Probing the Interactions of Thiazole Abietane Inhibitors with the Human Serine Hydrolases ABHD16A and ABHD12.

12-Thiazole abietanes are highly selective reversible inhibitors of hABHD16A that could potentially alleviate neuroinflammation. In this study, we used synthetic chemistry, competitive activity-based protein profiling, and computational methodologies to try to establish relevant structural determinants of activity and selectivity of this class of compounds for inhibiting ABHD16A over ABHD12. Five compounds significantly inhibited hABHD16A but also very efficiently discriminated between inhibition of hABHD16A and hABHD12, with compound 35 being the most effective, at 100 µM (55.1 ± 8.7%; p < 0.0001). However, an outstanding switch in the selectivity toward ABHD12 was observed in the presence of a ring A ester, if the C2' position of the thiazole ring possessed a 1-hydroxyethyl group, as in compound 28. Although our data were inconclusive as to whether the observed enzyme inhibition is allosteric or not, we anticipate that the structure-activity relationships presented herein will inspire future drug discovery efforts in this field.

A longitudinal study of restless legs symptoms among patients with depression.

BACKGROUND: The aim of this study was to analyse the relationship between depressive symptoms and clinical depression and restless legs symptoms in a longitudinal primary care setting. METHODS: The prevalence of restless legs symptoms at baseline and after a six-year follow-up was studied in 474 patients with depressive symptoms and 333 population-based control subjects without depressive symptoms. Depressive symptoms at the baseline and after the six-year follow-up were evaluated with the Beck Depression Inventory (BDI) Second Edition. A psychiatric diagnosis was confirmed with a diagnostic interview (M.I.N.I.). Statistical comparisons between groups were made using analysis of variance (ANOVA) for continuous variables and a chi-square test or logistic models for categorical variables. Repeated measures were analysed using generalizing estimating equations (GEE) models. RESULTS: At baseline the prevalence of restless legs symptoms was 24.3% in control subjects, 43.8% in the patients with depressive symptoms without a depression diagnosis, and 49.3% in clinically depressed patients. During the follow-up up the prevalence of restless legs symptoms declined significantly (p = 0.003). In addition to baseline restless legs symptoms, the prognostic factors for restless legs symptoms among patients with clinical depression were age and BDI score. In the control subjects, moderate and high leisure time physical activity was inversely associated with restless legs symptoms at the follow-up. CONCLUSIONS: A higher level of baseline depressive symptoms was a risk factor for restless legs symptoms in patients with clinical depression. In the prevention and treatment of restless legs symptoms among the patients with depression, the priority is the effective treatment of depression.

Mitoxantrone, pixantrone and mitoxantrone (2-hydroxyethyl)piperazine are toll-like receptor 4 antagonists, inhibit NF-κB activation, and decrease TNF-alpha secretion in primary microglia.

Toll-like receptor 4 (TLR4) recognizes various endogenous and microbial ligands and is an essential part in the innate immune system. TLR4 signaling initiates transcription factor NF-κB and production of proinflammatory cytokines. TLR4 contributes to the development or progression of various diseases including stroke, neuropathic pain, multiple sclerosis, rheumatoid arthritis and cancer, and better therapeutics are currently sought for these conditions. In this study, a library of 140 000 compounds was virtually screened and a resulting hit-list of 1000 compounds was tested using a cellular reporter system. The topoisomerase II inhibitor mitoxantrone and its analogues pixantrone and mitoxantrone (2-hydroxyethyl)piperazine were identified as inhibitors of TLR4 and NF-κB activation. Mitoxantrone was shown to bind directly to the TLR4, and pixantrone and mitoxantrone (2-hydroxyethyl)piperazine were shown to inhibit the production of proinflammatory cytokines such as tumor necrosis factor alpha (TNFα) in primary microglia. The inhibitory effect on NF-κB activation or on TNFα production was not mediated through cytotoxity at ≤ 1 µM concentration for pixantrone and mitoxantrone (2-hydroxyethyl)piperazine treated cells, as assessed by ATP counts. This study thus identifies a new mechanism of action for mitoxantrone, pixantrone, and mitoxantrone (2-hydroxyethyl)piperazine through the TLR4.

The influence of restless legs symptoms on musculoskeletal pain in depression.

Background and aims Restless legs syndrome is a sensorimotor disorder associated with mental health conditions notably depression. Restless legs symptoms and depression are commonly associated with pain. The study investigated the influence of restless legs symptoms on musculoskeletal pain in patients with depression or with increased depressive symptoms. Methods A cross-sectional study of primary care patients in the Central Finland Hospital District. The prevalence of restless legs symptoms was studied in patients with depressive symptoms (n = 695) and controls without a psychiatric diagnosis (n = 410) by using a structured questionnaire. The depressive symptoms were evaluated with the Beck Depression Inventory and the psychiatric diagnosis was confirmed by means of a diagnostic interview (MINI). The prevalence and intensity of musculoskeletal pain was captured with form-based questions. A single-question screen for restless legs symptoms was applied. Results There was a significant difference in the prevalence of continuous widespread musculoskeletal pain between the three study groups: the controls 4.6% (95% CI: 2.8-7.1), the patients with symptoms of depression without a diagnosis 16.0% (11.7-21.1), and the patients with diagnosed depression 22.1% (18.3-23.3) (p = 0.006 after being adjusted for age, sex, smoking, use of alcohol, education years, body mass index, use of antidepressants, and physical activity, after multiple corrections, all groups were significantly different from each other). Compared with those not having restless legs symptoms, subjects with restless legs symptoms had more often continuous widespread musculoskeletal pain in the control subjects (p = 0.001; 2.3% vs. 10.5%) and in the patients with depressive symptoms without a depression diagnosis (p = 0.024; 9.1 vs. 18.7%) but not in those with diagnosed depression (p = 0.98; 19.5 vs. 19.4%). The restless legs symptoms were associated with the intensity of pain in all groups (p < 0.001). Conclusions Restless legs symptoms were related to continuous widespread musculoskeletal pain in subjects without depressive symptoms and in patients with depressive symptoms without a depression diagnosis. Pain intensity was higher in the subjects with restless legs symptoms regardless of depressive symtoms or depression. Implications Clinical management of pain in patients with restless legs symptoms should include an increased focus on the prevention and treatment of either conditions.

Pyrazine-Fused Triterpenoids Block the TRPA1 Ion Channel in Vitro and Inhibit TRPA1-Mediated Acute Inflammation in Vivo.

TRPA1 is a nonselective cation channel, most famously expressed in nonmyelinated nociceptors. In addition to being an important chemical and mechanical pain sensor, TRPA1 has more recently appeared to have a role also in inflammation. Triterpenoids are natural products with anti-inflammatory and anticancer effects in experimental models. In this paper, 13 novel triterpenoids were created by synthetically modifying betulin, an abundant triterpenoid of the genus Betula L., and their TRPA1-modulating properties were examined. The Fluo 3-AM protocol was used in the initial screening, in which six of the 14 tested triterpenoids inhibited TRPA1 in a statistically significant manner. In subsequent whole-cell patch clamp recordings, the two most effective compounds (pyrazine-fused triterpenoids 8 and 9) displayed a reversible and dose- and voltage-dependent effect to block the TRPA1 ion channel at submicromolar concentrations. Interestingly, the TRPA1 blocking action was also evident in vivo, as compounds 8 and 9 both alleviated TRPA1 agonist-induced acute paw inflammation in mice. The results introduce betulin-derived pyrazine-fused triterpenoids as promising novel antagonists of TRPA1 that are potentially useful in treating diseases with a TRPA1-mediated adverse component.

Psychiatric diagnosis in primary care patients with increased depressive symptoms.

BACKGROUND: Screening of depression has been recommended in primary care and Beck's 21-item Depression Inventory (BDI-21) is a commonly used tool for screening. Depression has been shown to be frequently accompanied by comorbidities. AIMS: This study aimed to analyze the characteristics, psychiatric diagnoses, and psychiatric comorbidity of primary care patients who have been screened for depression and referred to a depression nurse. METHODS: The study subjects were primary care patients aged ≥ 35 years with depressive symptoms (BDI-21 > 9). Their psychiatric diagnosis were based on a diagnostic interview (Mini-International Neuropsychiatric Interview; M.I.N.I.) conducted by a trained study nurse. RESULTS: Of the 705 study subjects, 617 (87.5%) had at least one and 66.1% had at least two psychiatric diagnoses. The most common diagnosis was depression (63.4%). The next most common diagnoses were generalized anxiety disorder (GAD) (48.1%) and panic disorder (22.8%). Only 8.8% of the study subjects had depression without other psychiatric disorders. Ten percent of the subjects had both depression and a generalized anxiety disorder (GAD). Also other psychiatric comorbidities were common. Age was inversely associated with the psychiatric diagnosis in the M.I.N.I. CONCLUSIONS: This study suggests that most of the primary care patients with increased depressive symptoms have a psychiatric disorder. Although depression is the most common diagnosis, there are several other concurrent psychiatric comorbidities. Therefore, diagnostic assessment of primary care patients with a screening score over 9 in the BDI-21 should be reconsidered.

Elevation of tumor necrosis factor alpha levels is associated with restless legs symptoms in clinically depressed patients.

BACKGROUND: Restless legs syndrome is a sensorimotor disorder associated with several mental illnesses particularly depression. METHODS: A cross-sectional study of primary care patients. The prevalence of restless legs symptoms was studied in 706 patients with depressive symptoms and 426 controls without a psychiatric diagnosis by using a structured questionnaire. The depressive symptoms were evaluated with the BDI and the psychiatric diagnosis was confirmed by means of a diagnostic interview (M.I.N.I.). The subjects with elevated depressive symptoms were divided into two groups subjects with depressive symptoms with and without clinical depression. RESULTS: The prevalence of restless legs symptoms was 24.8% in the controls, 50.0% in the patients with clinical depression and 42.4% in the patients with depressive symptoms. CRP value was significantly higher (p = .003) in the clinically depressed patients than in the other groups. There was a higher concentration of TNF-α in the subjects with restless legs symptoms (7.4 ng/l ±â€¯3.2) compared with the subjects without symptoms (6.7 ng/l ±â€¯2.3)(p < .001). There was a significant difference in the TNF-α levels between the subjects with and without restless legs symptoms in the depression group (p < .001) and among the patients with depressive symptoms but no a depression diagnosis (p = .022). In these groups, restless legs symptoms were associated with elevated levels of TNF-α. CONCLUSIONS: TNF-α level was associated with restless legs symptoms only among subjects with depressive symptoms whether they had clinical depression or not. We suggest that TNF-α could be an underlying factor between restless legs symptoms and comorbidities.

Synthesis of 7ß-hydroxy-8-ketone opioid derivatives with antagonist activity at mu- and delta-opioid receptors.

Despite extensive years of research, the direct oxidation of the 7,8-double bond of opioids has so far received little attention and knowledge about the effects of this modification on activity at the different opioid receptors is scarce. We herein report that potassium permanganate supported on iron(II) sulfate heptahydrate can be used as a convenient oxidant in the one-step, heterogeneous conversion of Δ7,8-opioids to the corresponding 7ß-hydroxy-8-ketones. Details of the reaction mechanism are given and the effects of the substituent at position 6 of several opioids on the reaction outcome is discussed. The opioid hydroxy ketones prepared are antagonists at the mu- and delta-opioid receptors. Docking simulations and detailed structure-activity analysis revealed that the presence of the 7ß-hydroxy-8-ketone functionality in the prepared compounds can be used to gain activity towards the delta opioid receptor. The 7ß-hydroxy-8-ketones prepared with this method can also be regarded as versatile intermediates for the synthesis of other opioids of interest.

Discovery of 12-Thiazole Abietanes as Selective Inhibitors of the Human Metabolic Serine Hydrolase hABHD16A.

Screening of an in-house library of compounds identified 12-thiazole abietanes as a new class of reversible inhibitors of the human metabolic serine hydrolase. Further optimization of the first hit compound lead to the 2-methylthiazole derivative 18, with an IC50 value of 3.4 ± 0.2 µM and promising selectivity. ABHD16A has been highlighted as a new target for inflammation-mediated pain, although selective inhibitors of hABHD16A (human ABHD16A) have not yet been reported. Our study presents abietane-type diterpenoids as an attractive starting point for the design of selective ABHD16A inhibitors, which will contribute toward understanding the significance of hABHD16A inhibition in vivo.

Prevalence of restless legs symptoms according to depressive symptoms and depression type: a cross-sectional study.

BACKGROUND: Restless legs syndrome is a sensorimotor disorder and it is associated with several other diseases especially mental illnesses. AIMS: To analyze the relationship between the symptoms of restless legs syndrome and the severity of depressive symptoms and the prevalence of restless legs symptoms in depression subtypes. METHODS: A cross-sectional study of primary care patients in the Central Finland Hospital District. The prevalence of restless legs symptoms was studied in 706 patients with increased depressive symptoms and 426 controls without a psychiatric diagnosis by using a structured questionnaire. The depressive symptoms were evaluated with the Beck Depression Inventory (BDI) and the psychiatric diagnosis was confirmed by means of a diagnostic interview (Mini-International Neuropsychiatric Interview). The subjects with increased depressive symptoms were divided into three groups (subjects with depressive symptoms without a depression diagnosis, melancholic depression and non-melancholic depression). RESULTS: In the whole study population, the prevalence of restless legs symptoms increased with the severity of depressive symptoms. The prevalence of restless legs symptoms was highest in the melancholic and non-melancholic depressive patients (52 and 46%, respectively) and then in subjects with depressive symptoms without a depression diagnosis (43.4%), but the prevalence was also substantial (24.6%) in subjects without a psychiatric diagnosis. CONCLUSIONS: Restless legs symptoms are very common in primary care among subjects with depression, regardless of the depression type. The prevalence of restless legs symptoms increased with increasing severity of depressive symptoms, regardless of the diagnosis. These findings should be considered in clinical evaluation and treatment of patients visiting their physician due to restless legs or depressive symptoms.

Sex differences in the association of adiponectin and low-grade inflammation with changes in the body mass index from youth to middle age.

BACKGROUND: There are sex differences in low-grade inflammation markers in obesity-related disorders. Little is known, however, about a possible sex-specific association of relative weight change from youth to adulthood with actual low-grade inflammation. OBJECTIVE: The aim of this study was to identify possible sex differences in adiponectin, interleukin-1ß (IL-1ß), interleukin-1Ra (IL-1Ra), and high-sensitivity C-reactive protein (hs-CRP) levels with respect to the relative change in body mass index (BMI) from youth to middle age. METHODS: The study population consisted of 403 men and 500 women from 1 Finnish town. Weight, height, and adiponectin, IL-1ß, IL-1Ra, and hs-CRP levels were measured in 2003 at a mean age of 46 years. Self-reported weight at the age of 20 years was recorded. RESULTS: In women, even after adjustment for BMI in adulthood, a statistically significantly negative linear association was observed between the quartiles of relative change in BMI and adiponectin levels (P < 0.001 for linearity). Significantly positive linear associations were also observed between the change in BMI and IL-1Ra (P = 0.032 for linearity) and hs-CRP (P = 0.029 for linearity) levels. In men, there was no statistically significant association among the quartiles of relative change in BMI and measured inflammatory markers after adjustment for BMI in adulthood. CONCLUSIONS: A relative increase in weight may be more harmful in women than in men with respect to adiponectin and inflammatory markers.

Gender differences relating to metabolic syndrome and proinflammation in Finnish subjects with elevated blood pressure.

Fasting insulin, adiponectin, high-sensitivity C-reactive protein (hs-CRP), and interleukin-1 receptor antagonist (IL-1Ra) were determined in 278 men and 273 women with blood pressure > or = 130 and/or > or = 85 mmHg and/or with antihypertensive medication. Metabolic syndrome (MetS) with the National Cholesterol Education Program (NCEP) criteria was observed in 35% of men and 34% of women. Men with MetS had lower hs-CRP and IL-1Ra than women. The absolute gender difference in adiponectin was smaller and those in IL-1Ra and hs-CRP were greater in subjects with MetS compared to those without. After adjustment with body mass index the association between insulin and the odd's ratio (OR) for MetS remained significant in both genders, in females also the association between the OR for MetS and adiponectin. There are gender differences in subjects with elevated blood pressure and MetS with respect to inflammatory markers and the relationship between adiponectin levels and MetS.

Gender difference among smoking, adiponectin, and high-sensitivity C-reactive protein.

BACKGROUND: Subclinical inflammation is a novel risk factor of cardiovascular diseases and type 2 diabetes. An inverse association between plasma adiponectin and insulin resistance has been previously shown. Elevated levels of high-sensitivity C-reactive protein (hs-CRP) predict future cardiovascular events. Smoking has been proven to connect with inflammatory markers. There is also evidence of a difference between genders in pro-inflammation. This study aimed to examine the connections among adiponectin, hs-CRP, and smoking and to determine possible gender differences in these associations. METHODS: Included were 365 men and 476 women; all were nondiabetic and middle-aged. Daily smoking subjects were considered to be smokers. Adiponectin and hs-CRP were analyzed. Data were collected in 1997-1998, and cytokines were analyzed in 2003. RESULTS: Thirty-five percent of the men and 22% of the women were smokers. In women, the adiponectin level was significantly lower in smokers (6.94+/-3.27 microg/ml) compared to nonsmokers (8.27+/-4.72 microg/ml, p=0.0017). This association remained significant after adjustment for age and BMI (p=0.0061). The hs-CRP level was significantly higher in smoking men (1.59+/-1.71 pg/ml) compared to nonsmoking men (1.17+/-1.41 pg/ml, p=0.018). This result remained after adjustment for age and BMI (p=0.0056). When smokers were compared to nonsmokers, there was no difference in adiponectin among men or in hs-CRP among women. CONCLUSIONS: In the nondiabetic population, smoking associates differently with subclinical inflammation between genders, with a decreased adiponectin level in women and with an increased hs-CRP level in men.