Richard R. McNeer

Sialomucin Complex, a Heterodimeric Glycoprotein Complex EXPRESSION AS A SOLUBLE, SECRETABLE FORM IN LACTATING MAMMARY GLAND AND COLON

Ascites 13762 rat mammary adenocarcinoma cells express abundantly on their cell surfaces a heterodimeric glycoprotein complex composed of a sialomucin ascites sialoglycoprotein (ASGP)-1 and a transmembrane subunit ASGP-2. The latter, which contains two epidermal growth factor-like domains, binds the receptor tyrosine kinase p185neu, suggesting that the complex is bifunctional as well as heterodimeric. Immunoblot analyses using monoclonal antibodies prepared against the complex demonstrate high levels of expression in rat lactating mammary gland and colon. Immunolocalization studies with anti-ASGP-2 indicate that ASGP-2 is present in these two tissues in the apical regions of secretory epithelial cells. Both mammary gland and colon contain a soluble, secretable form of ASGP-2, which is not found in the ascites cells; milk and mammary gland also have the membrane form. Immunoblot analyses using a COOH-terminal-specific polyclonal antibody indicate that the soluble form of ASGP-2 is missing its COOH-terminal domains. Both the soluble and membrane forms of ASGP-2 are similar to the membrane-associated form from the 13762 adenocarcinoma with respect to Mr, antigenicity, and association with ASGP-1. The presence of ASGP-1 in milk suggests that it is a candidate for the uncharacterized high Mr milk mucin, MUCX. ASGP-2 expression is up-regulated in mammary gland during pregnancy, because it is undetectable in virgin and early pregnant rats but abundant in the gland from late pregnant and lactating animals. However, compared with the lactating mammary gland, the 13762 ascites cells overexpress ASGP-2 by more than 100-fold, which may contribute to their malignancy. These combined results indicate that sialomucin complex is a unique secreted product in the mammary gland and colon, whose behavior is different from that in the mammary ascites tumors, and which may play important roles in mammary and intestinal physiology.

Characterization of the expression and steroid hormone control of sialomucin complex in the rat uterus: implications for uterine receptivity.

The sialomucin complex (SMC), originally isolated as a heterodimeric glycoprotein complex from membranes of ascites sublines of a highly metastatic mammary adenocarcinoma, consists of a high Mr mucin subunit (ASGP‐1, ascites sialoglycoprotein) and a transmembrane subunit (ASGP‐2) with two epidermal growth factor‐like domains. SMC has been characterized in the mammary gland, where it is present in both membrane and nonmembrane (soluble) forms, the latter lacking its transmembrane domain. SMC in the mammary gland is observed during pregnancy and lactation, but not in the virgin gland, and is regulated by a posttranscriptional mechanism. Both membrane and nonmembrane forms of SMC are found in rat uterus, also as a complex of ASGP‐1 and ASGP‐2. Immunocytochemical analyses indicate that the primary site of expression is at the luminal surface of the endometrium. Approximately 40% of the SMC, corresponding to the nonmembrane fraction, is removed by rinsing uterine preparations with saline, indicating that the soluble form is adsorbed loosely to the cell surface. In contrast to mammary gland, SMC is most highly expressed in the virgin animal, and its expression varies during the estrous cycle with the steady state level of transcript. The complex is present in a location consistent with steric inhibition of blastocyst implantation and is abruptly lost at the beginning of the period of receptivity for implantation. Expression of SMC in the uterus is regulated by estrogen and progesterone and is inversely correlated with receptivity. Both implantation and loss of SMC expression can be blocked with RU486. We propose that the downregulation of SMC and its loss from the apical surface of the rat uterine lining contribute to the generation of the receptive state for uterine implantation. Furthermore, the presence of both membrane and soluble SMC at the luminal surface of the endometrium may provide a model for its proposed protective function in other accessible and vulnerable epithelia. J. Cell. Physiol. 176:110–119, 1998.

SIALOMUCIN COMPLEX IN TUMORS AND TISSUES

Sialomucin complex (SMC) is a high Mr glycoprotein heterodimer, originally discovered on the cell surfaces of ascites sublines of the highly metastatic 13762 rat mammary adenocarcinoma, and composed of mucin (ASGP-1) and transmembrane (ASGP-2) subunits. SMC is encoded by a single gene and synthesized as a large precursor protein which is cleaved into its subunits early in its transit to the cell surface. SMC exhibits behavior typical of both membrane and secreted mucins. In the ascites cells, it is found only in the membrane form, creating a protective barrier at the cell surface to reduce cell adhesiveness and protect the tumor cell from immune killing. Normal tissues express both the membrane formand a non-membrane form, which may be secreted by either constitutive or regulated, secretory granule mechanisms. This soluble form is proposed to contribute to multilayer mucus gels which protect epithelia, though it may also play other roles. ASGP-2 contains two EGF-like domains, one of which binds the receptor tyrosine kinase ErbB-2. Thus, SMC may be a bifunctional protein, the mucin serving a protective function and the transmembrane domain possibly playing a role in the proliferation of metastatic tumor cells or repair processes necessary for the maintenance of damaged epithelia.

A New Paradigm for the Design of Audible Alarms that Convey Urgency Information

ObjectiveThe current international standard (IEC 60601-1-8) stipulates that medical device audible alarms should be priority-encoded and validated for efficacy. Evidence suggests that the melodic alarms described in the standard are not functioning as originally intended. We present a multi-disciplinary, human factors paradigm for audible alarm development whereby urgency information is encoded via modulation of the physical characteristics of sounds. We also test the feasibility of this approach using information measures.MethodsWe designed series of experimental sounds that varied along controlled physical and acoustical dimensions. Subjects rated these sound series for perceived urgency. Based on these ratings, selected sounds from each series were assigned a␣priority category from ‹low’ to ‹high’ – we call these resulting sets of sounds ‹urgency-codecs’. The method of categorical judgments (based on information theory) was used to compare each urgency-codec for ability to convey urgency information.ResultsSubjects were consistent in their ratings of the three series of experimental sounds for perceived urgency. The urgency data pertaining to one of the series (harmonic interval) was successfully fit to a psychophysical empirical law. The urgency-codec derived from another sound series (melodic interval) was found to have the highest signal (correct interpretation of urgency level by subjects) transmission rate.ConclusionsThe proposed paradigm is feasible, and it offers an evidence-based strategy for alarm sound design and testing. This approach would be performed before implementation of new alarm sounds in clinical settings, and should result in development of alarm sounds that satisfy the requirements of priority-encoding and validation.

Expression and localization of Muc4/sialomucin complex (SMC) in the adult and developing rat intestine: Implications for Muc4/SMC function

Muc4/sialomucin complex (SMC) is a high molecular mass heterodimeric membrane mucin, encoded by a single gene, and originally discovered in a highly metastatic ascites rat mammary adenocarcinoma. Subsequent studies have shown that it is a prominent component of many accessible and vulnerable epithelia, including the gastrointestinal tract. Immunoblot and immunofluorescence analyses demonstrated that Muc4/SMC expression in the rat small intestine increases from proximal to distal regions and is located predominantly in cells at the base of the crypts. These cells were postulated to be Paneth cells, based on their location, morphology, and secretory granule content. Immunohistochemistry indicated the presence of Muc4/SMC in these granules. Muc4/SMC expression was higher in the rat colon than small intestine and was abundantly present in colonic goblet cells, but not in goblet cells in the small intestine. Immunohistochemistry also suggested the presence of MUC4 in human colonic goblet cells. Biochemical analyses indicated that rat colonic Muc4/SMC is primarily the soluble form of the membrane mucin. Analyses of Muc4/SMC during development of the rat gastrointestinal tract showed its appearance at embryonic day 14 of the esophagus and at day 15 at the surface of the undifferentiated stratified epithelium at the gastroduodenal junction, then later at cell surfaces in the more distal regions of the differentiated epithelium of the small intestine, culminating in expression as an intracellular form in the crypts of the small intestine at about day 21. Limited expression in the colon was observed during development before birth at cell surfaces, with expression as an intracellular form in the goblet cells arising during the second week after birth. These results suggest that membrane mucin Muc4/SMC serves different functions during development of the intestine in the rat, but is primarily a secreted product in the adult animal.

Influence of auditory stimulation rates on evoked potentials during general anesthesia: Relation between the transient auditory middle-latency response and the 40-Hz auditory steady state response

Background:The auditory middle-latency response (transient) and the 40-Hz auditory steady state response (ASSR) are modulated by anesthetics. However, the quantitative relation between these evoked responses is difficult to obtain because of technical limitations of the recording methods used to obtain transients at high stimulation rates. This study uses continuous-loop averaging deconvolution to fill this technical gap and to study the relation between the transient and ASSR waveform during general anesthesia. Methods:The authors recorded 5- and 40-Hz transients and 40-Hz ASSRs in 13 subjects during general anesthesia. The 5- and 40-Hz transients were used to predict the 40-Hz ASSR by linearly superimposing the transient waveforms. The predicted and recorded ASSRs were analyzed and compared using phasor and Hotelling T2 analyses. Results:Grand-averaged recordings revealed differences in the early middle-latency peaks between 5- and 40-Hz transients, e.g., the peak Px was present only in 5-Hz transient. Only the predicted 40-Hz ASSR derived from the 40-Hz transient matched the actual ASSR. Phasor analysis showed that the early peaks contribute significantly to the steady state waveform, and this explains why 5-Hz transient does not predict the 40-Hz ASSR. Oscillations in both the 5- and 40-Hz transients were observed during anesthesia. Discussion:The 40-Hz ASSR represents a composite waveform and arises when transient waveforms elicited with a 40-Hz stimulation rate are overlapped and superimposed. During general anesthesia, the morphology of the transient is dependent on the rate of stimulus presentation. The composite nature of the ASSR may explain nonmonotonic anesthetic dose–response relations observed by others.

Intraoperative noise increases perceived task load and fatigue in anesthesiology residents: A simulation-based study

BACKGROUND:Operating rooms are identified as being one of the noisiest of clinical environments, and intraoperative noise is associated with adverse effects on staff and patient safety. Simulation-based experiments would offer controllable and safe venues for investigating this noise problem. However, realistic simulation of the clinical auditory environment is rare in current simulators. Therefore, we retrofitted our operating room simulator to be able to produce immersive auditory simulations with the use of typical sound sources encountered during surgeries. Then, we tested the hypothesis that anesthesia residents would perceive greater task load and fatigue while giving simulated lunch breaks in noisy environments rather than in quiet ones. As a secondary objective, we proposed and tested the plausibility of a novel psychometric instrument for the assessment of stress. METHODS:In this simulation-based, randomized, repeated-measures, crossover study, 2 validated psychometric survey instruments, the NASA Task Load Index (NASA-TLX), composed of 6 items, and the Swedish Occupational Fatigue Inventory (SOFI), composed of 5 items, were used to assess perceived task load and fatigue, respectively, in first-year anesthesia residents. Residents completed the psychometric instruments after giving lunch breaks in quiet and noisy intraoperative environments (soundscapes). The effects of soundscape grouping on the psychometric instruments and their comprising items were analyzed with a split-plot analysis. A model for a new psychometric instrument for measuring stress that combines the NASA-TLX and SOFI instruments was proposed, and a factor analysis was performed on the collected data to determine the model’s plausibility. RESULTS:Twenty residents participated in this study. Multivariate analysis of variance showed an effect of soundscape grouping on the combined NASA-TLX and SOFI instrument items (P = 0.003) and the comparisons of univariate item reached significance for the NASA Temporal Demand item (P = 0.0004) and the SOFI Lack of Energy item (P = 0.001). Factor analysis extracted 4 factors, which were assigned the following construct names for model development: Psychological Task Load, Psychological Fatigue, Acute Physical Load, and Performance-Chronic Physical Load. Six of the 7 fit tests used in the partial confirmatory factor analysis were positive when we fitted the data to the proposed model, suggesting that further validation is warranted. CONCLUSIONS:This study provides evidence that noise during surgery can increase feelings of stress, as measured by perceived task load and fatigue levels, in anesthesiologists and adds to the growing literature pointing to an overall adverse impact of clinical noise on caregivers and patient safety. The psychometric model proposed in this study for assessing perceived stress is plausible based on factor analysis and will be useful for characterizing the impact of the clinical environment on subject stress levels in future investigations.

PT-SAFE: a software tool for development and annunciation of medical audible alarms.

BACKGROUND: Recent reports by The Joint Commission as well as the Anesthesia Patient Safety Foundation have indicated that medical audible alarm effectiveness needs to be improved. Several recent studies have explored various approaches to improving the audible alarms, motivating the authors to develop real-time software capable of comparing such alarms. We sought to devise software that would allow for the development of a variety of audible alarm designs that could also integrate into existing operating room equipment configurations. The software is meant to be used as a tool for alarm researchers to quickly evaluate novel alarm designs. METHODS: A software tool was developed for the purpose of creating and annunciating audible alarms. The alarms consisted of annunciators that were mapped to vital sign data received from a patient monitor. An object-oriented approach to software design was used to create a tool that is flexible and modular at run-time, can annunciate wave-files from disk, and can be programmed with MATLAB by the user to create custom alarm algorithms. The software was tested in a simulated operating room to measure technical performance and to validate the time-to-annunciation against existing equipment alarms. RESULTS: The software tool showed efficacy in a simulated operating room environment by providing alarm annunciation in response to physiologic and ventilator signals generated by a human patient simulator, on average 6.2 seconds faster than existing equipment alarms. Performance analysis showed that the software was capable of supporting up to 15 audible alarms on a mid-grade laptop computer before audio dropouts occurred. CONCLUSIONS: These results suggest that this software tool provides a foundation for rapidly staging multiple audible alarm sets from the laboratory to a simulation environment for the purpose of evaluating novel alarm designs, thus producing valuable findings for medical audible alarm standardization.

The use of pressure-tuning spectroscopy to distinguish between one- and two-electron transfer processes

The first use of pressure-tuning spectroscopy as a diagnostic tool in the evaluation of an intervalence transition band to determine the number of electrons transferred is described. The trinuclear complex [(CN)5FeCNPt(NH3)4NCFe(CN)5]4− was subjected in solution to hydrostatic pressures from ambient to 1.5 kbar giving a ΔV* = −5.7 ± 0.2 cm3 mol−1. Calculated values from various models ranged from −5.6 to −6.7 cm3 mol−1 assuming a one-electron transfer event and a negligible innersphere reorganizational energy compared with the apparently very large outer-sphere rearrangements that must occur.

Simulating environmental and psychological acoustic factors of the operating room.

In this study, an operating room simulation environment was adapted to include quadraphonic speakers, which were used to recreate a composed clinical soundscape. To assess validity of the composed soundscape, several acoustic parameters of this simulated environment were acquired in the presence of alarms only, background noise only, or both. These parameters were also measured for comparison from size-matched operating rooms at Jackson Memorial Hospital. The parameters examined included sound level, reverberation time, and predictive metrics of speech intelligibility in quiet and noise. It was found that the sound levels and acoustic parameters were comparable between the simulated environment and the actual operating rooms. The impact of the background noise on the perception of medical alarms was then examined, and was found to have little impact on the audibility of the alarms. This study is a first in kind report of a comparison between the environmental and psychological acoustical parameters of a hospital simulation environment and actual operating rooms.