Inhibitor Arc Research Articles

Sex and Cross-Sex Testosterone Treatment Alters Gamma-Hydroxybutyrate Acid Toxicokinetics and Toxicodynamics in Rats.

Γ-hydroxybutyric acid (GHB) is widely abused due to its sedative/hypnotic and euphoric effects. In recent years, GHB use has witnessed a notable rise within the LGBTQ+ community. GHB is a substrate of monocarboxylate transporters (MCTs) and exhibits nonlinear toxicokinetics, characterized by saturable metabolism, absorption, and renal reabsorption. This study investigates the impact of exogenous testosterone administration on GHB toxicokinetics and toxicodynamics, exploring the potential of MCT1 inhibition as a strategy to counteract toxicity. Ovariectomized (OVX) females and castrated (CST) male Sprague Dawley rats were treated with testosterone or placebo for 21 days. GHB was administered at two doses (1000 mg/kg or 1500 mg/kg i.v.), and the MCT1 inhibitor AR-C 155858 (1 mg/kg i.v.) was administered 5 min after GHB (1500 mg/kg i.v.) administration. Plasma and urine were collected up to 8 h post-dose, and GHB concentrations were quantified via a validated LC/MS/MS assay. Sleep time (sedative/hypnotic effect) was utilized as the toxicodynamic endpoint. Testosterone treatment significantly affected GHB toxicokinetics and toxicodynamics. Testosterone-treated CST rats exhibited significantly lower renal clearance, higher AUC, and increased sedative effect, while testosterone-treated OVX rats demonstrated higher metabolic clearance. AR-C 155858 treatment led to an increase in GHB renal and total clearance together with an improvement in sedative/hypnotic effect. In conclusion, exogenous testosterone treatment induces significant alterations in GHB toxicokinetics and toxicodynamics, and MCT inhibition can serve as a potential therapeutic strategy for GHB overdose in both cisgender and transgender male populations.

Failure Section Identification in Smart Distribution Network Based on IFIAPN Considering the Auto-reclosing and DG

This paper proposes a fault location method for smart distribution networks based on the intuitionistic fuzzy inhibitor arc Petri net (IFIAPN) technique that employs discrete evidence such as the status of protective devices and fault indicators (FIs) to estimate the faulted section. To eliminate the possible false fault section estimations and pinpoint the actual faulted location, an IFIAPN algorithm is improved. It amends the certainty and uncertainty of the information according to the action logic of the protection system and the direction of fault propagation. The authors further utilize the directional fault indicator to cope with the impact on the bidirectional power flows caused by distributed generation, and introduce the inhibitor arc to represent the action characteristics of auto-reclosing to optimize the diagnostic model. The proposed method can precisely determine the fault location in distribution systems with many laterals and sub-laterals in the presence of distributed generators (DGs). Finally, feasibility, fault tolerance, and effectiveness of the proposed method are verified by the IEEE 33 node feeder distribution system.

A Novel Intuitionistic Fuzzy Inhibitor Arc Petri Net With Error Back Propagation Algorithm and Application in Fault Diagnosis

The setting and adjustment of the weight parameters in the traditional fault diagnosis method depend entirely on personal experience, and the parameter setting lacks regularity. To reduce the fault diagnosis errors caused by human subjective factors and improve the speed and accuracy of power grid fault diagnosis, we propose a method for power grid fault diagnosis using intuitionistic fuzzy inhibitor arc Petri net (IFIAPN) with error back propagation (BP) algorithm. Firstly, according to the network topology analysis and relay protection configuration setting rules, the inhibitor arc (IA) tuple is introduced into the model structure of the intuitionistic fuzzy Petri net to reduce the ambiguity of protection and circuit breaker action. Then, the weight parameters in the model are trained using a BP neural network algorithm to enhance the objectivity of the parameters. Finally, a simulation of an IEEE-39 node system and a real case study using the Hou-zhong line local grid were used to verify the effectiveness of the fault diagnosis method. The results show that the method can effectively deal with the refusal and mis-operation of multiple circuit breakers and improve the diagnostic efficiency under complex data environment.

Optimal Petri Net Supervisors of Discrete Event Systems via Weighted and Data Inhibitor Arcs

This paper handles the optimal supervisory control problem of Petri nets (PNs) via two PN structures, namely, weighted and data inhibitor arcs. It is a two-stage method. In the first stage, for each transition that may lead to illegal markings, a set of observer places with weighted inhibitor arcs is used to optimally control the maximal number of marking/transition separation instances (MTSIs) through the proposed integer linear program. Then, the controlled MTSIs are removed from the set of MTSIs. In the second stage, at each iteration, for an MTSI that cannot be controlled at the first control stage, we design an optimal observer place with a data inhibitor arc. This process terminates after all the MTSIs are controlled. The first-stage can sharply lower the computational burden compared with the method by using data inhibitor arcs alone. Finally, a typical example is presented to shed light on this technique. The proposed control strategy can definitely yield an optimal supervisor for any bounded PN on the premise that such a supervisor exists.

Design of Optimal Petri Net Supervisors for Flexible Manufacturing Systems via Weighted Inhibitor Arcs

AbstractThis paper develops an approach to the design of an optimal Petri net supervisor that enforces liveness to flexible manufacturing systems. The supervisor contains a set of observer places with weighted inhibitor arcs. An observer place with a weighted inhibitor arc is used to forbid a net from yielding an illegal marking by inhibiting the firing of a transition at a marking while ensuring that all legal markings are preserved. A marking reduction technique is presented to decrease the number of considered markings, which can dramatically lower the computational burden of the proposed approach. An integer linear program is presented to simplify the supervisory structure by minimizing the number of observer places. Finally, several examples are used to shed light on the proposed approach which can lead to an optimal supervisor for the net models that cannot be optimally controlled via pure Petri net supervisors.

Compact Supervisory Control of Discrete Event Systems by Petri Nets With Data Inhibitor Arcs

This work proposes a novel structure in Petri nets, namely data inhibitor arcs, and their application to the optimal supervisory control of Petri nets. A data inhibitor arc is an arc from a place to a transition labeled with a set of integers. A transition is disabled by a data inhibitor arc if the number of tokens in the place is in the set of integers labeled on it. Its formal definitions and properties are given. Then, we propose a method to design an optimal Petri net supervisor with data inhibitor arcs to prevent a system from reaching illegal markings with respect to control specifications. Two techniques are developed to reduce the supervisor structure by compressing the number of control places. Finally, a number of examples are used to illustrate the proposed approaches and experimental results show that they can obtain optimal Petri net supervisors for the net models that cannot be optimally controlled by pure net supervisors. A significant result is that the proposed approach can always lead to an optimal supervisor with only one control place for bounded Petri nets on the premise that such a supervisor exists.

The Cell Death Inhibitor ARC Is Induced in a Tissue-Specific Manner by Deletion of the Tumor Suppressor Gene Men1, but Not Required for Tumor Development and Growth.

Multiple endocrine neoplasia type 1 (MEN1) is a genetic disorder characterized by tissue-specific tumors in the endocrine pancreas, parathyroid, and pituitary glands. Although tumor development in these tissues is dependent upon genetic inactivation of the tumor suppressor Men1, loss of both alleles of this gene is not sufficient to induce these cancers. Men1 encodes menin, a nuclear protein that influences transcription. A previous ChIP on chip analysis suggested that menin binds promoter sequences of nol3, encoding ARC, which is a cell death inhibitor that has been implicated in cancer pathogenesis. We hypothesized that ARC functions as a co-factor with Men1 loss to induce the tissue-restricted distribution of tumors seen in MEN1. Using mouse models that recapitulate this syndrome, we found that biallelic deletion of Men1 results in selective induction of ARC expression in tissues that develop tumors. Specifically, loss of Men1 in all cells of the pancreas resulted in marked increases in ARC mRNA and protein in the endocrine, but not exocrine, pancreas. Similarly, ARC expression increased in the parathyroid with inactivation of Men1 in that tissue. To test if ARC contributes to MEN1 tumor development in the endocrine pancreas, we generated mice that lacked none, one, or both copies of ARC in the context of Men1 deletion. Studies in a cohort of 126 mice demonstrated that, although mice lacking Men1 developed insulinomas as expected, elimination of ARC in this context did not significantly alter tumor load. Cellular rates of proliferation and death in these tumors were also not perturbed in the absence of ARC. These results indicate that ARC is upregulated by loss Men1 in the tissue-restricted distribution of MEN1 tumors, but that ARC is not required for tumor development in this syndrome.

New Petri Net Structure and Its Application to Optimal Supervisory Control: Interval Inhibitor Arcs

This paper presents a new Petri net structure, namely, an interval inhibitor arc, and its application to the optimal supervisory control of Petri nets. An interval inhibitor arc is an arc from a place to a transition labeled with an integer interval. The transition is disabled by the place if the number of tokens in the place is between the labeled interval. The formal definition and the firing rules of Petri nets with interval inhibitor arcs are developed. Then, an optimal Petri net supervisor based on the interval inhibitor arcs is designed to prevent a system from reaching illegal markings. Two techniques are developed to simplify the supervisory structure by compressing the number of control places. The proposed approaches are general since they can be applied to any bounded Petri net models. A marking reduction approach is also introduced if they are applied to Petri net models of flexible manufacturing systems. Finally, a number of examples are provided to demonstrate the proposed approaches and the experimental results show that they can obtain optimal Petri net supervisors for some net models that cannot be optimally controlled by pure net supervisors. Furthermore, the obtained supervisor is structurally simple.

Abstract 1181: The Apoptosis repressor with a CARD domain (ARC) is a direct HIF1 target gene and promotes survival and proliferation of VHL deficient renal cancer cells

Abstract The induction of hypoxia inducible factors (HIFs) is essential for the adaptation of tumor cells to a low oxygen environment. We found that the expression of the apoptosis inhibitor ARC was induced by hypoxia in a variety of cancer cell types and its induction is primarily HIF1 dependent. Chromatin immunoprecipitation (ChIP) and reporter assays also indicate that the ARC gene is regulated by direct binding of HIF1 to a hypoxia response element (HRE) located at -190 bp upstream of the transcription start site. HIFs play an essential role in the pathogenesis of renal cell carcinoma (RCC) under normoxic conditions, through the loss of the Von Hippel Lindau (VHL) gene. Accordingly, our results show that ARC is not expressed in normal renal tissue, but is highly expressed in 65% of RCC tumors, which also express high levels of Carbonic Anhydrase IX (CAIX), a HIF1-dependent gene. Compared to controls, ARC-deficient RCCs exhibited decreased colony formation and increased apoptosis in vitro. In addition, loss of ARC resulted in a dramatic reduction of RCC tumor growth in SCID mice in vivo. Thus, HIF-mediated increased expression of ARC in RCC can explain how loss of VHL can promote survival early in tumor formation. Citation Format: Olga V. Razorenova, Laura Castellini, Renata Colavitti, Laura E. Edgington, Monica Nicolau, Xin Huang, Barbara Bedogni, Edward M. Mills, Matthew Bogyo, Amato J. Giaccia. The Apoptosis repressor with a CARD domain (ARC) is a direct HIF1 target gene and promotes survival and proliferation of VHL deficient renal cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1181. doi:10.1158/1538-7445.AM2014-1181

Maximally Permissive Petri Net Supervisors with a Novel Structure

This paper presents a novel arc of Petri nets, namely data inhibitor arcs, and its application to deadlock control of discrete event systems. A data inhibitor arc is from a place to a transition labeled with a set of data. The transition is disabled by the place if the number of tokens in the place is in the set of data labeled on the arc. By using the data inhibitor arcs, a maximally permissive Petri net supervisor is designed to prevent a discrete event system from reaching illegal markings. Furthermore, a technique is proposed to find only one control place to make the system live with all legal markings. Finally, we provide examples to demonstrate the proposed approach.

The Apoptosis Inhibitor ARC Alleviates the ER Stress Response to Promote β-Cell Survival

Type 2 diabetes involves insulin resistance and β-cell failure leading to inadequate insulin secretion. An important component of β-cell failure is cell loss by apoptosis. Apoptosis repressor with caspase recruitment domain (ARC) is an inhibitor of apoptosis that is expressed in cardiac and skeletal myocytes and neurons. ARC possesses the unusual property of antagonizing both the extrinsic (death receptor) and intrinsic (mitochondria/endoplasmic reticulum [ER]) cell death pathways. Here we report that ARC protein is abundant in cells of the endocrine pancreas, including >99.5% of mouse and 73% of human β-cells. Using genetic gain- and loss-of-function approaches, our data demonstrate that ARC inhibits β-cell apoptosis elicited by multiple inducers of cell death, including ER stressors tunicamycin, thapsigargin, and physiological concentrations of palmitate. Unexpectedly, ARC diminishes the ER stress response, acting distal to protein kinase RNA-like ER kinase (PERK) and inositol-requiring protein 1α, to suppress C/EBP homologous protein (CHOP) induction. Depletion of ARC in isolated islets augments palmitate-induced apoptosis, which is dramatically rescued by deletion of CHOP. These data demonstrate that ARC is a previously unrecognized inhibitor of apoptosis in β-cells and that its protective effects are mediated through suppression of the ER stress response pathway.

ARC Synergizes with ABT-737 to Induce Apoptosis in Human Cancer Cells

Previously, we reported that the nucleoside analogue/transcriptional inhibitor ARC (4-amino-6-hydrazino-7-beta-D-ribofuranosyl-7H-pyrrolo(2,3-d)-pyrimidine-5-carboxamide) was able to induce p53-independent apoptosis in multiple cancer cell lines of different origins. This occurred, at least in part, by the suppression of short-lived, prosurvival member of the Bcl-2 family, Mcl-1. In contrast, we show here that treatment of human cancer cells with the pan-Bcl-2 inhibitor ABT-737 alone led to upregulation of Mcl-1 protein expression. Combination of subapoptotic concentrations of ABT-737 and ARC induced mitochondrial injury and potent caspase-3/caspase-9-dependent apoptosis in a wide variety of human cancer cell lines. These data suggest that the ABT-737/ARC combination, which simultaneously targets Bcl-2 and Mcl-1, may be efficient against human cancer.

Abstract 1036: ARC synergizes with ABT-737 to induce apoptosis in human cancer cells

Abstract Previously, we reported that the nucleoside analog/transcriptional inhibitor ARC was able to induce p53-independent apoptosis in multiple cancer cell lines of different origin. This occurred at least in part by the suppression of Mcl-1 expression, which a short-lived, pro-survival member of the Bcl-2 family. Interestingly, pan-caspase inibitor Z-VAD-FMK protected leukemia cells from ARC-induced mitochondrial damage and apoptosis, and Mcl-1 mRNA and protein from down-regulation by ARC. At the same time treatment of human cancer cells with the pan-Bcl-2 inhibitor ABT-737 alone led to up-regulation of Mcl-1 protein expression. Combination of sub-apoptotic concentrations of ABT-737 and ARC suppressed Mcl-1 expression, induced mitochondrial injury and potent apoptosis in wide variety of human cancer cell lines. In prostate cancer cells co-treatment with ARC and ABT-737 resulted in synergistic induction of cell death with combination index 0.7-0.8. These data suggest that ABT-737/ARC combination that simultaneously targets Bcl-2 and Mcl-1 may be efficient against human cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1036.

New potential anti‐cancer agents synergize with bortezomib and ABT‐737 against prostate cancer

We previously described the identification of a transcriptional inhibitor ARC and FoxM1 inhibitors, thiazole antibiotics, Siomycin A and thiostrepton that were able to induce potent p53-independent apoptosis in cancer cell lines of different origin. Here, we report the characterization of these drugs individually or in combination with ABT-737 and bortezomib on a panel of prostate cancer cell lines. DU 145, LNCaP and PC-3 prostate cancer cells were treated with ARC, Siomycin A and thiostrepton to evaluate their activity as single agents or in combination with ABT-737 and bortezomib to measure their synergistic potential in anti-proliferative and cell cycle assays. Chou-Talalay method was used to quantitate the synergistic interaction. Western blot method was used to determine Mcl-1 and FoxM1 expression and caspase-3 cleavage. We show that ARC inhibited the viability of prostate cancer cells and induced apoptosis in low nanomolar concentration. It potently downregulated the expression of Mcl-1 and showed synergistic combination effect with Bcl-2 inhibitor ABT-737. Thiazole antibiotics, Siomycin A and thiostrepton inhibited growth, FoxM1 expression and induced cell death in prostate cancer cells in low micromolar concentrations. In addition, thiostrepton and ARC synergistically induced apoptosis in prostate cancer cells following combination treatment with proteasome inhibitor bortezomib. Furthermore, we found that all tested drug combinations were able to induce apoptosis selectively in transformed, but not normal cells of the same origin. Based on their in vitro activity as single or combination agents, ARC, Siomycin A and thiostrepton represent potential candidates for drug development against prostate cancer.

Regulation of HIV‐1 transcription at 3% versus 21% oxygen concentration

HIV transcription is induced by the HIV-1 Tat protein, in concert with cellular co-factors including CDK9, CDK2, NF-kappaB, and others. The cells of most of the body's organs are exposed to approximately 3-6% oxygen, but most in vitro studies of HIV replication are conducted at 21% oxygen. We hypothesized that activities of host cell factors involved in HIV-1 replication may differ at 3% versus 21% O(2), and that such differences may affect HIV-1 replication. Here we show that Tat-induced HIV-1 transcription was reduced at 3% O(2) compared to 21% O(2). HIV-1 replication was also reduced in acutely or chronically infected cells cultured at 3% O(2) compared to 21% O(2). This reduction was not due the decreased cell growth or increased cellular toxicity and also not due to the induction of hypoxic response. At 3% O(2), the activity of CDK9/cyclin T1 was inhibited and Sp1 activity was reduced, whereas the activity of other host cell factors such as CDK2 or NF-kappaB was not affected. CDK9-specific inhibitor ARC was much less efficient at 3% compared to 21% O(2) and also expression of CDK9/cyclin T1-dependent IkappaB inhibitor alpha was repressed. Our results suggest that lower HIV-1 transcription at 3% O(2) compared to 21% O(2) may be mediated by lower activity of CDK9/cyclin T1 and Sp1 at 3% O(2) and that additional host cell factors such as CDK2 and NF-kappaB might be major regulators of HIV-1 transcription at low O(2) concentrations.

Reduction rules for reset/inhibitor nets

Reset/inhibitor nets are Petri nets extended with reset arcs and inhibitor arcs. These extensions can be used to model cancellation and blocking. A reset arc allows a transition to remove all tokens from a certain place when the transition fires. An inhibitor arc can stop a transition from being enabled if the place contains one or more tokens. While reset/inhibitor nets increase the expressive power of Petri nets, they also result in increased complexity of analysis techniques. One way of speeding up Petri net analysis is to apply reduction rules. Unfortunately, many of the rules defined for classical Petri nets do not hold in the presence of reset and/or inhibitor arcs. Moreover, new rules can be added. This is the first paper systematically presenting a comprehensive set of reduction rules for reset/inhibitor nets. These rules are liveness and boundedness preserving and are able to dramatically reduce models and their state spaces. It can be observed that most of the modeling languages used in practice have features related to cancellation and blocking. Therefore, this work is highly relevant for all kinds of application areas where analysis is currently intractable.

Reachability in Petri Nets with Inhibitor Arcs

We define 2 operators on relations over natural numbers such that they generalize the operators ‘+’ and ‘*’ and show that the membership and emptiness problem of relations constructed from finite relations with these operators and ∪ is decidable. This generalizes Presburger arithmetics and allows to decide the reachability problem for those Petri nets where inhibitor arcs occur only in some restricted way. Especially the reachability problem is decidable for Petri nets with only one inhibitor arc, which solves an open problem in [H. Kleine Büning, T. Lettmann, and E. W. Mayr. Projections of vector addition system reachability sets are semilinear. Theoret. Comput. Sci., 64:343–350, 1989]. Furthermore we describe the corresponding automaton having a decidable emptiness problem.

Novel anticancer compounds induce apoptosis in melanoma cells

We previously described the identification of a nucleoside analog transcriptional inhibitor ARC (4-amino-6-hydrazino-7-beta-D-ribofuranosyl-7H-Pyrrolo[2,3-d]-pyrimidine-5-carboxamide) and FoxM1 inhibitor, thiazole antibiotic Siomycin A that were able to induce apoptosis in cancer cell lines of different origin. Here, we report the characterization of these drugs on a panel of melanoma cell lines. We found that in contrast to the common anti-melanoma drug dacarbazine (DTIC), ARC and thiazole antibiotics, Siomycin A and thiostrepton, efficiently inhibited growth and induced cell death in melanoma cell lines in low concentrations. Overexpression of the antiapoptotic protein Mcl-1 protected melanoma cells from apoptosis induced by these compounds. Furthermore, we found that ARC and Siomycin A synergistically induce apoptosis in DM833 melanoma cell line suggesting that they may antagonize different anti-apoptotic pathways in melanoma cells. In general, these drugs may represent important candidates for anti-cancer drug development against melanoma.

Proapoptotic compound ARC targets Akt and N-myc in neuroblastoma cells

We have previously described the identification of a nucleoside analog transcriptional inhibitor ARC (4-amino-6-hydrazino-7-beta-D-ribofuranosyl-7H-pyrrolo[2,3-d]-pyrimidine-5-carboxamide) that was able to induce apoptosis in cancer cell lines of different origin. Here, we report the characterization of ARC on a panel of neuroblastoma cell lines. We found that these cell lines were more than 10-fold sensitive to ARC than to the well-known nucleoside analog DRB (5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole), and that ARC-induced apoptosis proceeds through mitochondrial injury. Also, we observed that ARC-mediated cell death was accompanied by caspase-3 cleavage and repression of antiapoptotic proteins such as Mcl-1 and survivin. Conversely, we found that overexpression of Mcl-1-protected neuroblastoma cell line NB-1691 from ARC-induced apoptosis. Furthermore, we found that while ARC inhibited the phosphorylation of Akt Ser-473 in multiple cancer cell lines, forced expression of myristoylated Akt promoted resistance to ARC-induced apoptosis in neuroblastoma cells. In addition, we observed that ARC was able to downregulate the protein levels of N-myc, a commonly amplified oncogene in neuroblastomas, and Akt protected N-myc from ARC-induced downregulation. These data suggest that ARC may antagonize different antiapoptotic pathways and induce apoptosis in neuroblastoma cells via multiple mechanisms. Overall, ARC could represent an attractive candidate for anticancer drug development against neuroblastomas.

A novel anticancer agent ARC antagonizes HIV-1 and HCV

Human immunodeficiency virus (HIV) and hepatitis C virus (HCV) pose major public health concerns worldwide. HCV is clearly associated with the occurrence of hepatocellular carcinoma, and recently HIV infection has also been linked to the development of a multitude of cancers. Previously, we identified a novel nucleoside analog transcriptional inhibitor ARC (4-amino-6-hydrazino-7-beta-D-ribofuranosyl-7H-pyrrolo[2,3-d]-pyrimidine-5-carboxamide) that exhibited proapoptotic and antiangiogenic properties in vitro. Here, we evaluated the effect of ARC on HIV-1 transcription and HCV replication. Using reporter assays, we found that ARC inhibited HIV-1 Tat-based transactivation in different cell systems. Also, using hepatoma cells that harbor subgenomic and full-length replicons of HCV, we found that ARC inhibited HCV replication. Together, our data indicate that ARC could be a promising candidate for the development of antiviral therapeutics against HIV and HCV.