Codeine influences the serum and urinary profile of endogenous androgens but does not interact with the excretion rate of administered testosterone.

Today's doping tests involve longitudinal monitoring of urinary steroids including the testosterone glucuronide and epitestosterone glucuronide ratio (T/E) in an Athlete Biological Passport (ABP). The aim of this study was to investigate the possible influence of short-term use of codeine on the urinary excretion of androgen metabolites included in the steroidal module of the passport prior to and after the co-administration with testosterone. The study was designed as an open study with the subjects being their own control. Fifteen healthy male volunteers received therapeutic doses of codeine (Kodein Meda) for 6 days. On Day 3, 500 mg or 125 mg of testosterone enanthate (Testoviron®-Depot) was administered. Spot urine samples were collected for 17 days, and blood samples were collected at baseline, 3, 6, and 14 days after codeine intake. The circulatory concentration of total testosterone decreased significantly by 20% after 3 days' use of codeine (p = 0.0002) and an atypical ABP result was noted in one of the subjects. On the other hand, the concomitant use of codeine and testosterone did not affect the elevated urinary T/E ratio. In 75% of the individuals, the concentration of urinary morphine (a metabolite of codeine) was above the decision limit for morphine. One of the participants displayed a morphine/codeine ratio of 1.7 after codeine treatment, indicative of morphine abuse. In conclusion, our study shows that codeine interferes with the endogenous testosterone concentration. As a result, the urinary steroid profile may lead to atypical findings in the doping test.

Effects of the peroxisome proliferator-activated receptor (PPAR)-δ agonist GW501516 on bone and muscle in ovariectomized rats.

Estrogen deficiency promotes bone loss and skeletal muscle dysfunction. Peroxisome proliferator-activated receptors (PPARs) have 3 subtypes (α, δ, and γ). PPARγ agonists induce bone loss, whereas PPARα agonists increase bone mass. Although PPARδ agonists are known to influence skeletal muscle metabolism, the skeletal effects are unsettled. This study investigated the musculoskeletal effects of the PPARδ agonist GW501516 in ovariectomized (OVX) rats. Female Sprague Dawley rats, 12 weeks of age, were allocated to a sham-operated group and 3 OVX groups; high-dose GW501516 (OVX-GW5), low-dose GW501516 (OVX-GW1), and a control group (OVX-CTR), respectively (n = 12 per group). Animals received GW501516 or vehicle (methylcellulose) daily for 4 months by gavage. Bone mineral density (BMD) was assessed by dual x-ray absorptiometry at the femur, spine, and whole body. Bone microarchitecture at the proximal tibia was assessed by microcomputed tomography, and dynamic histomorphometry was performed. Quadriceps muscle morphology and the relative expression of mitochondrial proteins were analyzed. Bone metabolism markers and metabolic markers were measured in plasma. After 4 months, the OVX-GW5 group displayed lower femoral BMD than OVX-CTR. Trabecular separation was higher in the GW-treated groups, compared with OVX-CTR. The OVX-GW5 group also exhibited lower cortical area fraction and a higher structure model index than OVX-CTR. These effects coincided with impaired bone formation in both GW groups. The OVX-GW5 group displayed elevated triglyceride levels and reduced adiponectin levels, whereas no effects on muscle morphology or mitochondrial gene expression appeared. In summary, the PPARδ agonist GW501516 negatively affected bone properties in OVX rats, whereas no effects were detected in skeletal muscle.

Human evidence for the involvement of insulin-induced gene 1 in the regulation of plasma glucose concentration.

AIMS/HYPOTHESIS: Insulin-induced gene 1 (INSIG1) is a protein that blocks proteolytic activation of sterol regulatory element-binding proteins (SREBPs), transcription factors that activate genes regulating cholesterol and fatty acid metabolism and possibly genes involved in glucose homeostasis. In search of genetic regulation of these processes we examined human INSIG1 for common polymorphisms and analysed their associations with biochemical parameters related to lipid and glucose metabolism. METHODS: Associations between common polymorphisms in INSIG1 and several biochemical parameters were analysed in a group of 618 healthy, 50-year-old men. A replication analysis was performed in a cohort of 472 healthy, middle-aged men. The impact of one promoter polymorphism on oral glucose tolerance was analysed in a subset of 181 subjects. Small interfering RNA (siRNA) inhibition was used to test the significance of INSIG1 for gene expression in human Huh7 hepatoma cells. RESULTS: A potentially functional polymorphism, a C to T substitution at position -169, was discovered in a highly conserved section of the promoter. Significant relationships between the -169C>T polymorphism and plasma glucose concentration were found in two cohorts of healthy, middle-aged men (p < 0.01 and p < 0.02, respectively). The -169T allele was associated with significantly lower post-load plasma glucose concentrations. A significant (p = 0.02) reduction in expression of phosphoenolpyruvate carboxykinase (PCK2) was observed following siRNA inhibition of INSIG1 in human Huh7 hepatoma cells. CONCLUSIONS/INTERPRETATION: Population studies demonstrate that INSIG1 plays a role in glucose homeostasis. Experiments with siRNA suggest that this action of INSIG1 is related to SREBP-mediated regulation of PCK2.

Long-term outcome of facial palsy in neuroborreliosis.

OBJECTIVES: The objectives of this study were to examine children with previous manifest neuroborreliosis and concommitant facial palsy to see whether there were any persisting symptoms and/or signs of persistent residual facial palsy. STUDY DESIGN: Open, controlled prospective study. SETTING: Tertiary referral center (University Hospital). PATIENTS: The study was conducted on twenty-four patients with clinically manifest neuroborreliosis and facial palsy 3 to 5 years prior to the investigation. MAIN OUTCOME MEASURES: Results of the clinical examination using the House-Brackmann scale were compared to results from two neuro-physiological examinations (qEMG and ENoG). RESULTS: Approximately one-half of the patients with reported subjective symptoms of residual facial palsy had signs of slight dysfunction in the clinical examination using the House-Brackmann scale. There was no correlation between the subjective feeling of facial dysfunction and presence of clinical signs. Likewise, about one-half of the subjective facial dysfunction group, as well as the control group, were found to demonstrate pathological values in their neurophysiological examinations using qEMG and ENoG. CONCLUSIONS: This study shows that the assumption is not true that all children who had neuroborreliosis with facial palsy will heal 100%. A small proportion of the children claim that several years after the infection, they have subjective symptoms of slight facial weakness on the affected side. Our study shows that some of these children, as well as some children without subjective symptoms of facial palsy, demonstrate a slight facial weakness when examined clinically. Likewise, signs of slight-to-moderate facial motor dysfunction were revealed in about half of the children with the two neurophysiological methods utilized in this study. It is interesting to note that there was no clear correlation between the presence of subjective symptoms, objective signs, and neurophysiological results.

Oral shedding of herpes simplex virus type 2.

OBJECTIVES: Herpes simplex virus (HSV) 1 and HSV-2 reactivate preferentially in the oral and genital area, respectively. We aimed to define frequency and characteristics associated with oral shedding of HSV-2. METHODS: Demographic, clinical and laboratory data of patients with documented HSV-2 infection and at least one oral viral culture obtained were selected from the University of Washington Virology Research Clinic database. RESULTS: Of 1388 people meeting the entry criteria, 44 (3.2%) had HSV-2 isolated at least once from their mouths. In comparison with the 1344 people who did not have HSV-2 isolated from their mouth, participants with oral HSV-2 were more likely to be male (OR = 1.9, 95% CI 1.0 to 3.7), HIV positive (OR = 2.9, 95% CI 1.4 to 6.0), and homosexual (OR = 2.2, 95% CI 1.1 to 4.2), and to have collected a larger number of oral specimens (median 32 v 4, p<0.001). Of the 58 days with oral HSV-2 isolation, 15 (25%) occurred during newly acquired HSV-2 infection, 12 (21%) during a recurrence with genital lesions, three (5%) during a recurrence with oral lesions, and three (5%) during a recurrence with oral and genital lesions; 25 (43%) occurred during asymptomatic shedding. Oral HSV-2 was found less frequently than oral HSV-1 (0.06% v 1%, p<0.001) in people with HSV-1 and HSV-2 antibody, and less frequently than genital HSV-2 (0.09% v 7%, p<0.001). CONCLUSIONS: Oral reactivation of HSV-2 as defined by viral isolation is uncommon and usually occurs in the setting of first episode of genital HSV-2 or during genital recurrence of HSV-2.

Depression predicts disability in long-term chronic pain patients.

PURPOSE: Investigators have examined factors that predict treatment outcome and disability status in chronic pain patients, including psychopathology and personality characteristics with equivocal results. The purpose of this study was to evaluate the usefulness of personality characteristics, depression, and personality disorders in predicting disability status in pain patients with long-term follow-up. The setting was a rehabilitation hospital in Southern Sweden. METHOD: Subjects were 184 pain patients (mean age = 43.4 (10.8) years; 72.8% female) who had no more than 365 sick leave days (Mean sick leave days = 132.7 (128.2)) prior to the baseline personality and psychiatric evaluation. The baseline evaluation consisted of a psychiatric interview that included the administration of the Structured Clinical Interview for DSM-IV Screen Questionnaire (SCID-II), the Montgomery-Asberg Depression Rating Scale (MADRS), and the Karolinska Scales of Personality (KSP). Disability status was assessed by insurance record review a minimum of two-and-a-half years after baseline evaluation. RESULTS: Multivariate logistic regression suggests that age (OR = 1.09, 95% CI = 1.02-1.18; p = 0.013), number of sick leave days prior to evaluation (OR= 1.01, 95% CI= 1.01-1.02; p = 0.018), and baseline diagnosis of depression significantly predicted subsequent disability status (OR = 7.04, 95% CI = 1.15-42.93; p = 0.034). Baseline personality traits and the diagnosis of a personality disorder were not useful predictors of disability status in our sample. CONCLUSIONS: These data suggest that depression, but not personality disorders characteristics, was an important disability predictor in chronic pain patients with extended follow-up.

Pharmacotherapy for obesity: a quantitative analysis of four decades of published randomized clinical trials.

AIM: This article provides the first comprehensive meta-analysis of randomized clinical trials of medications for obesity. METHOD: Based on stringent inclusionary criteria, a total of 108 studies were included in the final database. Outcomes are presented for comparisons of single and combination drugs to placebo and for comparisons of medications to one another. RESULT: Overall, the medications studied produced medium effect sizes. Four drugs produced large effect sizes (ie d>0.80; amphetamine, benzphetamine, fenfluramine and sibutramine). The placebo-subtracted weight losses for single drugs vs placebo included in the meta-analysis never exceeded 4.0 kg. No drug, or class of drugs, demonstrated clear superiority as an obesity medication. Effects of methodological factors are also presented along with suggestions for future research.

Long-lasting recall response of CD4+ and CD8+ alphabeta T cells, but not gammadelta T cells, to heat shock proteins of francisella tularensis.

Decades after recovery from tularemia, circulating alphabeta T cells are known to still recognize a variety of membrane proteins of Francisella tularensis. We studied the T cell response to 3 cytoplasmic heat shock proteins of the organism: DnaK, chaperone-60 (Cpn-60) and Cpn-10. Determination of subpopulations of responding T cells was of special interest as it has been suggested that homologs of these conserved proteins may be recognized by human gammadelta T cells. Compared with reference subjects with no history of tularemia or tularemia vaccination, subjects who had been infected with tularemia 10-30 y earlier showed a significantly (p = 0.01) higher proliferative T cell response to all 3 heat shock proteins. In general, the magnitude of responses of CD4 T cells was higher than that of CD8 T cells. By flow cytometry, blast cells were shown to express the alphabeta T cell receptor. Under conditions that allowed vigorous expansion of gammadelta T cells in response to a phosphorylated non-peptide antigen, no expansion of gammadelta T cells occurred in response to DnaK or Cpn60 of F. tularensis. In conclusion, a long-lasting recall response to heat shock proteins of F. tularensis was demonstrated in alphabeta T cells but not in gammadelta T cells. The results support the assumption that human alphabeta T cells recognize bacterial proteins irrespective of the nature or localization of the proteins in the bacterial cell and thereby contribute to the maintenance of a long-lasting broad T cell response based on a wide variety of specificities.

Differences in inhibition of replication between Coxsackie B4 virus strains in various cell lines by antibodies to some cell surface proteins.

Monoclonal antibodies that interact with the decay accelerating factor (DAF, CD55), the lymphocyte homing receptor (CD44) or the intercellular adhesion molecule I (ICAM- 1) were found to inhibit the replication of different strains of Coxsackievirus serotype B4 (CBV-4) to various extent. By adding antibodies to CD55 the replication of two (V345 and VD2921) of seven strains in HeLa cells, three (V89-4557, VD2921 and T318) of seven in A549-10C cells and one (VD2921) of five strains in RD cells was blocked totally. Consequently, the replication of one strain (VD2921) was blocked in all cells indicating that this strain uses CD55 as a receptor or as a co-receptor on all cell lines and is unable to use another cell surface protein. The binding of this strain to the cell surface was inhibited by the antibodies to CD55. None of the CBV-4 strains was blocked totally by adding antibodies to CD44 to HeLa and A549-10C cells, whereas in RD cells the replication of one (T318) of the CBV-4 strains was blocked totally. The antibodies to ICAM-1 did not inhibit totally the replication of any strain in HeLa and RD cells, but it blocked totally the replication of one strain (CBV-4-E) in A549-10C cells. In HeLa and A549-10C cells the degree of replication correlated highly with the degree of cytopathic effect (CPE). In RD cells, four of the strains replicated without CPE. The adding of antibodies to the integrin alpha(v)beta(3) led to slightly enhanced replication of three of the CBV-4 strains in all cell lines. It is concluded that the receptor usage by different strains of CBV-4 varies not only within the same cells but also between different cell lines.

Geometric phases for mixed states in interferometry.

We provide a physical prescription based on interferometry for introducing the total phase of a mixed state undergoing unitary evolution, which has been an elusive concept in the past. We define the parallel transport condition that provides a connection form for obtaining the geometric phase for mixed states. The expression for the geometric phase for mixed state reduces to well known formulas in the pure state case when a system undergoes noncyclic and unitary quantum evolution.

Sorting out bacterial viability with optical tweezers.

We have developed a method, using laser, optical tweezers and direct microscopic analysis of reproductive potential and membrane integrity, to assess single-cell viability in a stationary-phase Escherichia coli population. It is demonstrated here that a reduction in cell integrity, determined by using the fluorescent nucleic acid stain propidium iodide, correlated well with a reduction in cell proliferating potential during the stationary-phase period studied. Moreover, the same cells that exhibited reduced integrity were found to be the ones that failed to divide upon nutrient addition. A small but significant number of the intact cells (496 of 7,466 [6.6%]) failed to replicate. In other words, we did not find evidence for the existence of a large population of intact but nonculturable cells during the stationary-phase period studied but it is clear that reproductive ability can be lost prior to the loss of membrane integrity. In addition, about 1% of the stationary-phase cells were able to divide only once upon nutrient addition, and in a few cases, only one of the two cells produced by division was able to divide a second time, indicating that localized cell deterioration, inherited by only one of the daughters, may occur. The usefulness of the optical trapping methodology in elucidating the mechanisms involved in stationary-phase-induced bacterial death and population heterogeneity is discussed.

Does personality or psychopathology predict disability in chronic pain patients?

PURPOSE: The purpose of this study was to evaluate the usefulness of personality characteristics, depression and personality disorders in predicting disability status in pain patients one year later. METHOD: Subjects were 250 volunteer chronic pain patients. The baseline evaluation consisted of the Karolinska Scales of Personality and psychiatric evaluation of depression and personality disorders using standardized diagnostic instruments. Disability status was assessed by insurance record review one year after the evaluation. RESULTS: The results suggest that baseline personality traits and psychopathology (i.e. depression or personality disorders) were not useful predictors of disability status in pain patients with one-year follow-up. CONCLUSIONS: These data suggest that personality characteristics and psychopathology are probably not important disability predictors in chronic pain patients.

Dynamic microwave-assisted extraction.

An apparatus for extraction of solid matrices has been constructed which utilizes a microwave technique for heating in a dynamic mode. During the extraction, fresh solvent is continuously pumped through the extraction cell, which is maintained at a slight overpressure in order to keep the solvent in a liquid state. The extraction efficiency, which can be easily monitored, has been investigated in a factorial design and validated for polycyclic aromatic hydrocarbons in a reference sediment sample (EC-1). Important parameters were found to be temperature and duration of extraction. Flow-rate had no significant first-order effect on the recovery, but interaction effects with flow-rate were found to be significant. The dynamic microwave-assisted extraction apparatus was demonstrated to yield recoveries equivalent to Soxhlet extraction, but in a much shorter time. Each extraction of EC-1 typically takes 40 min.

Autonomic neuropathy--a complication less pronounced in patients with Type 1 diabetes mellitus who have lower blood glucose levels.

AIMS: Autonomic neuropathy is a serious diabetic complication, probably contributing to the death of many young people with Type 1 diabetes mellitus. It is often not diagnosed. METHODS: Patients with Type 1 diabetes from the Stockholm Diabetes Intervention Study were investigated with power spectral analysis (n = 88), heart rate and blood pressure reactions to tilting (n = 66), and heart rate variability during deep breathing (n = 70) a mean of 11.4 years after randomization to intensified conventional treatment (ICT) or standard treatment (ST), the treatment groups similar with regard to age, duration of diabetes and metabolic control at baseline (HbA1c 9.4 (1.3)%, mean (SD)). Blood glucose levels (mean of 29 HbA1c values) during the 10 years were lower in the patients from the ICT group (7.2 (0.6) vs. 8.3 (1.0)%, P = 0.001). RESULTS: Heart rate variability (HRV) in the high frequency range (P = 0.034), the expiration-inspiration ratio (P = 0.020), and the brake index during tilt (P = 0.044) were lower in the ST group, indicating more pronounced parasympathetic insufficiency. Systolic blood pressure fell by 10 (16) mmHg in the ST group, and by 2.5 (15) mmHg in the ICT group 8 min after rising from the supine to a 70 degrees upright position (P = 0.034). A decreased autonomic function was associated with age and higher HbA1c. CONCLUSION: Better autonomic nerve function is associated with lower HbA1c and lower age which were both the same in the intensively and the conventionally treatment groups at baseline. After a mean of 11.4 years autonomic function was better in the intensively treated group.

24-hour ECG frequency-domain measures in preeclamptic and healthy pregnant women during and after pregnancy.

OBJECTIVE: The aim of the present study was to evaluate the autonomic balance in women with preeclampsia and in healthy women during and after pregnancy by means of a 24-h ECG Holter recording combined with power spectral analysis. METHODS: Fifteen preeclamptic and 15 healthy women underwent 24-h Holter monitoring in the 32nd-36th week of gestation and 3-6 months postpartum. The power spectrum of the maternal electrocardiogram was analyzed with an autoregressive algorithm. MAIN OUTCOME MEASURES: The power spectrum contains two major components: a low-frequency peak, primarily attributed to sympathetic tone, and a high-frequency peak, reflecting vagal tone. RESULTS: The power spectrum of maternal heart rate variability did not differ between preeclamptic and normal women during pregnancy. After delivery, the amplitude of all components became significantly higher than those during pregnancy, with one exception: the high-frequency component in the patients who had been preeclamptic. In a comparison of the two groups, the high-frequency component after delivery was significantly lower in women who had preeclampsia than in normal healthy women (p = 0.03). CONCLUSIONS: During pregnancy, the power spectrum is reduced and cannot be used to distinguish between patients with preeclampsia and normal healthy women. Three to 6 months after delivery, the high-frequency component is still reduced in the preeclamptic group of women. This indicates an impaired vagal modulation even in the nonpregnant state in this group of women, unlike those who had a normotensive pregnancy.

The major Epstein-Barr virus (EBV) envelope glycoprotein gp340 when incorporated into Iscoms primes cytotoxic T-cell responses directed against EBV lymphoblastoid cell lines.

A recombinant form of the EBV envelope glycoprotein and vaccine candidate gp340, lacking its hydrophobic transmembrane region, was incorporated into Iscoms after coupling to phosphatidyl ethanolamine via carbohydrate residues. Coupling by partial oxidation of gp340 carbohydrate with sodium periodate partly denatured the incorporated gp340 as indicated by its reduced reactivity with monoclonal antibodies that recognise the major neutralising epitope. Immunisation of cottontop tamarins with these Iscoms elicited antibody responses to gp340, but these antibodies only poorly recognised the major neutralising epitope in a competition ELISA and were unable to neutralise EBV in vitro. Despite the lack of neutralising antibody, immunisation with these Iscoms primed significant in vitro proliferative responses to soluble gp340 in lymphocytes from the draining lymph nodes and spleen. T-cell lines were raised from both immunised and control animals by in vitro stimulation of peripheral blood lymphocytes or spleen cells with autologous EBV-transformed lymphoblastoid cell lines. The T-cell lines from control animals had higher numbers of CD4+ T-cells than CD8+ T-cells and were not cytotoxic for autologous lymphoblastoid cell lines (LCL). In contrast the lines from immunised animals contained more CD8+ T-cells than CD4+ T-cells and had marked cytotoxicity for autologous LCL.

Personality and the prediction of weight loss and relapse in the treatment of obesity.

OBJECTIVE: This prospective study examined whether stable personality traits, as measured by the Karolinska Scales of Personality (KSP), predicted initial weight loss or long-term maintenance in obesity patients. METHOD: The KSP was administered to 102 obese patients prior to entering an 8-week weight loss program. Patients were weighed again at the end of treatment and at 3- and 12-month follow-up. RESULTS: The KSP did not predict initial weight loss after the 8-week program. Several of the KSP scales (Muscle Tension, Monotony Avoidance, Suspicion, and Guilt) had weak associations with 12-month relapse status. Weight gain at the 3-month follow-up was the strongest predictor of 12-month relapse status (O.R. = 0.46; 95% C.I. = 0.32, 0.66). DISCUSSION: Personality traits, as measured by the KSP, do not appear to be important predictors of initial weight loss or 12-month relapse status. Personality assessment may not substantially contribute to predicting treatment outcome in obesity research.

IL-1 beta and IL-6 in mouse parotid acinar cells: characterization of synthesis, storage, and release.

Synthesis, storage, and secretion of the proinflammatory cytokine interleukin-1 beta (IL-1 beta) and the anti-inflammatory cytokine IL-6 have not been established in normal exocrine gland secretory cells. Parotid glands and isolated acinar cells prepared from BALB/c mice were homogenized for RNA isolation and reverse transcription-polymerase chain reaction (RT-PCR). IL-1 beta and IL-6 enzyme-linked immunosorbent assays (ELISAs) were done on supernatants prepared from mouse parotid acinar cell (MPAC) preparations unstimulated or stimulated between 0 and 10 min with 10(-5) M norepinephrine at 37 degrees C. MPACs were fixed in paraformaldehyde, frozen sectioned for light and electron microscopy, and labeled with antibodies to IL-1 beta and IL-6. Mouse specific riboprobes to IL-1 and IL-6 were used for in situ hybridization. RT-PCR yielded the expected IL-1 (336-bp) and IL-6 (614-bp) mRNA products. By ELISA, stimulated MPACs showed a significant increase in IL-1 beta (P < 0.03) and IL-6 (P < 0.01) release into supernatants by 10 min that paralleled the time course of amylase release. In situ hybridization showed the presence of transcripts for IL-1 and IL-6 in glandular epithelial cells. Gold-labeled IL-1 beta and IL-6 were significantly higher (P < 0.01) in granules than in the nucleus and cytoplasm. This study shows that MPACs synthesize IL-1 beta and IL-6 and release these cytokines from their granules after alpha- and beta-adrenergic stimulation.

D4 dopamine receptor gene exon III polymorphism and obesity risk.

Many genes have been identified that may play a role in increasing individual susceptibility to obesity. Reduced dopamine function appears to play a role in dysfunctional eating patterns and may predispose some individuals to obesity. The long version of the D4 dopamine receptor gene (D4DR) has been shown to alter receptor function and reduce intracellular response to dopamine. It also has been associated with novelty-seeking-related personality traits that are found with greater frequency in obese individuals. We examined the association between the long alleles of the D4DR and obesity in a sample of 115 obese patients participating in a weight management program. No direct relationship was found between the D4DR and body mass or novelty-seeking-related personality traits. We constructed four models of increased obesity risk that included combinations of traditional risk factors (i.e., long-term history of obesity, parental obesity, a body mass index > 40) and elevations on the novelty-seeking-related scales of the Karolinska Scales of Personality. There was a significant increase in the frequency of the D4DR long alleles in individuals defined as high risk using the combination of novelty-seeking-related personality traits, severe obesity (i.e., BMI > 40), and any other traditional risk factor, but not with the traditional risk factors alone. These preliminary data suggest a potential role for the D4DR gene in increasing obesity susceptibility.