Proteolytic activity of the mucosa-associated lymphoid tissue lymphoma translocation protein-1 (MALT1) paracaspase is required for survival of the activated B cell subtype of diffuse large B cell lymphoma (ABC-DLBCL). We have identified distinct derivatives of medicinal active phenothiazines, namely mepazine, thioridazine, and promazine, as small molecule inhibitors of the MALT1 protease. These phenothiazines selectively inhibit cleavage activity of recombinant and cellular MALT1 by a noncompetitive mechanism. Consequently, the compounds inhibit anti-apoptotic NF-κB signaling and elicit toxic effects selectively on MALT1-dependent ABC-DLBCL cells in vitro and in vivo. Our data provide a conceptual proof for a clinical application of distinct phenothiazines in the treatment of ABC-DLBCL.

These findings have important implications for classification and treatment of affective disorders. The greater stability of comorbid anxiety and depression than either disorder alone illustrates the importance of further investigation of comorbid states compared with noncomorbid states in etiologic and treatment research. The persistence of subthreshold-level depression and anxiety from early to mid adulthood also suggests the importance of characterizing the continuum of expression of depression and anxiety rather than adhering to strict diagnostic thresholds.

SUMMARY. Background -In order to minimise retrospective recall in developing estimates of the prevalence of mental disorders in the general population, we conducted a prospective study of a cohort of youth from Zurich, Switzerland. Method -A 20 year prospective study of a community-based cohort aged 19-20 from Zurich Switzerland. The sample was enriched by subjects scoring high on the Symptom Checklist 90 R (Derogatis, 1977). A semi-structured diagnostic interview was administered by clinically experienced psychologists and psychiatrists. The six interviews from 1979 to 1999 assessed diagnoses and sub-threshold manifestations of major diagnostic categories (with the exception of schizophrenia) for the past twelve months, depending on the current DSM versions (DSM-IH, DSM-HI R, DSM-IV). Additional information on symptoms and treatment were collected for the years between the interviews. The reported prevalence rates are weighted for stratified sampling and cumulate the one-year rates of the six interviews. Results -The cumulative weighted prevalence rates for any psychiatric disorder were 48.6% excluding, and 57.7% including tobacco dependence. In addition 29.2% and 21.8%, respectively manifested sub-diagnostic syndromes. Overall there were no significant gender differences. The corresponding treatment prevalence rates were 22.4% and 31.1%, respectively for the diagnostic subjects and 6.9% and 6.1 %, respectively for the sub-diagnostic groups. The total treatment prevalence rate was 37.2% of the population (males 30.0%, females 44.1%). Conclusions -Our findings reveal that psychiatric disorders are quite common in the general population. When the spectra of mental disorders are considered, nearly three quarters of the general population will have manifested at least one of the mental disorders across their lifetime. Limitations -The data are based on a relatively small sample; a single age cohort, and the study was conducted in Zurich, Switzerland. These study features may diminish the generalisability of the findings.

Wnt/b-catenin signaling is a highly conserved pathway essential for embryogenesis and tissue homeostasis. However, deregulation of this pathway can initiate and promote human malignancies, especially of the colon and head and neck. Therefore, Wnt/b-catenin signaling represents an attractive target for cancer therapy. We performed high-throughput screening using AlphaScreen and ELISA techniques to identify small molecules that disrupt the critical interaction between b-catenin and the transcription factor TCF4 required for signal transduction. We found that compound LF3, a 4-thioureido-benzenesulfonamide derivative, robustly inhibited this interaction. Biochemical assays revealed clues that the core structure of LF3 was essential for inhibition. LF3 inhibited Wnt/b-catenin signals in cells with exogenous reporters and in colon cancer cells with endogenously high Wnt activity. LF3 also suppressed features of cancer cells related to Wnt signaling, including high cell motility, cell-cycle progression, and the overexpression of Wnt target genes. However, LF3 did not cause cell death or interfere with cadherinmediated cell-cell adhesion. Remarkably, the self-renewal capacity of cancer stem cells was blocked by LF3 in concentrationdependent manners, as examined by sphere formation of colon and head and neck cancer stem cells under nonadherent conditions. Finally, LF3 reduced tumor growth and induced differentiation in a mouse xenograft model of colon cancer. Collectively, our results strongly suggest that LF3 is a specific inhibitor of canonical Wnt signaling with anticancer activity that warrants further development for preclinical and clinical studies as a novel cancer therapy. Cancer Res; 76(4); 891-901. Ó2015 AACR.

Rubredoxins (Rds) are essential electron transfer components of bacterial membrane-bound alkane hydroxylase systems. Several Rd genes associated with alkane hydroxylase or Rd reductase genes were cloned from gram-positive and gram-negative organisms able to grow on n-alkanes (Alk-Rds). Complementation tests in an Escherichia coli recombinant containing all Pseudomonas putida GPo1 genes necessary for growth on alkanes except Rd 2 (AlkG) and sequence comparisons showed that the Alk-Rds can be divided in AlkG1-and AlkG2-type Rds. All alkane-degrading strains contain AlkG2-type Rds, which are able to replace the GPo1 Rd 2 in n-octane hydroxylation. Most strains also contain AlkG1-type Rds, which do not complement the deletion mutant but are highly conserved among gram-positive and gram-negative bacteria. Common to most Rds are the two iron-binding CXXCG motifs. All Alk-Rds possess four negatively charged residues that are not conserved in other Rds. The AlkG1-type Rds can be distinguished from the AlkG2-type Rds by the insertion of an arginine downstream of the second CXXCG motif. In addition, the glycines in the two CXXCG motifs are usually replaced by other amino acids. Mutagenesis of residues conserved in either the AlkG1-or the AlkG2-type Rds, but not between both types, shows that AlkG1 is unable to transfer electrons to the alkane hydroxylase mainly due to the insertion of the arginine, whereas the exchange of the glycines in the two CXXCG motifs only has a limited effect.Rubredoxins (Rds) are the simplest of the iron-sulfur redox active proteins and usually contain a single Fe(S-Cys) 4 site. In the aerobic bacterium Pseudomonas putida GPo1, Rd is an essential component of the alkane hydroxylase system. It shuttles electrons from Rd reductase, a protein which reduces its flavin adenine dinucleotide at the expense of NADH, to alkane hydroxylase (19,21,26,28,39). The alkane hydroxylase belongs to a large class of integral membrane proteins (9, 42) that contain an oxobridged diiron cluster (32) and is of interest as a biocatalyst for the production of alcohols, fatty acids, and epoxides (24,44).Sequencing of the GPo1 alk genes, reviewed in reference 43, showed that the OCT plasmid of GPo1 encodes two Rds, AlkF and AlkG (14), the second of which has been studied in detail in earlier biochemical studies (see below). AlkG is unusual in that it is more than three times the size of other bacterial Rds. It consists of two Rd domains, AlkG1 and AlkG2, connected by a 70-amino-acid linker which shows no homology to other sequences in the Swiss-Prot database (3, 14, 27). AlkF has not been isolated or detected. The primary sequence of AlkF consists of an Rd-like N-terminal domain (AlkF1) and an 80-amino-acid C-terminal extension. AlkF1 and the N-terminal domain of AlkG (AlkG1) are more closely related to each other than to the C-terminal domain of AlkG (AlkG2) (see Fig. 3). Only plasmids which encode AlkG2 were able to restore growth on n-alkanes in P. putida carrying a CAM-OCT alkA mutant plasmid (14). Two Rd genes cloned ...

The skin is equipped with specialized mechanoreceptors that allow the perception of the slightest brush. Indeed, some mechanoreceptors can detect even nanometer-scale movements. Movement is transformed into electrical signals via the gating of mechanically activated ion channels at sensory endings in the skin. The sensitivity of Piezo mechanically gated ion channels is controlled by stomatin-like protein-3 (STOML3), which is required for normal mechanoreceptor function. Here we identify small-molecule inhibitors of STOML3 oligomerization that reversibly reduce the sensitivity of mechanically gated currents in sensory neurons and silence mechanoreceptors in vivo. STOML3 inhibitors in the skin also reversibly attenuate fine touch perception in normal mice. Under pathophysiological conditions following nerve injury or diabetic neuropathy, the slightest touch can produce pain, and here STOML3 inhibitors can reverse mechanical hypersensitivity. Thus, small molecules applied locally to the skin can be used to modulate touch and may represent peripherally available drugs to treat tactile-driven pain following neuropathy.

Success in small molecule screening relies heavily on the preselection of compounds. Here, we present a strategy for the enrichment of chemical libraries with potentially bioactive compounds integrating the collected knowledge of medicinal chemistry. Employing a genetic algorithm, substructures typically occurring in bioactive compounds were identified using the World Drug Index. Availability of compounds containing the selected substructures was analysed in vendor libraries, and the substructure-specific sublibraries were assembled. Compounds containing reactive, undesired functional groups were omitted. Using a diversity filter for both physico-chemical properties and the substructure composition, the compounds of all the sublibraries were ranked. Accordingly, a screening collection of 16,671 compounds was selected. Diversity and chemical space coverage of the collection indicate that it is highly diverse and well-placed in the chemical space spanned by bioactive com-

Combining tunable transcription with an enzyme-degradation tag affords an effective means to reduce intracellular enzyme concentrations from high to very low levels. Such fine-tuned control allows selection pressure to be systematically increased in directed-evolution experiments. This facilitates identification of mutants with wild-type activity, as shown here for an engineered chorismate mutase. Numerous selection formats and cell-based screening methodologies may benefit from the large dynamic range afforded by this easily implemented strategy.