This study comprised the first step in the psychometric development of a self-report screening instrument for risk of opioid medication misuse among chronic pain patients. A 26-item instrument, the Pain Medication Questionnaire (PMQ), was constructed based on suspected behavioral correlates of opioid medication misuse, which heretofore have received limited empirical investigation. The PMQ was administered to 184 patients at an interdisciplinary pain treatment center. Reliability coefficients for the PMQ were found to be of moderate but acceptable strength. Construct and concurrent validity were examined through correlation of PMQ scores to measures of substance abuse, physical and psychological functioning, and physicians' risk assessments. To explore high and low cutoff points for misuse risk, subgroups were formed according to the upper and lower thirds of PMQ scores and compared on validity measures. Higher PMQ scores were associated with history of substance abuse, higher levels of psychosocial distress, and poorer functioning. Future psychometric analyses will consider predictive validity and examine shortened versions of the instrument.

Mammalian TOR (mTOR) regulates cell growth, proliferation, and migration. Because mTOR knock-outs are embryonic lethal, we generated a viable hypomorphic mouse by neo-insertion that partially disrupts mTOR transcription and creates a potential physiologic model of mTORC1/ TORC2 inhibition. Homozygous knock-in mice exhibited reductions in body, organ, and cell size. Although reductions in most organ sizes were proportional to decreased body weight, spleens were disproportionately smaller. Decreases in the total number of T cells, particularly memory cells, and reduced responses to chemokines suggested alterations in T-cell homing/homeostasis. T-cell receptor-stimulated T cells proliferated less, produced lower cytokine levels, and expressed FoxP3. Decreased neutrophil numbers were also observed in the spleen, despite normal development and migration in the bone marrow. However, B-cell effects were most pronounced, with a partial block in B-cell development in the bone marrow, altered splenic populations, and decreases in proliferation, antibody production, and migration to chemokines. Moreover, increased AKT Ser473 phosphorylation was observed in activated B cells, reminiscent of cancers treated with rapamycin, and was reduced by a DNA-pk inhibitor. Thus, mTOR is required for the maturation and differentiation of multiple immune cell lineages. These mice provide a novel platform for studying the consequences of constitutively reduced mTORC1/TORC2 activity. IntroductionThe mammalian target of rapamycin (mTOR) is part of a conserved pathway regulating fundamental physiologic functions, including nutrient sensing and metabolism, and cell growth, proliferation, and migration. mTOR forms 2 protein complexes: one with RAPTOR, mLST8(G␤L), and PRAS40 to form TOR complex 1 (mTORC1) involved in phosphorylating S6K and 4EBP1, 1,2 and a second with RICTOR, mLST8(G␤L), SIN1, and PROTOR to form TOR complex 2 (mTORC2), which phosphorylates AKT on Ser473. [2][3][4] In yeast, TOR controls cell proliferation and size. 5,6 In Drosophila, inactivation of dTOR results in lethality and reduced embryo size. 7,8 Genetically targeting the kinase domain of murine mTOR for inactivation results in embryonic lethality, 9-11 although deletions in the C terminal portion yield mice that are normal and fertile. 11 ENU-mutagenesis screens uncovered an additional embryonic lethal mutation of mTOR, resulting in flat-top embryos lacking telencephalons resulting from limited neuroectodermal cell proliferation. 10,12 Knockouts of Raptor or Sin1 13 result in early embryonic lethality, whereas those of Rictor and mLST8(G␤L) lead to late embryonic lethality and defective vascular development. mTOR signaling/function has been deduced from studies with rapamycin, which associates with FKBP12, 14 and together binds mTOR to destabilize mTORC1. Although originally thought to affect only mTORC1, long-term treatment with rapamycin can also affect mTORC2. 15 Rapamycin and numerous rapalogs are potent immunosuppressants used in cancer chemotherapy and bone marro...

This study describes suicidal behavior in a cross-sectional sample of chronic pain patients and evaluates factors associated with increased risk for suicidal ideation. One hundred-fifty-three adults with nonmalignant pain (42% back pain) who were consecutively referred to a tertiary care pain center completed a Structured Clinical Interview for Suicide History, the McGill Pain Questionnaire, and the Beck Depression Inventory. Nineteen-percent reported current passive suicidal ideation (PSI), 13% had active thoughts of committing suicide (ASI), 5% had a current suicide plan, and 5% reported a previous suicide attempt. Drug overdose was the most commonly reported plan and method of attempt (75%). Thirteen-percent reported a family history of suicide attempt/completion. Pain-specific and traditional suicide risk factors were evaluated as predictors of current PSI and ASI. Logistic regression analyses revealed that a family history of suicide attempts/completions was associated with a 7.5 fold increase in risk of PSI (P=0.001) and a 6.6 fold increase in ASI (P=0.003), after adjusting for significant covariates. Having abdominal pain was associated with an adjusted 5.5 fold increase in PSI (P=0.05) and a 4.2 fold increase in ASI (P=0.10). Neuropathic pain significantly reduced risk for both PSI (P=0.002) and ASI (P=0.01). Demographics, pain severity, and depression severity were not associated with suicidal ideation in multivariate analyses. These findings highlight the need for routine evaluation and monitoring of suicidal behavior in chronic pain, especially for patients with family histories of suicide, those taking potentially lethal medications, and patients with abdominal pain.

We have synthesized three peptides from the mdm-2 binding domain of human p53, residues 12-26 (PPLSQETFSDLWKLL), residues 12-20, and 17-26. To enable transport of the peptides across the cell membrane and at the same time to maximize the active mdm-2 binding ␣-helical conformation for these peptides, each was attached at its carboxyl terminus to the penetratin sequence, KKWKMRRNQF-WVKVQRG, that contains many positively charged residues that stabilize an ␣-helix when present on its carboxyl terminal end. All three peptides were cytotoxic to human cancer cells in culture, whereas a control, unrelated peptide attached to the same penetratin sequence had no effect on these cell lines. The same three cytotoxic peptides had no effect on the growth of normal cells, including human cord blood-derived stem cells. These peptides were as effective in causing cell death in p53-null cancer cells as in those having mutant or normal p53. Peptide-induced cell death is not accompanied by expression of apoptosis-associated proteins such as Bax and waf p21 . Based on these findings, we conclude that the antiproliferative effects of these p53-derived peptides are not completely dependent on p53 activity and may prove useful as general anticancer agents.

The anticarcinogenicity of some flavonoids has been attributed to modulation of the cytochrome P450 enzymes, which metabolize procarcinogens to their activated forms. However, the mechanism by which flavonoids inhibit some P450-mediated activities while activating others is a longstanding, intriguing question. We employed flash photolysis to measure carbon monoxide binding to P450 as a rapid kinetic technique to probe the interaction of the prototype flavonoid ␣-naphthoflavone with human cytochrome P450s 1A1 and 3A4, whose benzo[a]pyrene hydroxylation activities are respectively inhibited and stimulated by this compound. This flavonoid inhibited P450 1A1 binding to benzo-[a]pyrene via a classical competitive mechanism. In contrast, ␣-naphthoflavone stimulated P450 3A4 by selectively binding and activating an otherwise inactive subpopulation of this P450 and promoting benzo-[a]pyrene binding to the latter. These data indicate that flavonoids enhance activity by increasing the pool of active P450 molecules within this P450 macrosystem. Activators in other biological systems may similarly exert their effect by expanding the population of active receptor molecules.Owing to their wide distribution in fruits, vegetables, and grain products, flavonoids are a regularly consumed component of the human diet (1, 2). Increased consumption of these phytochemicals is associated with decreased risk for colon, rectum, and lung cancers (3, 4). Anticarcinogenicity in rodents (5, 6) has been attributed to modulation of the cytochrome P450 enzymes that metabolize xenobiotic and endogenous compounds, including activation of procarcinogens to their carcinogenic forms (7-9). Numerous studies have shown that individual flavonoids may either activate or inhibit P450-mediated activities depending on the target P450 form (10 -12). The mechanism of flavonoid action is unclear yet of intense interest owing to the putative role of dietary flavonoid consumption in P450-mediated chemical carcinogenesis and the potential for development of therapeutic flavonoid modulators of specific P450s. ␣-Naphthoflavone (ANF) 1 is a prototype flavonoid whose inhibition of P450-mediated activities was first reported over 25 years ago (13) and that has subsequently been used to examine the mechanism of flavonoid action (14,15). Most of these studies have assessed the effect of ANF on P450-mediated hydroxylation of the polycyclic aromatic hydrocarbon benzo[a]pyrene (BP), an environmental pollutant present in cigarette smoke and polluted air that is carcinogenic in experimental animals (16).The P450 system metabolizes BP to a variety of products including activated metabolites that covalently bind to cellular macromolecules and initiate carcinogenesis (17,18). Among the human P450s primarily responsible for metabolizing BP (18) are P450s 3A4 and 1A1. P450 3A4 is a major liver P450 that also metabolizes many important drugs (19), whereas P450 1A1 is normally undetectable in human liver but present in lung (20), where it is induced by cigarette smoking and is ass...

Phosphorylation of the p53 tumor suppressor protein is known to modulate its functions. Using bacterially produced glutathione S-transferase (GST)-p53 fusion protein and baculovirus-expressed histidine-tagged p53 ( His p53), we have determined human p53 phosphorylation by purified forms of jun-N-kinase (JNK), protein kinase A (PKA), and ␤ subunit of casein kinase II (CKII␤) as well as by kinases present in whole cell extracts (WCEs). We demonstrate that PKA is potent p53 kinase, albeit, in a conformation-and concentration-dependent manner, as concluded by comparing full-length with truncated forms of p53. We further demonstrate JNK interaction with GST-p53 and the ability of JNK to phosphorylate truncated forms of GST-p53 or full-length His p53. Dependence of phosphorylation on conformation of p53 is further supported by the finding that the wild-type form of p53 (p53 wt ) undergoes better phosphorylation by CKII␤ and by WCE kinases than mutant forms of p53 at amino acid 249 (p53 249 ) or 273 (p53 273 ). Moreover, shifting the kinase reaction's temperature from 37؇C to 18؇C reduces the phosphorylation of mutant p53 to a greater extent than of p53 wt . Comparing truncated forms of p53 revealed that the ability of CKII␤, PKA, or WCE kinases to phosphorylate p53 requires amino acids 97-155 within the DNA-binding domain region. Among three 20-aa peptides spanning this region we have identified residues 97-117 that increase p53 phosphorylation by CKII␤ while inhibiting p53 phosphorylation by PKA or WCE kinases. The importance of this region is further supported by computer modeling studies, which demonstrated that mutant p53 249 exhibits significant changes to the conformation of p53 within amino acids 97-117. In summary, phosphorylation-related analysis of different p53 forms in vitro indicates that conformation of p53 is a key determinant in its availability as a substrate for different kinases, as for the phosphorylation pattern generated by the same kinase.

The proper medicinal use of opioids, in light of their notorious history and current relation to social ills, continues to be debated and remains unclear in several areas of medicine. This article will review several areas and points of controversy related to screening for potential problematic opioid behavior in chronic nonmalignant pain patients. Controversy over the prescription of opioids for chronic nonmalignant pain continues, despite the growing acceptance of this practice. Indeed, past research supports the beneficial use of opioids for noncancer pain. Unfortunately, traditional definitions of abuse and dependence, with their emphasis on tolerance and withdrawal, are inappropriate for chronic pain patients prescribed opioids. The component of traditional definitions of abuse and dependence that appears most applicable to chronic pain patients centers on the criterion that the patient continue to take the drug (in this case, the opioid) despite negative and harmful effects or despite any decrease in pain level. Although clinical observations exist about risk factors for opioid misuse in chronic pain patients, there is limited research. Further, the area of prescreening for problematic drug behavior is in its infancy. However, researchers have begun to delve into this challenging area and the application of rigorous empirical research will bring us closer to identifying those patients at risk so that their pain is managed without destructive outcomes in other areas of their life.

The large number of associations between SF-36 scores and outcome variables highlights the importance of assessing the health-related quality of life of patients, and supports the use of the SF-36 in accomplishing this task. Among the findings, perhaps the most significant was the value of assessing health-related quality of life, particularly the subjective physical components, after completion of a functional restoration program. Prediction of long-term socioeconomic outcomes is likely to be improved if assessment is conducted at the end of the treatment process. SF-36 is recommended for assessing general health status, and more spine-specific measures are recommended for assessing spinal pain and disability variables.