In vivo Hemodynamic Evaluation of an Implantable Left Ventricular Assist Device in a Long-term Anti-coagulation Regimen.

Left ventricular assist devices (LVADs) are used to treat patients with severe (New York Heart Association class IV) heart failure. Thrombosis and bleeding are severe LVAD-related complications; thus, an effective anticoagulation regimen is crucial for successful postoperative management. The CH-VAD™ (CH Biomedical, Inc.) is a small, implantable, full-support (>5 L/min) LVAD with a centrifugal flow pump that has a fully magnetically levitated rotor, which confers superior hemocompatibility. In this study, the CH-VAD™ was implanted in two calves to evaluate its hemocompatibility and to establish an anticoagulation regimen for future GLP (good laboratory practice) studies. Heparin infusion was used during the surgery, and during postoperative management, the proper dosage of warfarin was given orally to maintain an international normalized ratio (INR) between 2.0 and 3.0. Pump performance, animal condition, and hematology results were recorded throughout the study (approximately 60 days). The results show that under the established anticoagulation regimen, the CH-VAD™ was well tolerated in the bovine model, with no significant thrombus or thromboembolic lesion formation in distal end organs. Low plasma free hemoglobin levels suggest that the device did not cause hemolysis. These results and the experience gained pave the way for future GLP studies.

A natural indole alkaloid, norharmane, affects PIN expression patterns and compromises root growth in Arabidopsis thaliana.

Norharmane is an indole alkaloid that can be found in several terrestrial plants, as well as in some dinoflagellates and cyanobacteria. The aim of this study was to focus on the way this metabolite impacts the plant metabolism of the model species Arabidopsis thaliana. This metabolite caused increase of secondary and adventitious roots, as well as torsion, toxic effects, and a decrease in root length. Moreover, norharmane altered the cellular arrangement, resulting in unfinished cell walls, decreased auxin content and inhibited PIN proteins activity. All the alterations suggest that norharmane alters polar auxin transport by inhibiting PIN2, PIN3 and PIN7 transport proteins, thus causing a significant inhibitory effect on the growth of A. thaliana seedlings.

Rosmarinic acid induces programmed cell death in Arabidopsis seedlings through reactive oxygen species and mitochondrial dysfunction.

Phytotoxic potential of rosmarinic acid (RA), a caffeic acid ester largely found in aromatic species, was evaluated on Arabidopsis through metabolomic and microscopic approaches. In-vitro bioassays pointed out that RA affected root growth and morphology, causing ROS burst, ROS scavengers activity inhibition and consequently, an alteration on cells organization and ultrastructure. In particular, RA-treatment (175 µM) caused strong vacuolization, alteration of mitochondria structure and function and a consistent ROS-induced reduction of their transmembrane potential (ΔΨm). These data suggested a cell energy deficit also confirmed by the metabolomic analysis, which highlighted a strong alteration of both TCA cycle and amino acids metabolism. Moreover, the increase in H2O2 and O2- contents suggested that RA-treated meristems underwent oxidative stress, resulting in apoptotic bodies and necrotic cells. Taken together, these results suggest that RA inhibits two of the main ROS scavengers causing high ROS accumulation, responsible of the alterations on mitochondrial ultrastructure and activity through ΔΨm dissipation, TCA-cycle alteration, cell starvation and consequently cell death on Arabidopsis seedlings. All these effects resulted in a strong inhibition on root growth and development, which convert RA in a promising molecule to be explored for further use in weed management.

Auxin-like effects of the natural coumarin scopoletin on Arabidopsis cell structure and morphology.

The mode of action and phytotoxic potential of scopoletin, a natural compound belonging to the group of coumarins, has been evaluated in detail. Analysis conducted by light and electron transmission microscopy showed strong cell and tissue abnormalities on treated roots, such as cell wall malformations, multi-nucleated cells, abnormal nuclei and tissue disorganization. Scopoletin compromised root development by inducing wrong microtubule assembling, mitochondrial membrane depolarization and ultimate cell death, in a way similar to auxin herbicides. The structural similarities of the natural compound scopoletin and the auxin herbicide 2,4-D, as well as the ability of scopoletin to fit into the auxin-binding site TIR1, were analyzed, suggesting that the phytotoxic activity of scopoletin matches with that exhibited by auxinic herbicides.

The miniaturized pediatric continuous-flow device: Preclinical assessment in the chronic sheep model.

BACKGROUND: The Infant Jarvik 2015 is an implantable axial-flow ventricular assist device (VAD) that has undergone the major evolutionary design modifications to improve hemocompatibility. This study was conducted in anticipation of data submission to the US Food and Drug Administration to obtain Investigational Device Exemption approval. METHODS: The VAD was implanted via a left thoracotomy in Barbado sheep (n = 10, 26 (19-34] kg). Anticoagulation was maintained with coumadin, with a target international normalized ratio of greater than the individual sheep's baseline values. The VAD was managed at the highest possible speed as clinically tolerable. Complete necropsy was performed at the end of the study. RESULTS: There were 2 early mortalities: tension pneumothorax (n = 1) and shower emboli of the fragmented myocardium (n = 1). The remaining 8 sheep (2 with 30-day and 6 with 60-day protocols) completed the anticipated study duration in excellent condition, with the 6 completing 60-day sheep showing appropriate weight gain during support. There were no signs of clinically significant hemolysis, with the final plasma-free hemoglobin of 2 (1-17) mg/dL. Necropsy showed old renal infarction in 7 sheep. Although thromboembolism can be the potential etiology, given the mild anticoagulation regimen, other sources of emboli were identified in 2 sheep (graft coating material and fragmented myocardium). Flow study demonstrated favorable increase in flow (up to 3.0 L/min) in proportion to change in pump speed. CONCLUSIONS: This study has demonstrated that the Infant Jarvik 2015 VAD is capable of maintaining its functionality for an extended period of time with minimal hemolysis.

In Vivo Evaluation of a Pneumatic Extracorporeal Ventricular Assist Device for up to 90 Day Support.

In a previous study, we showed that the Vitalmex Extracorporeal Ventricular Assist Device-Pneumatic (EVAD-P)-a low-cost, pneumatically actuated, pulsatile blood pump-is easy to implant and safe for short-term in vivo support (30 ± 5 days). In the current study, we included additional 30 day experiments and assessed the safety and durability of the EVAD-P for up to 90 days of support. Using the same surgical procedure as in the previous study, we implanted the device into 14 healthy sheep. Group I subjects (n = 7) were evaluated for up to 30 days, and group II (n = 2) and group III (n = 5) subjects were evaluated for up to 49 and 93 days, respectively. After a system redesign, two of the five sheep in group III reached the scheduled end-point without device-related problems at a fixed beat rate of 56 bpm, a stroke volume (SV) of 58.0 ± 2.3 ml, and a flow of 3.5 ± 0.2 L/min. This study shows that the EVAD-P can provide safe pulsatile mechanical circulatory support (MCS) for up to 93 days. To further confirm that the system can consistently provide MCS for this duration, additional studies are recommended.

Ninety-day survival of a calf implanted with a continuous-flow total artificial heart.

We evaluated the effects of steady state flow and perfusion on end-organ function in a long-term calf model. The animal received a continuous-flow total artificial heart (CFTAH) that we created from two axial-flow ventricular assist devices. Pump flow, blood pressure, and other pump parameters were monitored throughout the study, as were arterial blood gas and hematologic values, including neurohormone levels. Some hematologic values were mildly abnormal transiently after surgery but returned to acceptable levels within the first week. During the 90-day study, the calf showed no signs of hemolysis or thrombosis. Its mental function remained normal, as evidenced by the animal's interest in its surroundings and response to stimuli. End-organ and vasomotor function was not adversely affected by 90 days of steady state flow. This was the first study in which CFTAH support of an animal model was maintained for this duration.

Elective tracheotomy facilitates total artificial heart implantation in calves.

Chronic ventilatory support may be required for survival after total artificial heart (TAH) implantation in calves. However, prolonged or repeated intubation may negatively affect a calf's ability to eat, drink, ruminate, and stand after surgery. To mitigate these limitations, we performed tracheotomies on 23 consecutive calves at the time of TAH implantation. The tracheostomies served as the primary route for ventilatory support and airway management. Tracheostomies were left in place for up to 25 days (mean, 7 +/- 7 days) and were well tolerated. Prolonged presence of a tracheostomy facilitated cleaning of the airway secretions and did not inhibit the calf's ability to stand, eat, or drink. The calves survived from 2 to 49 days (mean, 8 days). Twenty-two calves that maintained their tracheostomies died of causes unrelated to airway management. The calf that had the tracheostomy removed on day 25 died of respiratory arrest on day 49. At necropsy, we found that severe tracheal stenosis had developed at the tracheostomy site. In calves, an elective tracheotomy can be a valuable adjunct to TAH implantation or other procedures that might require prolonged ventilatory support or airway access, but care should be exercised if the tracheostomy is removed to ensure that the calf does not develop late tracheal stenosis.

Mechanical cardiac assistance improves outcome after prolonged hemorrhagic shock.

To examine the use of mechanical cardiac assist devices in prolonged hemorrhagic shock lasting up to 120 min. We induced hemorrhagic shock in anesthetized calves that were then treated 30 or 120 min later with either conventional fluid and blood resuscitation methods or the implantation of a mechanical assist device in addition to conventional fluid resuscitation. We measured hemodynamic and hematologic variables, inflammatory mediators, end-organ function via biochemical parameters, and survival time. Although cardiac output and blood flow in the left anterior descending artery decreased significantly in all calves at the end of the hemorrhage period, the drop was significantly less severe in calves who received mechanical assistance in addition to fluids. Furthermore, the biochemical profile, indicating liver and kidney function, and survival time were better after hemorrhage in device-treated calves than in conventionally treated calves. Levels of inflammatory mediators, which contribute to cell and organ dysfunction, were increased after hemorrhage, but calves with mechanical devices had less of an increase than did calves treated only with fluids. Our results indicate that the use of a mechanical cardiac assist device in combination with conventional fluid and blood resuscitation methods improves survival and end-organ recovery and decreases the myocardial inflammatory response after prolonged hemorrhagic shock when compared with the sole use of conventional fluid resuscitation techniques.

Circadian temperature rhythm of laboratory swine.

The circadian temperature rhythm (CTR) profile holds promise for monitoring the domestic pig's responses to stress and illness. In the present study we quantified the CTR profile of nine growing-finishing swine using a time-series, small-group design. Temperature was monitored using a probe implanted in the ear for 5 1/2 to 9 1/2 consecutive days while the unrestrained pigs were housed singly in pens. The dominant period of the temperature data was estimated with the autocorrelation function and then used in standard cosinor analysis to compute the amplitude (half of the distance between the highest and lowest value within the period), mesor (rhythm-adjusted mean), and acrophase (timing of the cosine maximum). To examine the effect of procedural stress on CTR, we compared data from the first 3 days with those from subsequent days. Eight of the nine (89%) pigs had CTR with a mean (+/- standard error) period of 23.6 (0.5) h, amplitude of 0.18 (0.02) degrees C, mesor of 38.7 (0.24) degrees C, and acrophase at 19:44 h. Mean mesor and acrophase were not different, but amplitude was lower (P = 0.03) during the first 3 days after instrumentation than during subsequent days. We conclude that: 1) laboratory-housed, unrestrained, growing-finishing swine have CTR; 2) our ear-based instrumentation protocol imposes acute stress as reflected in attenuated CTR amplitude during the first 3 days after instrumentation; and 3) CTR adaptation to stress appears to occur over time.

Feasibility of a porcine adult intensive care model.

A porcine adult ICU model would be useful for several avenues of investigation relevant to the care of critically ill patients. The purpose of the experiments reported here was to test the feasibility of such a model, using healthy swine. Swine (n = 4; body weight, 76 +/- 5 kg) were instrumented with endotracheal, bladder, and central arterial and venous catheters, and were admitted to the intensive care unit (ICU) while undergoing mechanical ventilation under the continuous care of nurses. Cardiopulmonary parameters were monitored continuously, and serum biochemical parameters were measured intermittently. Survival was seven days in subject 1 and five and a half days in subject 2. Subjects 3 and 4 survived an abbreviated protocol (44 and 41 h, respectively). Care of the subjects was complicated by iatrogenic hemorrhage (n = 3), pneumonia (n = 2), and acute respiratory distress syndrome (n = 1). One subject was free of complications. Critically ill swine > or = 70 kg can survive mechanical ventilation in the ICU for up to seven days. When iatrogenic injury occurs, swine respond well to clinical care protocols. Further testing is needed to develop a reproducible model and determine whether healthy swine can survive the ICU environment for longer than 41 h.