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While many areas of medicine have benefited from the development of objective assessment tools and biomarkers, there have been comparatively few improvements in techniques used to assess brain function and dysfunction. Brain functions such as perception, cognition, and motor control are commonly measured using criteria-based, ordinal scales which can be coarse, have floor/ceiling effects, and often lack the precision to detect change. There is growing recognition that kinematic and kinetic-based measures are needed to quantify impairments following neurological injury such as stroke, in particular for clinical research and clinical trials. This paper will first consider the challenges with using criteria-based ordinal scales to quantify impairment and recovery. We then describe how kinematic-based measures can overcome many of these challenges and highlight a statistical approach to quantify kinematic measures of behavior based on performance of neurologically healthy individuals. We illustrate this approach with a visually-guided reaching task to highlight measures of impairment for individuals following stroke. Finally, there has been considerable controversy about the calculation of motor recovery following stroke. Here, we highlight how our statistical-based approach can provide an effective estimate of impairment and recovery.

Introduction: Effective initial fluid resuscitation is the cornerstone intervention in the setting of severe burn injury. Critically, few major advances in burn resuscitation have been made since the 1970s, and since that time there has been only modest improvement in overall morbidity and mortality. Recently, investigations regarding the dynamic changes of vascular endothelium, and more specifically the vascular endothelial glycocalyx, in the setting of severe burn injury and resuscitation have offered insight into the possibility of more tightly controlling fluid shifts and understanding the consequences thereof during this critical period., Methods: We conducted a literature search of the PubMed database using the terms "burn", and "glycocalyx" limited to studies published in the English language over the past 10 y. A total of 31 articles were initially identified. Abstracts and full text were manually reviewed to identify suitable articles. Of the identified articles, 10 were deemed relevant and included within this review, along with additional articles necessary to provide background on glycocalyx structure and function as well as principles of burn injury management., Results: Glycocalyx shedding is a process known to occur early in the setting of severe burn injury and resuscitation. The degree of shedding tends to increase with age and severity of injury. Though the role and regulation of this shedding is incompletely understood, it has direct consequences on vascular unction and permeability and likely coagulation as well., Conclusions: Here in this research review, we examine what is known regarding the dynamic breakdown and reconstitution of the glycocalyx during burn injury and how it may be impacted by fluid resuscitation strategies. We further explore the need to more completely understand this mechanism and the consequences of its manipulation., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Competing Interests: The authors declare no conflicts of interest.

Background: Analysis of the costs associated with emergency department (ED) visits after discharge for violent injury could highlight subgroups for the development of cost-effective interventions to support healing and prevent treatment failures in violently injured patients., Methods: A retrospective cohort review was conducted of all patients with return ED visits within 90 days of discharge after treatment for a violent injury occurring between July 1, 2016, and June 30, 2018. Hospital costs were calculated for each incidence and analyzed against demographic and injury type variables to identify trends., Results: 218 return ED visits were identified. Hospital costs showed a high frequency of low-cost visits. For more complex visits, distinct cost patterns were observed for Black and LatinX males compared to White males as a function of age., Conclusions: Analysis of hospital cost per visit identified trends among different subgroups. Underlying etiologies presumably vary between groups, but hypothesis-driven further investigation and needs assessment is required. Understanding the driving forces behind these cost trends may aid in developing effective interventions., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Introduction: Firearms and motor vehicle collisions (MVC) are leading causes of mortality in children. We hypothesized that firearm injuries would have a higher mortality than MVCs in children and a higher level of resource utilization METHODS: Trauma patients <18 years old at a Level 1 pediatric trauma center sustaining gunshot wounds (GSW) or MVCs 2009-2019 were included. The primary outcome was mortality. The secondary outcome was immediate surgery. The California Department of Public Health's Overall Injury Surveillance tool was queried for patients <18 with GSW or MVC 2006-2015 to compare statewide case fatality rates (CFRs), and analyze proportions of GSWs by intent: assault, self-inflicted, and unintentional., Results: Of 13,840 pediatric trauma patients at our institution, 295 GSWs (2.1%) and 4467 MVCs (32.3%) were included. Mortality was higher for GSWs (7.5% vs. 0.8%, p<0.0001). GSW patients were more likely to require immediate surgery (34.4% vs. 11.2%, p<0.0001). On multivariable analysis, GSW patients were 7.8-times more likely to die than MVC patients (OR 7.83, 95% CI 3.68-16.66, p<0.0001), adjusted for age, sex, and injury severity. Statewide, there were 10,790 pediatric GSWs with 1586 deaths (CFR 14.7%) vs. 710 deaths in 261,363 children in MVCs (CFR 0.3%, p<0.0001). The GSW CFR rose (13.4% to 16.5%, p = 0.05) while the MVC CFR decreased (0.5% to 0.2%, p<0.0001) in 2015 vs. 2006., Conclusion: Firearm violence in pediatric patients is significantly more lethal than MVCs and is resource intensive. The case fatality rate for pediatric firearm violence is rising. Resources must be directed at preventing pediatric firearm injuries., Level of Evidence: Prognosis study, Level II., Competing Interests: Declaration of Competing Interest None, (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Thyroid storm is a rare, life-threatening endocrine emergency with a high mortality rate of up to 30%. We present a unique management challenge of a critically ill patient who developed thyroid storm in the setting of a duodenal perforation from amphetamine-associated non-occlusive mesenteric ischaemia. The diagnosis of 'thyroid storm' was made based on clinical criteria and a Burch-Wartofsky score of 100. During emergent exploratory laparotomy, a 1 cm duodenal perforation with surrounding friable tissue was found and repaired. Intraoperatively, a nasogastric tube was guided distal to the area of perforation to allow for enteric administration of medications, which was critical in the setting of thyroid storm. Therapeutic plasma exchange achieved biochemical control of our patient's thyroid storm but ultimately did not prevent in-hospital mortality., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Limited 2021. No commercial re-use. See rights and permissions. Published by BMJ.)

Introduction: Previously, in a murine model of blunt thoracic trauma, we provided evidence of primary pulmonary thrombosis associated with increased expression of the cell adhesion molecule, P-selectin. In this study, mice are treated with P-selectin blocking antibody after injury to investigate the clinical viability of this antibody for the prevention of pulmonary thrombosis. In addition, viscoelastic testing is performed to investigate if P-selectin inhibition has a detrimental impact on normal hemostasis., Methods: A murine model of thoracic trauma was used. Mice were divided into sham control and experimental injury groups. Thirty minutes after trauma, mice were treated with the following: P-selectin blocking antibody, isotype control antibody, low-dose heparin, high-dose heparin, or normal saline. At 90 minutes, whole blood was collected for characterization of coagulation by viscoelastic coagulation monitor (VCM Vet; Entegrion, Durham, NC). Mean clotting time, clot formation time, clot kinetics (α angle), and maximum clot firmness were compared between each treatment group., Results: Mice that received P-selectin antibody 30 minutes after blunt thoracic trauma had four- to fivefold less (p < 0.001) arterial fibrin accumulation than those that received the isotype control. In both sham and trauma groups, compared with vehicle (normal saline) alone, no statistical difference was noted in any coagulation parameters after injection with P-selectin antibody, isotype control, or low-dose heparin. In addition, blinded histopathological evaluation yielded no difference in hemorrhage scores between injured mice treated with P-selectin blocking antibody and those treated with isotype antibody control., Conclusion: This study supports the clinical use of P-selectin blocking antibody for the prevention of pulmonary thrombosis by confirming its efficacy when given after a blunt thoracic trauma. In addition, we demonstrated that the administration of P-selectin antibody does not adversely affect systemic coagulation as measured by viscoelastic testing, suggesting that P-selectin antibody can be safely given during the acute posttraumatic period., (Copyright © 2021 Wolters Kluwer Health, Inc. All rights reserved.)

Background: Combined burn and traumatic brain injury (TBI) treatment priorities may not align due to opposing fluid resuscitation paradigms used in treating burns and TBI. We developed a porcine model of combined thermal injury/TBI and compared an "aggressive" fluid resuscitation strategy using the Parkland formula and a "restrictive" resuscitation strategy using the modified Brooke formula., Methods: Twenty-eight swine were deeply anesthetized and received a 40% total body surface area full-thickness burn injury and TBI. Swine were then randomized to receive restrictive or aggressive resuscitation for 8 h after which time animals were euthanized and necropsy was performed. Volume of brain injury was assessed after analyzing segmental slices of brain tissue., Results: There were no differences between the restrictive and aggressive resuscitation groups in blood pressure, heart rate, central venous pressure, intra-cranial pressure (ICP), or serum lactate levels after 8 h of resuscitation. Urine output was higher in the aggressive resuscitation group. The restrictive group had a significantly higher serum blood urea nitrogen (BUN) compared with baseline and compared with the aggressive group. There was no significant difference in size of brain injury between groups., Conclusions: Both restrictive and aggressive resuscitation demonstrated adequate resuscitation at 8 h postinjury. Increased serum BUN in the restrictive group may be an indicator of early acute kidney injury, despite adequate urine output. Resuscitation strategy did not appear to affect ICP or the size of brain injury., Competing Interests: The authors report no conflicts of interest., (Copyright © 2020 by the Shock Society.)

Introduction: Viscoelastic coagulation tests are useful to assess coagulation status in the clinical setting and to aid in understanding underlying pathophysiological mechanisms that affect coagulation status. Such tests also are useful for coagulation research. Because mouse models are widely used to study molecular mechanisms in fine detail, a simple viscoelastic coagulation test requiring small blood volumes would be convenient for such studies in mice., Methods: We tested viscoelastic coagulation properties of normal healthy adult mice using a novel veterinary clinical point-of-care device, Viscoelastic Coagulation Monitor (VCM Vet™; Entegrion Corp.). Fresh whole blood was collected from 63 healthy mature adult C57 black 6N mice, with ultimately 54 mice, equal numbers of male and females, used to determine reference intervals (RIs) for VCM test parameters., Results: RIs were determined for equal numbers of male and female mice: clot time: 43.0-353.0 s; clot formation time: 49.4-137.6 s; alpha angle: 54.4-62.2°; A10: 25.0-49.6 VCM units; A20: 31.0-56.5 VCM units; maximum clot firmness: 37.6-62.8 VCM units; Lysis Index 30 (Li30): 99.8-100.0%; and Li45: 99.7-100.0%. Significant differences were found between male and female subgroups, where females had higher mean A10 and A20 and median MCF values, indicating greater clot firmness in female versus male mice., Conclusion: VCM Vet is a feasible viscoelastic coagulation test device for studies with mature adult mice, including studying inherent sex differences in coagulation parameters. Inherent differences in coagulability of male and female mice warrant further investigation to determine if such differences underlie greater coagulopathic, hemorrhagic, or thromboembolic risk during trauma or other pathophysiologic conditions., (© 2021 S. Karger AG, Basel.)

Background: Limited data are available on whether donor kidneys with diffuse glomerular fibrin thrombi (GFT) are safe to use. In this study, the clinicopathologic characteristics of allografts with diffuse donor-derived GFT were examined., Methods: All deceased donor kidney transplant implantation biopsies from our institution between July 2011 and February 2018 with diffuse GFT were included. A control group for comparison consisted of all cases with implantation biopsies obtained during the study period without diffuse GFT. Clinical data were extracted from electronic medical records for all study patients, including donor information., Results: Twenty-four recipients received kidneys with diffuse GFT from 16 deceased donors. All donors died from severe head trauma. On average, 79% of glomeruli contained fibrin thrombi. Nineteen cases had subsequent biopsy; all revealed resolution of GFT. Compared with the control group, kidneys with diffuse GFT had longer cold ischemia time (34 versus 27 h), were more frequently pumped using machine perfusion (100% versus 81%), and recipients experienced a higher frequency of delayed graft function (58% versus 27%). Only 2 grafts with diffuse GFT failed within the first year. Overall graft survival was similar between the diffuse GFT group and control group., Conclusions: Deceased donor kidneys with diffuse GFT appear to be safe to use given that nearly 92% of recipients in this cohort who received such allografts experienced good clinical outcomes. Histologically, GFT demonstrated rapid resolution following transplantation. Interestingly, diffuse GFT only occurred in donors who suffered severe head trauma in this cohort, which may be a predisposing factor.

Background: Thromboembolic events within the pulmonary arterial vasculature are a troublesome complication of severe blunt thoracic trauma. Mechanisms underlying these events are currently in question as pulmonary thromboembolic events in this particular trauma population tend to be diagnosed more rapidly, more frequently and without an associated systemic thrombosis. This study investigates the role of P-selectin in thrombus formation through the use of in vivo blocking antibodies. We hypothesize that P-selectin plays a pivotal role in de novo pulmonary arterial thrombosis following blunt thoracic trauma., Methods: A murine weight-drop model of lateral blunt thoracic trauma was used. Wild-type mice in the experimental group were given blocking antibodies against P-selectin prior to the trauma. All mice were euthanized at 24 hours for evaluation with hematoxylin-eosin staining or immunofluorescent staining for fibrin and P-selectin., Results: Injured mice that did not receive the P-selectin antibody showed a robust fourfold to fivefold increase in fibrin accumulation in both coup and contrecoup tissues (fluorescence per um of arterial wall) compared to uninjured sham mice. In contrast, mice pretreated with P-selectin blocking antibody showed no significant increase in fibrin accumulation on either side of the lungs after blunt thoracic trauma. No difference in mean fibrin deposition was found between sham controls that received the P-selectin-blocking antibody and those that received an isotype control antibody., Conclusion: P-selectin expression increases at the pulmonary arterial luminal surface following blunt thoracic trauma. In addition, P-selectin-blocking in vivo prevents pulmonary arterial fibrin accumulation after blunt thoracic trauma, confirming that P-selectin is necessary for de novo pulmonary arterial thrombosis after traumatic injury.

Pulmonary thromboembolic events cause significant morbidity and mortality after severe trauma. Clinically, these lesions are believed to be emboli arising secondary to deep venous thrombosis (DVT) in the lower extremities. Recently, this notion has been challenged by clinical studies, showing that pulmonary clots arise after trauma in the absence of DVT. This suggests that pulmonary blood clots arise in situ via de novo thrombosis. In the present study, we characterize a murine weight-drop model of lateral blunt thoracic trauma. Our model demonstrates severe unilateral lung contusion injury with low (10%) mortality in the absence of extrapulmonary injury, after impact with a 50-g weight dropped from 45 cm height (657 J/m). At 24 h after injury, immunofluorescence and histological evidence revealed early pulmonary arterial thrombosis in the form of eccentric accumulation of fibrin and CD41 positive eosinophilic proteinaceous material, on both coup and contrecoup lung lobes of injured mice, indicating early thrombotic events both within and outside of the area of primary lung injury. Our model is ideal in that lateral impact enables greater impact energy to be applied to achieve significant lung contusion without significant mortality or extrapulmonary injury, and the model has additional translational value in creating thrombosis analogous to pulmonary embolism observed clinically after blunt thoracic trauma. To our knowledge, this is the first demonstration of de novo pulmonary thrombosis in a clinically translational model of blunt thoracic trauma, and supports challenges to current assumptions about the origin of pulmonary blood clots in the wake of severe traumatic injury.

Background: Data suggest that methamphetamine may increase the risk of nonocclusive mesenteric ischemia (NOMI). We describe patterns of presentation and outcomes of patients with methamphetamine use who present with NOMI to a single institution., Methods: This is an observational study of patients from January 2015 to September 2017 with methamphetamine use who presented with NOMI at an academic medical center in Northern California. We summarize patient comorbidities, clinical presentation, operative findings, pathologic findings, hospital course, and survival., Results: Ten patients with methamphetamine use and severe NOMI were identified. One patient was readmitted with a perforated duodenal ulcer, for a total of 11 encounters. Most presented with acute (n = 3) or acute-on-chronic (n = 4) abdominal pain. Distribution of ischemia ranged from perforated duodenal ulcer (n = 3), ischemia of the distal ileum (n = 1), ischemia of entire small bowel (n = 2), and patchy necrosis of entire small bowel and colon (n = 5). Six patients died, three within 1 week of admission and three between 3 months and 8 months., Conclusion: Methamphetamine use may be associated with significant microvascular compromise, increasing the risk of mesenteric ischemia. Providers in areas with high prevalence of methamphetamine use should have a high index of suspicion for intestinal ischemia in this patient population. Patients with methamphetamine use admitted for trauma or other pathology may be at particular risk of ischemia and septic shock, especially in the setting of dehydration. Use of vasoconstrictors in this patient population may also exacerbate intestinal ischemia., Level of Evidence: Therapeutic Case series study, level V.

Introduction: Non-operative management is the standard of care for blunt solid organ injuries in stable patients. Angioembolization is an important technique that improves success rates of non-operative management. We aimed to provide a brief review of the indications, effectiveness and complications associated with angioembolization for solid organ injuries., Methods: We conducted a literature search of the PubMed database using the terms "trauma", "angioembolization", and "solid organ embolization" limited to studies published in the English language. Abstracts and full text were manually reviewed to identify suitable articles., Results: The current brief review is based on content from the more recently published articles related to angioembolization for solid organ injuries after trauma., Discussion: Angioembolization is appropriate for hemodynamically stable patients with contrast extravasation on CT scan or high-grade injury to a solid organ. Non-operative management success rates have improved with the adoption of angioembolization. The complications associated with angioembolization are acceptable in the context of avoiding a laparotomy, and are often related to the severity of the injury., Conclusion: Angioembolization is a natural extension of the move towards non-operative management for solid organ injuries. Randomized controlled trials are required to fully characterize the indications and efficacy of its use., (Copyright © 2015 IJS Publishing Group Ltd. Published by Elsevier Ltd. All rights reserved.)

Introduction: Pelvic stabilization with angioembolization (AE) is steadily supplanting operative management for the treatment of pelvic hemorrhage in trauma. We aimed to provide a brief review of the indications, effectiveness and complications associated with AE for pelvic injuries., Methods: We conducted a literature search using the terms "trauma," "angioembolization," and "pelvis" limited to studies published in the English language. Abstracts and full text were manually reviewed to identify suitable articles., Results: The current brief review is based on content from articles published in the last 10 years related to pelvic AE for retroperitoneal hemorrhage after trauma., Discussion: Pelvic injuries often require complex management because the high energy transfer causes concomitant injuries. Outcomes for hemodynamically unstable patients may be better with AE than with operative management., Conclusion: Pelvic AE is the most effective intervention for management of hemorrhage associated with pelvic fracture in both hemodynamically stable and unstable patients. It can be used as the primary definitive intervention or in conjunction with operative management in the setting of concomitant intra-abdominal injury., (Copyright © 2016 IJS Publishing Group Ltd. Published by Elsevier Ltd. All rights reserved.)

Purpose: Several clinical trials have highlighted general favorable outcomes of intravenous tissue type plasminogen activator (rt-PA) in acute ischemic stroke using different measures including, National Institutes of Health Stroke Scale (NIHSS) and modified Rankin Scale (mRS). Findings from most of these measures indicate that the benefits of rt-PA are time dependent, thus, supporting intensive efforts to fast-track hospital thrombolytic treatment in patients with stroke. Despite the widespread benefits of rt-PA, the effectiveness of this therapy on specific functions such as ambulatory performance of the recovering stroke patient is not fully investigated. We aim to investigate this issue in the current study., Methods: We analyzed data from a retrospective cohort of acute ischemic stroke patients admitted to Greenville Health System (GHS) between 2010-2013. We identified patients who received rt-PA within a 4.5 hour time frame following the onset of acute ischemic stroke symptoms. Our analysis compared ambulatory variables and hospital-level characteristics in proportions of patients receiving rt-PA with those not receiving rt-PA. This analysis determined whether early treatment with rt-PA is associated with favorable changes in ambulatory status from admission to discharge following acute ischemic stroke., Results: Among 663 patients with ischemic stroke who were eligible to receive rt-PA, 241 patients received rt-PA and 422 patients did not due to several risk factors. We found a statistically significant difference (P < 0.001) for changes in ambulation status from hospital admission to discharge between patients receiving rt-PA and patients who did not receive rt-PA. Among patients who received rt-PA, 27.8% improved in their ambulation status, 41.9% saw no change in their ambulation status, 0.4% worsened in their ambulation status, and 29.9% were unable to be determined. Of the patients who did not receive rt-PA, 20.1% improved in their ambulation status, 61.8% saw no change in their ambulation status, 1.4% worsened in their ambulation status, and 16.6% were unable to be determined., Conclusion: Our current study indicates that early treatment with rt-PA may be associated with favorable changes in ambulatory status from admission to discharge following acute ischemic stroke.

Weight loss is accompanied by several metabolic adaptations that work together to promote rapid, efficient regain. We employed a rodent model of regain to examine the effects of a regular bout of treadmill exercise on these adaptations. Obesity was induced in obesity-prone rats with 16 wk of high-fat feeding and limited physical activity. Obese rats were then weight reduced (approximately 14% of body wt) with a calorie-restricted, low-fat diet and maintained at that reduced weight for 8 wk by providing limited provisions of the diet with (EX) or without (SED) a daily bout of treadmill exercise (15 m/min, 30 min/day, 6 days/wk). Weight regain, energy balance, fuel utilization, adipocyte cellularity, and humoral signals of adiposity were monitored during eight subsequent weeks of ad libitum feeding while the rats maintained their respective regimens of physical activity. Regular exercise decreased the rate of regain early in relapse and lowered the defended body weight. During weight maintenance, regular exercise reduced the biological drive to eat so that it came closer to matching the suppressed level of energy expenditure. The diurnal extremes in fuel preference observed in weight-reduced rats were blunted, since exercise promoted the oxidation of fat during periods of feeding (dark cycle) and promoted the oxidation of carbohydrate (CHO) later in the day during periods of deprivation (light cycle) . At the end of relapse, exercise reestablished the homeostatic steady state between intake and expenditure to defend a lower body weight. Compared with SED rats, relapsed EX rats exhibited a reduced turnover of energy, a lower 24-h oxidation of CHO, fewer adipocytes in abdominal fat pads, and peripheral signals that overestimated their adiposity. These observations indicate that regimented exercise altered several metabolic adaptations to weight reduction in a manner that would coordinately attenuate the propensity to regain lost weight.

Purpose: To investigate the antitumor efficacy of T-cell anergy reversal through homeostatic proliferation and regulatory T-cell (Treg) depletion in a clinically relevant murine adoptive immunotherapy model., Experimental Design: B16 melanoma cells were engineered to express the model SIYRYYGL (SIY) antigen to enable immune monitoring. Tumor-specific T cells expanded in tumor-challenged wild-type hosts but became hyporesponsive. To examine whether lymphopenia-induced homeostatic proliferation could reverse tumor-induced T-cell anergy, total splenic T cells were transferred into lymphopenic RAG2-/- mice or control P14/RAG2-/- mice. Tumor growth was measured, and SIY-specific immune responses were monitored using ELISPOT and SIY/K(b) tetramers. To determine whether Treg depletion could synergize with homeostatic proliferation, RAG2-/- mice received total or CD25-depleted T cells, followed or preceded by B16.SIY challenge. This approach was further investigated in wild-type mice lymphodepleted with sublethal total body irradiation., Results: Adoptive transfer of total splenic T cells into RAG2-/- mice moderately affected the growth rate of B16.SIY. As Treg expansion occurred in tumor-bearing mice, CD25+ T cells were depleted from total T cells before adoptive transfer. Interestingly, transfer of CD25-depleted T cells into RAG2-/- mice resulted in potent rejection of B16 melanoma in both prophylactic and short-term preimplanted tumor settings and was associated with maintained T-cell effector function. Using a clinically applicable approach, wild-type mice were lymphodepleted using sublethal total body irradiation, which similarly supported tumor rejection upon transfer of CD25-depleted T cells., Conclusions: Our results indicate that combined CD25 depletion and homeostatic proliferation support a potent antitumor immune response--an approach with potential for clinical translation.

A new robotic exoskeleton for the upper-limb has been designed and constructed. Its primary purpose is to act as a proof-of-concept prototype for a more sophisticated rehabilitation and assessment device that is currently in development. Simultaneously, it is intended to extend the capabilities of an existing planar exoskeleton device. The robot operates in the horizontal plane and provides independent control of a user's shoulder, elbow and wrist joints using a cable-driven actuation system. The novel component of the design is a curved track and carriage which allows the mechanism that drives the shoulder joint to be located away from the user, underneath their arm. This paper describes the design of the robot, and provides an initial indication of its performance.

Absence of CD4(+) T cell help has been suggested as a mechanism for failed anti-tumor cytotoxic T lymphocytes (CTL) response. We examined the requirement for CD4(+) T cells to eliminate an immunogenic murine fibrosarcoma (6132A) inoculated into the peritoneal cavity. Immunocompetent C3H mice eliminated both single and repeat intraperitoneal (IP) inoculums, and developed high frequency of 6132A-specific interferon-gamma (IFNgamma)-producing CTL in the peritoneal cavity. Adoptive transfer of peritoneal exudate cells (PEC) isolated from control mice, protected SCID mice from challenge with 6132A. In contrast, CD4 depleted mice had diminished ability to eliminate tumor and succumbed to repeat IP challenges. Mice depleted of CD4(+) T cells lacked tumor-specific IFNgamma producing CTL in the peritoneal cavity. Adoptive transfer of PEC from CD4 depleted mice failed to protect SCID mice from 6132A. However, splenocytes isolated from same CD4 depleted mice prevented tumor growth in SCID mice, suggesting that 6132A-specific CTL response was generated, but was not sustained in the peritoneum. Treating CD4 depleted mice with agonist anti-CD40 antibody, starting on days 3 or 8 after initiating tumor challenge, led to persistence of 6132A-specific IFNgamma producing CTL in the peritoneum, and eliminated 6132A tumor. The findings suggest that CTL can be activated in the absence of CD4(+) T cells, but CD4(+) T cells are required for a persistent CTL response at the tumor site. Exogenous stimulation through CD40 can restore tumor-specific CTL activity to the peritoneum and promote tumor clearance in the absence of CD4(+) T cells.

We constructed a physiologically realistic model of a lower-limb, mammalian muscle spindle composed of mathematical elements closely related to the anatomical components found in the biological spindle. The spindle model incorporates three nonlinear intrafusal fiber models (bag(1), bag(2), and chain) that contribute variously to action potential generation of primary and secondary afferents. A single set of model parameters was optimized on a number of data sets collected from feline soleus muscle, accounting accurately for afferent activity during a variety of ramp, triangular, and sinusoidal stretches. We also incorporated the different temporal properties of fusimotor activation as observed in the twitchlike chain fibers versus the toniclike bag fibers. The model captures the spindle's behavior both in the absence of fusimotor stimulation and during activation of static or dynamic fusimotor efferents. In the case of simultaneous static and dynamic fusimotor efferent stimulation, we demonstrated the importance of including the experimentally observed effect of partial occlusion. The model was validated against data that originated from the cat's medial gastrocnemius muscle and were different from the data used for the parameter determination purposes. The validation record included recently published experiments in which fusimotor efferent and spindle afferent activities were recorded simultaneously during decerebrate locomotion in the cat. This model will be useful in understanding the role of the muscle spindle and its fusimotor control during both natural and pathological motor behavior.

Although recent work has suggested that lymphopenia-induced homeostatic proliferation may improve T cell-mediated tumor rejection, there is little direct evidence isolating homeostatic proliferation as an experimental variable, and the mechanism by which improved antitumor immunity occurs via homeostatic proliferation is poorly understood. An adoptive transfer model was developed in which tumor-specific 2C/RAG2(-/-) TCR transgenic CD8+ T cells were introduced either into the lymphopenic environment of RAG2(-/-) mice or into P14/RAG2(-/-) mice containing an irrelevant CD8+ TCR transgenic population. RAG2(-/-), but not P14/RAG2(-/-) recipients supported homeostatic proliferation of transferred T cells as well as tumor rejection. Despite absence of tumor rejection in P14/RAG2(-/-) recipients, 2C cells did become activated, as reflected by CFSE dilution and CD44 up-regulation. However, these cells showed poor IFN-gamma and IL-2 production upon restimulation, consistent with T cell anergy and similar to the hyporesponsiveness induced by administration of soluble peptide Ag. To determine whether homeostatic proliferation could uncouple T cell anergy, anergic 2C cells were transferred into RAG(-/-) recipients, which resulted in vigorous homeostatic proliferation, recovery of IL-2 production, and acquisition of the ability to reject tumors. Taken together, our data suggest that a major mechanism by which homeostatic proliferation supports tumor rejection is by maintaining and/or re-establishing T cell responsiveness.

Central nervous system (CNS) tumors are thought to be poorly immunogenic. However, whether defective anti-tumor immunity is a consequence of a relative failure of T cell priming versus a deficient effector phase of the anti-tumor immune response is not clear. We utilized a well-defined model system of B16 melanoma expressing the model antigen SIY-GFP to evaluate tumor antigen cross-priming and tumor rejection from the CNS versus subcutaneous compartments. We observed that B16-SIY cells implanted in the CNS were capable of inducing T cell priming as measured by IFN-gamma ELISPOT in the spleen. Cross-priming occurred in the absence of detectable systemic dissemination of the tumor. Despite the induction of a T cell response, CNS tumors grew progressively and were fatal, whereas the same tumor implanted in the flank was rejected. To study the effector phase of the immune response in more detail, in vitro primed 2C/RAG2-/- TCR transgenic CD8+ cells, which recognize the SIY peptide, were adoptively transferred. In addition, the CNS microenvironment was modulated by intracranial delivery of IL-2. While mice that received primed 2C cells alone showed an increase in survival, co-administration of intracranial IL-2 led to a marked prolongation of survival, with 20% of mice surviving at least 120 days. Our results demonstrate that CD8+ T cell cross-priming does indeed occur in response to a CNS tumor, but that manipulation of the brain tumor microenvironment may be necessary to support the effector phase of the anti-tumor immune response.

Maintenance of a sufficient population of naive CD8+ T cells in the peripheral lymphoid compartment is critical for immunocompetence. Peripheral T cell number is a function of T cell generation, survival, and death. Homeostasis, a critical balance between survival and death, must exist to prevent either lymphopenia or lymphocytosis. In the current review, we discuss known requirements for the survival of naive peripheral CD8+ T cells as well as mechanisms of death when survival signals are lost. We also discuss associations between survival and homeostasis-driven proliferation, and highlight the gaps in our knowledge of these critical processes.

Realistic models of neuromusculoskeletal systems can provide a safe and convenient environment for the design and evaluation of controllers for functional electrical stimulation (FES) prior to clinical trials. We have developed a set of integrated musculoskeletal modeling tools to facilitate the model building process. Simulink models of musculoskeletal systems are created using two software packages developed in our laboratory, Musculoskeletal Modeling in Simulink (MMS) and virtual muscle, in addition to one software package available commercially, SIMM (Musculographics Inc., USA). MMS converts anatomically accurate musculoskeletal models generated by SIMM into Simulink(R) blocks. It also removes run-time constraints on kinetic simulations in SIMM, and allows the development of complex musculoskeletal models without writing a line of code. Virtual muscle builds realistic Simulink models of muscles responding to either natural recruitment or FES. Models of sensorimotor control systems can be developed using various Matlab (Mathworks Inc., USA) toolboxes and integrated easily with these musculoskeletal blocks in the graphical environment of Simulink.

Enzyme-catalyzed beta-elimination of sugar uronic acids, exemplified by the degradation of plant cell wall pectins, plays an important role in a wide spectrum of biological processes ranging from the recycling of plant biomass through to pathogen virulence. The three-dimensional crystal structure of the catalytic module of a "family PL-10" polysaccharide lyase, Pel10Acm from Cellvibrio japonicus, solved at a resolution of 1.3 A, reveals a new polysaccharide lyase fold and is the first example of a polygalacturonic acid lyase that does not exhibit the "parallel beta-helix" topology. The "Michaelis" complex of an inactive mutant in association with the substrate trigalacturonate/Ca2+ reveals the catalytic machinery harnessed by this polygalacturonate lyase, which displays a stunning resemblance, presumably through convergent evolution, to the tetragalacturonic acid complex observed for a structurally unrelated polygalacturonate lyase from family PL-1. Common coordination of the -1 and +1 subsite saccharide carboxylate groups by a protein-liganded Ca2+ ion, the positioning of an arginine catalytic base in close proximity to the alpha-carbon hydrogen and numerous other conserved enzyme-substrate interactions, considered in light of mutagenesis data for both families, suggest a generic polysaccharide anti-beta-elimination mechanism.

This report describes the temporal relationship between the latency of responses to peripheral stimulation in primary somatosensory (SI) cerebral cortex and the timing of climbing fiber inputs to the lateral hemispheres of the rat cerebellum. Examined in the tactilely responsive regions of crus IIa in the rat, the results show that SI influences the timing of both evoked and spontaneous climbing fiber activity in these cerebellar regions without affecting the rate or probability of complex spike discharge. By reversibly blocking SI activity, we demonstrate that the absence of cortical input results in a lengthening of climbing fiber response latency to peripheral stimuli. Similarly, enhancing the cortical input by subthreshold electrical stimulation of SI results in a shortening of climbing fiber response latency. These results provide a new explanation for the tendency of the inferior olive to oscillate at 7-12 Hz and is consistent with the hypothesis that the inferior olive provides the cerebellum information about the timing of cortical computational cycles. Results are discussed in the context of previous and current hypotheses concerning the physiology and function of the inferior olive/climbing fiber system and are interpreted to provide additional evidence of a role for the cerebellum in the tactile somatosensory system.

Natural engagement of CTLA-4 on host B7 limits T cell activation. We hypothesized that therapeutic cross-linking of CTLA-4 in vivo may further inhibit T cell function and prevent allograft rejection. However, none of the currently available CTLA-4-binding reagents have ligating properties when injected in vivo. The observation that surface-immobilized anti-CTLA-4 mAb inhibits T cell activation in vitro prompted us to develop a membrane-bound single-chain anti-CTLA-4 Ab (7M). To model whether tissue expression of 7M could suppress allograft rejection, we examined the ability of H-2L(d)-specific TCR-transgenic T cells to reject 7M-expressing allogeneic tumor cells injected s.c. Expression of 7M significantly inhibited allogeneic rejection in mice that received CTLA-4(+/+) but not CTLA-4(-/-) T cells. Furthermore, CTLA-4(+/+) T cells that had encountered 7M-expressing tumors in vivo acquired defects in cytokine production and cytotoxicity. Thus, deliberate ligation of CTLA-4 in vivo potently inhibits allogeneic T cell responses.

When neuroscientists gather to discuss "Movement and Sensation", they tend to discuss neurons rather than muscles and bones. Neurons may be more interesting, but their roles in motor control depend on the mechanical properties of the system to be controlled. Understanding of those properties has been surprisingly elusive, despite the well-developed disciplines of biomechanics and muscle physiology. Each experimental field has its favorite, often unique preparation. Mathematical models range in scale from individual cross-bridges to articulated limbs, usually written in different computer languages. The shortcomings of such fragmented knowledge become particularly apparent when biomedical engineers must design safe and effective control systems for real limbs, such as for functional electrical stimulation (FES) of reach and grasp in quadriplegic patients. We are addressing the question of how to model neuromusculoskeletal systems so that they are sufficiently complete, valid and accessible to be useful in both basic and applied sensorimotor research.

Biological recycling of plant material is essential for biosphere maintenance. This perpetual task involves a complex array of enzymes, including extracellular polysaccharide hydrolases and lyases. Whilst much is known about the structure and function of the hydrolases, relatively little is known about the structures and mechanisms of the corresponding lyases. To this end, crystals of the catalytic module of a novel family 10 pectate lyase, Pel10A from Pseudomonas cellulosa, were obtained using polyethylene glycol 2000 monomethylether as a precipitant. They belong to space group P2(1), with unit-cell parameters a = 47.7, b = 106.1, c = 55.4 A, beta = 92.0 degrees, and have two molecules in the asymmetric unit. The crystals diffract beyond 1.5 A using synchrotron radiation.

Pectate lyase 10A (Pel10A) enzyme from Pseudomonas cellulosa is composed of 649 residues and has a molecular mass of 68.5 kDa. Sequence analysis revealed that Pel10A contained a signal peptide and two serine-rich linker sequences that separate three modules. Sequence similarity was seen between the 9.2 kDa N-terminal module of Pel10A and family 2a carbohydrate-binding modules (CBMs). This N-terminal module of Pel10A was shown to encode an independently functional module with affinity to crystalline cellulose. A high sequence identity of 66% was seen between the 14.2 kDa central module of Pel10A and the functionally uncharacterized central modules of the xylan-degrading enzymes endoxylanase 10B, arabinofuranosidase 62C and esterase 1D, also from P. cellulosa. The 35.8 kDa C-terminal module of Pel10A was shown to have 30 and 36% identities with the family 10 pectate lyases from Azospirillum irakense and an alkaliphilic strain of Bacillus sp. strain KSM-P15, respectively. This His-tagged C-terminal module of the Pel10A was shown to encode an independent catalytic module (Pel10Acm). Pel10Acm was shown to cleave pectate and pectin in an endo-fashion and to have optimal activity at pH 10 and in the presence of 2 mM Ca2+. Highest enzyme activity was detected at 62 degrees C. Pel10Acm was shown to be most active against pectate (i.e. polygalacturonic acid) with progressively less activity against 31, 67 and 89% esterified citrus pectins. These data suggest that Pel10A has a preference for sequences of non-esterified galacturonic acid residues. Significantly, Pel10A and the P. cellulosa rhamnogalacturonan lyase 11A, in the accompanying article [McKie, Vincken, Voragen, van den Broek, Stimson and Gilbert (2001) Biochem. J. 355, 167-177], are the first CBM-containing pectinases described to date.

We have examined the spatial relationship between the mossy fiber and climbing fiber projections to crus IIa in the lateral hemispheres of the rat cerebellum. Experiments were performed in ketamine/xylazine anesthetized rats using extracellular recordings and high-density micromapping techniques. Responses were elicited using small, tactile stimuli applied to the perioral and forelimb regions at a rate of 0.5 Hz. In our first series of experiments we demonstrate that the primary (i.e., strongest) receptive field for a single Purkinje cell's complex spike is similar to the primary receptive field of the granule cells immediately subjacent to that Purkinje cell. In our second series of experiments we demonstrate that the granule cell region most strongly activated by a particular peripheral stimulus is immediately subjacent to the Purkinje cells whose complex spikes are also activated most strongly by the same stimulus. The region of climbing fibers activated by a localized peripheral stimulus is "patchy"; it clearly does not conform to the notion of a continuous microzone. These results support original observations first reported in the 1960s using evoked potential recording techniques that the mossy fiber and climbing fiber pathways converge in cerebellar cortex. However, we extend this earlier work to show that the two pathways converge at the level of single Purkinje cells. Many cerebellar theories assume that mossy fiber and climbing fiber pathways carry information from different peripheral locations or different modalities to cerebellar Purkinje cells. Our results appear to contradict this basic assumption for at least the tactile regions of the lateral hemispheres., (Copyright 2001 Wiley-Liss, Inc.)

This paper describes a computational approach to modeling the complex mechanical properties of muscles and tendons under physiological conditions of recruitment and kinematics. It is embodied as a software package for use with Matlab and Simulink that allows the creation of realistic musculotendon elements for use in motor control simulations. The software employs graphic user interfaces (GUI) and dynamic data exchange (DDE) to facilitate building custom muscle model blocks and linking them to kinetic analyses of complete musculoskeletal systems. It is scalable in complexity and accuracy. The model is based on recently published data on muscle and tendon properties measured in feline slow- and fast-twitch muscle, and incorporates a novel approach to simulating recruitment and frequency modulation of different fiber-types in mixed muscles. This software is distributed freely over the Internet at http://ami.usc.edu/mddf/virtualmuscle.

Stretch-induced force enhancement and shortening-induced force depression were examined in fast-twitch feline caudofemoralis muscle at 37 degrees C. These phenomena were induced by applying ramp length changes during the first 100--200 ms of an otherwise isometric contraction. The effects of various stimulus frequencies ranging from 30 to 120 pps were investigated over lengths ranging from 0.85 to 1.15 L0. Distributed asynchronous stimulation of bundles of ventral roots was employed to produce smooth contractions at sub-tetanic stimulus frequencies in whole muscle. Of the two components of force enhancement identified by Noble (1992) we observed only the transient component that decays with time; we did not observe residual force enhancement. The force depression that we observed was symmetrical in almost all respects to the transient force enhancement, and was unlike the shortening-induced de-activation and residual force depression identified by Edman (Edman. 1975; Edman et al., 1993). Both transient force enhancement and depression were independent of work, load and activation. Reversals in the direction of ramp length changes following either an initial stretch or initial shortening were shown to cancel the effects of both transient force enhancement and transient force depression. The distances over which these cancellations could be achieved were different for the lengthening and shortening effects. This asymmetry can be reconciled with the predictions of Huxley's original cross-bridge mechanism by incorporating the recent suggestion that myosin heads can interact with multiple actin binding sites during a single 'working' stroke. We conclude that the types of force enhancement/ depression that are most likely to be encountered under physiological conditions are the transient effects observed here, but that even these will have relatively little effect on force production during most natural behaviors.

The interactive effects of length and stimulus frequency on rise and fall times and on sag were investigated in fast-twitch feline caudofemoralis at normal body temperature. The length and stimulus frequency ranges studied were 0.8 1.2 L0 and 15 60 pps. Isometric rise times were shortest under two sets of conditions: short lengths + low stimulus frequencies and long lengths + high stimulus frequencies. In contrast the isometric fall time relationship showed a single minimum at short lengths + low stimulus frequencies. Velocity was shown to have an additional effect on fall time, but only at higher stimulus frequencies (40 60 pps): fall times were shorter during movement in either direction as compared to isometric. The effects of sag were greatest at shorter lengths and lower stimulus frequencies during isometric stimulus trains. Potential mechanisms underlying this last effect were investigated by comparing isometric twitches elicited prior to and immediately following a sag-inducing stimulus train. Post-sag twitches produced less force, reached peak force earlier and initially decayed more quickly compared to pre-sag twitches. However, the final rate of force decay and the initial rate of force rise (during the first 15 ms) were unaffected by sag. We construct a logical argument based on these findings to hypothesize that the predominant mechanism underlying sag is an increase in the rate of sarcoplasmic calcium ion removal. All of the above findings were used to construct a model of activation dynamics for fast-twitch muscle, which was then extrapolated to slow-twitch muscle. When coupled with a previous model of kinematic dynamics, the complete model produced accurate predictions of the forces actually recorded during experiments in which we applied concurrent dynamic changes in length. velocity and stimulus frequency.

Interactions between physiological stimulus frequencies, fascicle lengths and velocities were analyzed in feline caudofemoralis (CF), a hindlimb skeletal muscle composed exclusively of fast-twitch fibers. Split ventral roots were stimulated asynchronously to produce smooth contractions at sub-tetanic stimulus frequencies. As described previously, the peak of the sub-tetanic force-length relationship was found to shift to longer lengths with decreases in stimulus frequency, indicating a length dependence for activation that is independent of filament overlap. The sub-tetanic force-velocity (FV) relationship was affected strongly both by stimulus frequency and by length; decreases in either decreased the slope of the FV relationship around isometric. The shapes of the force transients following stretch or shortening revealed that these effects were not due to a change in the instantaneous FV relationship; the relative shape of the force transients following stretch or shortening was independent of stimulus frequency and hardly affected by length. The effects of stimulus frequency and length on the sub-tetanic FV relationship instead appear to be caused by a time delay in the length-dependent changes of activation. In contrast to feline soleus muscle, which is composed exclusively of slow-twitch fibers, CF did not yield at sub-tetanic stimulus frequencies for the range of stretch velocities tested (up to 2 L0/s). The data presented here were used to build a model of muscle that accounted well for all of the effects described. We extended our model to account for slow twitch muscle by comparing our fast-twitch model with previously published data and then changing the necessary parameters to fit the data. Our slow-twitch model accounts well for all previous findings including that of yielding.

Activation of mammalian fast-twitch skeletal muscle induces a persistent effect known as post-activation potentiation (PAP), classically defined as an increase in force production at sub-maximal levels of activation. The underlying mechanism is thought to be phosphorylation of the myosin regulatory light chain (MRLC), which leads to an increase in the rate constant for cross-bridge attachment (Sweeney et al., 1993). If true, this suggests the hypothesis that other contractile properties should be affected during PAP. Using a feline fast-twitch whole-muscle preparation (caudofemoralis) at 37 degrees C, we observed that PAP greatly increased tetanic forces during active lengthening decreased isometric tetanic rise times and delayed isometric tetanic force relaxation. The first two of these effects were length dependent with a greater effect occurring at shorter lengths. These findings confirmed that PAP has other functionally important effects beyond a simple increase in sub-maximal isometric forces. Furthermore, length was found to have an effect independent of PAP on the shortening half of the FV relationship (less force was produced at longer lengths) and on the rate of force relaxation during the later stages of isometric tetanic force decay (slower relaxation at longer lengths). All of these findings can be explained with a simplified, two-state model of cross-bridge dynamics that accounts for the interaction of both interfilament spacing and MRLC phosphorylation on the apparent rate constants for cross-bridge attachment and detachment. These findings are largely consistent with data collected previously from reduced preparations such as skinned fibers at cold, unphysiological temperatures (e.g. 5 degrees C). One finding that could not be explained by our model was that twitch fall times in the dispotentiated state were parabolically correlated with length, whereas in the potentiated state the relationship was linear. The time course of decay of this effect did not follow the time course of force dispotentiation, suggesting that there are other activation-dependent processes occurring in parallel with MRLC phosphorylation.

Successful performance of a sensorimotor task arises from the interaction of descending commands from the brain with the intrinsic properties of the lower levels of the sensorimotor system, including the dynamic mechanical properties of muscle, the natural coordinates of somatosensory receptors, the interneuronal circuitry of the spinal cord, and computational noise in these elements. Engineering models of biological motor control often oversimplify or even ignore these lower levels because they appear to complicate an already difficult problem. We modeled three highly simplified control systems that reflect the essential attributes of the lower levels in three tasks: acquiring a target in the face of random torque-pulse perturbations, optimizing fusimotor gain for the same perturbations, and minimizing postural error versus energy consumption during low- versus high-frequency perturbations. The emergent properties of the lower levels maintained stability in the face of feedback delays, resolved redundancy in over-complete systems, and helped to estimate loads and respond to perturbations. We suggest a general hierarchical approach to modeling sensorimotor systems, which better reflects the real control problem faced by the brain, as a first step toward identifying the actual neurocomputational steps and their anatomical partitioning in the brain.

Feline caudofemoralis (CF) is a promising preparation in which to study the properties of mammalian fast-twitch skeletal muscle, but little is known about its muscle fiber properties, architecture, and motor innervation. We used histochemical techniques to confirm that it contained predominantly type IIB fibers (95+/-2%, n=8, with six of eight muscles composed exclusively of type IIA and IIB fibers), but physiological experiments showed less fatiguability than for the type IIB component of medial gastrocnemius. This may be related to the surprisingly strong and regular recruitment of CF during repetitive tasks such as walking and trotting, which we demonstrated electromyographically. We measured muscle length over the anatomical range of motion for CF (approximately 0.6-1.2 L0) and estimated working length during walking and trotting (approximately 0.95-1.15 L0). The specific tension was similar to that of the exclusively slow-twitch soleus muscle (31.2+/-4.7 N/cm2 compared with 31.8+/-4.1 N/cm2; P>0.8). Single fiber dissections of CF revealed a series-fibered architecture with a mean of 2.3 fibers, each 2.5 cm long, required to span the fascicle length. We identified two neuromuscular compartments in CF by cutting one of the two nerve branches innervating CF and depleting the glycogen stores in the intact motor units. These compartments were in parallel and extended the length of the muscle; their electromyographic activity was similar during various natural behaviors. CF and gluteus maximus motoneurons were labeled concurrently with a combination of fluorescent, retrograde tracers including Fluororuby, Fluorogold and Fast Blue. The CF motor nucleus was located in L7-S1, overlapping and intermingling extensively with the nucleus of the adjacent gluteus maximus muscle. Distributions of CF motoneuron diameter revealed one large peak around 50-55 microm, with relatively few small-diameter (less than 35 microm) cells. Using estimates of the total number of fibers in three muscles and the estimated number of alpha-motoneurons for those same muscles, we calculated a mean innervation ratio of approximately 270, which is at the low end of the innervation ratios for type IIB motor units from other feline muscles and more similar to type IIA motor units. In general, CF appears to be a useful preparation in which to study the properties of fast-twitch muscle, but these properties may vary somewhat from type IIB fibers from different muscles.

The location of triads within a mammalian skeletal muscle sarcomere has traditionally been defined as 'at the A-I junction'. We attempted to verify this statement by examining systematically the location of triads within the sarcomere over the physiological range of sarcomere lengths. This study was conducted using intact feline muscle fibres from caudofemoralis and exclusively fast-twitch muscle from the hindlimb. Our results intact fibres indicate that the distance between the Z-band and triad (ZT) is relatively constant over the range of sarcomere lengths (SLs)examined in this study (1.8-3.4 micron). The slope between ZT and SL was measured to be 0.06 +/- 0.01 (r= 0.36, p < 0.001) while the slope between the M-line to triad distance (MT) and SL was measured to be 0.44 +/- 0.01 (r > 0.9, p < 0.001). The mean ZT was 0.52 +/- 0.07 micron, which corresponds to a triad location approximately halfway along the thin filaments. These results do not support the traditional statement regarding triad location. Nor do these results support a similar recent study conducted using chemically skinned muscle fibres from rat extensor digitorum longus (also a homogeneously fast-twitch muscle of the hindlimb), in which a slope of 0.25 was observed between ZT and SL (r > 0.9, p < 0.01). These results are, however, in qualitative agreement with results using intact fibres from fast-twitch rat semitendinosus. Based upon known morphology, we suggest that the only structure supporting triad position is the SR itself, and that a non-homogeneous distribution of the SR within the sarcomere might be responsible for maintaining triad location near the mid-region of the thin filaments. We also suggest that there might be optimal design reasons for locating the triads at the mid-region of the thin filaments.

The relationships between range of motion, optimal length for force production (lo), and passive force provide useful insights into the structure and function of muscles but are unknown for most individual muscles. We measured these values and examined their relationships in five strap-like muscles of the cat hind limb: caudofemoralis, semitendinosus, sartorius anterior, tenuissimus, and biceps femoris anterior. The range of motion relative to lo was found to vary significantly between different muscles and even between different specimens of the same muscle. The passive force-length (FL) curve was found to be correlated with both lo and lmax (maximal in situ muscle length) but was correlated more strongly with lmax. The mean passive force produced by these muscles at lmax was less than 7% of estimated maximal isometric force, suggesting that passive force may not be important in these muscles during normal activation patterns. The variance in passive FL curves between specimens of the same muscle was found to be significantly lower when length was scaled by lmax as opposed to lo. These results suggest that lmax may provide a more useful scaling factor for generic models of muscle. However, the passive length-tension properties of mammalian muscle appear to reflect a complex mix of structures at both the myofilament and connective tissue levels that may differ depending on muscle-fiber architecture and perhaps on the history of trophic influences on a particular specimen.

We have developed a mathematical model to describe force production in cat soleus during steady-state activation over a range of fascicle lengths and velocities. The model was based primarily upon a three element design by Zajac but also considered the many different features present in other previously described models. We compared quantitatively the usefulness of these features and putative relationships to account for a set of force and length data from cat soleus wholemuscle described in a companion paper. Among the novel features that proved useful were the inclusion of a short-length passive force resisting compression, a new normalisation constant for connective-tissue lengths to replace the potentially troublesome slack length, and a new length dependent term for lengthening velocities in the force-velocity relationship. Each feature of this model was chosen to provide the most accurate description of the data possible without adding unneeded complexity. Previously described functions were compared with novel functions to determine the best description of the experimental data for each of the elements in the model.

The aim of the present study was to quantify how fascicle length and velocity modify force production in cat soleus. A computerized muscle puller controlled the length and velocity of the whole-muscle. We recorded the force output at the tendon and the length of muscle fascicles using sonomicrometry during whole-muscle isometric and isokinetic contractions. Peak muscle stress was estimated as 31.8 +/- 4.1 N cm-2 (mean and SD) and optimal fascicle length, Lo, was estimated as 3.8 +/- 0.6 cm which corresponds to an optimal sarcomere length of 2.49 +/- 0.08 microns. The isometric force-length data followed closely the expected force-length relationship for cat sarcomeres. The force-velocity relationship was found to be similar in shape between cats, but the per cent increment of force over isometric levels for lengthening contractions was highly variable. Estimates of the kinematics of the fascicles based on whole-muscle length were systematically incorrect; whole-muscle velocity was 21% greater than fascicle velocity. The force-velocity data demonstrated consistent dependencies on fascicle length. At lengths below 0.7 Lo (1.74 microns), the shape of the force-velocity relationship was altered by the inclusion of a passive, repulsive force in the estimate of active isometric force. The shape of the force-velocity relationship changed at lengths greater than 0.7 Lo, but was restricted to lengthening velocities where the increment of force with respect to isometric levels was found to increase with fascicle length. This change in shape in the force-velocity relationship for lengthening contractions reveals a systematic, but previously unknown interdependence between fascicle length and velocity on muscle force production.

A histologic method has been developed to assess the migration of chronically-implanted intramuscular devices. Bullet-shaped glass devices with varying tip configurations and glass-encapsulated microstimulators of a similar size were loaded with the fluorescent tracer Procion Yellow dissolved in molten glucose. Dissolution of the hardened glucose soon after contact with body fluids released the Procion Yellow, which binds irreversibly to local tissues, thereby marking the initial site of implantation with a localized fluorescent spot. After survival times of 2-7 weeks, histologic analysis usually showed a close physical relationship between the fluorescent spot and the connective-tissue capsule from which the device was extracted. In one case, migration of a sharply pointed device from the deep surface of an implanted muscle was recognized by differences in the location of the dye spot and the site of explantation from nearby fascia. Results suggested that this method could measure migratory distances as small as 5-10 mm.

Pyramidal cells in the mammalian neocortex do not normally contain detectable levels of the enzyme nitric oxide synthase. However one region of the human neocortex contains pyramidal neurones that express neuronal nitric oxide synthase activity. These neurons are mainly located in layer V of the precentral gyrus and frontal cortex and are predominantly Betz cells. The proportion of Betz cells stained in the eight brains examined varied from 5 to 80%. The brains of eight rats that had received a stab wound to the parietal cortex were also examined. Following a survival period of 7 or 14 days, small groups of pyramidal neurones surrounding the lesion contained moderate levels of neuronal nitric oxide synthase. We suggest that human pyramidal neurones may start expressing nitric oxide synthase as a response to damage or age-related stress and that the nitric oxide released may have a neuroprotective role.

The effects of sleep deprivation on medical personnel have received much attention. This study evaluates the effects of sleep loss on divergent-thinking (creative or innovative) processes as measured by the Torrance Test of Creative Thinking (TTCT). Anesthesia residents who had approximately 30 minutes sleep while being on-call were evaluated. These physicians had similar caffeine and nicotine consumption before and after the test. The results reported here demonstrate that postcall residents had TTCT scores that were appreciably below those scores of rested residents. Postcall verbal fluency was less among the on-call group than among the rested group (94.0 +/- 9.7 vs 101.8 +/- 9.8) as was figural originality (89.9 +/- 22.1 vs 113.3 +/- 20.3). These study results suggest that sleep deprivation affects divergent, or creative, thinking. Divergent-thinking processes are usually innovative and are used during complex problem-solving tasks. Further studies are needed on the effects of sleep deprivation. This information can then be used to help improve residents' working conditions and patient care.

We requested information concerning the job description for the chief resident in anesthesiology from 50 different programs. Thirty-two responses were returned, with 9 responses that no such job description existed at their institutions. Eighteen of the remaining 23 respondents had a written job description for the position. Considerable variation existed in the various aspects of the position among institutions. Differences were found in the selection process, administrative duties, number of committee memberships, and educational responsibilities. We present a suggested description of the responsibilities for a chief resident in anesthesiology.