We identified peak annual incidence rates for medical and nonmedical use of prescription opioid analgesics, stimulants, sedatives, and anxiolytics (controlled medication), and explored cohort effects ...on age of initiation.
Data were gathered retrospectively between 2009 and 2012 from Detroit area students (n = 5185). Modal age at the last assessment was 17 years. A meta-analytic approach produced age-, year-, and cohort-specific risk estimates of first-time use of controlled medication. Cox regression models examined cohort patterns in age of initiation for medical and nonmedical use with any of 4 classes of controlled medication (opioid analgesics, stimulants, sedatives, or anxiolytics).
Peak annual incidence rates were observed at age 16 years, when 11.3% started medical use, and 3.4% started using another person's prescription for a controlled medication (ie, engaged in nonmedical use). In the more recent birth cohort group (1996-2000), 82% of medical users and 76% of nonmedical users reported initiating such use by age 12 years. In contrast, in the less recent birth cohort group (1991-1995), 42% of medical users and 35% of nonmedical users initiated such use by age 12 years. Time to initiation was 2.6 times less in the more recent birth cohort group (medical use: adjusted hazard ratio aHR = 2.57 95% confidence interval {CI} = 2.32-2.85; nonmedical use: aHR = 2.57 95% CI = 2.17-3.03).
Peak annual incidence rates were observed at age 16 years for medical and nonmedical use. More recent cohorts reported initiating both types of use at younger ages. Earlier interventions may be needed to prevent adolescent nonmedical use of controlled medication.
Cells of hematopoietic origin express high levels of the immunoproteasome, a cytokine‐inducible proteasome variant comprising the proteolytic subunits LMP2 (β1i), MECL‐1 (β2i), and LMP7 (β5i). ...Targeting the immunoproteasome in pre‐clinical models of autoimmune diseases with the epoxyketone inhibitor ONX 0914 has proven to be effective. ONX 0914 was previously described as a selective LMP7 inhibitor. Here, we show that PRN1126, developed as an exclusively LMP7‐specific inhibitor, has limited effects on IL‐6 secretion, experimental colitis, and experimental autoimmune encephalomyelitis (EAE). We demonstrate that prolonged exposure of cells with ONX 0914 leads to inhibition of both LMP7 and LMP2. Co‐inhibition of LMP7 and LMP2 with PRN1126 and LMP2 inhibitors LU‐001i or ML604440 impairs MHC class I cell surface expression, IL‐6 secretion, and differentiation of naïve T helper cells to T helper 17 cells, and strongly ameliorates disease in experimental colitis and EAE. Hence, co‐inhibition of LMP2 and LMP7 appears to be synergistic and advantageous for the treatment of autoimmune diseases.
Synopsis
Simultaneous targeting of the immunoproteasome subunits LMP2 and LMP7 is required for optimal therapeutic efficacy in the treatment of autoimmunity. These findings will promote the design of novel immunoproteasome inhibitors with optimal therapeutic efficacy.
Simultaneously targeting the immunoproteasome subunits LMP2 and LMP7 blocks autoimmunity.
Co‐inhibition of LMP2 and LMP7 is required to reduce MHC‐I surface expression, IL‐ 6 production, and Th17 differentiation.
Co‐inhibition strongly ameliorates disease phenotypes of experimental colitis and EAE.
Simultaneous targeting of the immunoproteasome subunits LMP2 and LMP7 is required for optimal therapeutic efficacy in the treatment of autoimmunity. These findings will promote the design of novel immunoproteasome inhibitors with optimal therapeutic efficacy.
Background The benefit of radical prostatectomy in patients with early prostate cancer has been assessed in only one randomized trial. In 2005, we reported that radical prostatectomy improved ...prostate cancer survival compared with watchful waiting after a median of 8.2 years of follow-up. We now report results after 3 more years of follow-up. Methods From October 1, 1989, through February 28, 1999, 695 men with clinically localized prostate cancer were randomly assigned to radical prostatectomy (n = 347) or watchful waiting (n = 348). Follow-up was complete through December 31, 2006, with histopathologic review and blinded evaluation of causes of death. Relative risks (RRs) were estimated using the Cox proportional hazards model. Statistical tests were two-sided. Results During a median of 10.8 years of follow-up (range = 3 weeks to 17.2 years), 137 men in the surgery group and 156 in the watchful waiting group died (P = .09). For 47 of the 347 men (13.5%) who were randomly assigned to surgery and 68 of the 348 men (19.5%) who were not, death was due to prostate cancer. The difference in cumulative incidence of death due to prostate cancer remained stable after about 10 years of follow-up. At 12 years, 12.5% of the surgery group and 17.9% of the watchful waiting group had died of prostate cancer (difference = 5.4%, 95% confidence interval CI = 0.2 to 11.1%), for a relative risk of 0.65 (95% CI = 0.45 to 0.94; P = .03). The difference in cumulative incidence of distant metastases did not increase beyond 10 years of follow-up. At 12 years, 19.3% of men in the surgery group and 26% of men in the watchful waiting group had been diagnosed with distant metastases (difference = 6.7%, 95% CI = 0.2 to 13.2%), for a relative risk of 0.65 (95% CI = 0.47 to 0.88; P = .006). Among men who underwent radical prostatectomy, those with extracapsular tumor growth had 14 times the risk of prostate cancer death as those without it (RR = 14.2, 95% CI = 3.3 to 61.8; P < .001). Conclusion Radical prostatectomy reduces prostate cancer mortality and risk of metastases with little or no further increase in benefit 10 or more years after surgery.
Nuclear receptors are transcription factors with important functions in a variety of physiological and pathological processes. Targeting glucocorticoid receptor (GR) activity using glucocorticoids is ...a cornerstone in the treatment of patients with T cell acute lymphoblastic leukemia (T‐ALL), and resistance to GC‐induced cell death is associated with poor outcome and a high risk for relapse. Next to ligand‐binding, heterodimerization with other transcription factors presents an important mechanism for the regulation of GR activity. Here, we describe a GC‐induced direct association of the Liver Receptor Homolog‐1 (LRH‐1) with the GR in the nucleus, which results in reciprocal inhibition of transcriptional activity. Pharmacological and molecular interference with LRH‐1 impairs proliferation and survival in T‐ALL and causes a profound sensitization to GC‐induced cell death, even in GC‐resistant T‐ALL. Our data illustrate that direct interaction between GR and LRH‐1 critically regulates glucocorticoid sensitivity in T‐ALL opening up new perspectives for developing innovative therapeutic approaches to treat GC‐resistant T‐ALL.
Synopsis
Physical interaction of liver receptor homolog‐1 (LRH‐1) with the glucocorticoid receptor contributes to glucocorticoid (GC) resistance. Treatment with the LRH‐1 inhibitor 3d2 reverses resistance, thereby re‐sensitizing leukemic T cells towards GC‐induced apoptosis.
LRH‐1 inhibits the glucocorticoid receptor (GR) and regulates GC responsiveness of leukemic T cells.
The LRH‐1 inhibitor 3d2 promotes GR induction and upregulation of the pro‐apoptotic BCL‐2 homolog Bim.
Combined treatment of primary patient‐derived T‐ALL cells with GCs and LRH‐1 inhibitor results in synergistic cell death.
LRH‐1 is a critical regulator of T‐ALL cell proliferation and survival.
Physical interaction of liver receptor homolog‐1 (LRH‐1) with the glucocorticoid receptor contributes to glucocorticoid (GC) resistance. Treatment with the LRH‐1 inhibitor 3d2 reverses resistance, thereby re‐sensitizing leukemic T cells towards GC‐induced apoptosis.
Diadenosine polyphosphates (ApnAs) are non‐canonical nucleotides whose cellular concentrations increase during stress and are therefore termed alarmones, signaling homeostatic imbalance. Their ...cellular role is poorly understood. In this work, we assessed ApnAs for their usage as cosubstrates for protein AMPylation, a post‐translational modification in which adenosine monophosphate (AMP) is transferred to proteins. In humans, AMPylation mediated by the AMPylator FICD with ATP as a cosubstrate is a response to ER stress. Herein, we demonstrate that Ap4A is proficiently consumed for AMPylation by FICD. By chemical proteomics using a new chemical probe, we identified new potential AMPylation targets. Interestingly, we found that AMPylation targets of FICD may differ depending on the nucleotide cosubstrate. These results may suggest that signaling at elevated Ap4A levels during cellular stress differs from when Ap4A is present at low concentrations, allowing response to extracellular cues.
Ap4A was investigated for its potential applicability as a cosubstrate in AMPylation mediated by FICD, instead of the common cosubstrate ATP. Proficient AMPylation capability of Ap4A was observed, and new potential target proteins were found by chemical proteomics. Interestingly, AMPylation targets of FICD may differ depending on the cosubstrate.
New treatment options and drug targets for colorectal carcinoma are a pressing medical need. Inflammation and pro-inflammatory cytokines produced by Th1 and Th17 cells like IL-6, TNF, IL-17 and IL-23 ...promote the development and growth of colorectal cancer (CRC). The immunoproteasome is a proteasome subtype highly expressed in immune cells but also in the intestine. Since the immunoproteasome promotes Th1 and Th17 differentiation and pro-inflammatory cytokine production, we investigated here whether deficiency or inhibition of the immunoproteasome subunit LMP7 would interfere with CRC development and exacerbation in preventive and therapeutic mouse models. Treatment with the LMP7 inhibitor ONX 0914 blocked tumor initiation and progression in either chemically-induced (AOM/DSS) or transgenic mouse models (Apc
) of colon carcinogenesis. ONX 0914 treatment strongly reduced tumor numbers and CRC-associated loss of body weight while the survival rates were significantly enhanced. Moreover, genetic LMP7 deficiency markedly reduced the tumor burden in AOM/DSS induced wild type and Apc
mice. In conclusion, we show that the immunoproteasome is involved in CRC development and progression and we identify LMP7 as a new potential drug target for the treatment of CRC.
The extracellular matrix (ECM) represents the natural environment of cells in tissue and therefore is a promising biomaterial in a variety of applications. Depending on the purpose, it is necessary ...to equip the ECM with specific addressable functional groups for further modification with bioactive molecules, for controllable cross‐linking and/or covalent binding to surfaces. Metabolic glycoengineering (MGE) enables the specific modification of the ECM with such functional groups without affecting the native structure of the ECM. In a previous approach (S. M. Ruff, S. Keller, D. E. Wieland, V. Wittmann, G. E. M. Tovar, M. Bach, P. J. Kluger, Acta Biomater. 2017, 52, 159–170), we demonstrated the modification of an ECM with azido groups, which can be addressed by bioorthogonal copper‐catalyzed azide‐alkyne cycloaddition (CuAAC). Here, we demonstrate the modification of an ECM with dienophiles (terminal alkenes, cyclopropene), which can be addressed by an inverse‐electron‐demand Diels‐Alder (IEDDA) reaction. This reaction is cell friendly as there are no cytotoxic catalysts needed. We show the equipment of the ECM with a bioactive molecule (enzyme) and prove that the functional groups do not influence cellular behavior. Thus, this new material has great potential for use as a biomaterial, which can be individually modified in a wide range of applications.
Metabolic glycoengineering was employed to incorporate dienophiles (terminal alkenes and a cyclopropene) into the extracellular matrix (ECM) of adipose‐derived stem cells (ASCs). A subsequent inverse‐electron‐demand Diels‐Alder (IEDDA) reaction allowed the equipment of the ECM with bioactive molecules, opening numerous applications.
Human peripheral neuropathies are poorly understood, and the availability of experimental models limits further research. The PeriTox test uses immature dorsal root ganglia (DRG)-like neurons, ...derived from induced pluripotent stem cells (iPSC), to assess cell death and neurite damage. Here, we explored the suitability of matured peripheral neuron cultures for the detection of sub-cytotoxic endpoints, such as altered responses of pain-related P2X receptors. A two-step differentiation protocol, involving the transient expression of ectopic neurogenin-1 (NGN1) allowed for the generation of homogeneous cultures of sensory neurons. After >38 days of differentiation, they showed a robust response (Ca2+-signaling) to the P2X3 ligand α,β-methylene ATP. The clinical proteasome inhibitor bortezomib abolished the P2X3 signal at ≥5 nM, while 50−200 nM was required in the PeriTox test to identify neurite damage and cell death. A 24 h treatment with low nM concentrations of bortezomib led to moderate increases in resting cell intracellular Ca2+ concentration but signaling through transient receptor potential V1 (TRPV1) receptors or depolarization-triggered Ca2+ influx remained unaffected. We interpreted the specific attenuation of purinergic signaling as a functional cell stress response. A reorganization of tubulin to form dense structures around the cell somata confirmed a mild, non-cytotoxic stress triggered by low concentrations of bortezomib. The proteasome inhibitors carfilzomib, delanzomib, epoxomicin, and MG-132 showed similar stress responses. Thus, the model presented here may be used for the profiling of new proteasome inhibitors in regard to their side effect (neuropathy) potential, or for pharmacological studies on the attenuation of their neurotoxicity. P2X3 signaling proved useful as endpoint to assess potential neurotoxicants in peripheral neurons.
Eps15 homology (EH) domains are universal interaction domains to establish networks of protein–protein interactions in the cell. These networks mainly coordinate cellular functions including ...endocytosis, actin remodeling, and other intracellular signaling pathways. They are well characterized in structural terms, except for the internal EH domain from human γ‐synergin (EHγ). Here, we complete the family of EH domain structures by determining the solution structure of the EHγ domain. The structural ensemble follows the canonical EH domain fold and the identified binding site is similar to other known EH domains. But EHγ differs significantly in the N‐ and C‐terminal regions. The N‐terminal α‐helix is shortened compared to known homologues, while the C‐terminal one is fully formed. A significant proportion of the remaining N‐ and C‐terminal regions are well structured, a feature not seen in other EH domains. Single mutations in both the N‐terminal and the C‐terminal structured extensions lead to the loss of the distinct three‐dimensional fold and turn EHγ into a molten globule like state. Therefore, we propose that the structural extensions in EHγ function as a clamp and are undoubtedly required to maintain its tertiary fold.
PDB Code(s): 2MX7;
The aim of this study was to establish the manner in which the LEPR 223, 1019, 492, and 976 gene polymorphisms influence child obesity.We performed a prospective case-control study on 264 ...hospitalized children from Romania (Nutrichild study) whom we divided into 2 groups: Group I -143 controls and Group II-121 obese children.The 2 groups were evaluated regarding the anthropometry (MUAC, TST, H/L, hip, and abdominal circumference), paraclinical results (protein, leptin, adiponectin, TNF alfa, IL 6, IL 8, VEGF, protein, albumin) and LEPR 223, 1019, 492, and 976 gene polymorphisms. We noticed that the most frequent genotypes in obese children were AG+GG for LEPR 223 gene (P = 0.0001) and GA+AA for LEPR 1019 gene (P = 0.0001), whereas LEPR 492 and LEPR 976 gene polymorphisms did not correlate with obesity. MUAC, TST, H/L, leptin, and adiponectin were correlated with the GG genotype of the LEPR 223 gene, whereas the AG genotype correlated with TNF alpha and serum IL 8. Hip and abdominal perimeters were higher in LEPR 1019 AA genotype carriers, whereas TNF alpha and IL 6 correlated with the GG genotype of the same gene. Obesity did not correlate with protein serum levels.We observed that obesity is more frequent in children with LEPR 223 AG+GG and LEPR 1019 GA+AA genotypes. In obese children LEPR 223/492/1019 AG/GG/GA, GG/GG/GA and AA/GG/GA combined genotypes are more frequent.
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