A surgeon's guide to the universe.
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Biomedical subjects
Publications and source records attributed to Scott A Dulchavsky.
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We have previously demonstrated that TNF-alpha gene therapy with myeloid progenitor cells inhibits the progression of 32Dp210 myeloid leukemia in mice. Because TNF-alpha has been shown to induce the activation and maturation of dendritic cells (DCs), we investigated the efficacy of DC-based leukemia vaccine for eradication of residual disease when administered following cytoreductive therapy. Immunization with DC cells loaded with 32Dp210 myeloid leukemia cells (32Dp210 vaccine) was far more effective in preventing the development of leukemia compared to immunization with irradiated leukemia cells alone. The resistance to leukemia could be adoptively transferred to naïve mice with the spleen cells of mice immunized with DC-32Dp210 vaccine, and splenic cells responsible for adoptive transfer of resistance were identified as CD90 + T lymphocytes. Development of immunity in vaccinated mice was associated with the generation of leukemia specific cytotoxic T lymphocytes (CTLs) and secretion of cytokines TNF-alpha and IFN-gamma. Further, immunization with DC-32Dp210 vaccine following cytoreductive therapy with Cytoxan was effective in eradicating residual disease in approximately 50 percent of the animals. However, eradication of residual disease was significantly improved (approximately 74%) when animals were treated with DC-32Dp210 vaccine in which DCs were activated with TNF-alpha prior to loading of 32Dp210 leukemia cells. Cured mice were in molecular remission since Bcr/Abl oncogene could not be amplified from the DNA isolated from the marrow, spleen, or liver of cured mice. Taken together, these data demonstrate the efficacy of DC-based leukemia vaccine for eradication of residual disease and prevention of relapse.
BACKGROUND: Focused assessment with sonography for trauma (FAST) examination has been proved accurate for diagnosing trauma when performed by nonradiologist physicians. Recent reports have suggested that nonphysicians also may be able to perform the FAST examination reliably. A multipurpose ultrasound system is installed on the International Space Station as a component of the Human Research Facility. Nonphysician crew members aboard the International Space Station receive modest training in hardware operation, sonographic techniques, and remotely guided scanning. This report documents the first FAST examination conducted in space, as part of the sustained effort to maintain the highest possible level of available medical care during long-duration space flight. METHODS: An International Space Station crew member with minimal sonography training was remotely guided through a FAST examination by an ultrasound imaging expert from Mission Control Center using private real-time two-way audio and a private space-to-ground video downlink (7.5 frames/second). There was a 2-second satellite delay for both video and audio. To facilitate the real-time telemedical ultrasound examination, identical reference cards showing topologic reference points and hardware controls were available to both the crew member and the ground-based expert. RESULTS: A FAST examination, including four standard abdominal windows, was completed in approximately 5.5 minutes. Following commands from the Mission Control Center-based expert, the crew member acquired all target images without difficulty. The anatomic content and fidelity of the ultrasound video were excellent and would allow clinical decision making. CONCLUSIONS: It is possible to conduct a remotely guided FAST examination with excellent clinical results and speed, even with a significantly reduced video frame rate and a 2-second communication latency. A wider application of trauma ultrasound applications for remote medicine on earth appears to be possible and warranted.
BACKGROUND: Ultrasound imaging is a successful modality in a broad variety of diagnostic applications including trauma. Ultrasound has been shown to be accurate when performed by non-radiologist physicians; recent reports have suggested that non-physicians can perform limited ultrasound examinations. A multipurpose ultrasound system is installed on the International Space Station (ISS) as a component of the Human Research Facility (HRF). This report documents the first ocular ultrasound examination conducted in space, which demonstrated the capability to assess physiologic alterations or pathology including trauma during long-duration space flight. METHODS: An ISS crewmember with minimal sonography training was remotely guided by an imaging expert from Mission Control Center (MCC) through a comprehensive ultrasound examination of the eye. A multipurpose ultrasound imager was used in conjunction with a space-to-ground video downlink and two-way audio. Reference cards with topological reference points, hardware controls, and target images were used to facilitate the examination. Multiple views of the eye structures were obtained through a closed eyelid. Pupillary response to light was demonstrated by modifying the light exposure of the contralateral eye. RESULTS: A crewmember on the ISS was able to complete a comprehensive ocular examination using B- and M-mode ultrasonography with remote guidance from an expert in the MCC. Multiple anteroposterior, oblique, and coronal views of the eye clearly demonstrated the anatomic structures of both segments of the globe. The iris and pupil were readily visualized with probe manipulation. Pupillary diameter was assessed in real time in B- and M-mode displays. The anatomic detail and fidelity of ultrasound video were excellent and could be used to answer a variety of clinical and space physiologic questions. CONCLUSIONS: A comprehensive, high-quality ultrasound examination of the eye was performed with a multipurpose imager aboard the ISS by a non-expert operator using remote guidance. Ocular ultrasound images were of diagnostic quality despite the 2-second communication latency and the unconventional setting of a weightless spacecraft environment. The remote guidance techniques developed to facilitate this successful NASA research experiment will support wider applications of ultrasound for remote medicine on Earth including the assessment of pupillary reactions in patients with severe craniofacial trauma and swelling.
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INTRODUCTION: The performance of complex tasks on the International Space Station (ISS) requires significant preflight crew training commitments and frequent skill and knowledge refreshment. This report documents a recently developed "just-in-time" training methodology, which integrates preflight hardware familiarization and procedure training with an on-orbit CD-ROM-based skill enhancement. This "just-in-time" concept was used to support real-time remote expert guidance to complete ultrasound examinations using the ISS Human Research Facility (HRF). METHODS: An American and Russian ISS crewmember received 2 h of "hands on" ultrasound training 8 mo prior to the on-orbit ultrasound exam. A CD-ROM-based Onboard Proficiency Enhancement (OPE) interactive multimedia program consisting of memory enhancing tutorials, and skill testing exercises, was completed by the crewmember 6 d prior to the on-orbit ultrasound exam. The crewmember was then remotely guided through a thoracic, vascular, and echocardiographic examination by ultrasound imaging experts. RESULTS: Results of the CD-ROM-based OPE session were used to modify the instructions during a complete 35-min real-time thoracic, cardiac, and carotid/jugular ultrasound study. Following commands from the ground-based expert, the crewmember acquired all target views and images without difficulty. The anatomical content and fidelity of ultrasound video were adequate for clinical decision making. CONCLUSIONS: Complex ultrasound experiments with expert guidance were performed with high accuracy following limited preflight training and multimedia based in-flight review, despite a 2-s communication latency. In-flight application of multimedia proficiency enhancement software, coupled with real-time remote expert guidance, facilitates the successful performance of ultrasound examinations on orbit and may have additional terrestrial and space applications.
Malignant glioma cells are generally resistant or only weakly sensitive to tumor necrosis factor family of cell death-inducing ligands, including TNF-related apoptosis-inducing ligand (TRAIL)/Apo2L. The chemopreventive activity of polyphenolic compounds present in plant-derived food products has been well recognized in epidemiological studies; however, the mechanism of chemoprevention by these dietary constituents largely remains unknown. Curcumin, the yellow pigment in the spice turmeric, has profound anti-inflammatory activity and exhibits chemopreventive and tumor growth inhibitory activity. In the present study, we investigated whether curcumin sensitizes malignant glioma cell lines U251MG and U87MG to TRAIL-induced apoptosis. Treatment with low concentrations (5-20 microM) of curcumin alone had no effect on the viability of either cell line. At low concentration (5 ng/ml) TRAIL induced cytotoxicity in U251MG cells but not in U87MG cells. Whereas curcumin at subtoxic concentration sensitized U87MG cells to TRAIL-induced cytotoxicity, it had no effect on TRAIL-mediated cytotoxicity in U251MG cells. The combined curcumin and TRAIL treatment enhanced accumulation of hypo-diploid U87MG cells in sub G1 cell cycle phase and induced the cleavage of procaspases-3, -8, -9 and release of cytochrome c from mitochondria. These data indicate that curcumin differentially sensitizes glioma cells to TRAIL-induced apoptosis through the activation of both extrinsic (receptor-mediated) and intrinsic (chemical-induced) pathways of apoptosis. These results define a potential use of curcumin to sensitize glioma cells for TRAIL-mediated immunotherapy.
Failure to undergo apoptosis has been implicated in the resistance of tumor cells to anticancer therapies. Promotion of apoptosis in tumor cells could potentially increase the efficacy of conventional treatment regimens and improve prognosis. Prostate cancer cells are generally resistant to induction of apoptosis by anticancer agents and death ligands. We investigated the sensitization of prostate cancer cell lines by curcumin (diferuloyl-methane) to TNF-related apoptosis inducing ligand (TRAIL)-induced apoptosis. Prostate cancer cells treated with curcumin or TRAIL or curcumin and TRAIL together were assessed for induction of apoptosis and pathway of apoptosis was determined from the activation of procaspases and release of cytochrome c from mitochondria. Curcumin sensitized LNCaP, DU145 and PC3 tumor cell lines to TRAIL. Combined curcumin and TRAIL treatment produced the most loss of viable cells by inducing apoptosis as revealed by accumulation of hypodiploid cells in sub-G1 phase, enhanced annexin V binding, DNA fragmentation, cleavage of procaspases-3, -8, and 9, truncation of proapoptotic Bid, and release of cytochrome c from mitochondria. Tumor cells expressed constitutively active NF-kappaB and sensitization to TRAIL involved inhibition of NF-kappaB by curcumin. These findings suggest that combined curcumin/TRAIL chemo-immunotherapy may be a beneficial adjunct to the standard therapeutic regimens for prostate cancer.
Investigative procedures were approved by Henry Ford Human Investigation Committee and NASA Johnson Space Center Committee for Protection of Human Subjects. Informed consent was obtained. Authors evaluated ability of nonphysician crewmember to obtain diagnostic-quality musculoskeletal ultrasonographic (US) data of the shoulder by following a just-in-time training algorithm and using real-time remote guidance aboard the International Space Station (ISS). ISS Expedition-9 crewmembers attended a 2.5-hour didactic and hands-on US training session 4 months before launch. Aboard the ISS, they completed a 1-hour computer-based Onboard Proficiency Enhancement program 7 days before examination. Crewmembers did not receive specific training in shoulder anatomy or shoulder US techniques. Evaluation of astronaut shoulder integrity was done by using a Human Research Facility US system. Crew used special positioning techniques for subject and operator to facilitate US in microgravity environment. Common anatomic reference points aided initial probe placement. Real-time US video of shoulder was transmitted to remote experienced sonologists in Telescience Center at Johnson Space Center. Probe manipulation and equipment adjustments were guided with verbal commands from remote sonologists to astronaut operators to complete rotator cuff evaluation. Comprehensive US of crewmember's shoulder included transverse and longitudinal images of biceps and supraspinatus tendons and articular cartilage surface. Total examination time required to guide astronaut operator to acquire necessary images was approximately 15 minutes. Multiple arm and probe positions were used to acquire dynamic video images that were of excellent quality to allow evaluation of shoulder integrity. Postsession download and analysis of high-fidelity US images collected onboard demonstrated additional anatomic detail that could be used to exclude subtle injury. Musculoskeletal US can be performed in space by minimally trained operators by using remote guidance. This technique can be used to evaluate shoulder integrity in symptomatic crewmembers after strenuous extravehicular activities or to monitor microgravity-associated changes in musculoskeletal anatomy. Just-in-time training, combined with remote experienced physician guidance, may provide a useful approach to complex medical tasks performed by nonexperienced personnel in a variety of remote settings, including current and future space programs.
Curcumin (diferuloylmethane), a major curcumanoid found in the spice turmeric, exhibits anti-inflammatory, anti-oxidant, and chemopreventive activities. However, the effect of curcumin on the development of T cell-mediated immunological responses largely remains unknown. In this study we have investigated the effect of curcumin on mitogen/antigen induced proliferation of splenic lymphocytes, induction of cytotoxic T lymphocytes (CTLs), lymphokine activated killer (LAK) cells, and the production of cytokines by T lymphocytes and macrophages. We found that mitogen, interleukin-2 (IL-2) or alloantigen induced proliferation of splenic lymphocytes, and development of cytotoxic T lymphocytes is significantly suppressed at 12.5-30 micromol/L curcumin. The generation of LAK cells at similar concentrations was less sensitive to the suppressive effect of curcumin compared to the generation of antigen specific CTLs. Curcumin irreversibly impaired the production of these immune functions, since lymphoid cells failed to respond to the activation signals following 8h pretreatment with curcumin. Curcumin also inhibited the expression/production of IL-2 and interferon-gamma (IFN-gamma) by splenic T lymphocytes and IL-12 and tumor necrosis factor-alpha (TNF-alpha) by peritoneal macrophages irreversibly. Curcumin inhibited the activation of the transcription factor nuclear factor kappaB (NF-kappaB) without affecting the levels of constitutively expressed NF-kappaB. The latter result suggests that curcumin most likely inhibits cell proliferation, cell-mediated cytotoxicity (CMC), and cytokine production by inhibiting NF-kappaB target genes involved in induction of these immune responses.
BACKGROUND: New portable ultrasound (US) systems are capable of detecting fractures in the remote setting. However, the accuracy of ultrasound by physicians with minimal ultrasound training is unknown. METHODS: After one hour of standardized training, physicians with minimal US experience clinically evaluated patients presenting with pain and trauma to the upper arm or leg. The investigators then performed a long-bone US evaluation, recording their impression of fracture presence or absence. Results of the examination were compared with routine plain or computer aided radiography (CT). RESULTS: 58 patients were examined. The sensitivity and specificity of US were 92.9% and 83.3%, and of the physical examination were 78.6% and 90.0%, respectively. US provided improved sensitivity with less specificity compared with physical examination in the detection of fractures in long bones. CONCLUSION: Ultrasound scans by minimally trained clinicians may be used to rule out a long-bone fracture in patients with a medium to low probability of fracture.
INTRODUCTION: An intrathoracic injury may be disastrous to a crew-member aboard the International Space Station (ISS) if the diagnosis is missed or delayed. Symptomatic or clinically suspicious thoracic trauma is treated as a surgical emergency on Earth, usually with immediate stabilization and rapid transport to a facility that is able to deliver the appropriate medical care. A similar approach is planned for the ISS; however, an unnecessary evacuation would cause a significant mission impact and an exorbitant expense. HYPOTHESIS: The use of ultrasound imaging for the detection of pneumothorax and hemothorax in microgravity is both possible and practical. METHODS: Sonography was performed on anesthetized pigs in a ground-based laboratory (n = 4) and microgravity conditions (0 G) during parabolic flight (n = 4). Aliquots of air (50-500 ml) or saline (10-200 ml) were introduced into the pleural space to simulate pneumothorax and hemothorax, respectively. RESULTS: The presence of "lung sliding" excluded pnemothorax. In microgravity, a loss of "lung sliding" was noted simultaneously in the anterior and posterior sonographic windows after 100 ml of air was introduced into the chest, indicating pneumothorax. The presence of the fluid layer in simulated hemothorax was noted in the anterior and posterior sonographic windows after 50 ml of fluid was injected into the pleural space. During the microgravity phase, the intrapleural fluid rapidly redistributed so that it could be detected using either anterior or posterior sonographic windows. CONCLUSION: Modest to severe pneumothorax and hemothorax can be diagnosed using ultrasound in microgravity.
Epidemiologic studies suggest that diet rich in plant-derived foods plays an important role in the prevention of prostate cancer. Curcumin, the yellow pigment in the spice turmeric, has been shown to exhibit chemopreventive and growth inhibitory activities against multiple tumor cell lines. We have shown previously that curcumin and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)/Apo2L interact to induce cytotoxicity in the LNCaP prostate cancer cell line. In this study, we investigated the mechanism by which curcumin augments TRAIL-induced cytotoxicity in LNCaP cells. Subtoxic concentrations of the curcumin-TRAIL combination induced strong apoptotic response in LNCaP cells as demonstrated by the binding of Annexin V-FITC and cleavage of procaspase-3. Furthermore, LNCaP cells express constitutively active nuclear factor-kappaB (NF-kappaB), which is inhibited by curcumin. Because NF-kappaB has been shown to mediate resistance to TRAIL-induced apoptosis in tumor cells, we investigated whether there is a relationship between NF-kappaB activation and resistance to TRAIL in LNCaP prostate cancer cells. Pretreatment with curcumin inhibited the activation of NF-kappaB and sensitized LNCaP cells to TRAIL. A similar increase in the sensitivity of LNCaP cells to TRAIL-induced apoptosis was observed following inhibition of NF-kappaB by dominant negative mutant IkappaBalpha, an inhibitor of NF-kappaB. Finally, curcumin was found to inhibit NF-kappaB by blocking phosphorylation of IkappaBalpha. We conclude that NF-kappaB mediates resistance of LNCaP cells to TRAIL and that curcumin enhances the sensitivity of these tumor cells to TRAIL by inhibiting NF-kappaB activation by blocking phosphorylation of IkappaBalpha and its degradation.
BACKGROUND: Secretory immunoglobulin A (SIgA) is the principal immune defense against luminal pathogens at gut mucosal surfaces. It also has anti-inflammatory activities that may be important for the maintenance of mucosal surface integrity. Enterocyte apoptosis (Apo) is increased after challenge with invasive bacteria and ischemia-reperfusion insults. Increased Apo also has been associated with impaired intestinal barrier function. However, the impact of SIgA on enterocyte apoptosis and mucosal barrier integrity after challenge with commensal bacteria and ischemia-reperfusion is unknown. METHODS: Caco2 intestinal epithelial cell monolayers were subjected to 21% O(2) (control) or 95% N(2)/15% CO(2) (hypoxic) conditions for 90 minutes, followed by 21% O(2). Escherichia coli and SIgA were added in subsets. Caco2 cell Apo was identified by flow cytometry and barrier function indexed by permeability to dextran-fluorescein isothiocyanate. RESULTS: There were no differences in the percentage of Apo Caco2 cells after exposure to either bacteria or hypoxic-reoxygenation versus control. There was a significant increase in Apo after the combined bacteria/hypoxia-reoxygenation challenge. SIgA abrogated the Apo response and preserved barrier function after this combined challenge. CONCLUSION: Modulation of enterocyte Apo by SIgA may serve to maintain intestinal barrier function and thereby decrease the systemic inflammatory response after clinical conditions associated with gut ischemia-reperfusion insults.
BACKGROUND: The Focused Assessment with Sonography for Trauma (FAST) examines for fluid in gravitationally dependent regions. There is no prior experience with this technique in weightlessness, such as on the International Space Station, where sonography is currently the only diagnostic imaging tool. STUDY DESIGN: A ground-based (1 g) porcine model for sonography was developed. We examined both the feasibility and the comparative performance of the FAST examination in parabolic flight. Sonographic detection and fluid behavior were evaluated in four animals during alternating weightlessness (0 g) and hypergravity (1.8 g) periods. During flight, boluses of fluid were incrementally introduced into the peritoneal cavity. Standardized sonographic windows were recorded. Postflight, the video recordings were divided into 169 20-second segments for subsequent interpretation by 12 blinded ultrasonography experts. Reviewers first decided whether a video segment was of sufficient diagnostic quality to analyze (determinate). Determinate segments were then analyzed as containing or not containing fluid. A probit regression model compared the probability of a positive fluid diagnosis to actual fluid levels (0 to 500 mL) under both 0-g and 1.8-g conditions. RESULTS: The in-flight sonographers found real-time scanning and interpretation technically similar to that of terrestrial conditions, as long as restraint was maintained. On blinded review, 80% of the recorded ultrasound segments were considered determinate. The best sensitivity for diagnosis in 0 g was found to be from the subhepatic space, with probability of a positive fluid diagnosis ranging from 9% (no fluid) to 51% (500 mL fluid). CONCLUSIONS: The FAST examination is technically feasible in weightlessness, and merits operational consideration for clinical contingencies in space.
BACKGROUND: Clinical data indicate that gut perfusion deficits must be rectified within 24 hours after traumatic injury to decrease organ failure and death. Ischemia/reperfusion injury to the gut causes enterocyte apoptosis (Apo), which may contribute to intestinal barrier failure. The temporal response of enterocyte Apo to acidosis and hypoxia/reoxygenation (H/R) in vitro is unknown. The purpose of this study was to examine the effect of various time points of acidosis or H/R on enterocyte apoptosis and monolayer integrity in an in vitro model. METHODS: Caco-2 cell monolayers were made acidic (Dulbecco's modified Eagle's medium, pH 6.9) by hydrochloric acid or exposed to 95% nitrogen/5% carbon dioxide (hypoxia) and then 21% oxygen (reoxygenation). Escherichia coli C-25 were added to the apical media in subsets. Apo and necrosis were quantified by flow cytometry. Permeability was determined by fluorescein isothiocyanate-dextran. Transepithelial electrical resistance (TEER) indexed monolayer. RESULTS: Extracellular acidosis and C-25 significantly increased apoptosis of Caco-2 cells at 18 hours (extracellular acidosis [EC] + C-25, 14.5 +/- 3.0; control, 3.8 +/- 0.8; p < 0.001 by analysis of variance). Similarly, the H/R + C-25 group showed a significant increase in apoptosis at 12 hours (H/R + C-25 vs. control, 22.86 +/- 2.12 vs. 3.74 +/- 0.7; p < 0.001 by analysis of variance). The permeability difference was not significant for EC + C-25 versus control at 18 hours (0.68 +/- 0.25 vs. 0.43 +/- 0.0.0.36, respectively; p > 0.05). The H/R + C-25 group had a profound increase in permeability over control at 12 hours (10.8 +/- 0.5 vs. 2.1 +/- 0.3, respectively; p < 0.001). The TEER was significantly lowered for EC versus control at 18 hours (458 +/- 1.5 vs. 468 +/- 8.2) and at 0, 6, and 18 hours for EC + C-25 (409 +/- 28.1, 443 +/- 16.8, and 438 +/- 8.9 vs. 455 +/- 6.5, 467 +/- 6.5, and 469 +/- 8.2, respectively). There was no significant change in the H/R and H/R + C-25 groups. CONCLUSION: Synergism of H/R or tissue acidosis and bacteria caused increased Apo, TEER, and permeability in vitro.
Tissue oxygenation is a critical factor in host defense against bacteria. Gut mucosal tissue oxygenation (partial pressure of O2) is normally low putting the gut at risk of invasion by luminal microbes. Secretory immunoglobulin (Ig) A (sIgA) is the principal immune defense at mucosal surfaces. The protective effect of IgA under low oxygen conditions is unknown. We studied the interaction of varying O2 environments and sIgA on protection against bacterial invasion in our in vitro model. Cell monolayers of Madin-Darby canine kidney (MDCK) cells transfected with the cDNA for polymeric immunoglobulin receptor were established in a two-chamber cell culture system. A commensal strain of Escherichia coli (10(8) colony-forming units) was added to the apical medium and cell cultures were placed in either a 5, 21, or 95 per cent O2 environment at 37 degrees C. Polyclonal sIgA (100 microg/mL) was added to the apical chamber in subsets. Basal medium was sampled at intervals and bacterial translocation quantitated. The cell monolayers of MDCK transfected cells then had 100 microg/mL IgA added to the basal compartment at 4 degrees C for 2 hours followed by various oxygen environments for 90 minutes. Afterwards apical medium was removed at one, 3, and 12 (overnight) hours. The bacterial translocation data showed a significance increase in translocation with hypoxia. Both increased oxygen and IgA abrogated these effects significantly. The transcytosis of IgA was increased during hypoxic conditions. Normal and hyperoxic conditions did not produce any significant difference in IgA transcytosis. We conclude that O2 and sIgA are protective against bacterial invasion at epithelial surfaces. Effects to either boost O2 delivery to the gut or enhance mucosal IgA production and delivery may be protective in the critically ill surgical patient.
Orthopedic fractures are a common injury in operational activities, which are injuries that often occur in isolated or hostile environments. Clinical ultrasound devices have become more user-friendly and lighter, allowing them to be easily transported with forward medical teams. The bone-soft tissue interface has a very large acoustic impedance with a high reflectance that can be used to visualize breaks in contour including fractures. Herein reported is a case of an ulnar fracture that was quickly visualized in the early phase of a multisystem trauma resuscitation with a hand-held ultrasound device. The implications for operational medicine are discussed.