Effects of Policy for Controlling Agricultural Non-Point Source Pollution in China: From a Perspective of Regional and Policy Measures Differences.

The concerns about the contradiction between agricultural production and Agricultural Non-Point (ANPS) pollution has become increased with economic development in China. Government interventions are key to controlling ANPS pollution through the establishment of laws and policies. This paper uses the entropy method to calculate the emissions amount of ANPS pollution and policy strength of 31 provinces from 2010 to 2019 in China. The dynamic panel data model with system generalized moment is used to estimate the impacts of policies with different measures on ANPS pollution emission. According to our findings, China's policies have been helpful in controlling ANPS pollution though there are important regional differences. Moreover, four types of policy measures all contribute to the reduction in ANPS pollution. These findings improve our understanding of the relationship between policies and ANPS pollution in the analyzed period, thus providing support for the formulation of pollution management strategies in the next stage.

Real-Time Adaptation of an Artificial Neural Network for Transfemoral Amputees Using a Powered Prosthesis.

OBJECTIVE: We evaluated a two-step method to improve control accuracy for a powered prosthetic leg using machine learning and adaptation, while reducing subject training time. METHODS: First, information from three transfemoral amputees was grouped together, to create a baseline control system that was subsequently tested using data from a fourth subject (user-independent classification). Second, online adaptation was investigated, whereby the fourth subject's data were used to improve the baseline control system in real-time. Results were compared for user-independent classification and for user-dependent classification (data collected from and tested in the same subject), with and without adaptation. RESULTS: The combination of a user-independent classifier with real-time adaptation based on a unique individual's data produced a classification error rate as low as 1.61% [0.15 standard error of the mean (SEM)] without requiring collection of initial training data from that individual. Training/testing using a subject's own data (user-dependent classification), combined with adaptation, resulted in a classification error rate of 0.9% [0.12 SEM], which was not significantly different (P 0.05) but required, on average, an additional 7.52 hours [0.92 SEM] of training sessions. CONCLUSION AND SIGNIFICANCE: We found that the combination of a user-independent dataset with adaptation resulted in error rates that were not significantly different from using a user-dependent dataset. Furthermore, this method eliminated the need for individual training sessions, saving many hours of data collection time.

Has Carbon Emissions Trading Reduced PM2.5 in China?

China today has an urgent need to develop low-carbon policies that also address local air pollution. This study uses a difference-in-differences model to estimate the extent to which China's Emissions Trading Scheme (ETS), which directly addresses CO2 emissions, has also generated cobenefits by reducing PM2.5. Using monthly PM2.5 concentration and weather data for 297 Chinese cities from January 2005 to December 2017, we find that (1) China's ETS has reduced PM2.5 concentrations by 4.8%, and this reduction effect is strongest in summer. The results are confirmed by robustness tests including propensity score matching, mahalanobis distance matching, two placebo tests, and controlling atmospheric policies. (2) The ETS pilots have positive spillovers on neighboring nonpilot cities within 300 km of the pilot cites, especially for those downwind neighboring nonpilot cities, and the ETS's cobenefits are primarily a result of firms adopting abatement activities and adjusting industrial structure. (3) The cobenefits of China's ETS pilots are greater in regions with larger carbon trading volume and higher carbon trading price. (4) The reduction in PM2.5 concentration caused by China's ETS may have avoided 23,363 deaths and saved $41.38 billion annually in GDP.

Feasibility of removable balloon implant for simultaneous magnetic nanoparticle heating and HDR brachytherapy of brain tumor resection cavities.

AIM: Hyperthermia (HT) has been shown to improve clinical response to radiation therapy (RT) for cancer. Synergism is dramatically enhanced if HT and RT are combined simultaneously, but appropriate technology to apply treatments together does not exist. This study investigates the feasibility of delivering HT with RT to a 5-10mm annular rim of at-risk tissue around a tumor resection cavity using a temporary thermobrachytherapy (TBT) balloon implant. METHODS: A balloon catheter was designed to deliver radiation from High Dose Rate (HDR) brachytherapy concurrent with HT delivered by filling the balloon with magnetic nanoparticles (MNP) and immersing it in a radiofrequency magnetic field. Temperature distributions in brain around the TBT balloon were simulated with temperature dependent brain blood perfusion using numerical modeling. A magnetic induction system was constructed and used to produce rapid heating (>0.2°C/s) of MNP-filled balloons in brain tissue-equivalent phantoms by absorbing 0.5 W/ml from a 5.7 kA/m field at 133 kHz. RESULTS: Simulated treatment plans demonstrate the ability to heat at-risk tissue around a brain tumor resection cavity between 40-48°C for 2-5cm diameter balloons. Experimental thermal dosimetry verifies the expected rapid and spherically symmetric heating of brain phantom around the MNP-filled balloon at a magnetic field strength that has proven safe in previous clinical studies. CONCLUSIONS: These preclinical results demonstrate the feasibility of using a TBT balloon to deliver heat simultaneously with HDR brachytherapy to tumor bed around a brain tumor resection cavity, with significantly improved uniformity of heating over previous multi-catheter interstitial approaches. Considered along with results of previous clinical thermobrachytherapy trials, this new capability is expected to improve both survival and quality of life in patients with glioblastoma multiforme.

Segmenting Mechanomyography Measures of Muscle Activity Phases Using Inertial Data.

Electromyography (EMG) is the standard technology for monitoring muscle activity in laboratory environments, either using surface electrodes or fine wire electrodes inserted into the muscle. Due to limitations such as cost, complexity, and technical factors, including skin impedance with surface EMG and the invasive nature of fine wire electrodes, EMG is impractical for use outside of a laboratory environment. Mechanomyography (MMG) is an alternative to EMG, which shows promise in pervasive applications. The present study used an exerting squat-based task to induce muscle fatigue. MMG and EMG amplitude and frequency were compared before, during, and after the squatting task. Combining MMG with inertial measurement unit (IMU) data enabled segmentation of muscle activity at specific points: entering, holding, and exiting the squat. Results show MMG measures of muscle activity were similar to EMG in timing, duration, and magnitude during the fatigue task. The size, cost, unobtrusive nature, and usability of the MMG/IMU technology used, paired with the similar results compared to EMG, suggest that such a system could be suitable in uncontrolled natural environments such as within the home.

Adapting myoelectric control in real-time using a virtual environment.

BACKGROUND: Pattern recognition technology allows for more intuitive control of myoelectric prostheses. However, the need to collect electromyographic data to initially train the pattern recognition system, and to re-train it during prosthesis use, adds complexity that can make using such a system difficult. Although experienced clinicians may be able to guide users to ensure successful data collection methods, they may not always be available when a user needs to (re)train their device. METHODS: Here we present an engaging and interactive virtual reality environment for optimal training of a myoelectric controller. Using this tool, we evaluated the importance of training a classifier actively (i.e., moving the residual limb during data collection) compared to passively (i.e., maintaining the limb in a single, neutral orientation), and whether computational adaptation through serious gaming can improve performance. RESULTS: We found that actively trained classifiers performed significantly better than passively trained classifiers for non-amputees (P < 0.05). Furthermore, collecting data passively with minimal instruction, paired with computational adaptation in a virtual environment, significantly improved real-time performance of myoelectric controllers. CONCLUSION: These results further support previous work which suggested active movements during data collection can improve pattern recognition systems. Furthermore, adaptation within a virtual guided serious game environment can improve real-time performance of myoelectric controllers.

Robust Pattern Recognition Myoelectric Training for Improved Online Control within a 3D Virtual Environment.

It has been shown that maintaining a neutral arm position during collection of pattern recognition training data for myoelectric prosthesis control results in high offline classification accuracies; however, that precision does not translate to real-time applications, when the arm is used in different positions. Previous studies have shown that collecting training data with the arm in a variety of positions can improve pattern recognition control systems. In this work, we extended these findings to real-time myoelectric control in an immersive testing environment using virtual reality. We show that collecting training data for a pattern recognition algorithm under dynamic conditions, where the user moves their arm, significantly improves control efficiency and achievement of testing metrics.

A coupled recreational anglers' decision and fish population dynamics model.

The effective management of fish populations requires understanding of both the biology of the species being managed and the behavior of the humans who harvest those species. For many marine fisheries, recreational harvests represent a significant portion of the total fishing mortality. For such fisheries, therefore, a model that captures the dynamics of angler choices and the fish population would be a valuable tool for fisheries management. In this study, we provide such a model, focusing on red drum and spotted seatrout, which are the two of the main recreational fishing targets in the Gulf of Mexico. The biological models are in the form of vector autoregressive models. The anglers' decision model takes the discrete choice approach, in which anglers first decide whether to go fishing and then determine the location to fish based on the distance and expected catch of two species of fish if they decide to go fishing. The coupled model predicts that, under the level of fluctuation in the abundance of the two species experienced in the past 35 years, the number of trips that might be taken by anglers fluctuates moderately. This fluctuation is magnified as the cost of travel decreases because the anglers can travel long distance to seek better fishing conditions. On the other hand, as the cost of travel increases, their preference to fish in nearby areas increases regardless of the expected catch in other locations and variation in the trips taken declines. The model demonstrates the importance of incorporating anglers' decision processes in understanding the changes in a fishing effort level. Although the model in this study still has a room for further improvement, it can be used for more effective management of fish and potentially other populations.

Comparing the Effects of Signal Noise on Pattern Recognition and Linear Regression-Based Myoelectric Controllers.

Myoelectric pattern recognition using linear discriminant analysis (LDA) classifiers has been a wellestablished control method for upper limb prostheses for many years. More recently, linear regression (LR) controllers have been proposed as an alternative solution due to their ability to control multiple degrees of freedom (DOF) simultaneously. The aim of this experiment was to compare the online performance of LDA and LR control systems under three electromyographic (EMG) signal conditions: baseline, noise in all channels, and noise in a single channel. To simulate the last two conditions, different levels of Gaussian noise were added to the EMG signals. Completion rate, path efficiency, dwelling time, and completion time were computed after virtual Fitts' Law tasks. While both controllers were significantly affected by the lowest noise levels, we found no significant differences between the controllers under the baseline and all-channel noise conditions. However, the LDA controller outperformed the LR controller in the single-channel noise condition. Therefore, while both controllers are comparable in most cases, the added complexity of simultaneous control affects an LR controller's performance under certain noise conditions. Based on these results, neither control system should be dismissed in future developments.

Performance of a wearable acoustic system for fetal movement discrimination.

Fetal movements (FM) are a key factor in clinical management of high-risk pregnancies such as fetal growth restriction. While maternal perception of reduced FM can trigger self-referral to obstetric services, maternal sensation is highly subjective. Objective, reliable monitoring of fetal movement patterns outside clinical environs is not currently possible. A wearable and non-transmitting system capable of sensing fetal movements over extended periods of time would be extremely valuable, not only for monitoring individual fetal health, but also for establishing normal levels of movement in the population at large. Wearable monitors based on accelerometers have previously been proposed as a means of tracking FM, but such systems have difficulty separating maternal and fetal activity and have not matured to the level of clinical use. We introduce a new wearable system based on a novel combination of accelerometers and bespoke acoustic sensors as well as an advanced signal processing architecture to identify and discriminate between types of fetal movements. We validate the system with concurrent ultrasound tests on a cohort of 44 pregnant women and demonstrate that the garment is capable of both detecting and discriminating the vigorous, whole-body 'startle' movements of a fetus. These results demonstrate the promise of multimodal sensing for the development of a low-cost, non-transmitting wearable monitor for fetal movements.

Teprotumumab for Thyroid-Associated Ophthalmopathy.

BACKGROUND: Thyroid-associated ophthalmopathy, a condition commonly associated with Graves' disease, remains inadequately treated. Current medical therapies, which primarily consist of glucocorticoids, have limited efficacy and present safety concerns. Inhibition of the insulin-like growth factor I receptor (IGF-IR) is a new therapeutic strategy to attenuate the underlying autoimmune pathogenesis of ophthalmopathy. METHODS: We conducted a multicenter, double-masked, randomized, placebo-controlled trial to determine the efficacy and safety of teprotumumab, a human monoclonal antibody inhibitor of IGF-IR, in patients with active, moderate-to-severe ophthalmopathy. A total of 88 patients were randomly assigned to receive placebo or active drug administered intravenously once every 3 weeks for a total of eight infusions. The primary end point was the response in the study eye. This response was defined as a reduction of 2 points or more in the Clinical Activity Score (scores range from 0 to 7, with a score of ≥3 indicating active thyroid-associated ophthalmopathy) and a reduction of 2 mm or more in proptosis at week 24. Secondary end points, measured as continuous variables, included proptosis, the Clinical Activity Score, and results on the Graves' ophthalmopathy-specific quality-of-life questionnaire. Adverse events were assessed. RESULTS: In the intention-to-treat population, 29 of 42 patients who received teprotumumab (69%), as compared with 9 of 45 patients who received placebo (20%), had a response at week 24 (P<0.001). Therapeutic effects were rapid; at week 6, a total of 18 of 42 patients in the teprotumumab group (43%) and 2 of 45 patients in the placebo group (4%) had a response (P<0.001). Differences between the groups increased at subsequent time points. The only drug-related adverse event was hyperglycemia in patients with diabetes; this event was controlled by adjusting medication for diabetes. CONCLUSIONS: In patients with active ophthalmopathy, teprotumumab was more effective than placebo in reducing proptosis and the Clinical Activity Score. (Funded by River Vision Development and others; ClinicalTrials.gov number, NCT01868997 .).

User intent prediction with a scaled conjugate gradient trained artificial neural network for lower limb amputees using a powered prosthesis.

Powered lower limb prostheses have the ability to provide greater mobility for amputee patients. Such prostheses often have pre-programmed modes which can allow activities such as climbing stairs and descending ramps, something which many amputees struggle with when using non-powered limbs. Previous literature has shown how pattern classification can allow seamless transitions between modes with a high accuracy and without any user interaction. Although accurate, training and testing each subject with their own dependent data is time consuming. By using subject independent datasets, whereby a unique subject is tested against a pooled dataset of other subjects, we believe subject training time can be reduced while still achieving an accurate classification. We present here an intent recognition system using an artificial neural network (ANN) with a scaled conjugate gradient learning algorithm to classify gait intention with user-dependent and independent datasets for six unilateral lower limb amputees. We compare these results against a linear discriminant analysis (LDA) classifier. The ANN was found to have significantly lower classification error (P<;0.05) than LDA with all user-dependent step-types, as well as transitional steps for user-independent datasets. Both types of classifiers are capable of making fast decisions; 1.29 and 2.83 ms for the LDA and ANN respectively. These results suggest that ANNs can provide suitable and accurate offline classification in prosthesis gait prediction.

Identification and Characterization of GAL-021 as a Novel Breathing Control Modulator.

BACKGROUND: The authors describe the preclinical pharmacological properties of GAL-021, a novel peripheral chemoreceptor modulator. METHODS: The ventilatory effects of GAL-021 were characterized using tracheal pneumotachometry (n = 4 to 6), plethysmography (n = 5 to 6), arterial blood gas analyses (n = 6 to 11), and nasal capnography (n = 3 to 4) in naive animals and those subjected to morphine-induced respiratory depression. Morphine analgesia in rats was evaluated by tail-flick test (n = 6). Carotid body involvement in GAL-021 ventilatory effects was assessed by comparing responses in intact and carotid sinus nerve-transected rats. Hemodynamic effects of GAL-021 were evaluated in urethane-anesthetized rats (n = 7). The pharmacological profile of GAL-021 in vitro was investigated using radioligand binding, enzyme inhibition, and cellular electrophysiology assays. RESULTS: GAL-021 given intravenously stimulated ventilation and/or attenuated opiate-induced respiratory depression in rats, mice, and nonhuman primates, without decreasing morphine analgesia in rats. GAL-021 did not alter mean arterial pressure but produced a modest increase in heart rate. Ventilatory stimulation in rats was attenuated by carotid sinus nerve transection. GAL-021 inhibited KCa1.1 in GH3 cells, and the evoked ventilatory stimulation was attenuated in Slo1 mice lacking the pore-forming α-subunit of the KCa1.1 channel. CONCLUSIONS: GAL-021 behaved as a breathing control modulator in rodents and nonhuman primates and diminished opioid-induced respiratory depression without compromising opioid analgesia. It acted predominantly at the carotid body, in part by inhibiting KCa1.1 channels. Its preclinical profile qualified the compound to enter clinical trials to assess effects on breathing control disorders such as drug (opioid)-induced respiratory depression and sleep apnea.

Practical precautionary resource management using robust optimization.

Uncertainties inherent in fisheries motivate a precautionary approach to management, meaning an approach specifically intended to avoid bad outcomes. Stochastic dynamic optimization models, which have been in the fisheries literature for decades, provide a framework for decision making when uncertain outcomes have known probabilities. However, most such models incorporate population dynamics models for which the parameters are assumed known. In this paper, we apply a robust optimization approach to capture a form of uncertainty nearly universal in fisheries, uncertainty regarding the values of model parameters. Our approach, developed by Nilim and El Ghaoui (Oper Res 53(5):780-798, 2005), establishes bounds on parameter values based on the available data and the degree of precaution that the decision maker chooses. To demonstrate the applicability of the method to fisheries management problems, we use a simple example, the Skeena River sockeye salmon fishery. We show that robust optimization offers a structured and computationally tractable approach to formulating precautionary harvest policies. Moreover, as better information about the resource becomes available, less conservative management is possible without reducing the level of precaution.

Pharmacological characterization of recombinant N-type calcium channel (Cav2.2) mediated calcium mobilization using FLIPR.

The N-type voltage-gated calcium channel (Ca(v)2.2) functions in neurons to regulate neurotransmitter release. It comprises a clinically relevant target for chronic pain. We have validated a calcium mobilization approach to assessing Ca(v)2.2 pharmacology in two stable Ca(v)2.2 cell lines: alpha1(B), alpha2delta, beta(3)-HEK-293 and alpha1(B), beta(3)-HEK-293. Ca(v)2.2 channels were opened by addition of KCl and Ca(2+) mobilization was measured by Fluo-4 fluorescence on a fluorescence imaging plate reader (FLIPR(96)). Ca(v)2.2 expression and biophysics were confirmed by patch-clamp electrophysiology (EP). Both cell lines responded to KCl with adequate signal-to-background. Signals from both cell lines were inhibited by omega-conotoxin (ctx)-MVIIa and omega-conotoxin (ctx)-GVIa with IC(50) values of 1.8 and 1nM, respectively, for the three-subunit stable, and 0.9 and 0.6nM, respectively, for the two-subunit stable. Other known Ca(v)2.2 blockers were characterized including cadmium, flunarizine, fluspirilene, and mibefradil. IC(50) values correlated with literature EP-derived values. Novel Ca(v)2.2 pharmacology was identified in classes of compounds with other primary pharmacological activities, including Na(+) channel inhibitors and antidepressants. Novel Na(+) channel compounds with high potency at Ca(v)2.2 were identified in the phenoxyphenyl pyridine, phenoxyphenyl pyrazole, and other classes. The highest potency at Ca(v)2.2 tricyclic antidepressant identified was desipramine.

Pharmacology of 2-[4-(4-chloro-2-fluorophenoxy)phenyl]-pyrimidine-4-carboxamide: a potent, broad-spectrum state-dependent sodium channel blocker for treating pain states.

Voltage-gated Na(+) channels may play important roles in establishing pathological neuronal hyperexcitability associated with chronic pain in humans. Na(+) channel blockers, such as carbamazepine (CBZ) and lamotrigine (LTG), are efficacious in treating neuropathic pain; however, their therapeutic utility is compromised by central nervous system side effects. We reasoned that it may be possible to gain superior control over pain states and, in particular, a better therapeutic index, by designing broad-spectrum Na(+) channel blockers with higher potency, faster onset kinetics, and greater levels of state dependence than existing drugs. 2-[4-(4-Chloro-2-fluorophenoxy)phenyl]-pyrimidine-4-carboxamide (PPPA) is a novel structural analog of the state-dependent Na(+) channel blocker V102862 [4-(4-fluorophenoxy)benzaldehyde semicarbazone]. Tested on recombinant rat Na(v)1.2 channels and native Na(+) currents in cultured rat dorsal root ganglion neurons, PPPA was approximately 1000 times more potent, had 2000-fold faster binding kinetics, and > or =10-fold higher levels of state dependence than CBZ and LTG. Tested in rat pain models against mechanical endpoints, PPPA had minimal effective doses of 1 to 3 mg/kg p.o. in partial sciatic nerve ligation, Freund's complete adjuvant, and postincisional pain. In all cases, efficacy was similar to clinically relevant comparators. Importantly, PPPA did not produce motor deficits in the accelerating Rotarod assay of ataxia at doses up to 30 mg/kg p.o., indicating a therapeutic index >10, which was superior to CBZ and LTG. Our experiments suggest that high-potency, broad-spectrum, state-dependent Na(+) channel blockers will have clinical utility for treating neuropathic, inflammatory, and postsurgical pain. Optimizing the biophysical parameters of broad-spectrum voltage-gated Na(+) channel blockers may lead to improved pain therapeutics.

State-dependent compound inhibition of Nav1.2 sodium channels using the FLIPR Vm dye: on-target and off-target effects of diverse pharmacological agents.

Voltage-gated sodium channels (NaChs) are relevant targets for pain, epilepsy, and a variety of neurological and cardiac disorders. Traditionally, it has been difficult to develop structure-activity relationships for NaCh inhibitors due to rapid channel kinetics and state-dependent compound interactions. Membrane potential (Vm) dyes in conjunction with a high-throughput fluorescence imaging plate reader (FLIPR) offer a satisfactory 1st-tier solution. Thus, the authors have developed a FLIPR Vm assay of rat Nav1.2 NaCh. Channels were opened by addition of veratridine, and Vm dye responses were measured. The IC50 values from various structural classes of compounds were compared to the resting state binding constant (Kr)and inactivated state binding constant (Ki)obtained using patch-clamp electrophysiology (EP). The FLIPR values correlated with Ki but not Kr. FLIPRIC50 values fell within 0.1-to 1.5-fold of EP Ki values, indicating that the assay generally reports use-dependent inhibition rather than resting state block. The Library of Pharmacologically Active Compounds (LOPAC, Sigma) was screened. Confirmed hits arose from diverse classes such as dopamine receptor antagonists, serotonin transport inhibitors, and kinase inhibitors. These data suggest that NaCh inhibition is inherent in a diverse set of biologically active molecules and may warrant counterscreening NaChs to avoid unwanted secondary pharmacology.

V102862 (Co 102862): a potent, broad-spectrum state-dependent blocker of mammalian voltage-gated sodium channels.

1. 4-(4-Fluorophenoxy)benzaldehyde semicarbazone (V102862) was initially described as an orally active anticonvulsant with robust activity in a variety of rodent models of epilepsy. The mechanism of action was not known. We used whole-cell patch-clamp techniques to study the effects of V102862 on native and recombinant mammalian voltage-gated Na+ channels. 2. V102862 blocked Na+ currents (I(Na)) in acutely dissociated cultured rat hippocampal neurons. Potency increased with membrane depolarization, suggesting a state-dependent mechanism of inhibition. There was no significant effect on the voltage dependence of activation of I(Na). 3. The dissociation constant for the inactivated state (K(I)) was approximately 0.6 microM, whereas the dissociation constant for the resting state (K(R)) was >15 microM. 4. The binding to inactivated channels was slow, requiring a few seconds to reach steady state at -80 mV. 5. The mechanism of inhibition was characterized in more detail using human embryonic kidney-293 cells stably expressing rat brain type IIA Na+ (rNa(v)1.2) channels, a major Na+ channel alpha subunit in rat hippocampal neurons. Similar to hippocampal neurons, V102862 was a potent state-dependent blocker of rNa(v)1.2 channels with a K(I) of approximately 0.4 microM and K(R) approximately 30 microM. V102862 binding to inactivated channels was relatively slow (k(+) approximately = 1.7 microM(-1) s(-1)). V102862 shifted the steady-state availability curve in the hyperpolarizing direction and significantly retarded recovery of Na+ channels from inactivation. 6. These results suggest that inhibition of voltage-gated Na+ channels is a major mechanism underlying the anticonvulsant properties of V102862. Moreover, understanding the biophysics of the interaction may prove to be useful in designing a new generation of potent Na+ channel blocker therapeutics.

3-(4-phenoxyphenyl)pyrazoles: a novel class of sodium channel blockers.

A series of 3-(4-phenoxyphenyl)-1H-pyrazoles were synthesized and characterized as potent state-dependent sodium channel blockers. A limited SAR study was carried out to delineate the chemical requirements for potency. The results indicate that the distal phenyl group is critical for activity but will tolerate lipophilic (+pi) electronegative (+sigma) substituents at the ortho and/or para position. Substitution at the pyrazole nitrogen with a H-bond donor improves potency. Compound 18 showed robust activity in the rat Chung neuropathy paradigm.

MX1013, a dipeptide caspase inhibitor with potent in vivo antiapoptotic activity.

1. Caspases play a critical role in apoptosis, and are considered to be key targets for the design of cytoprotective drugs. As part of our antiapoptotic drug-discovery effort, we have synthesized and characterized Z-VD-fmk, MX1013, as a potent, irreversible dipeptide caspase inhibitor. 2. MX1013 inhibits caspases 1, 3, 6, 7, 8, and 9, with IC50 values ranging from 5 to 20 nm. MX1013 is selective for caspases, and is a poor inhibitor of noncaspase proteases, such as cathepsin B, calpain I, or Factor Xa (IC50 values >10 microm). 3. In several cell culture models of apoptosis, including caspase 3 processing, PARP cleavage, and DNA fragmentation, MX1013 is more active than tetrapeptide- and tripeptide-based caspase inhibitors, and blocked apoptosis at concentrations as low as 0.5 microm. 4. MX1013 is more aqueous soluble than tripeptide-based caspase inhibitors such as Z-VAD-fmk. 5. At a dose of 1 mg kg-1 i.v., MX1013 prevented liver damage and the lethality caused by Fas death receptor activation in the anti-Fas mouse-liver apoptosis model, a widely used model of liver failure. 6. At a dose of 20 mg kg-1 (i.v. bolus) followed by i.v. infusion for 6 or 12 h, MX1013 reduced cortical damage by approximately 50% in a model of brain ischemia/reperfusion injury. 7. At a dose of 20 mg kg-1 (i.v. bolus) followed by i.v. infusion for 12 h, MX1013 reduced heart damage by approximately 50% in a model of acute myocardial infarction. 8. Based on these studies, we conclude that MX1013, a dipeptide pan-caspase inhibitor, has a good combination of in vitro and in vivo properties. It has the ability to protect cells from a variety of apoptotic insults, and is systemically active in three animal models of apoptosis, including brain ischemia.