiation has not been well studied and responsible risk factors remain elusive. A viral etiology (other than HPV) has been hypothesized. METHODS: First we analyzed outcomes from 748 head and neck cancer patients with locoregionally advanced stage tumors treated with curative-intent chemoradiation by anatomic site. Second, we analyzed seven oral tongue (OT) tumors from young, non-smokers/non-drinkers for the presence of viral mRNA using short-read massively-parallel sequencing (RNA-Seq) in combination with a newly-developed digital subtraction method followed by viral screening and discovery algorithms. For positive controls we used an HPV16-positive HNC cell line, a cervical cancer, and an EBV-LMP2A transgene lymphoma. RESULTS: Younger patients with oral cavity tumors had worse outcomes compared to non-oral cavity patients. Surprisingly none of the seven oral tongue cancers showed significant presence of viral transcripts. In positive controls the expected viral material was identified. CONCLUSION: Oral cavity tumors in younger patients have a poor prognosis and do not appear to be caused by a transcriptionally active oncovirus.

Functional genomics and the study of development, variation and evolution.

It has rarely been possible to connect the developmental and evolutionary branches of genetics, particularly with regard to the precise changes in the molecular control of development that are responsible for phenotypic variation and evolution. Making such connections will require a high-resolution molecular description of the genetic networks that underlie development and an understanding of their responses to genetic and environmental variation. Functional genomics approaches to development and evolution promise to accelerate the research necessary to accomplish these goals.

Classification, epidemiology, and natural history of fibromyalgia.

Fibromyalgia (FM), also known as fibromyalgia syndrome (FMS) and fibrositis, is a common form of nonarticular rheumatism that is associated with chronic generalized musculoskeletal pain, fatigue, and a long list of other complaints. Some have criticized the classification of FM as a distinct medical entity, but existing data suggest that individuals meeting the case definition for FM are clinically somewhat distinct from those with chronic widespread pain who do not meet the full FM definition. Clinic studies have found FM to be common in countries worldwide; these include studies in specialty and general clinics. The same is true of general population studies, which show the prevalence of FM to be between 0.5% and 5%. Knowledge about risk factors for FM is limited. Females are at greater risk, and risk appears to increase through middle age, then decline. Although some authors claim that an epidemic of FM has been fueled by an over-generous Western compensation system, there are no data that demonstrate an increasing incidence or prevalence of FM; moreover, existing data refute any association between FM prevalence and compensation. Claims that the FM label itself causes illness behavior and increased dependence on the medical system also are not supported by existing research. This article reviews the classification, epidemiology, and natural history of FM.

Arteriovenous ratios of angiotensin II during acute infusion experiments: a model-based analysis.

The hormone angiotensin II (AII) is a vasoconstrictor known to participate in the natural regulation of blood pressure via the renin-angiotensin system. A third-order model was developed which describes the dynamics of venous and arterial plasma AII concentrations (PAC) and mean arterial blood pressure (BP) during acute constant rate AII infusion experiments. The model is calibrated using approximate blood circulation rates and steady-state PAC and BP data for published experiments in sheep. Analysis of the dynamic model demonstrates that local changes in PAC during the first several minutes of acute infusion are characterized by the comparatively rapid distribution of exogenous AII making its forward passage across the blood circulation, combined with the more gradual elevation of exogenous AII recycled through the circulation. This analysis explains the observed divergence in physiological levels of venous and arterial PAC at steady state in terms of the monotonic net clearance of elevated levels of circulating AII along the circulatory path between the point of infusion and the two sites at which the PAC measurements are taken. The model suggests that the differing arteriovenous AII concentration ratios and differing PAC and BP relationships reported for different dose-response experiments may be explained in part by differences in the specific infusion and measurement sites employed in those experiments.

The occurrence and impact of generalized pain.

A major problem with estimating the impact of chronic generalized pain is that the term remains undefined. It appears to encompass several distinct clinical entities, including rheumatoid arthritis and fibromyalgia, which can exist alone or together in a given individual. Nonetheless, chronic generalized pain appears to have a sizable impact on both the individual and society. Although little is known about causal relationships, demographic risk factors for chronic generalized pain are female sex, age in the forties and fifties, lower income, lower education, and being divorced or separated. Chronic generalized pain affects the individual in several ways, including physical and psychological distress, losses of function, quality of life, employment and income, and prolonged litigation for many. Its impact on society includes increased utilization of health care resources, loss of work productivity, disability and insurance costs, costs of litigation and social policy. Future research into the impact of chronic generalized pain must begin by defining this term in a way that is both valid in construct and convenient to use. Research is also warranted to develop and validate diagnostic tools that may better distinguish various subsets of chronic generalized pain, both to better understand the pathological processes involved and to allow for estimates of the relative contribution of each subset to societal costs.

Lumbar pannus presenting as cauda equina syndrome in a patient with longstanding rheumatoid arthritis.

Relatively little attention has been paid to lumbar spine involvement in rheumatoid arthritis (RA), and indeed it is generally considered to be an uncommon and usually clinically minor manifestation of the disease. We describe a case of acute right lower extremity weakness secondary to compression of multiple lumbar nerve roots by a large interforaminal rheumatoid pannus, and review the literature on this complication and other lumbar spine involvement in RA.

Cognitive behavioral treatment of fibromyalgia syndrome: a followup assessment.

OBJECTIVE: In a previous study, we showed that cognitive behavioral therapy (CBT) was effective in the short term treatment of fibromyalgia (FMS). We tested the hypothesis that a pooled index of 10 target psychometric outcomes would show overall improved outcome at longterm followup versus pretreatment. METHODS: Using a quasiexperimental design and validated psychometric instruments, we compared pretreatment psychometric variables with longterm outcomes in the 25 subjects who had completed the earlier study. A distinction was made between target variables the program had specifically addressed and nontarget variables not specifically addressed. RESULTS: Twenty-two subjects participated at a mean of 30 months after discharge from the program. As at short term followup, no nontarget variable changed from pretreatment to longterm assessment. The short term improvement that had been observed in all 10 target variables was maintained in 3 variables at a Bonferroni corrected alpha level of 0.005: worry, p < 0.001; observed pain behavior, p < 0.001; and control over pain, p < 0.002. All 10 target variables changed in the direction of improvement. A pooled index of these 10 variables showed statistically significant improvement from baseline (p < 0.002). CONCLUSION: Our results suggest that CBT may have a role in the longterm treatment of FMS.

Electromechanical dissociation following verapamil and propranolol ingestion: a physiologic profile.

We present a case of combined verapamil and propranolol ingestion in a young woman with a DDD pacer from whom hemodynamic, echocardiographic, and toxicologic data were obtained simultaneously. Analysis of the data from these combined sources suggests a progression of (1) recovery of myocardial contractility, (2) return of systemic vascular resistance, and (3) return of intrinsic electrical activity of the heart. That contractility returns before intrinsic electrical activity strongly for the early use of transvenous temporary pacing wires. Furthermore, our patient's complete recovery after a prolonged electromechanical dissociation arrest would again argue for very aggressive resuscitation efforts in this setting.

A general population study of fibromyalgia tender points in noninstitutionalized adults with chronic widespread pain.

OBJECTIVE: To assess the distribution and predictive ability of fibromyalgia (FM) tender points (TP) in adults with chronic widespread pain. METHODS: Using published classification criteria, we confirmed 100 FM cases and 76 controls with widespread pain not meeting the 1990 American College of Rheumatology (ACR) classification criteria for FM (pain controls) in a survey of 3,395 adults screened for widespread musculoskeletal pain in a general population survey. RESULTS: At each of the 18 FM tender points, FM cases were more likely than pain controls to have tenderness, and the likelihood ratio (LR) was statistically greater than 1.0 for 13 of 18 points. However, the LR for individual points ranged from 4.0 to as low as 1.2. Females were more likely to have TP, especially at lower body points; however, lower body points were more discriminatory in males. CONCLUSION: TP differ in their ability to predict FM among adults in the general population with chronic widespread pain.