Adenosine Receptor Agonist HE-NECA Enhances Antithrombotic Activities of Cangrelor and Prasugrel in vivo by Decreasing of Fibrinogen Density in Thrombus

Blood platelets’ adenosine receptors (AR) are considered to be a new target for the anti-platelet therapy. This idea is based on in vitro studies which show that signaling mediated by these receptors leads to a decreased platelet response to activating stimuli. In vivo evidence for the antithrombotic activity of AR agonists published to date were limited, however, to the usage of relatively high doses given in bolus. The present study was aimed at verifying if these substances used in lower doses in combination with inhibitors of P2Y₁₂ could serve as components of dual anti-platelet therapy. We have found that a selective A_2A agonist 2-hexynyl-5’-N-ethylcarboxamidoadenosine (HE-NECA) improved the anti-thrombotic properties of either cangrelor or prasugrel in the model of ferric chloride-induced experimental thrombosis in mice. Importantly, HE-NECA was effective not only when applied in bolus as other AR agonists in the up-to-date published studies, but also when given chronically. In vitro thrombus formation under flow conditions revealed that HE-NECA enhanced the ability of P2Y₁₂ inhibitors to decrease fibrinogen content in thrombi, possibly resulting in their lower stability. Adenosine receptor agonists possess a certain hypotensive effect and an ability to increase the blood–brain barrier permeability. Therefore, the effects of anti-thrombotic doses of HE-NECA on blood pressure and the blood–brain barrier permeability in mice were tested. HE-NECA applied in bolus caused a significant hypotension in mice, but the effect was much lower when the substance was given in doses corresponding to that obtained by chronic administration. At the same time, no significant effect of HE-NECA was observed on the blood–brain barrier. We conclude that chronic administration of the A_2A agonist can be considered a potential component of a dual antithrombotic therapy. However, due to the hypotensive effect of the substances, dosage and administration must be elaborated to minimize the side-effects. The total number of animals used in the experiments was 146.     

Michell cantilevers constructed within a half strip. Tabulation of selected benchmark results

The paper delivers the benchmark results for the Michell cantilevers constructed within a half strip, for selected values of the σ _T /σ _C ratio, σ _T , σ _C being the admissible stresses in tension and compression, respectively.     

A micromechanical flow sensor for microfluidic applications

We fabricated a microfluidic flow meter and measured its response to fluid flow in a microfluidic channel. The flow meter consisted of a micromechanical plate, coupled to a laser deflection system to measure the deflection of the plate during fluid flow. The 100 /spl mu/m square plate was clamped on three sides and elevated 3 /spl mu/m above the bottom surface of the channel. The response of the flow meter was measured for flow rates, ranging from 2.1 to 41.7 /spl mu/L/min. Several fluids, with dynamic viscosities ranging from 0.8 to 4.5/spl times/10/sup -3/ N/m, were flowed through the channels. Flow was established in the microfluidic channel by means of a syringe pump, and the angular deflection of the plate monitored. The response of the plate to flow of a fluid with a viscosity of 4.5/spl times/10/sup -3/ N/m was linear for all flow rates, while the plate responded linearly to flow rates less than 4.2 /spl mu/L/min of solutions with lower dynamic viscosities. The sensitivity of the deflection of the plate to fluid flow was 12.5/spl plusmn/0.2 /spl mu/rad/(/spl mu/L/min), for a fluid with a viscosity of 4.5/spl times/10/sup -3/ N/m. The encapsulated plate provided local flow information along the length of a microfluidic channel.     

Optimization of reinforcement layout of scissor‐type bridge using differential evolution algorithm

Scissors mechanisms are commonly used in safety engineering during the construction of temporary structures, owing to their inherent advantages of foldability, transformability, and reusability. We effectively utilized these scissors mechanism features to develop a lightweight, deployable emergency Mobile Bridge (MB) based on optimization, and control of the folding structure. Here, we discuss the problems of optimal reinforcement layout for the MB by formulating and solving three optimization problems, namely: (a) the load capacity maximization problem, (b) the weight minimization problem, and (c) coupling the load capacity maximization problem and the weight minimization problem. The potential benefits resulting from the application of reinforcement were evaluated using a combination of finite element analysis and an optimization algorithm based on the differential evolution method. The results demonstrate the significant positive influence of the additional reinforcing members. In particular, the limit load was increased by over 10 times, while the weight was decreased to approximately half. The proposed methodology enabled the development of a substantially improved version of the MB characterized by a higher load capacity and lower weight in comparison to the initial bridge design.     

An electrospray ionization source for integration with microfluidics

We have demonstrated a new electrospray ionization (ESI) device incorporating a tip made from a shaped thin film, bonded to a microfluidic channel, and interfaced to a time-of-flight mass spectrometer (TOFMS). A triangular-shaped thin polymer tip was formed by lithography and etching. A microfluidic channel, 20 microm wide and 10 microm deep, was embossed in a cyclo olefin substrate using a silicon master. The triangular tip was aligned with the channel and bonded between the channel plate and a flat plate to create a microfluidic channel with a wicking tip protruding from the end. This structure aided the formation of a stable Taylor cone at the apex of the tip, forming an electrospray ionization source. This source was tested by spraying several solutions for mass spectrometric analysis. Because the components are all made by lithographic approaches with high geometrical fidelity, an integrated array system with multiple channels can be formed with the same method and ease as a single channel. We tested a multichannel system in a multiplexed manner and showed reliable operation with no significant cross contamination between closely spaced channels.     

A Soft-Landing Waveform for Actuation of a Single-Pole Single-Throw Ohmic RF MEMS Switch

A soft-landing actuation waveform was designed to reduce the bounce of a single-pole single-throw (SPST) ohmic radio frequency (RF) microelectromechanical systems (MEMS) switch during actuation. The waveform consisted of an actuation voltage pulse, a coast time, and a hold voltage. The actuation voltage pulse had a short duration relative to the transition time of the switch and imparted the kinetic energy necessary to close the switch. After the actuation pulse was stopped, damping and restoring forces slowed the switch to near-zero velocity as it approached the closed position. This is referred to as the coast time. The hold voltage was applied upon reaching closure to keep the switch from opening. An ideal waveform would close the switch with near zero impact velocity. The switch dynamics resulting from an ideal waveform were modeled using finite element methods and measured using laser Doppler vibrometry. The ideal waveform closed the switch with an impact velocity of less than 3 cm/s without rebound. Variations in the soft-landing waveform closed the switch with impact velocities of 12.5 cm/s with rebound amplitudes ranging from 75 to 150 nm compared to impact velocities of 22.5 cm/s and rebound amplitudes of 150 to 200 nm for a step waveform. The ideal waveform closed the switch faster than a simple step voltage actuation because there was no rebound and it reduced the impact force imparted on the contacting surfaces upon closure     

A randomized phase-II study of BB-10010 (macrophage inflammatory protein- 1alpha) in patients with advanced breast cancer receiving 5-fluorouracil, adriamycin, and cyclophosphamide chemotherapy

BB-10010 is a variant of the human form of macrophage inflammatory protein-1alpha (MIP-1alpha), which has been shown in mice to block the entry of hematopoietic stem cells into S-phase and to increase their self-renewal capacity during recovery from cytotoxic damage. Its use may constitute a novel approach for protecting the quality of the stem cell population and its capacity to regenerate after periods of cytotoxic treatment. Thirty patients with locally advanced or metastatic breast cancer were entered into the first randomized, parallel group controlled phase II study. This was designed to evaluate the potential myeloprotective effects of a 7-day regimen of BB-10010 administered to patients receiving six cycles of 5-fluorouracil (5-FU), adriamycin, and cyclophosphamide (FAC) chemotherapy. Patients were randomized, 10 receiving 100 microgram/kg BB-10010, 11 receiving 30 microgram/kg BB-10010, and nine control patients receiving no BB-10010. BB-10010 was well-tolerated in all patients with no severe adverse events related to the drug. Episodes of febrile neutropenia complicated only 4% of the treatment cycles and there was no difference in incidence between the treated and nontreated groups. Studies to assess the generation of progenitor cells in long-term bone marrow cultures were performed immediately preceding chemotherapy and at the end of six dosing cycles in 18 patients. Circulating neutrophils, platelets, CD 34(+) cells, and granulocyte/macrophage colony-forming cell (GM-CFC) levels were determined at serial time points in cycles 1, 3, and 6. The results showed similar hemoglobin and platelet kinetics in all three groups. On completion of the six treatment cycles, the average pretreatment neutrophil levels were reduced from 5.3 to 1.7 x 10(9)/L in the control patients and from 4.3 to 1.9 and 4.5 to 2.5 x 10(9)/L in the 30/100 microgram/kg BB-10010 groups, respectively. Relative to their pretreatment values, 50% of the patients receiving BB-10010 completed the treatment with neutrophil values significantly higher than any of the controls (P = .02). Mobilization of GM-CFC was enhanced by BB-10010 with an additional fivefold increase over that generated by chemotherapy alone, giving a maximal 25-fold increase over pretreatment values. Bone marrow progenitor assays before and after this standard regimen of chemotherapy indicated little long-term cumulative impairment to recovery from chemotherapy. Despite the limited cumulative damage to the bone marrow, which may have minimized the protective value of BB-10010 during this regimen of chemotherapy, better recovery of neutrophils in the later treatment cycles with BB-10010 was indicated in a number of patients.     

Prediction of the structures of proteins with the UNRES force field, including dynamic formation and breaking of disulfide bonds

The presence of disulfide bonds is essential for maintaining the structure and function of many proteins. The disulfide bonds are usually formed dynamically during folding. This process is not accounted for in present algorithms for protein-structure prediction, which either deduce the possible positions of disulfide bonds only after the structure is formed or assume fixed disulfide bonds during the course of simulated folding. In this work, the conformational space annealing (CSA) method and the UNRES united-residue force field were extended to treat dynamic formation of disulfide bonds. A harmonic potential is imposed on the distance between disulfide-bonded cysteine side-chain centroids to describe the energetics of bond distortion and an energy gain of 5.5 kcal/mol is added for disulfide-bond formation. Formation, breaking and rearrangement of disulfide bonds are included in the CSA search by introducing appropriate operations; the search can also be carried out with a fixed disulfide-bond arrangement. The algorithm was applied to four proteins: 1EI0 (alpha), 1NKL (alpha), 1L1I (beta-helix) and 1ED0 (alpha + beta). For 1EI0, a low-energy structure with correct fold was obtained both in the runs without and with disulfide bonds; however, it was obtained as the lowest in energy only with the native disulfide-bond arrangement. For the other proteins studied, structures with the correct fold were obtained as the lowest (1NKL and 1L1I) or low-energy structures (1ED0) only in runs with disulfide bonds, although the final disulfide-bond arrangement was non-native. The results demonstrate that, by including the possibility of formation of disulfide bonds, the predictive power of the UNRES force field is enhanced, even though the disulfide-bond potential introduced here rarely produces disulfide bonds in native positions. To the best of our knowledge, this is the first algorithm for energy-based prediction of the structure of disulfide-bonded proteins without any assumption as to the positions of native disulfides or human intervention. Directions for improving the potentials and the search method are suggested.