Paul A. Iaizzo

Muscle strength in overwintering bears

Unlike humans, bears retain their muscle tone when moribund for long periods.

Temperature-modulated pressure ulcers: A porcine model

OBJECTIVE A reliable porcine model was developed to facilitate investigations of pressure ulcer formation, healing, and prevention. In the present study, it was specifically used to study the relationship between applied temperature, applied pressure, and time of application in the formation of cutaneous and deep tissue injuries. DESIGN An apparatus and procedure were created to simultaneously apply 12 metal discs (each with a diameter of 51mm) on the dorsal aspect of the swine, all at an equal pressure of 100mmHg, for a 5-hour period, while servo-controlling disc temperatures at either 25, 35, 40, or 45 degrees C. RESULTS The severity of the resultant tissue injuries correlated with an increase in applied temperature. No damage was observed in the superficial or deep tissues underlying the sites of the 25 degrees C pressure applicators. In general, only deep tissue damage resulted from the application of a 35 degrees C temperature, whereas the application of higher temperatures caused both cutaneous and subdermal damage (the extent of necrosis being greater at the 45 degrees C sites). There was a high degree of reproducibility of these results among a large population of sites per temperature (n = 64) and number of animals investigated (n = 16). Furthermore, subsequent healing (monitored up to 4 weeks) was uniform for the degree of induced damage. Insights into pressure ulcer formation were also sought via systematic examination of histological sides and postmortem visual assessment over the 4-week period. CONCLUSION It was concluded that this animal model of temperature-modulated pressure ulcers has the potential for significant use in all major areas of this field, ie, wound formation, healing, and prevention.

Isolated four-chamber working swine heart model

BACKGROUND Isolated heart models separate cardiac characteristics from systemic characteristics with subsequent findings used in cardiac research, including responses to pharmacologic, mechanical, and electrical components. The model objective was to develop the ability to represent in situ physiologic cardiac function ex vivo. METHODS Swine hearts were chosen over rat or guinea pig models due to their notably greater anatomical and physiologic similarities to humans. An in vitro apparatus was designed to work all four chambers under simulated in situ physiologic conditions. Using standard cardiac surgical techniques, 12 porcine hearts (mean weight 331 +/- 18 g) were explanted into the apparatus. Preload and afterload resistances simulated in situ input and output physiologic conditions. Hemodynamic characterizations, including cardiac output, max +/- dP/dt, and heart rate, were used to determine in situ function leading to explantation (prethoracic operation, postmedial sternotomy, and postperidectomy) and during in vitro function (t = 0, 60, 120, and 240 minutes). RESULTS In vitro performance decayed with time, with statistical differences from base line (t = 0) function at t = 240 minutes (p > 0.05). CONCLUSIONS An isolation and in vitro explantation protocol has been improved to aid in the study of isolated cardiac responses, and to determine cardiac hemodynamic function during open chest operation, transplantation, and in vitro reanimation with a crystalloid perfusate. The resulting model offers similar working physiologic function, with real-time imaging capabilities. The resulting model is advantageous in representing human cardiac function with regard to anatomic and physiologic functions, and can account for atrial and ventricular interactions.

Fura-2 detected myoplasmic calcium and its correlation with contracture force in skeletal muscle from normal and malignant hyperthermia susceptible pigs

Fura-2 was used to estimate myoplasmic [Ca2+] in intact intercostal muscle fibers from normal and malignant hyperthermia susceptible (MHS) pigs. Small muscle bundles (20–50 fibers) were loaded with the membrane-permeant form of the dye. Resting myoplasmic [Ca2+] were not significantly different in normal and MHS muscles. Halothane produced increases in myoplasmic Ca2+ with associated contractures in MHS muscles, but not in normal muscles. These halothane effects were reversible. Caffeine produced increases in myoplasmic Ca2+ and contractures in both MHS and normal muscles. The threshold concentrations were lower in the MHS muscles. The correlations between myoplasmic [Ca2+] and force in MHS and normal muscles were similar.

The Cerebral and Systemic Effects of Movement in Response to a Noxious Stimulus in Lightly Anesthetized Dogs Possible Modulation of Cerebral Function by Muscle Afferents

BackgroundAfferentation theory predicts that agents or maneuvers that stimulate muscle stretch receptors (i.e., muscle afferents) will produce cerebral stimulation. From this theory it follows that, regardless of the source (e.g., drug effect, active muscle movement), increases in stretch receptor activity should result in a similar effect on the brain. The present study tested the hypothesis that active muscle movement in lightly anesthetized subjects would result in cerebral stimulation. MethodsStudies were conducted in six dogs who were lightly anesthetized with halothane (0.70% end-expired). The following physiologic variables were quantified before and for 6 min after the initiation of a standardized (1-min duration) noxious stimulus to the trachea and the skin overlying the hind limb: cerebral blood flow, cerebral metabolic rate for oxygen (CMRO2), cerebral perfusion pressure, cerebral vascular resistance, electroencephalogram activity, electromyogram activity, arterial carbon dioxide partial pressure (PaCO2), central venous pressure, and serum epinephrine and norepincphrine concentrations. Response to stimulation was evaluated initially in unparalyzed dogs and later was evaluated in the same dogs after they were paralyzed with intravenous pancuronium (0.2 mg/kg). ResultsIn unparalyzed dogs, stimulation produced episodes of coughing plus head and limb movement during the 6-min study period. Accompanying the movement was activation of the electromyogram, an increase in electroencephalogram frequency, and a reduction in electroencephalogram amplitude. There also was a 35% increase in cerebral blood flow, a 25% decrease in cerebral vascular resistance, and a 7% increase in CMRO2 versus the baseline values for each variable. There were no significant increases in either cerebral perfusion pressure, central venous pressure, PaCO2, or serum norepinephrine concentration to account for the cerebral effects; however, serum epinephrine concentrations increased by 61%. In pancuronium-paralyzed dogs, noxious stimulation resulted in a 5% increase in cerebral blood flow, a 7% decrease in cerebral vascular resistance, and an 5% increase in CMRO2 versus baseline levels. Electroencephalogram frequency was increased, but amplitude was unchanged. Central venous pressure, electromyogram activity, and serum norepinephrine concentration were unaffected. The serum epinephrine response was similar to that observed when the dogs were not paralyzed. ConclusionsThese data support the hypothesis that active muscle movement in lightly anesthetized subjects has an effect on the brain that is mediated in part by muscle afferent receptors. This cerebral response was manifested as electroencephalogram activation, cerebral vasodilation unrelated to central venous pressure changes, and an increase in cerebral blood flow greater than that required to meet metabolic demands. Paralysis with pancuronium abolished movement induced by stimulation (and, thus, the muscle afferent response) and also attenuated the cerebral blood flow, cerebral vascular resistance, and electroencephalogram responses.

Wound status evaluation using color image processing

An accurate diagnosis of burns and pressure ulcers in the early stages can be made by computerized image processing. This study describes a critical assessment of potential methodologies for noninvasive wound evaluation using a color imaging system. The authors also developed a method for quantifying histological readings and applied these techniques to a porcine animal model of wound formation. Differences in calibrated hue between injured and noninjured skin provided a repeatable differentiation of wound severity for situations when the time of injury was known. This color analysis allowed statistically significant differentiation of mild, moderate, and severe injuries within 30 minutes after the application of the injury. It was more difficult to distinguish wound severity one to four days later, however the correlation re-emerged when the wounds were five to seven days old. This technique could be adapted for assessing and tracking wound severity in humans in a clinical setting.

Characteristics of Na+ channels and Cl- conductance in resealed muscle fibre segments from patients with myotonic dystrophy.

1. Electrical and contractile properties of resealed fibre segments were investigated by a variety of in vitro techniques. The preparations were removed from skeletal muscles of normal subjects and of eight patients with myotonic dystrophy. 2. Several hours after removal, fibre segments from normal subjects and those patients in whom myotonia was the primary symptom had resting membrane potentials of approximately ‐80 mV. In contrast, fibre segments obtained from patients in whom muscle dystrophy was more expressed were depolarized (‐60 to ‐70 mV). 3. Contractions induced in fibre segments of myotonic muscle which had normal potentials were characterized by slowed relaxation which was due to electrical after‐activity. 4. After single stimuli, long‐lasting (3‐100) runs of action potentials were recorded intracellularly from the myotonic muscle. In some of these fibre segments complex repetitive discharges were observed: multiple sites of locally gated currents were identified. 5. The three‐electrode voltage clamp was used to determine the total membrane conductance, gm, and the ion component conductances. All fibres of a particular patient had similar conductances. However, the Cl‐ conductance varied from patient to patient from normal (74% of gm) to low values (30% of gm). The K+ conductance was normal in all fibres of all patients. 6. The patch‐clamp technique was used to record currents through single Na+ channels of the sarcolemma. After treatment of the fibre segments with collagenase gigaohm seals were routinely obtained. The rate of success was greater when using the cell‐attached mode than the inside‐out mode. 7. Sodium channel currents were elicited by depolarizing voltage steps which produced an initial burst of Na+ channel openings. Up to ten channels were activated simultaneously when the patch was depolarized to potentials more positive than ‐30 mV. The Na+ channels re‐opened very rarely in controls. The macroscopic sodium current, INa, was reconstructed by averaging depolarizing pulses. The time constant of rapid decay of INa reflecting macroscopic inactivation, the onset of INa and the amplitude of INa were voltage dependent. The mean amplitude of the current produced by re‐openings was on average only 0.11 +/‐ 0.04% of the amplitude of the peak current. 8. Late openings of the Na+ channels were frequent in patches on the myotonic fibre segments. The amplitude of the current produced by re‐openings was as high as about 0.75 +/‐ 0.11% of the amplitude of the peak current.(ABSTRACT TRUNCATED AT 400 WORDS)

Investigation of the thermal and tissue injury behaviour in microwave thermal therapy using a porcine kidney model

Minimally invasive microwave thermal therapies are being developed for the treatment of small renal cell carcinomas (RCC, d<3 cm). This study assessed the thermal history and corresponding tissue injury patterns resulting from microwave treatment of the porcine renal cortex. Three groups of kidneys were evaluated: (1) in vitro treated, (2) in vivo with 2-h post-treatment perfusion (acute) and (3) in vivo with 7-day post-treatment perfusion (chronic). The kidneys were treated with an interstitial water-cooled microwave probe (Urologix, Plymouth, MN) that created a lesion centered in the renal cortex (50 W for 10 min). The thermal histories were recorded at 0.5 cm radial intervals from the probe axis for correlation with the histologic cellular and vascular injury. The kidneys showed a reproducible 2 cm chronic lesion with distinct histologic injury zones identified. The thermal histories at the edge of these zones were found using Lagrangian interpolation. The threshold thermal histories for microvascular injury and stasis appeared to be lower than that for renal epithelial cell injury. The Arrhenius kinetic injury models were fit to the thermal histories and injury data to determine the kinetic parameters (i.e. activation energy and frequency factor) for the thermal injury processes. The resultant activation energies are consistent in magnitude with those for thermally induced protein denaturation. A 3-D finite element thermal model based on the Pennes bioheat equation was developed and solved using ANSYS (V7.0). The real geometry of the kidneys studied and temperature dependent thermal properties were used in this model. The specific absorption rate (SAR) of the microwave probe required for the thermal modelling was experimentally determined. The results from the thermal modelling suggest that the complicated change of local renal blood perfusion with temperature and time during microwave thermal therapy can be predicted, although a first order kinetic model may be insufficient to capture blood flow changes. The local blood perfusion was found to be a complicated function of temperature and time. A non-linear model based on the degree of vascular stasis was introduced to predict the blood perfusion. In conclusion, interstitial microwave thermal therapy in the normal porcine kidney results in predictable thermal and tissue injury behaviour. Future work in human kidney tissue will be necessary to confirm the clinical significance of these results.

Delayed onset of malignant hyperthermia induced by isoflurane and desflurane compared with halothane in susceptible swine

Background:Desflurane (difluoromethyl 1-fluoro 2,2,2-trifluoroethyl ether) is a new inhalational anesthetic currently under investigation for use in humans. Recently, the authors showed that desflurane is a trigger of malignant hyperthermia (MH) in susceptible swine. To date, there has been no in vivo comparison of the relative ability of inhalational anesthetics to trigger MH. The effects of desflurane, isoflurane, and halothane on six MH-susceptible purebred and six MH-susceptible mixed-bred Pietrain swine were examined. Methods:The animals were exposed to 1 MAC and 2 MAC (if MH was not triggered after 1 MAC hour) doses of each of the three volatile anesthetics in random sequence at 7-10·day intervals and changes in end-tidal CO2, arterial blood gases, serum lactate, core and muscle temperature, blood pressure, and heart rate were measured. Results:There was a statistical difference between anesthetics in the time required to trigger MH; halothane exposure resulted in the fastest onset of an MH episode (20 ± 5 min), compared with isoflurane (48 ± 24 min) and desflurane (65 ± 28 min), both of which required significantly longer exposures. There was no statistical difference between the MH purebred and mixed-bred swine in the time required to trigger MH (defined as a Paco2 of 70 mmHg) with a given agent, and time to triggering was also independent of the order of exposure to the three anesthetics. Malignant hyperthermia susceptibility was confirmed in ten surviving animals, by both in vivo succinylcholine challenge and in vitro contracture testing. Conclusions:Although all three volatile anesthetics triggered MH, exposure to halothane resulted in significantly shorter times to MH triggering when compared with desflurane and isoflurane.

Hibernating Bears Conserve Muscle Strength and Maintain Fatigue Resistance

Black bears spend several months each winter confined to a small space within their den without food or water. In nonhibernating mammals, these conditions typically result in severe muscle atrophy, causing a loss of strength and endurance. However, an initial study indicated that bears appeared to conserve strength while denning. We conducted an in vivo, nonsubjective measurement of strength, resistance to fatigue, and contractile properties on the tibialis anterior muscle of six hibernating bears during both early and late winter using a rigid leg brace and foot force plate. After 110 d of anorexia and confinement, skeletal muscle strength loss in hibernating bears was about one‐half that in humans confined to bed rest. Bears lost 29% of muscle strength over 110 d of denning without food, while humans on a balanced diet but confined to bed for 90 d have been reported to lose 54% of their strength. Additionally, muscle contractile properties, including contraction time, half‐relaxation time, half–maximum value time, peak rate of development and decay, time to peak force development, and time to peak force decay did not change, indicating that no small‐scale alterations in whole‐muscle function occurred over the winter. This study further supports our previous findings that black bears have a high resistance to atrophy despite being subjected to long‐term anorexia and limited mobility.