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Characterization of the distribution of matter in hybrid liver support devices where cells are cultured in a 3-D membrane network or on flat substrata.

Bioreactors for liver assist tested on small animal models are generally scaled-up to treat humans by increasing their size to host a given liver cell mass. In this process, liver cell function in different culture devices is often established based on the metabolite concentration difference between the bioreactor inlet and outlet irrespective of how matter distributes in the bioreactor. In this paper, we report our investigation aimed at establishing whether bioreactor design and operating conditions influence the distribution of matter in two bioreactors proposed for liver assist. We investigated a clinical-scale bioreactor where liver cells are cultured around a three-dimensional network of hollow fiber membranes and a laboratory-scale bioreactor with cells adherent on collagen-coated flat substrata. The distribution of matter was characterized under different operating modes and conditions in terms of the bioreactor residence time distribution evaluated by means of tracer experiments and modeled as a cascade of N stirred tanks with the same volume. Under conditions recommended by the manufacturers, matter distributed uniformly in the clinical-scale bioreactor as a result of the intense backmixing (N=1) whereas axial mixing was negligible in the laboratory-scale bioreactor (N=8). Switching from recycle to single-pass operation definitely reduced axial mixing in the clinical-scale bioreactor (N=2). Increasing feed flow rate significantly enhanced axial mixing in the laboratory-scale bioreactor (N=4). The effects of design, operating mode and conditions on matter distribution in bioreactors for liver cell culture suggest that characterization of the distribution of matter is a necessary step in the scale-up of bioreactors for liver assist and when function of liver cells cultured in different bioreactors is evaluated and compared.

Bioreactors↗

Methylenedioxymethamphetamine (MDMA, 'ecstasy') serves as a robust positive reinforcer in a rat runway procedure.

Although 'ecstasy' (3,4-methylenedioxymethamphetamine, MDMA) is, after marijuana, the second most prevalent illegal drug of abuse in European adolescents, animal experimental evidence of MDMA's reinforcing effect has remained scarce, particularly in the rodent model, raising questions about the robustness of MDMA's reinforcing effect under controlled laboratory conditions. In the present rat runway study, Sprague-Dawley and Long-Evans rats were given the opportunity to run for intravenous injections of saline or MDMA (1 mg/kg). MDMA significantly decreased runtimes in both rat strains. Thus, MDMA's positive reinforcing effect can be demonstrated not only across rat strains but also across operant conditioning paradigms. These findings should reassure the drug abuse research community that the investigation of MDMA's reinforcing effect in the inexpensive and widely used rodent model is indeed feasible.

Animals↗

Implicit operant learning of pain sensitization.

Operant conditioning mechanisms have been demonstrated to be important in the development of chronic pain behavior, but it is not clear whether and how this extends to pain perception itself. The fear-avoidance theory suggests that hypersensitivity may be induced by anticipatory pain avoidance learned through negative reinforcement by acute reductions of pain and fear. But the precise mechanism of the assumed 'sensory decalibration' has not been specified. The present study with healthy subjects investigated whether operant learning of enhanced short-term sensitization may provide the 'proximal' mechanism and whether gradual learning of hypersensitivity can take place without subjects' awareness. We used an experimental model of implicit learning based on a behavioral adjustment method of sensitization measurement developed and validated previously, combining it with standard methods of operant response shaping of increased sensitization or habituation. Results indicated that operant discrimination training with reinforcement of short-term sensitization in the seconds range can produce gross up or down changes in sensitivity within an hour without subjects' awareness of reinforcement contingencies. Consequently, implicit learning of enhanced pain sensitization may be a suitable model to investigate operant plasticity of pain perception in addition to basic sensory and neuronal mechanisms and to link these with the clinical construct of pain-fear avoidance.

Adaptation, Physiological↗

A mass balance study to assess the extent of contaminant removal achieved in the operations for the primary recovery of plasmid DNA from Escherichia coli cells.

Mass balances were performed on an alkaline lysis operation for the primary recovery of supercoiled plasmid DNA as part of a process for plasmid gene preparation. Escherichia coli DH5alpha/pSVbeta was cultured in defined medium by fed-batch fermentation and harvested at the end of the exponential phase. Alkaline lysis of the recombinant cells was performed at fixed shear rates ranging between 46 and 461 s(-1), with neutralization 100 and 300 s after the initiation of the lysis. Mass balance calculations were used to optimize the operating conditions for carrying out the alkaline lysis operation. The results indicated that a plasmid yield of 75% and purity with respect to total DNA of 60% were achievable during the primary recovery operation. The influences of key contaminants, including the soluble proteins and the suspended solids, as they bear on the subsequent purification operations, were evaluated and discussed.

Bacteriolysis↗

The production and generalization of large-magnitude heart rate deceleration by contingently faded biofeedback.

Eight subjects were taught to decrease their heart rates via biofeedback training. Four of these received contingently faded, beat-by-beat analogue feedback and contingent reinforcement each time their performance met a specified and adjusting criterion. The other four received continuous, beat-by-beat analogue feedback, but not the contingent reinforcement. Subjects in the two groups were yoked to ensure equal densities of reinforcement. Subjects in the first group were asked to decrease heart rates 15% from baseline and were then trained using only 75%, 50% and 25% of beat-by-beat feedback. It was hypothesized that the immediate reinforcement of appropriate behavior and the contingent fading (following mastery) of feedback would aid in the generalization of the response. Following completion of all criterion steps or 10 training sessions, whichever came first, all subjects were tested with no feedback and no contingent reinforcement. The group receiving contingently faded feedback training showed a significantly greater heart rate decrease in the training sessions and also the test session. These results were interpreted as indicating that biofeedback can be conceptualized as an operant conditioning paradigm, and that the use of operant techniques may help subjects produce clinically significant changes.

Biofeedback, Psychology↗

Dual nebulizer sample introduction system for simultaneous determination of volatile elemental hydrides and other elements.

A dual sample introduction system was explored for volatile hydride generation in inductively coupled plasma-optical emission spectrometry (ICP-OES) performed in the radially viewed mode. The system consists of two pneumatic nebulizers connected to the conventional spray chamber of the instrument via a simple adaptor. This configuration permits hydride generation but still allows other elements to be determined by pneumatic nebulization. This work was focused on the optimization of the plasma operating conditions for the determination of As, Hg, Sb and Se and other elements. The excitation conditions of the ICP-OES instrument operated with the dual sample introduction system were also explored. Results showed that the analytical performance of the dual system for the determination of As, Hg, Sb and Se was superior to those of conventional nebulization systems. The dual system also enabled the determination of elements that do not form volatile hydrides, but with less sensitivity than conventional nebulization systems. An evaluation of the plasma robustness showed that the gases generated in the hydride reactions did not significantly affect the plasma discharge. Similar to conventional hydride generation techniques, the analysis was susceptible to nonspectroscopic interferences produced by transition metals. Finally, the applicability of the dual nebulization system to practical ICP-OES studies was demonstrated by determining the trace elements in an oyster tissue standard reference material.

Journal Article↗

Sugar-dependent rats show enhanced responding for sugar after abstinence: evidence of a sugar deprivation effect.

Studies have shown that intermittent sugar availability (12 h/day) produces signs of dependence in rats, including escalation of intake, mu-opioid and dopamine receptor changes, behavioral and neurochemical indices of withdrawal, and cross-sensitization with amphetamine. "Deprivation-effect" paradigms, whereby abstinence from a substance results in enhanced intake, are often used to measure "craving" for drugs of abuse, such as alcohol. The present study used operant conditioning to investigate consumption of sugar after abstinence in rats selected for glucose avidity. The experimental group was trained on a fixed ratio (FR-1) schedule for 25% glucose for 30 min/day for 28 days and also had glucose access in the home cages for an additional 11.5 h daily. The control group had only the 30-min/day access to glucose in the operant chambers. Then, both groups were deprived of glucose for 2 weeks. After this period of abstinence, animals were put back in the operant chambers. The experimental group responded significantly more than ever before, and significantly more than the control group. In conclusion, daily 12-h access to sugar, in the paradigm used, can result in an altered neural state that lasts throughout 2 weeks of abstinence, leading to enhanced intake. Together with previous results, this deprivation effect supports the theory that animals may become dependent on sugar under selected dietary circumstances.

Amphetamine-Related Disorders↗

Subject variation in caprine anterior cruciate ligament reconstruction.

We studied the subject and treatment contributions to anterior cruciate ligament (ACL) reconstruction biomechanics by reexaming the results of two bilateral reconstruction studies. Bilateral reconstruction allows a comparison between treatments exposed to the same subject related healing factors. The studies examined the effects of gamma irradiation and the effects of initial graft size and initial graft laxity. In both studies different treatments were applied to contralateral limbs. We found that the subject was the best predictor of outcome, while the surgical treatments had little influence on outcome. There was a large variation between subjects despite similar treatments, and little difference between contralateral limbs despite different surgical treatments. At 26 weeks, the graft cross sectional area and modulus were most strongly influenced (p < 0.002) by the subject. We interpret this as a subject related factor is regulating the quantity and quality of the healing tissue. Potential sources of subject related factors include the subject's pre-operative condition, the activity during the post-operative period, and an intrinsic biologic response. By better understanding the source of subject variation, more successful and consistent ACL reconstructions might be achieved.

Animals↗

Prospective evaluation of topical versus retrobulbar anesthesia: a converting surgeon's experience.

PURPOSE: To evaluate and compare the efficacy of topical versus retrobulbar anesthesia for cataract surgery performed by a surgeon newly converting to the topical technique. SETTING: Department of Ophthalmology, Moran Eye Center, University of Utah, Salt Lake City, Utah, USA. METHODS: Ninety patients were prospectively assigned by permuted block restricted randomization to receive topical (Group 1; n = 45) or retrobulbar (Group 2; n = 45) anesthesia. Group 1 received topical bupivacaine 0.75% and intravenous midazolam and fentanyl for anesthesia. Group 2 received intravenous methohexital followed by retrobulbar block with an equal mixture of lidocaine 2% and bupivacaine 0.75% plus hyaluronidase 150 units. A visual pain analog scale was used to assess the degree of pain during anesthesia administration and surgery and postoperatively. The degree to which eye movement, touch, and light caused patient discomfort was assessed. Intraoperative conditions and complications were recorded. RESULTS: Intraoperative operating conditions were significantly better in Group 2 (P < .05). There was a small but statistically significant difference in the degree of discomfort during anesthesia administration and surgery (P < .05). There was no difference in postoperative discomfort. Chemosis, subconjunctival hemorrhage, and eyelid hemorrhage occurred only in Group 2, in which there was one retrobulbar hemorrhage. Although eyelid squeezing and ocular motility were present more frequently in Group 1, neither was a problem to the surgeon. CONCLUSION: Cataract surgery was safely performed by a surgeon converting to topical anesthesia. After a distinct learning curve, the procedure was performed with minimal patient discomfort. Surgical training and patient preparation are the key to safe use of topical anesthesia.

Administration, Topical↗

Polycyclic aromatic hydrocarbon and particulate emissions from two-stage combustion of polystyrene: the effect of the primary furnace temperature.

A study is presented on laboratory-scale combustion of polystyrene (PS) to identify staged-combustion conditions that minimize emissions. Batch combustion of shredded PS was conducted in fixed beds placed in a bench-scale electrically heated horizontal muffle furnace. In most cases, combustion of the samples occurred by forming gaseous diffusion flames in atmospheric pressure air. The combustion effluent was mixed with additional air, and it was channeled to a second muffle furnace (afterburner) placed in series. Further reactions took place in the secondary furnace at a residence time of 0.7 s. The gas temperature of the primary furnace was varied in the range of 500-1,000 degrees C, while that of the secondary furnace was kept fixed at 1,000 degrees C. Sampling for CO, CO2, O2, soot, and unburned hydrocarbon emissions (volatile and semivolatile, by GC-MS) was performed at the exits of the two furnaces. Results showed that the temperature of the primary furnace, where PS gasifies, is of paramount importance to the formation and subsequent emissions of organic species and soot. Atthe lowesttemperatures explored, mostly styrene oligomers were identified at the outlet of the primary furnace, but they did not survive the treatment in the secondary furnace. The formation and emission of polycyclic aromatic hydrocarbons (PAH) and soot were suppressed. As the temperature in the first furnace was raised, increasing amounts of a wide range of both unsubstituted and substituted PAH containing up to at least seven condensed aromatic rings were detected. A similar trend was observed for total particulate yields. The secondary furnace treatment reduced the yields of total PAH, but it had an ambiguous effect on individual species. While most low molecular mass PAH were reduced in the secondary furnace, concentrations of some larger PAH increased under certain conditions. Thus, care in the selection of operating conditions of both the primary furnace (gasifier/ burner) and the secondary furnace (afterburner) must be exercised to minimize the emission of hazardous pollutants. The emissions of soot were also reduced in the afterburner but not drastically. This indicates that soot is indeed resistant to oxidation; thus, it would be best to avoid its formation in the first place. An oxidative pyrolysis temperature of PS in the vicinity of 600 degrees C appears to accomplish exactly that. An additional afterburner treatment at a sufficiently high temperature (1,000 degrees C) may be a suitable setting for minimization of most pollutants. To obtain deeper understanding of chemical processes, the experimental results were qualitatively compared with preliminary predictions of a detailed kinetic model that describes formation and destruction pathways of chemical species including most PAH observed in the present work. The modeling was performed forthe secondary furnace assuming plug-flow conditions therein. The experimentally determined chemical composition at the outlet of the primary furnace was part of the input parameters of the model calculation.

Air Movements↗

Thermal lesions produced by CO2 laser beams: new findings to improve the quality of minimally invasive and transmyocardial laser revascularization protocols.

OBJECTIVE: The objective of this study was to investigate the possible existence of a set of critical combinations among medical CO2 laser device set-ups and to compare the resigns of diameter and depth of irradiated tissue. These data are useful to the surgeon to identify a general operative protocol that allows both to forecast the beam behavior on a large variety of operative conditions accurately and to identify the related safety margins. Two methods have been addressed: the more traditional free-air beam delivery to the tissue and the more modern requirements of the minimally invasive surgery concepts, which use fiberoptic-based delivery systems for the CO2 laser beam. BACKGROUND DATA: In the past many articles have been written about the interaction between laser beams and biological media. However, the CO2 laser beam at 10.6 microm has always challenged research institutions and manufacturers to find the ideal combinations between a suitable fiberoptic-based delivery system concept and the injury threshold conditions recommended for minimally invasive surgery (MIS) and transmyocardial laser revascularization (TMLR) applications. One of the main challenges is to deliver the radiation at 10.6 microm without serious damage to the fiberoptic-based scalpel and the irradiated tissue by the burn of the fiber in use. METHODS: Five rabbits weighing 3 to 3.5 kg were sacrificed and 60 samples of trachea, myocardium, aorta, and esophagus were immediately excised, trimmed free of the adherent connective tissue, and irradiated in the intima portion of the wall. A commercial TEM medical CO2 laser was coupled to a silver halide fiber (0.9 mm in diameter) and subsequently to regular focusing heads (2.5-inch and 5-inch focal length) for pulsed laser beam delivery. The parameters chosen were: 33, 50, 65, and 100 mJ per pulse, 10 and 20 ms pulse width, delivered in two sets of experiments, one with all the pulsed beam frequencies below 5 Hz, the other between 15 Hz and 20 Hz. The same tests were conducted on 10 blocks of 3 different plastic types to simulate in one single procedure the laser radiation in responses to both hard, low-water content tissues such as bone and of common plastic compounds (such as polymethylmethacrylates [PMMA]) routinely used in orthopedic surgery. An optical microscope was used to measure all the lesions (diameter and depth in millimeters) in all the samples and to identify the smallest and the largest one against which similar thermal injuries found on the other media were compared. RESULTS: This study demonstrates that for power densities between about 520 and 790 W/cm2 per pulse achieved with the silver halide fiber generated a minimal lesion on the myocardium and aortic tissues. This can be used as reference threshold for MIS and TMLR. The minimal injury threshold on PMMA has been reached at 393 W/cm2 per pulse at 1 Hz. This approached the conditions of a pulsed beam delivered in air on the same medium via a focal spot of an 8.7 inch focal. Surprisingly, all the treated media show lesions that follow similar patterns within a well-defined and limited range of both diameter and depth. This effect was obtained by using power densities ranging from 393 to 6310 W/cm2 per pulse regardless of all the other parameters, including power delivery method, the type of irradiated tissue and the frequency between 0 and 20 Hz. Only the combination power density-type of tissue appears to be decisive. The geometrical convergence of the diameter shows a much smoother pattern than the one of the depth, due to two different irradiation modalities. CONCLUSIONS: The selection of the CO2 laser beam parameters and the irradiated media reported in this article have allowed identification of a critical set of ablative conditions to be further used in the operating room.

Animals↗

[The application study of mimic operation with 3D-cut in maxillofacial bone tumor].

OBJECTIVE: To study the feasibility of guiding operation planning and complication prevention. METHODS: Ten patients suffer from Maxillofacial bone tumour were examined with spiral CT. Firstly, to get three-dimensional reconstructive image of tumour and other tissues. Secondly, use 3D-cut software to cut off every layer of normal tissue and tumour mass. Lastly, to compare virtual operation with mimic operation in virtual operation process. RESULTS: Three-dimensional mimic operation clearly displays the relationship between tumor and surrounding tissues, reflect the actual operation condition. CONCLUSIONS: It is very useful designing operation program and predicting the potential complications before the actual operation.

Adult↗

[Clinical studies on the use of neuroleptanalgesia in Wertheim's abdominal radical surgery].

The application of the Neurolept-Analgesia to great gynaecological operations was tested in 50 Wertheim-radical-operations with removal of the glands, in comparison to a group of 50 patients whose operations were executed under the usual narcosis. There were controlled: blood-pressure, pulse, respiration, common dosing of the narcotica, dosing of the analgetica, and the patients' post-operative condition. Under NLA, the course of the operation was characterized by stability of circulation and a considerably reduced post-operative recovery, the comparative group, however, did not show so favourable results that can be proved mathematically. The application of the Neurolept-Analgesia as described thoroughly in this paper, has to be considered as especially suitable for executing gynaecological operations.

Adult↗

Electrochemical pilot scale study for reduction of 2,4-DNT.

An electrochemical pilot scale reactor was used to treat simulated munitions wastewater containing 100 mg/L of 2,4-dinitrotoluene (DNT). Experiments were conducted by using a glassy carbon (zero porosity) coated graphite cylinder as the cathode and a platinum wire as the anode. All experiments were conducted under dissolved oxygen concentration of less than 1.5 mg/L. Initially, simulating batch conditions were conducted to obtain the optimum operating conditions for the reactor. During this batch-mode study, the effects of various parameters such as applied current, electrolyte concentration, and type of electrolyte on the reduction of DNT were evaluated. Results obtained showed that the rates of reduction of DNT increased with an increase in current or concentration of electrolyte. Based on the results obtained from the batch simulation experiments, continuous flow experiments were conducted at three different currents. The ionic strength of the feed solution was maintained at 0.027 M. A current of 200 mA was found to provide a stable reduction of DNT at the 80% level for a period of 14 days after which reactor cleaning is necessary for removal of solids that were formed within the reactor. End products determined for the continuous flow experiments showed 100% molar balance conversion.

Carcinogens↗

Flow analysis of the Cleveland Clinic centrifugal pump.

An implantable ventricular assist blood pump is being developed by the Cleveland Clinic Foundation in cooperation with the NASA Lewis Research Center. At the nominal design condition, the pump provides blood flow at the rate of 5 L/min at a pressure rise of 100 mmHg and a rotation speed of 3000 RPM. Bench testing of the centrifugal pump in a water/glycerin mixture has provided flow and pressure data at several rotative speeds. A one-dimensional empirically based pump flow analysis computer code developed at NASA Lewis Research Center has been used in the design process to simulate the flow in the primary radial pump stage. The computer model was used to size key impeller and volute geometric parameters that influence pressure rise and flow. Input requirements to the computer model include a simple representation of the pump geometry. The model estimates the flow conditions under design and off-design operating conditions at the impeller leading and trailing edges, and the volute inlet and exit. Output from the computer model is compared to flow and pressure data obtained from bench testing.

Biomedical Engineering↗

Sensitization to cocaine in pigeons: interaction with an operant contingency.

A 2-part study with pigeons investigated the role of an explicit operant contingency in determining how cocaine interacts with locomotor activity. In Part 1, pigeons pecked on a fixed-ratio-20 schedule of food presentation. In Part 2, different pigeons were studied without opportunity to peck for food. After determination of cocaine's initial effects, pigeons were exposed to daily administrations of a locomotion-increasing dose of cocaine. Locomotor sensitization was evident in the pigeons of Part 2, and tolerance developed to cocaine's effects on key pecking in the pigeons of Part 1. Locomotor sensitization was generally not evident in the pigeons of Part 1. These results suggest that explicitly conditioned operant behavior may compete with behavior sensitized by prolonged exposure to cocaine.

Animals↗

Reinforcement enhances hippocampal acetylcholine release in rats: an in vivo microdialysis study.

Rats were trained to press a lever under a 'Multiple FI-60s and Extinction' schedule with food reinforcements. After learning the task, an in vivo microdialysis probe was inserted into the dentate gyrus and CA3 regions of the hippocampus, and the sequential changes in the dialysate acetylcholine (ACh) concentration were analyzed. In the session, two fixed-interval of reinforcement (FI) components (for 20 min) and two extinction (EXT) components (for 30 min) were alternated to examine the correlation between behavioral and neurochemical outcomes. The dialysate ACh concentration increased during the FI component and returned to the baseline during the EXT component of the schedule. Next, in order to dissociate the effect of discrimination from the effect of rewarding on the neurochemical changes in the hippocampus, we used a final TEST period (for 20 min) during which the actual schedule was extinction but the discriminative stimulus was on, i.e. the manifest condition of the test period was reinforcement. In the TEST period, the animals pressed the lever with almost the same frequency as during the FI component; however, the dialysate ACh concentration did not increase above the baseline concentration. These results suggest that ACh release in the rat hippocampus is associated with reinforcement but not with discrimination in operant conditioning.

Acetylcholine↗

Effects of chronic epileptic foci on control of pyramidal tract neurons in monkeys.

Five Macaca mulatta monkeys were operantly conditioned to control the firing patterns of single precentral pyramidal tract neurons. The accuracy with which the monkeys could control normal PTNs from within the focus was significantly poorer than PTNs from contralateral, homotopic cortex. In comparison to nonepileptic monkeys, there was no significant difference in the accuracy with which PTNs from cortex contralateral to interictal foci were controlled. By contrast, comparison of the time necessary to gain accurate control over individual PTNs from contralateral cortex showed the epileptic monkeys to be significantly encumbered when compared to nonepileptic monkeys. These data suggest that interictal foci produce "noise" in remote regions of brain that are involved in an operant task requiring a high degree of discrimination.

Aluminum Hydroxide↗