Inhibition of PARP1 improves cardiac function after myocardial infarction via up-regulated NLRC5.

The incidence and mortality rate of myocardial infarction are increasing per year in China. The polarization of macrophages towards the classically activated macrophages (M1) phenotype is of utmost importance in the progression of inflammatory stress subsequent to myocardial infarction. Poly (ADP-ribose) polymerase 1(PARP1) is the ubiquitous and best characterized member of the PARP family, which has been reported to support macrophage polarization towards the pro-inflammatory phenotype. Yet, the role of PARP1 in myocardial ischemic injury remains to be elucidated. Here, we demonstrated that a myocardial infarction mouse model induced cardiac damage characterized by cardiac dysfunction and increased PARP1 expression in cardiac macrophages. Inhibition of PARP1 by the PJ34 inhibitors could effectively alleviate M1 macrophage polarization, reduce infarction size, decrease inflammation and rescue the cardiac function post-MI in mice. Mechanistically, the suppression of PARP1 increase NLRC5 gene expression, and thus inhibits the NF-κB pathway, thereby decreasing the production of inflammatory cytokines such as IL-1ß and TNF-α. Inhibition of NLRC5 promote infection by effectively abolishing the influence of this mechanism discussed above. Interestingly, inhibition of NLRC5 promotes cardiac macrophage polarization toward an M1 phenotype but without having major effects on M2 macrophages. Our results demonstrate that inhibition of PARP1 increased NLRC5 gene expression, thereby suppressing M1 polarization, improving cardiac function, decreasing infarct area and attenuating inflammatory injury. The aforementioned findings provide new insights into the proinflammatory mechanisms that drive macrophage polarization following myocardial infarction, thereby introducing novel potential targets for future therapeutic interventions in individuals affected by myocardial infarction.

Degradation of Chloroquine by Ammonia-Oxidizing Bacteria: Performance, Mechanisms, and Associated Impact on N2O Production.

Since the mass production and extensive use of chloroquine (CLQ) would lead to its inevitable discharge, wastewater treatment plants (WWTPs) might play a key role in the management of CLQ. Despite the reported functional versatility of ammonia-oxidizing bacteria (AOB) that mediate the first step for biological nitrogen removal at WWTP (i.e., partial nitrification), their potential capability to degrade CLQ remains to be discovered. Therefore, with the enriched partial nitrification sludge, a series of dedicated batch tests were performed in this study to verify the performance and mechanisms of CLQ biodegradation under the ammonium conditions of mainstream wastewater. The results showed that AOB could degrade CLQ in the presence of ammonium oxidation activity, but the capability was limited by the amount of partial nitrification sludge (∼1.1 mg/L at a mixed liquor volatile suspended solids concentration of 200 mg/L). CLQ and its biodegradation products were found to have no significant effect on the ammonium oxidation activity of AOB while the latter would promote N2O production through the AOB denitrification pathway, especially at relatively low DO levels (≤0.5 mg-O2/L). This study provided valuable insights into a more comprehensive assessment of the fate of CLQ in the context of wastewater treatment.

Standing in customers' shoes: How responsible leadership inhibits unethical pro-organizational behavior.

Although the negative impact of responsible leadership on employees' unethical pro-organizational behavior has been documented in the literature, little is known about its underlying processes and boundaries. Drawing on social information processing theory and social learning theory, we built a moderated mediation model to explain why and when unethical pro-organizational behavior could be inhibited by responsible leadership. We conducted a two-phase questionnaire survey to collect data. The empirical results based on the sample of 557 Chinese salespeople showed that customer-oriented perspective taking partially mediated the negative link between responsible leadership and unethical pro-organizational behavior and that leader competence strengthened the direct effects of responsible leadership on customer-oriented perspective taking and unethical pro-organizational behavior as well as the indirect effect of responsible leadership on unethical pro-organizational behavior via customer-oriented perspective taking. These findings enrich the current understanding of how responsible leadership relates to unethical pro-organizational behavior, extend the limited literature on customer-oriented perspective taking, and offer some suggestions that managers can follow to inhibit unethical pro-organizational behavior. Limitations and future research directions are also discussed.

A contingency perspective of pro-organizational motives, unethical pro-organizational behavior, and organizational citizenship behavior.

Although the effects of pro-organizational motives on pro-organizational behaviors [i.e., unethical pro-organizational behavior (UPB) and organizational citizenship behavior (OCB)] and their boundaries have been explored to some extent, extant studies are rather piecemeal and in need of synthesis and extension. Based on prior motivational research on pro-organizational behaviors, we developed a comprehensive contingent model in which moral identity and impression management motives would moderate the links between pro-organizational motives, UPB, and OCB. Adopting a time-lagged design, we collected data from 218 salespeople in an internet technology service company in China. Results showed that pro-organizational motives were positively related to UPB and OCB. Moral identity weakened the impact of pro-organizational motives on UPB but strengthened the influence of pro-organizational motives on OCB. Furthermore, we found that impression management motives strengthened the effects of pro-organizational motives on UPB and OCB, and the interaction of impression management motives and pro-organizational motives was stronger on UPB than on OCB. Theoretical and practical implications, limitations, and future directions are discussed.

Can leader self-sacrificial behavior inhibit unethical pro-family behavior? A personal identification perspective.

As a kind of deviant and unethical behavior in the workplace, unethical pro-family behavior (UPFB) has recently received increased attention. Yet, the question of how to reduce UPFB remains less well understood. From the personal identification perspective, we hypothesize that leader self-sacrificial behavior (LSSB) inhibits employees' UPFB through the mediation of identification with the leader. We further argue that employees' perceived insider status enhances this hypothesized relationship. Our analysis of two-wave data collected from 236 Chinese employees indicated that identification with the leader partially mediated the negative relationship between LSSB and UPFB. Moreover, the effect of LSSB on identification with the leader and the aforementioned mediating relationship were stronger for employees who perceived themselves as insiders than outsiders. These findings provide theoretical implications for research on UPFB and LSSB and offer some suggestions that managers can follow to inhibit UPFB. Limitations and future research directions are also discussed.

NLRP3 Inflammasome: A Potential Target in Isoflurane Pretreatment Alleviates Stroke-Induced Retinal Injury in Diabetes.

Ischemic stroke remains a devastating disease which is the leading cause of death worldwide. Visual impairment after stroke is a common complication which may lead to vision loss, greatly impacting life quality of patients. While ischemic stroke is traditionally characterized by a blockage of blood flow to the brain, this may coincide with reduced blood flow to the eye, resulting in retinal ischemia and leading to visual impairment. Diabetes increases the risk of ischemic stroke and induces diabetic retinopathy; the latter may be more sensitive to the ischemic retinal injury. In diabetic status, the underlying mechanism in stroke-induced retinal injury has not been fully clarified. The NLR pyrin domain containing 3 (NLRP3) inflammasome is an important activator of inflammation, which may play a critical role in catalyzing and forming certain pro-inflammatory cytokines in both cerebral and retinal ischemia. Isoflurane has been demonstrated to inhibit the activation of the NLRP3 inflammasome and show neuroprotective effects. In this study, we established a diabetic mouse model and performed the middle cerebral artery occlusion procedure to induce ischemic stroke. Our results revealed that cerebral ischemia-induced retinal injury in the diabetic model. Isoflurane pretreatment alleviated the cerebral and retinal injury after ischemic stroke. Of note, isoflurane pretreatment inhibited the NLRP3 inflammasome activation in the retina, indicating that isoflurane pretreatment may provide substantial retinal protection in stroke-induced retinal injury in diabetes.

Transcriptome Co-expression Network Analysis Identifies Key Genes Regulating Conchosporangia Maturation of Pyropia haitanensis.

Conchosporangia maturation is crucial for the yield of Pyropia/Porphyra. However, the molecular mechanisms underlying this process are poorly understood. In this study, we selected two strains of Pyropia haitanensis that show significant differences in conchosporangia maturation as materials to produce RNA-Seq libraries. Then, we identified key molecular pathways and genes involved in conchosporangia maturation by conducting a weighted gene co-expression network analysis. Two specific modules were identified, and included functions such as phosphorus metabolism, lipid metabolism, and the phosphatidylinositol signaling system. The hub genes that responded positively during conchosporangia maturation encoded diacylglycerol kinase (DGK) and phosphatidylinositol-3-phosphate-5-kinase, which are involved in the synthesis of phosphatidic acid, a key component of lipid metabolism. A full-length DGK sequence of P. haitanensis, designated as PhDGK1, was obtained by rapid-amplification of cDNA ends. Conserved motif and phylogenetic tree analyses showed that PhDGK1 belongs to DGK Cluster II. The transcript level of PhDGK1 increased during conchosporangia maturation in both strains, but increased earlier, and to higher levels, in the early-maturing strain than in the late-maturing strain. This pattern of gene expression was consistent with the patterns of maturity and changes in pigment contents. These results indicate that lipid metabolism plays a key role in regulating conchosporangia maturation in Pyropia spp., and that PhDGK1 might be a useful molecular marker for breeding new early-maturing strains.

Proteomic analysis of healthy and atretic porcine follicular granulosa cells.

Follicular atresia is initiated with the apoptosis of granulosa cells (GCs) after birth in mammals. The molecular mechanisms underlying GC apoptosis during follicular selection are unclear at present. The objective of this study is to identify the proteins and pathways that may be involved in porcine follicular atresia. Proteins isolated from GCs collected from healthy and atretic follicles were detected by tandem mass tag (TMT) protein labeling and LC-MS/MS. A total of 4591 proteins in the healthy follicle granulosa cell (HFGC) and atretic follicle granulosa cell (AFGC) groups were identified, and 399 differentially abundant proteins were found between the HFGC and AFGC groups; of which 262 proteins were significantly up-regulated and 137 proteins were significantly down-regulated. Differential protein enrichment analysis showed that proteins involved in proteolysis, protein destabilization, phagocytosis, and engulfment were more abundant in the AFGC group. Also, these proteins were mainly involved in the lysosome, phagosome, autophagy, and apoptosis pathways. Specially, PTGFRN is potential important regulated protein in the development of the antral follicle, and down-regulation of PTGFRN in GCs may lead to follicular atresia. Our study shows that the identified proteins and their related signaling pathways may play crucial roles during health follicle develop to atretic follicle. SIGNIFICANCE: Follicular atresia during 'selection' reduces the reproductive potential of sows. In this study, we found 399 proteins differentially abundant. between the HFGC and AFGC groups. These results establish a foundation for elucidating the mechanism of follicular atresia in swine.

Remediation of persistent organic pollutants in aqueous systems by electrochemical activation of persulfates: A review.

Sulfate-radical-based advanced oxidation processes (SR-AOPs) have been widely applied in environmental remediation during the past decade, especially in the degradation of refractory organic contaminants. The electrochemical method, which is considered as one of the most efficient ways to generate sulfate radical, has been extensively investigated for the activation of persulfate recently. This work presented a thorough assessment towards the performance of electrochemically activated persulfate for the removal of persistent organic pollutants (POPs) in aqueous systems. The mechanism and superiority of electrochemically activated persulfates were revealed accordingly. Some major factors (e.g., electrode material, pH, current density, and persulfate concentration) influencing the electrochemical activation of persulfates to remove POPs were also discussed. Considering the increasing quantity of publications on this subject, it is significant to broader guidelines such as the efficiency for practical application, quantization of organic by-products, and cost-effectiveness of the electrochemical method to optimize active persulfate in the water treatment processes.

Insight into transketolase of Pyropia haitanensis under desiccation stress based on integrative analysis of omics and transformation.

BACKGROUND: Pyropia haitanensis, distributes in the intertidal zone, can tolerate water losses exceeding 90%. However, the mechanisms enabling P. haitanensis to survive harsh conditions remain uncharacterized. To elucidate the mechanism underlying P. haitanensis desiccation tolerance, we completed an integrated analysis of its transcriptome and proteome as well as transgenic Chlamydomonas reinhardtii carrying a P. haitanensis gene. RESULTS: P. haitanensis rapidly adjusted its physiological activities to compensate for water losses up to 60%, after which, photosynthesis, antioxidant systems, chaperones, and cytoskeleton were activated to response to severe desiccation stress. The integrative analysis suggested that transketolase (TKL) was affected by all desiccation treatments. Transgenic C. reinhardtii cells overexpressed PhTKL grew better than the wild-type cells in response to osmotic stress. CONCLUSION: P. haitanensis quickly establishes acclimatory homeostasis regarding its transcriptome and proteome to ensure its thalli can recover after being rehydrated. Additionally, PhTKL is vital for P. haitanensis desiccation tolerance. The present data may provide new insights for the breeding of algae and plants exhibiting enhanced desiccation tolerance.

Inhibitory Effect on the Hepatitis B Cells through the Regulation of miR-122-MAP3K2 signal pathway.

The aim of this study was to investigate the inhibitory effect of regulation of miR-122-MAP3K2 signal pathway on the hepatitis B cells. We detected the content of MAP3K2 from patients with HBV blood serum samples and analyzed the correlation between content of MAP3K2 and copies of HBV-DNA. Wound healing and Transwell assays were used to detect the function of cells from control group (wild type) and observer group (overexpresses miR-122). Secretion levels of HBsAg and MAP3K2 in the supernatant and level of MAP3K2 in cells were detected by ELISA and western blot, respectively. The results showed that there was a positive correlation between the copies of HBV-DNA and MAP3K2 in serum. In the assays involving detection of the number of HBV-DNA copies, the supernatant levels of HBsAg and MAP3K2, and the level of MAP3K2 in the cells, the rate of increase of these indicators significantly slowed as culture time. In conclusion, overexpression of miR-122 could inhibit the migration of hepatoblastoma cells; however, following transfection with miR-122, DNA synthesis and the secretion of HBsAg were inhibited. Overexpression of miR-122 can also downregulate MAP3K2. Consequently, we concluded that regulating the miR-122-MAP3K2 signaling pathway exerts an inhibitory effect in hepatitis B cells.

Investigation of miR-490-3p Expression in Hepatocellular Carcinoma Based on Reverse Transcription-Polymerase Chain Reaction (RT-qPCR) and a Meta-Analysis of 749 Cases.

BACKGROUND miR-490-3p could play vital roles in multiple cancers. However, the role of miR-490-3p in hepatocellular carcinoma (HCC) remains uncertain. In this study, we sought to explore the underlying role of miR-490-3p in HCC. MATERIAL AND METHODS In this study, we explored the clinical role of miR-490-3p in HCC via quantitative reverse transcription-polymerase chain reaction (RT-qPCR) and The Cancer Genome Atlas (TCGA) database. Then, a meta-analysis was performed to evaluate the expression trend and diagnostic value of miR-490-3p in HCC. Furthermore, 12 miRNA prediction algorithms were applied to predict the potential target genes of miR-490-3p. The differentially expressed genes in HCC in the Gene Expression Profiling Interactive Analysis (GEPIA) database were also selected. Additionally, bioinformatics analyses were utilized to investigate the possible functions and pathways of the target genes. RESULTS miR-490-3p was clearly down-regulated in HCC based on RT-qPCR (P=0.002). Consistent with the results of RT-qPCR, miR-490 was more highly expressed in normal liver tissue than in HCC (P<0.001). Additionally, the meta-analysis confirmed the results from RT-qPCR and TCGA. Furthermore, based on the prediction algorithms and GEPIA, a total of 113 genes were selected. According to the bioinformatics analyses, we found that the most remarkably enriched functional terms included protein transport, poly(A) RNA binding, and intracellular organelle part. Additionally, the miR-490-3p target genes were significantly related to the pathways in cancer. CONCLUSIONS We found that miR-490-3p is down-regulated in HCC and is related to genes that have potential tumoral functions. However, the exact mechanism should be confirmed by functional experiments.

Transcriptomic study to understand thermal adaptation in a high temperature-tolerant strain of Pyropia haitanensis.

Pyropia haitanensis, a high-yield commercial seaweed in China, is currently undergoing increasing levels of high-temperature stress due to gradual global warming. The mechanisms of plant responses to high temperature stress vary with not only plant type but also the degree and duration of high temperature. To understand the mechanism underlying thermal tolerance in P. haitanensis, gene expression and regulation in response to short- and long-term temperature stresses (SHS and LHS) was investigated by performing genome-wide high-throughput transcriptomic sequencing for a high temperature tolerant strain (HTT). A total of 14,164 differential expression genes were identified to be high temperature-responsive in at least one time point by high-temperature treatment, representing 41.10% of the total number of unigenes. The present data indicated a decrease in the photosynthetic and energy metabolic rates in HTT to reduce unnecessary energy consumption, which in turn facilitated in the rapid establishment of acclimatory homeostasis in its transcriptome during SHS. On the other hand, an increase in energy consumption and antioxidant substance activity was observed with LHS, which apparently facilitates in the development of resistance against severe oxidative stress. Meanwhile, ubiquitin-mediated proteolysis, brassinosteroids, and heat shock proteins also play a vital role in HTT. The effects of SHS and LHS on the mechanism of HTT to resist heat stress were relatively different. The findings may facilitate further studies on gene discovery and the molecular mechanisms underlying high-temperature tolerance in P. haitanensis, as well as allow improvement of breeding schemes for high temperature-tolerant macroalgae that can resist global warming.

Discovery of a 3-(4-Pyrimidinyl) Indazole (MLi-2), an Orally Available and Selective Leucine-Rich Repeat Kinase 2 (LRRK2) Inhibitor that Reduces Brain Kinase Activity.

Leucine-rich repeat kinase 2 (LRRK2) is a large, multidomain protein which contains a kinase domain and GTPase domain among other regions. Individuals possessing gain of function mutations in the kinase domain such as the most prevalent G2019S mutation have been associated with an increased risk for the development of Parkinson's disease (PD). Given this genetic validation for inhibition of LRRK2 kinase activity as a potential means of affecting disease progression, our team set out to develop LRRK2 inhibitors to test this hypothesis. A high throughput screen of our compound collection afforded a number of promising indazole leads which were truncated in order to identify a minimum pharmacophore. Further optimization of these indazoles led to the development of MLi-2 (1): a potent, highly selective, orally available, brain-penetrant inhibitor of LRRK2.

Discovery of Novel 3,3-Disubstituted Piperidines as Orally Bioavailable, Potent, and Efficacious HDM2-p53 Inhibitors.

A new subseries of substituted piperidines as p53-HDM2 inhibitors exemplified by 21 has been developed from the initial lead 1. Research focused on optimization of a crucial HDM2 Trp23-ligand interaction led to the identification of 2-(trifluoromethyl)thiophene as the preferred moiety. Further investigation of the Leu26 pocket resulted in potent, novel substituted piperidine inhibitors of the HDM2-p53 interaction that demonstrated tumor regression in several human cancer xenograft models in mice. The structure of HDM2 in complex with inhibitors 3, 10, and 21 is described.

MLi-2, a Potent, Selective, and Centrally Active Compound for Exploring the Therapeutic Potential and Safety of LRRK2 Kinase Inhibition.

Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are the most common genetic cause of familial and sporadic Parkinson's disease (PD). That the most prevalent mutation, G2019S, leads to increased kinase activity has led to a concerted effort to identify LRRK2 kinase inhibitors as a potential disease-modifying therapy for PD. An internal medicinal chemistry effort identified several potent and highly selective compounds with favorable drug-like properties. Here, we characterize the pharmacological properties of cis-2,6-dimethyl-4-(6-(5-(1-methylcyclopropoxy)-1H-indazol-3-yl)pyrimidin-4-yl)morpholine (MLi-2), a structurally novel, highly potent, and selective LRRK2 kinase inhibitor with central nervous system activity. MLi-2 exhibits exceptional potency in a purified LRRK2 kinase assay in vitro (IC50 = 0.76 nM), a cellular assay monitoring dephosphorylation of LRRK2 pSer935 LRRK2 (IC50 = 1.4 nM), and a radioligand competition binding assay (IC50 = 3.4 nM). MLi-2 has greater than 295-fold selectivity for over 300 kinases in addition to a diverse panel of receptors and ion channels. Acute oral and subchronic dosing in MLi-2 mice resulted in dose-dependent central and peripheral target inhibition over a 24-hour period as measured by dephosphorylation of pSer935 LRRK2. Treatment of MitoPark mice with MLi-2 was well tolerated over a 15-week period at brain and plasma exposures >100× the in vivo plasma IC50 for LRRK2 kinase inhibition as measured by pSer935 dephosphorylation. Morphologic changes in the lung, consistent with enlarged type II pneumocytes, were observed in MLi-2-treated MitoPark mice. These data demonstrate the suitability of MLi-2 as a compound to explore LRRK2 biology in cellular and animal models.

Acne patients frequently associated with abnormal plasma lipid profile.

This study was designed to investigate the relationship between plasma lipid profile and acne. Acne patients (n = 181) and healthy volunteers (n = 130) matched in terms of both age and sex were enrolled. Plasma total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol, low-density lipoprotein cholesterol (LDL-C) and lipoprotein (LP)(a) levels were measured. TC, LDL-C and LP(a) levels in male and female patients with severe acne were significantly higher than in the healthy control group (P < 0.05). TG in male patients with severe and moderate acne was significantly higher than in the healthy control group (P < 0.05). LP(a) in male and female patients with mild, moderate and severe acne was significantly higher than in the healthy control group (P < 0.05). The constituent ratio of male and female patients with TC, TG, LDL-C and LP(a) over the normal range was significantly higher than in the healthy control group. In this study, acne patients were frequently associated with abnormal lipid profile, providing a new basis for further exploration of the pathogenesis, as well as new treatments, of acne vulgaris.

Reactive postural control deficits in patients with posterior parietal cortex lesions after stroke and the influence of auditory cueing.

OBJECTIVE: The purpose of this study was to investigate the ways in which stroke-induced posterior parietal cortex (PPC) lesions affect reactive postural responses and whether providing auditory cues modulates these responses. DESIGN: Seventeen hemiparetic patients after stroke, nine with PPC lesions (PPCLesion) and eight with intact PPCs (PPCSpared), and nine age-matched healthy adults completed a lateral-pull perturbation experiment under noncued and cued conditions. The activation rates of the gluteus medius muscle ipsilateral (GMi) and contralateral to the pull direction, the rates of occurrence of three types of GM activation patterns, and the GMi contraction latency were investigated. RESULTS: In noncued pulls toward the paretic side, of the three groups, the PPCLesion group exhibited the lowest activation rate (56%) of the GMi (P < 0.05), which is the primary postural muscle involved in this task, and the highest rate of occurrence (33%) of the gluteus medius muscle contralateral-activation-only pattern (P < 0.05), which is a compensatory activation pattern. In contrast, in cued pulls toward the paretic side, the PPCLesion group was able to increase the activation rate of the GMi to a level (81%) such that there became no significant differences in activation rate of the GMi among the three groups (P > 0.05). However, there were no significant differences in the GM activation patterns and GMi contraction latency between the noncued and cued conditions for the PPCLesion group (P > 0.05). CONCLUSIONS: The PPCLesion patients had greater deficits in recruiting paretic muscles and were more likely to use the compensatory muscle activation pattern for postural reactions than the PPCSpared patients, suggesting that PPC is part of the neural circuitry involved in reactive postural control in response to lateral perturbations. The auditory cueing used in this study, however, did not significantly modify the muscle activation patterns in the PPCLesion patients. More research is needed to explore the type and structure of cueing that could effectively improve patterns and speed of postural responses in these patients.

Synthesis and spectroscopic properties of some novel BODIPY dyes.

BODIPY dyes have some unique properties including high fluorescence quantum yield, large extinction coefficiency, narrow absorption and emission band. However, most of BODIPY dyes display short emission wavelength and small Stokes shift, which limits their applications in biosensing and bioimaging in vivo. For bioimaging application, a fluorescent dye with long emission wavelength and large Stokes shift is highly desired. To push the absorption and emission spectrum of BODIPY to red and even far-red region, a COOEt group was introduced to the meso position, and some aromatic group was attached to the 3, 5 position of BODIPY core. The structure of resulting compounds were comfirmed by 1H NMR, 13C NMR and HR-MS. Dye-1 displays a strong UV-Vis absorption band centered at 536 nm and a sharp emission band is located at 592 nm, which is significantly red-shifted (80 nm) compared to ordinary BODIPY analogs. In addition, the meso-COOEt substituted BODIPYs exhibit high quantum yield and red to far-red emission. Notably surprisingly, the meso-COOEt substituted BODIPYs display almost separated UV-Vis absorption and emission spectra with a large Stokes shift (-60 nm). Time-dependent density functional theory calculations were conducted to understand the structure-optical properties relationship, and it was revealed that the large Stokes shift was resulted from the geometric change from the ground state to the first excited singlet state. The spectroscopic properties of these BODIPY dyes display very subtle solvent-dependence effect. Furthermore, BODIPY was tested for its ability of imaging in living cells. The results indicate that Dye-1 is a water-soluble and membrane-permeable probe. Therefore, these BODIPYs are a new family dyes with excellent spectroscopic properties and can be good candidates for bioimaging in living cells.

The JAMM motif of human deubiquitinase Poh1 is essential for cell viability.

Poh1 deubiquitinase activity is required for proteolytic processing of polyubiquitinated substrates by the 26S proteasome, linking deubiquitination to complete substrate degradation. Poh1 RNA interference (RNAi) in HeLa cells resulted in a reduction in cell viability and an increase in polyubiquitinated protein levels, supporting the link between Poh1 and the ubiquitin proteasome pathway. To more specifically test for any requirement of the zinc metalloproteinase motif of Poh1 to support cell viability and proteasome function, we developed a RNAi complementation strategy. Effects on cell viability and proteasome activity were assessed in cells with RNAi of endogenous Poh1 and induced expression of wild-type Poh1 or a mutant form of Poh1, in which two conserved histidines of the proposed catalytic site were replaced with alanines. We show that an intact zinc metalloproteinase motif is essential for cell viability and 26S proteasome function. As a required enzymatic component of the proteasome, Poh1 is an intriguing therapeutic drug target for cancer.