Target Hits Research Articles

Otubain 2 is a novel promoter of beta cell survival as revealed by siRNA high-throughput screens of human pancreatic islets

Pro-inflammatory cytokines induce death of beta cells and hamper engraftment of transplanted islet mass. Our aim was to reveal novel genes involved in this process, as a platform for innovative therapeutic approaches. Small interfering RNA (siRNA) high-throughput screening (HTS) of primary human islets was employed to identify novel genes involved in cytokine-induced beta cell apoptosis. Dispersed human islets from nine human donors, treated with a combination of TNF-α, IL-1β and IFN-γ were transfected with ∼730 different siRNAs. Caspase-3/7 activity was measured, results were analysed and potential anti- and pro-apoptotic genes were identified. Dispersed human pancreatic islets appeared to be suitable targets for performance of siRNA HTS. Using this methodology we found a number of potential pro- and anti-apoptotic target hits that have not been previously associated with pancreatic beta cell death. One such hit was the de-ubiquitinating enzyme otubain 2 (OTUB2). OTUB2 knockdown increased caspase-3/7 activity in MIN6 cells and primary human islets and inhibited insulin secretion and increased nuclear factor-κB (NF-κB) activity both under basal conditions and following cytokine treatment. Use of dispersed human islets provides a new platform for functional HTS in a highly physiological system. Employing this technique enabled the identification of OTUB2 as a novel promoter of viability and insulin secretion in human beta cells. OTUB2 acts through the inhibition of NF-κB signalling, which is deleterious to beta cell survival. siRNA screens of human islets may therefore identify new targets, such as OTUB2, for therapeutic intervention in type 1 diabetes and islet transplantation.

Identification of inhibitors against interaction between pro-inflammatory sPLA2-IIA protein and integrin αvβ3

Increased concentrations of secreted phospholipase A2 type IIA (sPLA2-IIA), have been found in the synovial fluid of patients with rheumatoid arthritis. It has been shown that sPLA2-IIA specifically binds to integrin αvβ3, and initiates a signaling pathway that leads to cell proliferation and inflammation. Therefore, the interaction between integrin and sPLA2-IIA could be a potential therapeutic target for the treatment of proliferation or inflammation-related diseases. Two one-bead-one-compound peptide libraries were constructed and screened, and seven target hits were identified. Herein we report the identification, synthesis, and biological testing of two pyrazolylthiazole-tethered peptide hits and their analogs. Biological assays showed that these compounds were able to suppress the sPLA2-IIA–integrin interaction and sPLA2-IIA-induced migration of monocytic cells and that the blockade of the sPLA2-IIA–integrin binding was specific to sPLA2-IIA and not to the integrin.

Optimalizace odhodové vzdálenosti pro test přesnosti hodu na cíl u mládeže věkové kategorie 11–14 let

Throw accuracy is not being currently quite developed within school PE lessons; it is being developed only as part of skills of a particular sports discipline. Th is shortcoming may stem from the lack of a suitable test assessing this skill. In the literature we have seen only three throw accuracy tests intended for school youngsters (Bös 2001, Mecner 1975, Měkota; Blahuš 1983). Th ese tests consist of a few repeated series of throws, failing to distinguish a throwing distance for boys and girls and even age groups, and requiring a rather complicated graphic record of successful target hits that cannot be assessed easily and quickly during a PE lesson. Th e purpose of the study is to create a test of measuring throw accuracy at a target eliminating the aforementioned shortcomings. Th e optimisation of a throwing distance is based on the condition of a normal Gaussian distribution of data related to individual throwing distances, and on the condition that an average number of target hits within a concrete throwing distance will be close to 7.5 target hits. To assess the importance of the diff erence of the observed throwing distances one-factor ANOVA (One Way Analysis of Variance) was used. Th e assessed group consisted of 105 boys and 114 girls of three randomly selected schools in Ostrava between 11 and 14 years old. Th ree throwing distances were determined (boys 6; 8; 10 m, girls 3.5; 5.5; 7.5 m) in combination with a vertical round target of 0.7 m in diameter. Th e tested person used an overarm throwing technique and made 20 throws for each throwing distance. Th e test score was the number of the target hits from the 6th - 20th throw (training: 1st -5th throw). Th e optimum throwing distances were identifi ed for the accuracy test of a target throw for boys of the 11-12 year category (6 m), 13-14 year category (8 m), for girls of the 11-12 year category (5.5 m), 13-14 year category (7.5 m) while using a target of 0.7 m in diameter.

MicroRNA expression profiles of porcine skeletal muscle

MicroRNAs (miRNAs) are endogenous non-coding RNAs of ∼22 nucleotides in length that play important roles in multiple biological processes by degrading targeted mRNAs or repressing mRNA translation. To evaluate the roles of miRNA in porcine skeletal muscle, miRNA expression profiles were investigated using longissimus muscle tissue from pigs at embryonic day 90 (E90) and postpartum day 120 (PD120). First, we used previously known miRNA sequences from humans and mice to perform blast searches against the porcine expressed sequence tag (EST) database; 98 new miRNA candidates were identified according to a range of filtering criteria. These miRNA candidates and 73 known miRNAs (miRBase 13.0) from pigs were chosen for porcine miRNA microarray analysis. A total of 16 newly identified miRNAs and 31 previously known miRNAs were detected in porcine skeletal muscle tissues. During later foetal development at E90, miR-1826, miR-26a, miR-199b and let-7 were highly expressed, whilst miR-1a, miR-133a, miR-26a and miR-1826 showed highest abundance during the fast growing stage at PD120. Using the 47 miRNAs detected by the microarray assay, we performed further investigations using the publicly available porcine mRNA database from NCBI and computed potential target hits using the software rnahybrid. This study identified 16 new miRNA candidates, computed potential target hits for 18 miRNA families and determined the miRNA expression profiles in porcine skeletal muscle tissues at different developmental stages. These results provide a valuable resource for investigators interested in post-transcriptional gene regulation in pigs and related animals.

Computational identification of new porcine microRNAs and their targets

MicroRNAs (miRNAs) represent a newly identified class of non-protein-coding approximately 22 nt small RNAs which play important roles in multiple biological processes by degrading targeted mRNAs or repressing mRNA translation. Here we present an expressed sequence tag (EST)-based combined approach for the detection of novel porcine miRNAs. This was initiated by using previously known miRNA sequences from Homo sapiens (human) and Mus musculus (mouse) to blast the databases of Sus scrofa (pig) EST. A total of 65 new miRNAs were detected following a range of filtering criteria. Using these new potential miRNA sequences, we further obtained the publicly available porcine mRNA database from NCBI and detected 48 586 potential target hits using a software RNA hybrid. So far, compared to human and mouse, fewer miRNAs (only 54 miRNAs) were identified in Sus scrofa species. These 65 new miRNAs and their targets in pig have been run through miRHelper to yield data that may help us better understand the possible role of miRNAs in regulating the growth and development of pigs. These findings suggest that EST analysis is a good alternative strategy for identifying new miRNA candidates, their targets and other genes.

Normalized Movement Quality Measures for Therapeutic Robots Strongly Correlate With Clinical Motor Impairment Measures

In this paper, we analyze the correlations between four clinical measures (Fugl-Meyer upper extremity scale, Motor Activity Log, Action Research Arm Test, and Jebsen-Taylor Hand Function Test) and four robotic measures (smoothness of movement, trajectory error, average number of target hits per minute, and mean tangential speed), used to assess motor recovery. Data were gathered as part of a hybrid robotic and traditional upper extremity rehabilitation program for nine stroke patients. Smoothness of movement and trajectory error, temporally and spatially normalized measures of movement quality defined for point-to-point movements, were found to have significant moderate to strong correlations with all four of the clinical measures. The strong correlations suggest that smoothness of movement and trajectory error may be used to compare outcomes of different rehabilitation protocols and devices effectively, provide improved resolution for tracking patient progress compared to only pre- and post-treatment measurements, enable accurate adaptation of therapy based on patient progress, and deliver immediate and useful feedback to the patient and therapist.

Performance comparisons of the kicking of stationary and rolling balls in a futsal context

Angular kinematics of the hip, knee, and ankle joints, as well as ball velocity and accuracy for stationary and rolling balls were compared in a futsal (Five-a-Side Indoor soccer) context. Ten futsal athletes performed five kicks each on stationary and rolling futsal balls. Six digital cameras (120 Hz) recorded the kicks. For both kick types, angles for hip, knee, and ankle joints were calculated using Euler angle conventions. Angular velocity, ball velocity, foot linear velocity, relative velocity, and accuracy also were analyzed. The kicking of both stationary and rolling balls showed similarities for ball velocity (24.2 ± 2.2 m/s and 23.8 ± 2.7 m/s, respectively), foot velocity (17.6 ± 1.8 m/s and 17.2 ± 2.2 m/s, respectively), and accuracy (26% and 24% target hits, respectively). We observed few differences in angular kinematics and angular joint velocities between kick types. Elite players can make online adjustments in the preparatory phase so that kicking a rolling ball is almost exactly like kicking a stationary ball.

Expertise-Based Performance Measures in a Virtual Training Environment

This paper introduces and validates quantitative performance measures for a rhythmic target-hitting task. These performance measures are derived from a detailed analysis of human performance during a month-long training experiment where participants learned to operate a 2-DOF haptic interface in a virtual environment to execute a manual control task. The motivation for the analysis presented in this paper is to determine measures of participant performance that capture the key skills of the task. This analysis of performance indicates that two quantitative measures—trajectory error and input frequency—capture the key skills of the target-hitting task, as the results show a strong correlation between the performance measures and the task objective of maximizing target hits. The performance trends were further explored by grouping the participants based on expertise and examining trends during training in terms of these measures. In future work, these measures will be used as inputs to a haptic guidance scheme that adjusts its control gains based on a real-time assessment of human performance of the task. Such guidance schemes will be incorporated into virtual training environments for humans to develop manual skills for domains such as surgery, physical therapy, and sports.

Energy deposition and relative frequency of hits of cylindrical nanovolume in medium irradiated by ions: Monte Carlo simulation of tracks structure

Radiation track structure simulations have been used for many years to study the DNA damage caused by heavy ions. These studies are highly relevant for treatment planning of heavy ion radiotherapy and space radiation risk assessment. Measurements of the frequency of delta-rays hits, mean specific energy per target hits and per ion, and the frequency of dose distribution in a cylindrical target volume placed at various radial distances from 4He 2+, 12C 6+ and 16O 8+ tracks have been performed by Schmollack et al. (in Radiat Res 153:469-478, 2000). In the present work, Monte Carlo simulation of radiation tracks has been performed with the RITRACKS and the RETRACKS codes along with a target volume to simulate the experiment of Schmollack et al. The results of these simulations are compared to those of previous deterministic models of the radial dependence of the mean specific energy. Our Monte Carlo simulations are consistent with the experimental data both in the core and in the penumbra of the beam, and are shown to provide a better description of the experimental data than deterministic codes.

ProbeMatch: rapid alignment of oligonucleotides to genome allowing both gaps and mismatches

We have developed a tool, called ProbeMatch, for matching a large set of oligonucleotide sequences against a genome database using gapped alignments. Unlike most of the existing tools such as ELAND which only perform ungapped alignments allowing at most two mismatches, ProbeMatch generates both ungapped and gapped alignments allowing up to three errors including insertion, deletion and mismatch. To speedup sequence alignment, ProbeMatch uses gapped q-grams and q-grams of various patterns to identify target hits to a query sequence. This approach results in fewer initial sequences to examine with no loss in sensitivity. ProbeMatch has been used to align 169,095 Illumina GAII reads against the human genome, which could not be mapped by ELAND, and found alignments for 28,625 reads of the 169,095 reads in less than 3 h. Source code is freely available at (http://www.cs.wisc.edu/~jignesh/probematch/).

Extensive Analysis of the Cytoplasmic Proteome of Human Erythrocytes Using the Peptide Ligand Library Technology and Advanced Mass Spectrometry

The erythrocyte cytoplasmic proteome is composed of 98% hemoglobin; the remaining 2% is largely unexplored. Here we used a combinatorial library of hexameric peptides as a capturing agent to lower the signal of hemoglobin and amplify the signal of low to very low abundance proteins in the cytoplasm of human red blood cells (RBCs). Two types of hexapeptide library beads have been adopted: amino-terminal hexapeptide beads and beads in which the peptides have been further derivatized by carboxylation. The amplification of the signal of low abundance and suppression of the signal of high abundance species were fully demonstrated by two-dimensional gel maps and nano-LC-MSMS analysis. The effect of this new methodology on quantitative information also was explored. Moreover using this approach on an LTQ-Orbitrap mass spectrometer, we could identify with high confidence as many as 1578 proteins in the cytoplasmic fraction of a highly purified preparation of RBCs, allowing a deep exploration of the classical RBC pathways as well as the identification of unexpected minor proteins. In addition, we were able to detect the presence of eight different hemoglobin chains including embryonic and newly discovered globin chains. Thus, this extensive study provides a huge data set of proteins that are present in the RBC cytoplasm that may help to better understand the biology of this simplified cell and may open the way to further studies on blood pathologies using targeted approaches.

Functional Cell-Based Assays in Microliter Volumes for Ultra-High Throughput Screening

Functional cell-based assays have gained increasing importance for microplate-based high throughput screening (HTS). The use of high-density microplates, most prominently 1536-well plates, and miniaturized assay formats allow screening of comprehensive compound collections with more than 1 million compounds at ultra-high throughput, i.e. in excess of 100,000 samples per day. uHTS operations with numerous campaigns per year should generally support this throughput at all different steps of the process, including the underlying compound logistics, the (automated) testing of the corporate compound collection in the bioassay, and the subsequent follow-up studies for hit confirmation and characterization. A growing number of reports document the general feasibility of cell-based uHTS in microliter volumes. In addition, full automation with integrated robotic systems allows the realization of also complex assay protocols with multiple liquid handling and signal detection steps. For this review, cell-based assays are categorized based on the kinetics of the cellular response to be quantified in the test and the readout method employed. Thus, assays measuring fast cellular responses with high temporal resolution, e.g., receptor mediated calcium signals or changes in membrane potential, are at one end of this spectrum, while tests quantifying cellular transcriptional responses mark the opposite end. Trends for cell-based uHTS assays developed at Bayer-Schering Pharma are, first, to incorporate assay integral reference signals allowing the experimental differentiation of target hits from non-specifically acting compounds, and second, to make use of kinetic, real-time readouts providing additional information on the mode-of-action of test compounds.

Eye movements and prospective memory: What the eyes can tell us about prospective memory

In this study we used eye tracking methodology in combination with multi-element displays to examine the processes underlying event-based prospective memory in a visual search paradigm. In the task individuals searched for a different target stimulus (i.e., a letter) on each trial that could be present or absent, and made prospective responses to the letters D or M. The response accuracy data revealed that target hits were more frequent than prospective hits, and that there was no difference in response time for target and prospective responses. The eye tracking data revealed that both first and total fixation durations increased from distractors (stimuli that were neither targets nor prospective cues) to targets to prospective cues when a target or prospective response was made. These measures also revealed that the presence of a target in the display served to disrupt prospective memory. In addition, prospective memory misses resulted from a combination of failures to fixate the prospective cues and failures to engage in strategic processing of the prospective cues. These data demonstrate the utility of eye tracking methods in examining the processes underlying event-based prospective memory.

Controlling reaching movements with predictable and unpredictable target motion in 10-year-old children and adults

We investigated interception behavior in adults and 10-year-old children. Participants had to intercept virtual targets moving on either a predictable (linear) or unpredictable (non-linear) path (with random direction changes). Targets moved at two different velocities, which varied randomly from trial to trial. Participants reached for the targets via a force-feedback device. Reaching movements for linearly moving targets in a block of linearly moving targets were compared with reaching movements for linearly moving targets in the context of non-linearly moving targets. Movement direction and maximum speed of the first sub-movement were analyzed as well as frequency of target hits and number of sub-movements. Unpredictable target motion caused faster movement speeds than predictable target motion for both children and adults. Additionally, unpredictable target motion caused children and adults to gear their initial movement direction further towards the current position of the target, while with predictable target motion, they geared their initial movement direction further ahead of the target towards an anticipated interception position. Together, these results suggest differential processing of predictable and unpredictable object behavior in an interception task, and that this differential processing is already in place in 10-year-olds.

Shared Control in Haptic Systems for Performance Enhancement and Training

This paper presents a shared-control interaction paradigm for haptic interface systems, with experimental data from two user studies. Shared control, evolved from its initial telerobotics applications, is adapted as a form of haptic assistance in that the haptic device contributes to execution of a dynamic manual target-hitting task via force commands from an automatic controller. Compared to haptic virtual environments, which merely display the physics of the virtual system, or to passive methods of haptic assistance for performance enhancement based on virtual fixtures, the shared-control approach offers a method for actively demonstrating desired motions during virtual environment interactions. The paper presents a thorough review of the literature related to haptic assistance. In addition, two experiments were conducted to independently verify the efficacy of the shared-control approach for performance enhancement and improved training effectiveness of the task. In the first experiment, shared control is found to be as effective as virtual fixtures for performance enhancement, with both methods resulting in significantly better performance in terms of time between target hits for the manual target-hitting task than sessions where subjects feel only the forces arising from the mass-spring-damper system dynamics. Since shared control is more general than virtual fixtures, this approach may be extremely beneficial for performance enhancement in virtual environments. In terms of training enhancement, shared control and virtual fixtures were no better than practice in an unassisted mode. For manual control tasks, such as the one described in this paper, shared control is beneficial for performance enhancement, but may not be viable for enhancing training effectiveness.

Optimal Compensation for Changes in Task-Relevant Movement Variability

Effective movement planning should take into account the consequences of possible errors in executing a planned movement. These errors can result from either sensory uncertainty or variability in movement planning and production. We examined the ability of humans to compensate for variability in sensory estimation and movement production under conditions in which variability is increased artificially by the experimenter. Subjects rapidly pointed at a target region that had an adjacent penalty region. Target and penalty hits yielded monetary rewards and losses. We manipulated the task-relevant variability by perturbing visual feedback of finger position during the movement. The feedback was shifted in a random direction with a random amplitude in each trial, causing an increase in the task-relevant variability. Subjects were unable to counteract this form of perturbation. Rewards and penalties were based on the perturbed, visually specified finger position. Subjects rapidly acquired an estimate of their new variability in <120 trials and adjusted their aim points accordingly. We compared subjects' performance to the performance of an optimal movement planner maximizing expected gain. Their performance was consistent with that expected from an optimal movement planner that perfectly compensated for externally imposed changes in task-relevant variability. When exposed to novel stimulus configurations, aim points shifted in the first trial without showing any detectable trend across trials. These results indicate that subjects are capable of changing their pointing strategy in the presence of externally imposed noise. Furthermore, they manage to update their estimate of task-relevant variability and to transfer this estimate to novel stimulus configurations.

Experience-dependent behavioral plasticity is disturbed following traumatic injury to the immature brain

Traumatic brain injury (TBI) is most prevalent in children and young adults. The long-term effects of pediatric TBI include cognitive and behavioral impairments; however, over time, it is difficult to distinguish individual variability in intellect and behavior from sequelae of early injury. Postnatal day (PND) 19 rats underwent lateral fluid percussion (FP) injury, followed by rearing in either standard (STD) or enriched environment (EE) conditions. The hypothesis was that the traditional enhancement of cognitive functioning following EE rearing would be attenuated when this rearing is preceded by TBI at PND19. Thirty days after injury, Morris water maze (MWM) acquisition and subsequent probe trial retention were used to assess the behavioral effects of injury on experience-dependent plasticity induced by housing in EE at two different time windows. MWM acquisition demonstrated improvements following early EE rearing in both sham and injured animals; however, the degree of improvement was greater for uninjured animals. When EE rearing was delayed for 2 weeks after injury, the injury effect was absent and the effect of rearing even stronger. Memory testing in the early EE groups using a delayed probe trial showed an effect of injury and housing, with the sham EE animals benefiting the most. After the delayed EE, sham EE animals again showed more probe target hits, while FPEE animals did not, demonstrating an enduring memory deficit. These data confirm that early TBI has effects on experience-dependent plasticity resulting in long-term neurobehavioral deficits. In addition, the ability to benefit from environmental stimulation following TBI is dependent upon time after injury.

Numerical Estimation of Munitions Payload using Random Distributions

Abstract This mathematical research involves formulating mathematical explanations and solving discrete probabilistic problems related to the numerical estimation of target hits. These estimation formulas enable the cost estimation of operating and support funding for weapons payloads. Two of the three analytical problems involve estimating the probability that a target is hit after a tactical munitions is dispensed from an aircraft or missile launcher and the submunitions are ejected to clear a targeted area. The first problem entails determining the probability of hitting a target if a munitions may fall within a circle of radius r with a landing spread resembling a Gaussian or bivariate normal distribution, i.e., a two-dimensional probability distribution given jointly by (X,Y) ∼N(0, [sgrave]2); the second problem generalizes the first and assumes that a mine's true position and a munitions’ targeting are subject to random errors whose distributions are circular Gaussian; this problem is solved using d...

The effect of recovery duration on running speed and stroke quality during intermittent training drills in elite tennis players

The aim of this study was to assess the effect of the recovery duration in intermittent training drills on metabolism and coordination in sport games. Ten nationally ranked male tennis players (age 25.3±3.7 years, height 1.83±0.8 m, body mass 77.8±7.7 kg; mean ±s x ) participated in a passing-shot drill (baseline sprint with subsequent passing shot) that aimed to improve both starting speed and stroke quality (speed and precision). Time pressure for stroke preparation was individually adjusted by a ball-machine and corresponded to 80% of maximum running speed. In two trials (T10, T15) separated by 2 weeks, the players completed 30 strokes and sprints subdivided into 6 2 5 repetitions with a 1 min rest between series. The rest between each stroke-and-sprint lasted either 10 s (T10) or 15 s (T15). The sequence of both conditions was randomized between participants. Post-exercise blood lactate concentration was significantly elevated in T10 (9.04±3.06 vs 5.01±1.35 mmol·l-1, P ≪ 0.01). Running time for stroke preparation (1.405±0.044 vs 1.376±0.045 s, P ≪ 0.05) and stroke speed (106±12 vs 114±8 km·h-1, P ≪ 0.05) were significantly decreased in T10, while stroke precision - that is, more target hits ( P ≪ 0.1) and fewer errors (P ≪ 0.05) - tended to be higher. We conclude that running speed and stroke quality during intermittent tennis drills are highly dependent on the duration of recovery time. Optimization of training efficacy in sport games (e.g. combined improvement of conditional and technical skills) requires skilful fine-tuning of monitoring guidelines.

An Investigation of Learning during Propofol Sedation and Anesthesia Using the Process Dissociation Procedure

Many studies have shown that patients may remember words learned during apparently adequate anesthesia. Performance on memory tests may be influenced by explicit and implicit memory. We used the process dissociation procedure to estimate implicit and explicit memory for words presented during sedation or anesthesia. We investigated intraoperative learning in 72 women undergoing pervaginal oocyte collection during propofol and alfentanil infusion. One word list was played once before infusion, another was played 10 times during surgery. Venous blood was taken for propofol assay at the end of the intraoperative list. Behavioral measures of anesthetic depth (eyelash reflex, hand squeeze response to command) were recorded and used to adjust the dose of anesthetic where clinically appropriate. On recovery, memory was assessed using an auditory word stem completion test with inclusion and exclusion instructions. The mean blood propofol concentration was 2.5 microg/ml (median, 2.3 microg/ml; range, 0.7-6.1 microg/ml). Mean alfentanil dose was 2.1 mg (median, 2.0 mg; range, 1.2-3.4 mg). Comparison of target and distractor hits in the inclusion condition showed memory for preoperative words only. However, the process dissociation procedure estimates showed explicit (mean, 0.18; P < 0.001) and implicit (mean, 0.05; P < 0.05) memory for the preoperative words, and a small amount of explicit memory for the intraoperative words (mean, 0.06; 95% confidence interval, 0.01-0.10). Memory performance did not differ between the 17 patients who consistently responded to command and eyelash reflex and the 32 patients who remained unresponsive. Blood propofol concentration and alfentanil dose did not correlate with memory for the intraoperative list. There was no unprompted recall of surgery, but the process dissociation procedure showed memory for words presented during surgery. This memory was apparently explicit but did not correlate with the measures of depth of anesthesia used.