CD4 expression is differentially required for deletion of MLS-1a-reactive T cells. Clonal deletion of thymocytes expressing potentially self-reactive T cell receptors (TCRs) occurs during thymocyte ontogeny. Mice deficient for CD4 expression provide a unique model system to study the contribution of the CD4 molecule in negative selection of T cells reactive against the major histocompatibility complex class II-associated retroviral self-superantigen, Mls-1a. In the presence of Mls-1a determinants, mature CD8+ T cells expressing V beta 6, 8.1, and 9 were deleted in CD4-deficient mice, thus demonstrating that TCR affinity for Mls-1a is sufficient for deletion and that a signal through CD4 was not required. However, in instances where the TCR affinity for Mls-1a is low, as in the case of V beta 7+ T cells, CD4 expression was required for clonal deletion. These results demonstrate that for Mls-1a-mediated clonal deletion of T cells, the requirement for the accessory or coreceptor function of CD4 depends on the affinity of the TCR.


Reactivation of a major histocompatibility complex class II gene in mouse plasmacytoma cells and mouse T cells. Terminally differentiated plasma cells and mouse T cells do not express major histocompatibility complex (MHC) class II genes although class II gene expression is observed in pre-B and mature B cells as well as in activated human T cells. Transient heterokaryons were prepared and analyzed to investigate the mechanisms of inactivation of MHC class II gene in mouse plasmacytoma cells and mouse T cells. The endogenous MHC class II genes in both mouse plasmacytoma cells and mouse T cells can be reactivated by factors present in B cells. This reactivation of class II gene is also observed by fusion with a human T cell line which expresses MHC class II genes, but not with a class II negative human T cell line. It appears that the loss of MHC class II gene expression during the terminal differentiation of B cells or T cell lineage is due to absence of positive regulatory factor(s) necessary for class II transcription.


Characterization and cloning of a novel glycoprotein expressed by stromal cells in T-dependent areas of peripheral lymphoid tissues. A novel glycoprotein (gp) expressed by stromal cells of peripheral lymphoid tissue has been characterized immunohistochemically, biochemically, and at the molecular level. This molecule, gp38, was identified with a monoclonal antibody (mAb) (clone 8.1.1) previously shown to react with a subpopulation of thymic epithelium. This mAb generated a reticular labeling pattern in medullary and paracortical areas of lymph nodes and in splenic white pulp. At the ultrastructural level, labeling by the 8.1.1 mAb was restricted to fibroblastic reticular stromal cells. Serial sections of lymph node and spleen labeled with anti-CD3, anti-B220, and 8.1.1 mAbs clearly showed that the 8.1.1+ cells were associated with T cell-dependent areas. In severe combined immunodeficiency (SCID) or Nu/Nu mice, splenic white pulp also exhibited reticular labeling with the 8.1.1 mAb in the absence of detectable numbers of T cells, indicating that the appearance of 8.1.1-reactive stromal cells in discrete areas of peripheral lymphoid tissue was T cell independent. The cDNA encoding this stromal cell molecule was cloned by direct expression in COS cells and found to encode a 172 amino acid sequence with the typical features of a type I integral membrane protein. COS cells transfected with the gp38 clone direct the expression of an approximately 38-kD protein that reacts with the 8.1.1 mAb but not with isotype-matched controls. Comparison of the predicted amino acid sequence of 8.1.1 mAb but not with isotype-matched controls. Comparison of the predicted amino acid sequence of 8.1.1 with proteins in the National Biomedical Research Foundation (NBRF) data base showed that gp38 is very closely related to the early response protein OTS-8 obtained from a cDNA library of tumor promoting agent (TPA)-induced murine osteoblastic cell line, MC3T3-E1.


Primacy and recency effects in nonhuman primates. The reports of primacy and recency memory effects in nonhuman primates have been criticized because they have all used an initiating response. That is, the presentation of the to-be-remembered list of items was always contingent on a response being initiated by the nonhuman primate. It has been argued that this initiating response improves performance for early items in the list, resulting in the occurrence of the primacy effect, independent of any memory processing mechanism. This criticism was addressed in the present study by not using an initiating response prior to the presentation of the list. Nevertheless, both a primacy and a recency effect were observed in all 6 rhesus monkeys evaluated using a serial probe recognition task. Thus, the results are similar to those for humans, in that both primacy and recency effects can be obtained in nonhuman primates. A brief literature review is included, and it is proposed that the primacy and recency effects observed in humans, nonhuman primates, and infraprimates can be explained within the context of the configural-association theory.


Choice behavior in transition: development of preference with ratio and interval schedules. In Experiment 1, the choice responses of 8 pigeons were observed during 50 periods of transition. Each condition began with equal probabilities of reinforcement on 2 response keys and switched to unequal probabilities. With the ratio of the 2 probabilities held constant, preference for the higher probability developed more rapidly when the 2 probabilities were high than when they were low. In Experiment 2, each condition began with 2 equal variable-interval schedules, but later 1 key delivered 60%, 75%, or 90% of the reinforcers. The rate of approach to asymptotic performance was roughly the same with all 3 reinforcement percentages. These and previous results pose difficulties for some well-known models of acquisition, but the results are well described by a simple model that states that the strength of each response is independently increased by reinforcement and decreased by nonreinforcement.


Effects of discrimination training on responding during a compound conditioned stimulus. Four experiments examined responding in the presence of a triple-element compound ABC after discrimination training in which 2 compounds, AB and BC, signaled the delivery of food and 1 element alone, B, signaled the absence of food. In Experiments 1 and 2, using rats, responding during ABC was more vigorous than in a control group for which A and C but not B had been individually paired with food. This finding was replicated in Experiment 3, which used pigeons, and in Experiment 4, where all 3 stimuli of the control condition were individually paired with food. The results are more consistent with a configural than with an elemental theory of learning.


Discrimination of contour-deleted images by pigeons. Three experiments attempted to determine which properties of pictorial representations of objects control their discrimination by pigeons. A particular focus was whether the representation mediating such discriminations could be described by the simple viewpoint-invariant primitive volumes of Biederman's (1987) recognition-by-components theory of object recognition or by Cerella's (1990) particulate features. In all 3 experiments, pigeons were first trained to discriminate drawings of 4 stimulus objects with half of the contour deleted but with the component geons postulated by Biederman's theory recoverable. Discrimination accuracy was then compared for test items containing the original particulate features, affording the retrieval of the original component geons, or having neither of these properties of the training stimuli. Although response accuracy was significantly greater when the component geons of the original objects were retrievable, measurable control over recognition by the particulate features of the objects and by their specific locations was also found. The results are consistent with the idea of component geon recognition as one of the important factors in object discrimination.


Associative regulation of Pavlovian fear conditioning: unconditional stimulus intensity, incentive shifts, and latent inhibition. Conditional stimuli (CS) associated with painful unconditional stimuli (US) produce a naloxone-reversible analgesia. The analgesia serves as a negative-feedback regulation of fear conditioning that can account for the impact of US intensity and CS predictiveness on Pavlovian fear conditioning. In Experiment 1 training under naloxone produced learning curves that approached the same high asymptote despite US intensity. Shifting drug treatment during acquisition had effects that paralleled US intensity shifts. In Experiment 3 naloxone reversed Hall-Pearce (1979) negative transfer using a contextual CS, indicating that conditional analgesia acquired during the CS-weak-footshock phase retards acquisition in the CS-strong-footshock phase. Experiment 5 used a tone CS in both a latent-inhibition and a negative-transfer procedure. Only negative transfer was blocked by naloxone. Therefore, negative transfer but not latent inhibition is mediated by a reduction of US processing.


Measuring recognition memory.

Measuring recognition memory. Recent years have seen an expanded interest in recognition memory tasks. This resurgence of interest has also renewed concerns with measurement problems. Comparing 4 models of recognition memory, Snodgrass and Corwin (1988) found that measures of bias from the distribution-free (nonparametric) model were inadequate. However, their analysis was based on bias measures that can be shown a priori to be nonindependent of discrimination. This article traces the history of the nonparametric model and develops a better measure of bias. The consequence of developing this better measure is that the nonparametric model deserves serious consideration.


Similarity-scaling studies of dot-pattern classification and recognition. Classification performance in the dot-pattern, prototype-distortion paradigm (e.g., Posner & Keele, 1968) was modeled within a multidimensional scaling (MDS) framework. MDS solutions were derived for sets of dot patterns that were generated from prototypes. These MDS solutions were then used in conjunction with exemplar, prototype, and combined models to predict classification and recognition performance. Across 3 experiments, an MDS-based exemplar model accounted for the effects of several fundamental learning variables, including level of distortion of the patterns, category size, delay of transfer phase, and item frequency. Most important, the model quantitatively predicted classification probabilities for individual dot patterns in the sets, not simply general trends of performance. There was little evidence for the existence of a prototype-abstraction process that operated above and beyond pure exemplar-based generalization.


Comparative assessment of psychomotor performance: target prediction by humans and macaques (Macaca mulatta). Although nonhuman primates such as rhesus monkeys (Macaca mulatta) have been useful models of many aspects of cognition and performance, it has been argued that, unlike humans, they may lack the capacity to respond as predictor-operators. Data from the present series of experiments undermine this claim, suggesting instead a continuity of predictive competency between humans and nonhuman primates. A prediction coefficient was devised to examine the degree to which each subject's response path approximated the optimal predictive strategy. Whereas human subjects (N = 30) generally predicted more accurately, rhesus monkeys (N = 10) also significantly anticipated the movements of the target in all conditions. It appears that humans and rhesus monkeys both exhibit the capacity to respond to where a stimulus is going.


Memory for day of the week: a 5 + 2 day cycle. People use a variety of schemes to keep track of time. One such scheme is the week, with its 7 distinctly named days. This article examines memory for day of the week and its relation to memory for time of day and for elapsed time (number of days). Data are presented from a study in which people answered a set of interview questions about the time of occurrence of a particular event. Two issues are addressed. The first issue concerns the way different temporal schemes are organized in relation to each other in memory. Memory for day of the week is independent of other aspects of temporal memory. The second issue concerns whether the week is organized hierarchically into either weekday periods or weekend periods or both. The weekday period forms a distinct 5-day unit within the 7-day weekly cycle. The present data, together with those from the authors' earlier work, suggest that the time an event occurred is encoded in relation to a set of separate temporal scripts (e.g., daily and weekly) and that such scripts may be hierarchically organized.


The use of indirect memory tests to assess malingered amnesia: a study of metamemory. Many techniques have been suggested for identifying criminal suspects who are simulating amnesia for the events surrounding a crime. The present research focuses on indirect memory tests as a potential means of discriminating between those who genuinely suffer from amnesia and those who are simulating. Subjects studied a list of words and subsequently performed either a word completion or a fragment completion task. Under normal indirect test instructions, typical priming effects were observed. When subjects were motivated to simulate amnesia for the list, target completion rates were consistently, and sometimes reliably, below baseline completion rates. This finding is contrary to the performance of genuine amnesics, whose performance on indirect tests typically mirrors that of normal subjects. Indirect tests may prove useful in discriminating genuine and simulating amnesics.


Selectivity, scope, and simplicity of models: a lesson from fitting judgments of perceived depth. When comparing psychological models a researcher should assess their relative selectivity, scope, and simplicity. The third of these considerations can be measured by the models' parameter counts or equation length, the second by their ability to fit random data, and the first by their differential ability to fit patterned data over random data. These conclusions are based on exploration of integration models reflecting depth judgments. Replication of Massaro's (1988a) results revealed an additive model (Bruno & Cutting, 1988), and Massaro's fuzzy-logical model of perception (FLMP) fit data equally well, but further exploration showed that the FLMP fit random data better. The FLMP's successes may reflect not its sensitivity in capturing psychological process but its scope in fitting any data and its complexity as measured by equation length.


The picture superiority effect in categorization: visual or semantic? Two experiments are reported whose aim was to replicate and generalize the results presented by Snodgrass and McCullough (1986) on the effect of visual similarity in the categorization process. For pictures, Snodgrass and McCullough's results were replicated because Ss took longer to discriminate elements from 2 categories when they were visually similar than when they were visually dissimilar. However, unlike Snodgrass and McCullough, an analogous increase was also observed for word stimuli. The pattern of results obtained here can be explained most parsimoniously with reference to the effect of semantic similarity, or semantic and visual relatedness, rather than to visual similarity alone.


Serial pattern learning by event observation. Serial pattern learning was investigated in a variation of the task introduced by Nissen and Bullemer (1987). We presented an asterisk at 1 of 4 spatial locations on each trial, and Ss either responded with a keypress or observed the event. The first 4 blocks contained 10 repetitions of a 10- or 16-element pattern, and the 5th block contained a random sequence. The difference in response time on the 5th random block and the previous patterned block served as an indirect measure of pattern learning. A direct measure was obtained in a final test block in which Ss predicted the next asterisk position. Equivalent learning occurred for responding and observing with indirect measures, but observation was superior with direct measures. These findings indicate that knowledge of serial order can develop through simple perceptual experience, and this is more available to deliberate recall than is knowledge acquired while responding.


Framing and conflict: aspiration level contingency, the status quo, and current theories of risky choice. The effect of positive versus negative frames on risky choice was examined for a variety of scenarios and risks. Preferences in the positive domain were strong and mainly risk averse, with notable exceptions. Preferences in the negative domain, however, were marked by their inconsistency, shown both by an overwhelming lack of significant majority preferences and a surprisingly strong tendency of individual subjects to vacillate in their negatively framed choices across presentations. This finding is accounted for by a proposed aspiration level contingency in which aspiration levels are systematically set to be more difficult to achieve in the face of a perceived loss than a gain. The implications of the results, and the aspiration level contingency, are explored with respect to current theories of risky choice, including Kahneman and Tversky's (1979) prospect theory and Lopes's (1987, 1990) security-potential/aspiration theory.


Time course of movement planning: selection of handgrips for object manipulation. A goal of research on the cognitive control of movement is to determine how movements are chosen when many movements are possible. We addressed this issue by studying how subjects reached for a bar to be moved as quickly as possible from a home location to a target location. Ss generally grabbed the bar in a way that afforded a comfortable posture at the target location (the end-state comfort effect) and with the thumb toward the end of the bar that would be aligned with the target (the thumb-toward bias). The data suggested that subjects chose handgrips by retrieving instances of previous reaches, not by carrying out computations that treated candidate reaches as new behavioral events.


Cue familiarity but not target retrievability enhances feeling-of-knowing judgments. Two hypotheses concerning people's ability to predict later memory performance for unrecalled items were investigated. The target retrievability hypothesis states that feeling-of-knowing judgments (FKJs) are based on partial target information; and the cue familiarity hypothesis asserts that they are based on recognition of the cues. In Experiments 1 and 2, subjects either generated or read the targets of paired associates. Half of the cues had been primed in a pleasantness-rating task. The generation manipulation increased recall but had no effect on FKJs. Cue priming had no effect on recall but increased FKJs. In Experiment 3, using general information questions, primed after the initial recall attempt, both cue and target priming increased FKJs. Experiment 4, which remedied difficulties in Experiment 3, showed no effect of target priming whereas cue priming increased FKJs. The results favor the cue familiarity hypothesis.


Mental animation: inferring motion from static displays of mechanical systems. Reaction-time and eye-fixation data are analyzed to investigate how people infer the kinematics of simple mechanical systems (pulley systems) from diagrams showing their static configuration. It is proposed that this mental animation process involves decomposing the representation of a pulley system into smaller units corresponding to the machine components and animating these components in a sequence corresponding to the causal sequence of events in the machine's operation. Although it is possible for people to make inferences against the chain of causality in the machine, these inferences are more difficult, and people have a preference for inferences in the direction of causality. The mental animation process reflects both capacity limitations and limitations of mechanical knowledge.


Relational properties and memory-based category construction. Although many experiments have investigated factors that constrain perceptual category construction, there have been no investigations of factors that constrain memory-based (MB) category construction. Six experiments examined the extent to which perceptual and MB sorting were influenced by correlated dimensions, family resemblance principles, and conceptual knowledge. Sensitivity to many types of relational information (e.g., correlated features, causal relations, interactive properties of objects, and family resemblance relations) was observed with perceptual sorting, but these properties were rarely used to organize information in MB sorting conditions. Instead, there was a clear preference to organize categories around single dimensions. Even when perfectly correlated features were causally related, Ss in memory conditions did not use correlations to construct categories. The strengths and limitations of MB analyses and categorizations are discussed.


Evaluating hypnotic memory enhancement (hypermnesia and reminiscence) using multitrial forced recall. Two experiments investigated whether hypnosis enhances memory retrieval per se or merely increases a person's willingness to report recollections. Both experiments assessed immediate and delayed (i.e., 1 week) recall for pictorial stimuli. In Experiment 1, following an initial waking baseline recall, subjects of high or low hypnotic ability completed a series of recall trials conducted either in hypnosis or in the walking condition. The classic hypermnesia effect was obtained, but with no supplemental contribution of hypnosis. In Experiment 2, hypnosis was introduced only after 6 waking-recall trials. Hypnosis again failed to enhance retrieval of new correct items, although it increased the production of new incorrect recall among hypnotizable individuals. The findings provide no evidence for alleged hypermnesic properties of hypnosis.


Shapes of reaction-time distributions and shapes of learning curves: a test of the instance theory of automaticity. The instance theory assumes that automatic performance is based on single-step direct-access retrieval from memory of prior solutions to present problems. The theory predicts that the shape of the learning curve depends on the shape of the distribution of retrieval times. One can deduce from the fundamental assumptions of the theory that (1) the entire distribution of reaction times, not just the mean, will decrease as a power function of practice; (2) asymptotically, the retrieval-time distribution must be a Weibull distribution; and (3) the exponent of the Weibull, which is the parameter that determines its shape, must be the reciprocal of the exponent of the power function. These predictions were tested and mostly confirmed in 12 data sets from 2 experiments. The ability of the instance theory to predict the power law is contrasted with the ability of other theories to account for it.


Auditory priming: implicit and explicit memory for words and voices. Five experiments explore priming effects on auditory identification and completion tasks as a function of semantic and nonsemantic encoding tasks and whether speaker's voice is same or different at study and test. Auditory priming was either unaffected by the study task manipulation (Experiments 2, 4, and 5) or was less affected than was explicit memory (Experiments 1 and 3). Study-to-test changes of speaker's voice had nonsignificant effects on priming when white noise masked target items on the identification test (Experiments 1 and 2) or the stem-completion test (Experiment 5). However, significant voice change effects were observed on priming of completion performance when stems were spoken clearly (Experiments 3 and 4). Results are consistent with the idea that a presemantic auditory perceptual representation system plays an important role in the observed priming. Alternative explanations of the presence or absence of voice change effects under different task conditions are considered.


Short-term memory for the timing of auditory and visual signals. Short-term memory for the timing of irregular sequences of signals has been said to be more accurate when the signals are auditory than when they are visual. No support for this contention was obtained when the signals were beeps versus flashes (Experiments 1 and 3) nor when they were sets of spoken versus typewritten digits (Experiments 4 and 5). On the other hand, support was obtained both for beeps versus flashes (Experiments 2 and 5) and for repetitions of a single spoken digit versus repetitions of a single typewritten digit (Experiment 6) when the subjects silently mouthed a nominally irrelevant item during sequence presentation. Also, the timing of sequences of auditory signals, whether verbal (Experiment 7) or nonverbal (Experiments 8 and 9), was more accurately remembered when the signals within each sequence were identical. The findings are considered from a functional perspective.


The remembering of auditory event durations. Three experiments examined the incidental remembering of event durations. In each study, Ss engaged in an initial learning phase in which they performed a set of perceptual ratings on events for a varying number of trials. These events consisted of tonal sequences or ecological sounds that varied in their internal structure and ending. Ss were then given a surprise memory task in which they were asked to recognize the duration of each event (Experiments 1 and 3) or extrapolate its completion (Experiment 2). Results showed that in contrast to irregularly timed events, those filled with regularly timed or continuous pitch information yielded high levels of accuracy that increased with greater learning experience. In addition, durations marked by a nonarbitrary ending were more accurately remembered than those marked by an arbitrary ending which, in fact, were misremembered as shorter than their actual duration. These findings are discussed in terms of an approach that emphasizes the role of event structure on perceiving and remembering activities.


Individual differences in working memory and comprehension: a test of four hypotheses. A relationship has consistently been found between measures of working memory and reading comprehension. Four hypotheses for this relationship were tested in 3 experiments. In the first 2 experiments, a moving window procedure was used to present the operation-word and reading span tasks. High- and low-span subjects did not differentially trade off time on the elements of the tasks and the to-be-remembered word. Furthermore, the correlation between span and comprehension was undiminished when the viewing times were partialed out. Experiment 3 compared a traditional experimenter-paced simple word-span and a subject-paced span in their relationship with comprehension. The experimenter-paced word-span correlated with comprehension but the subject-paced span did not. The results of all 3 experiments support a general capacity explanation for the relationship between working memory and comprehension.


Persistence of negative priming: II. Evidence for episodic trace retrieval. Responses to recently ignored stimuli may be slower or less accurate than to new stimuli. This negative priming effect decays over time when delay is randomized within subjects, but not when delay varies between subjects. In Experiment 1, response-stimulus intervals (RSI) of 500 and 4,000 ms were randomized within subjects in a target localization task. Negative priming of ignored locations diminished with longer delay. However, no significant decay was obtained when RSI and the preceding RSI were equal. Similar results were obtained when RSI and preceding RSI were deliberately confounded by blocking (Experiment 2). Negative priming appears to depend on temporal discriminability of the priming episode.


Tools, teamwork, and tenacity: an office system for cancer prevention. Despite national priorities in cancer control, the number of people with established ongoing medical care who do not receive indicated preventive services is substantial. Proven strategies to optimize preventive care in community practice are limited. In the Cancer Prevention in Community Practice Project (CPCP), 50 primary care providers were randomly assigned to receive an "office system" intervention. The intervention led to reorganization of office operations based on four functional core components: identifying patients' needs for services; monitoring their status over time; providing positive reinforcement to patients; and establishing an internal feedback component consisting of a brief audit to assess how the system is operating. Implementation of the CPCP system in each practice was accomplished using trained facilitators, and involved incorporating one or more tools developed to meet the functional components of the practice. One hundred percent of the practices were successful in implementing some changes in their office operations that met CPCP office system functional criteria. All study practices implemented customized flow sheets, while use of other office system tools were incorporated at between 32% to 75% of study sites. Identifying patients in need of preventive services was performed most often by the clinical staff (39%), whereas monitoring patients' receipt of preventive services over time and reinforcing positive patient behavior were performed most often by physicians (63% and 46%, respectively). Changes made in practices were maintained for at least 12 months. Primary care practices in community settings can implement significant and lasting changes in their practice environment that will improve their performance of preventive activities. The functional components of the CPCP office system design proposed and tested here are applicable to a wide variety of practice settings.


Anabolic steroid use among adolescents in a rural state. Anabolic-androgenic steroid use is an increasing problem among high school students. Previous reports have been mainly from metropolitan areas. We report the first study of anabolic-androgenic steroid use to concentrate on rural communities. The study was conducted using an anonymous survey of a random sample of male high school students (N = 3900) in grades 10 through 12 encompassing 31 high schools in a predominantly rural state. Two-hundred five (5.3%) students reported using steroids. The prevalence of illicit drug use was significantly higher (P < .05) in steroid users (74%) than in nonusers (31%) (P < .001). The association was between anabolic-androgenic steroid use and illicit drug use rather than between sports participation of any type and illicit drug use (P > .2). Comparison of the prevalence of illicit drug use among athletic (63.2%) and nonathletic (36.8%) steroid users found no significant difference. Findings were similar with cigarette use. There was no difference in the rate of steroid use by school enrollment (69 to 1495) or by city population size (< 200 to 64,000). The predominant reason for steroid use was to improve appearance (43%). This study found the prevalence of steroid use throughout a predominantly rural state to be similar to that found by previous studies conducted in metropolitan areas; prevalence was not affected by city or school size. Steroid use was closely associated with illicit drug and cigarette use, a new finding that deserves further examination.


Determinants of delayed pregnancy testing among adolescents. Pregnant teenagers often prolong the interval between suspecting and confirming that they are pregnant. Prior studies suggest a number of potential determinants for this delay but do not specify which ones are most salient. In a cross-sectional survey, 123 pregnant teenagers, 64 of whom maintained their pregnancies and 59 of whom had abortions, completed a short version of the Center for Epidemiologic Studies Depression Scale, the Family APGAR test, and a study-specific questionnaire. Significant bivariate determinants of delayed pregnancy testing included young maternal age, black race, lower educational attainment, lack of pregnancy symptoms, continuing the pregnancy, and denial. Only denial, however, retained a significant net effect on delayed testing (P < .05) when the effects of these six variables were modeled using multiple linear regression. These results suggest that psychological barriers are the most salient determinants of delayed pregnancy testing among the teenagers surveyed in this study. Some teenagers may not volunteer information about a suspected pregnancy. Providers, therefore, should directly question teenagers about sexual activity and discuss the importance of early testing when pregnancy is suspected. Findings also suggest further research that would increase understanding of adolescent health behavior in pregnancy and identify effective clinical and educational interventions.


Comparative validity of two hearing loss screening questionnaires. Hearing loss is one of the most common of all physical impairments, but physicians seldom screen adults for it, and patients often overlook or deny hearing problems. This study was designed to validate the use of two self-administered hearing loss questionnaires. Two self-administered screening questionnaires and a hearing screening evaluation using the Welch Allyn Audioscope 3 instrument were given to 409 consecutive family practice patients over the age of 18 years. Correlational, discriminate, sensitivity, and specificity analyses were conducted on the data. Neither of the existing questionnaires was clinically sensitive enough to be recommended for use. A new tool based on a discriminate function analysis of the existing questionnaires was developed. In contrast, the audioscope proved to be a sensitive screening tool. Of those patients who were identified, 88% did not follow recommendations to obtain further evaluation. Existing self-administered questionnaires cannot be recommended for use. A controlled clinical study using the newly derived questionnaire, the Smith Hearing Screening, should be conducted.


Attitudes toward and experience in research among family medicine chairs. Productive research environments are important for the development of academic family medicine, yet many of the current family medicine chairs have had little research training or experience and have rated research skills as a low priority for themselves. The younger chairs, however, representing the next generation of academic leadership, may have more traditional academic values, including the promotion of research. The 106 active and interim chairs of family medicine academic units were surveyed by mail to determine their characteristics and attitudes toward their work responsibilities. We compared chairs 50 years of age or younger with those over 50 years of age. Before attaining their positions, younger chairs, in general, were more likely than older chairs to have received formal training in management, patient care, and academic skills, but they shared similar work experiences. Specifically, younger chairs were more likely to have had formal research training but did not have a great deal more research experience. Younger chairs were more likely to consider research skills to be essential in their present work activities and to identify faculty with formal training and extensive experience in research as potential chair replacements. Younger chairs appear to have a greater appreciation for the importance of research, having received more formal training and valuing research skills in themselves and potential replacements. With the impending large turnover in family medicine leadership, there will be an opportunity to recruit chair replacements with similar viewpoints toward research, thus improving the outlook for research in academic family medicine.


Diagnosis of acute pelvic pain.

Diagnosis of acute pelvic pain. The diagnosis of acute pelvic pain in the woman of reproductive age represents a major clinical challenge. In approaching such a patient, the clinician must differentiate between pregnancy-related causes, gynecologic disorders, and nonreproductive tract causes. A careful history and physical examination, along with selective and knowledgeable use of diagnostic tests and procedures, are essential to the diagnostic process. Diagnostic laparoscopy represents the reference standard for diagnosis of many of its possible causes and can obviate the need for exploratory laparotomy. Once competing diagnoses have been adequately excluded, an empiric trial of antibiotic therapy for acute pelvic inflammatory disease, coupled with close clinical follow-up, should be considered in patients with acute pelvic pain found to have cervical motion tenderness and bilateral adnexal tenderness on examination.


Coding and reimbursement of primary care biopsy and destruction procedures. Current medical practice requires physicians to accurately report services provided to patients. Billing for destruction of benign and malignant lesions and for surgical, needle, and endoscopic biopsy procedures involves the selection of specific 1992 Current Procedural Terminology (CPT) codes. Payment for these procedures by third-party payers often requires the International Classification of Diseases, 9th Revision, Clinical Modification (ICD-9-CM) coding for neoplastic lesions. This review explains the proper codes to use in identifying common biopsy and destruction procedures performed by primary care physicians. The Health Care Financing Administration's relative value units and one state's published Medicaid payment rates are included for each procedure code. Instructions for selecting site-specific biopsy and destruction codes are provided.


Acute leukostasis pulmonary distress syndrome. A 75-year-old woman in accelerated-phase chronic myeloid leukemia with hyperleukocytosis presented with acute respiratory distress syndrome. Despite early and aggressive pulmonary support and cytoreductive chemotherapy, the patient died. Autopsy confirmed the presence of the leukostasis syndrome. The clinical, radiologic, pathophysiologic, and therapeutic aspects of this entity are reviewed.


Hyperemesis, hyperthyroidism, or both?

Hyperemesis, hyperthyroidism, or both? Nausea and vomiting are common during pregnancy and, when severe enough to require intervention, may develop into the syndrome known as hyperemesis gravidarum. When the diagnosis of hyperemesis is considered, a careful search for secondary causes is necessary. The list of secondary causes includes hyperthyroidism, a relatively uncommon condition during pregnancy. Because many of the signs and symptoms of hyperthyroidism are common, and thyroid function tests are more difficult to interpret during normal pregnancy, making the diagnosis of hyperemesis gravidarum is a challenge. The decision to treat or to await spontaneous resolution depends on the severity of the illness and the likelihood of the presence of true Graves' disease. The case summarized here demonstrates these issues, and includes treatment options for hyperemesis-associated hyperthyroidism.


Osmotic and ionic regulation during hypoxia in the medicinal leech, Hirudo medicinalis L. The concentrations of inorganic and organic ions and osmolality in the blood of the medicinal leech, Hirudo medicinalis, were determined during normoxia and hypercapnic and hypocapnic hypoxia. In normoxic animals, the blood sodium concentration was 124.5 +/- 4.2 mmol/l and the total cation concentration was 132.2 +/- 4.3 mEq/l (mean +/- S.D.). Major anionic compounds were chloride (40.8 +/- 1.6 mmol/l), bicarbonate (8.4 +/- 1.3 mmol/l), and organic anions (42.5 +/- 2.3 mEq/l). Among the latter, malate accounts for 30.4 +/- 2.2 mEq/l. The nature of the remaining anion fraction, which balances cation and anion concentrations in leech blood, remains unknown. Within 96 h of hypercapnic hypoxia, the amount of organic osmolytes in leech tissue increased from the control level of 56.6 +/- 9.1 to 158.3 +/- 19.5 mumol/g dry weight. An even higher amount of organic acids was accumulated within 96 h of hypocapnic hypoxia (218.0 +/- 53.7 mumol/g dry weight). A possible reason for this is that lactate, which is a major end-product of hypocapnic hypoxia, cannot be excreted to the external medium as easily as propionate. The accumulation of blood organic acids generating osmotic stress in the animals was compensated by an equimolar decrease in sodium and chloride ion concentrations. In hypercapnic animals these changes resulted in a constant osmotic concentration of the blood (200 mosmol/kg H2O) during the experimental period. Between 24 and 96 h of hypocapnic hypoxia, however, the increase in the osmotic gradient between animal and medium was correlated with further net water uptake and the obvious deterioration of the volume- and ion-regulatory mechanisms in these animals.


Mouse limb bud cells respond to retinoic acid in vitro with reduced growth. Retinoic acid (RA) has dramatic effects on the pattern of developing and regenerating vertebrate limbs. These effects are considered to result from RA-induced changes in the positional identity of limb cells, and involve the formation of extra structures. Whether the growth required to form the supernumerary parts of the pattern is a primary effect of RA treatment or a secondary effect that follows after a change in positional identity is not at present known. In this paper we have investigated the effects of RA treatment on the growth of cells from anterior and posterior halves of mouse limb buds in vitro. We observed that under our culture conditions, limb bud cells treated with 1 nM to 1 microM RA (0.3 ng/ml to 300 ng/ml) continue to grow but do so at a significantly slower rate than control cultures. There is a maximum inhibition of growth (50% of controls) between 10 nM and 100 nM RA, which corresponds to the measured range of concentrations of RA in vivo. Our observation of a significant decrease in growth rate over a wide range of RA concentrations is consistent with comparable reports of growth inhibition for a large number of other cell types in vitro as well as with the observation that exogenous RA inhibits blastemal growth in amphibians during the period of exposure to RA. We propose that the effects of RA on growth, either enhancement in vivo or reduction in vitro, can be seen as consequences of the ability of RA to alter positional identity.(ABSTRACT TRUNCATED AT 250 WORDS)


Epibolic extension of the presumptive ectodermal layer of embryos of the newt Cynops pyrrhogaster before and during gastrulation. Epibolic extension of the presumptive ectodermal layer (PEL) was investigated in embryos of the newt Cynops pyrrhogaster before and during gastrulation. The PEL was composed of only one layer of columnar cells at all stages examined. The cells of the PEL became elongated from the blastula to the early gastrula stage. They were most elongated at the early gastrula stage and then shortened during gastrulation. Present observations suggest that changes in cell shape of the PEL play an important role in the control of the epibolic extension of the newt embryos. The morphology and movement of the isolated cells from the PEL were examined in an attempt to elucidate the role of cell movement in epibolic extension of the PEL. Blebbing and vermiform cells which showed active cell movement appeared at the early blastula stage. The blebbing cells, which formed large hyaline blebs that moved around the circumference of each cell, appeared in large numbers at the early blastula stage. The frequency of the blebbing cells decreased from the early blastula to the early gastrula stage and increased again during gastrulation. The vermiform cells, which had an elongated cell body and moved in a worm-like manner, increased in frequency from the early blastula to the early gastrula stage. The relative number of such vermiform cells was maximal at the early gastrula stage and decreased abruptly during gastrulation. These results suggest that the elongation of the cells of the PEL is controlled by the active cell movement which resembles that of a worm.


Retinoic acid respecifies limb bud cells in vitro. Retinoic acid (RA) is known to have dramatic effects on limb pattern formation and has been shown to exert its effects on limbs by converting anterior limb bud cells into cells with posterior positional properties. In this study we find that dissociated posterior limb bud cells from chick and mouse embryos cultured at high density (micromass cultures) are able to stimulate the formation of supernumerary digits when grafted into developing wing buds and that the positional identity of both chick and mouse limb bud cells can be maintained for finite periods of time in vitro. Furthermore, using this assay system we have tested whether anterior cells from mouse and chick limb buds can be converted into cells with posterior identity by exposure to RA in vitro. We find that anterior limb bud cells acquire posterior properties after culture in the presence of RA.


Body temperature rhythm of the tree shrew, Tupaia belangeri. The circadian rhythm of body temperature of the tree shrew Tupaia belangeri was studied by telemetry. The amplitude of the daily (or circadian) variation was found to be much larger than that of most endotherms (amplitude approximately 5 degrees C) and the bimodal shape of the rhythm differed from the cosine waveform that characterizes the temperature rhythms of most other species. In free-running conditions, as well as in the entrained state, the temperature rhythm remained synchronized to the rhythm of locomotor activity.


Micromanipulation of cleaved embryos cultured in protein-free medium: a mouse model for assisted hatching. A mouse model for studying anomalies of human embryonic hatching following micromanipulation is proposed. Initiation and completion of mouse blastocyst hatching was severely impaired (34/292; 12% and 28/292; 10%, respectively) with protein deprivation, resembling the situation in human in vitro fertilization. Hatching ability was restored when an artificial gap was introduced in the zona pellucida by micromanipulation at the cleaved embryo stage. This enabled 77% (285/371) and 36% (134/371) of the embryos to initiate and complete hatching in protein-free medium. No differences were found in overall cell counts between the two groups of embryos. Transfer of micromanipulated blastocysts to pseudopregnant females resulted in development of healthy fetuses.


Hyoid fracture and strangulation.

Hyoid fracture and strangulation. Observation of hyoid fracture in skeletonized remains offers potentially valuable information on the history of the skeleton or evidence of foul play, or both. Perimortem hyoid fracture frequently indicates manual strangulation, although ligature strangulation, hanging, and other forms of trauma to the neck cannot be ruled out without additional evidence. Such fractures are rare in children and infants, since the hyoid components are not fully ossified and are more flexible than in adults. Both antemortem and postmortem origins of the fractures must also be considered.


Estimation of adult stature from fragmentary tibias. Linear-regression equations derived from measurements of tibial condyles from 100 individuals in the Hamann-Todd Collection retrodicted known stature with a level of confidence comparable to many of the existing stature-estimation techniques. Statures of an independent control group were estimated with similar success. The strong linear relationship that exists between the length of the tibia and the size of the condyles allows adult stature (of American whites and blacks) to be estimated from the proximal tibia. Since complete tibial length is not required, this technique could prove useful in forensic science and archaeological cases where less-than-intact elements are recovered.


Modification of the Trotter and Gleser female stature estimation formulae. Stature-estimation formulae in common use are those of Trotter and Gleser. Their formulae for females are based on Terry collection skeletons. These skeletons are from people who died in the early 1990s. Because there has been considerable change in body size since then, it is possible that the Trotter and Gleser formulae are inappropriate for modern forensic-science application. The Trotter and Gleser female formulae are tested using data from the Forensic Data Bank at the University of Tennessee. For whites, the femur and tibia yield stature estimates differing from one another by about 3 cm. Using femur and tibia lengths from modern forensic cases and modern height data from anthropometric surveys, new regression intercepts are calculated for Trotter and Gleser's female formulae. The new intercepts improve the performance of the formulae on modern individuals. The Trotter and Gleser formulae for black females require no adjustment. Both blacks and whites have experienced a secular increase in bone length, but whites have experienced a change in proportions as well.


Time since death determinations of human cadavers using soil solution. This study was conducted to collect data on specific volatile fatty acids (produced from soft tissue decomposition) and various anions and cations (liberated from soft tissue and bone), deposited in soil solution underneath decomposing human cadavers as an aid in determining the "time since death." Seven nude subjects (two black males, a white female and four white males) were placed within a decay research facility at various times of the year and allowed to decompose naturally. Data were amassed every three days in the spring and summer, and weekly in the fall and winter. Analyses of the data reveal distinct patterns in the soil solution for volatile fatty acids during soft tissue decomposition and for specific anions and cations once skeletonized, when based on accumulated degree days. Decompositional rates were also obtained, providing valuable information for estimating the "maximum time since death." Melanin concentrations observed in soil solution during this study also yields information directed at discerning racial affinities. Application of these data can significantly enhance "time since death" determinations currently in use.


Surveillance of human immunodeficiency virus (HIV) antibodies in medicolegal autopsies in Finland--monitoring early changes in HIV-seropositivity among risk groups and average population. In order to cooperate with voluntary screening programs aimed at the surveillance of the HIV epidemic in Finland, we have studied medicolegal autopsies for HIV antibodies since 1986 using an enzyme immunoassay on postmortem sera. The investigation covered 47.4% and 39.2%, respectively, of all deaths under the age of 65 years in the metropolitan areas of Helsinki and Turku--two cities on the densely populated southern coast of Finland from which most HIV infections have thus far been detected. Nine HIV-positive cases (0.12%) were detected among the 7305 medicolegal autopsies tested in 1986 to 1990. This figure is higher than the prevalence of 0.01 to 0.03% in voluntary screening programs for the general population would suggest. Seven of our cases had previously tested positive, and two were previously unknown cases, indicating that people at high risk are clustered in the medicolegal autopsy series. Of the six cases in an early stage of infection, three committed suicide suggesting the importance of HIV-screening in suicide cases in tracing symptomless HIV carriers. Five of the cases were detected in 1990, a year when the number of new HIV infections had more than doubled compared to the previous two years. This suggests that testing of medicolegal autopsies as surrogate tests for the population gives useful information even in low-prevalence areas like Finland. Such testing has none of the ethical problems of many other back-up surveys, and may be particularly sensitive to early changes in epidemiology.


ABO genotyping by polymerase chain reaction. ABO blood group system's genotyping by polymerase chain reaction in genomic DNA level is developed. The positions of nucleotide 258 and 700 of cDNA from A transferase were used to distinguish A, B, and O alleles by restriction enzyme digestion. To identify the 258th nucleotide, a 199- or 200-bp DNA fragment was amplified by PCR and digested with Kpn I. For the 700th nucleotide, a 128-bp PCR amplified fragment was designed and digested with Alu I. By examining the DNA fragment digested patterns, ABO genotypes were easily determined. Results obtained using this method on 20 ABO-known peripheral blood samples showed that this new technique could provide accurate ABO genotype. Biologic forensic samples, such as, blood stains, saliva stains, semen stains, hair, bone tissue, and semen contaminated with vaginal secretion were also successfully typed. This rapid, sensitive and reliable method should be applicable not only in forensic identification but also in medical examination.


Robotic method for the analysis of morphine and codeine in urine. A totally automated procedure has been developed for the detection and quantitation of morphine and codeine in urine case samples. The samples were initially screened for these drugs by a Syva EMIT Toxicology System (ETS). A Zymate laboratory robotic system confirms positive samples from Syva ETS by performing the hydrolysis, extraction, and derivatization of morphine and codeine. The derivatized morphine and codeine were detected using gas chromatography/mass spectrometry (GC/MS). Enzymatic hydrolysis conditions were experimentally optimized during method development. The automation of these procedures has proven to be reliable and efficient.


A psychiatric study of persons charged with arson. A total of 29 court-referred individuals charged with arson were psychiatrically studied. From this pre-trial cohort from a large heterogeneous urban population base, a higher rate of psychosis was found than in other recent studies. However, consistent with these studies was the rarity of the diagnosis of pyromania. An important finding of this study was the substantial number of fires set by individuals who are homeless mentally disordered or substance abusing, or both.


Criminals' explanations of their criminal behavior, Part I: The contribution of criminologic variables. The author reviewed the literature concerning criminals' explanations of their crimes and then studied the explanations given by 100 incarcerated men. He found no significant associations between juvenile or adult arrest histories, alias use, age at time of the crime, trial plea, sentence length, duration of incarceration, and explanation types used. Only murderers significantly used a specific explanation type. These observations suggest that explanations are largely independent of traditional criminological attributes; that prolonged confinement to prison does not result in offenders admitting to their crimes; and, that killers have an especially difficult time accepting responsibility for taking the life of another human being.


Criminals' explanations of their criminal behavior, Part II: A possible role for psychopathy. The author reviewed the literature concerning the effect of criminals' current psychopathology on their explanations about their crimes. He then studied 100 incarcerated men, examining the associations between their explanations and various historically documented aspects of psychopathy. Previously and independently documented patterns of pathological lying, lack of remorse or guilt, callousness or lack of empathy, and failure to accept responsibility for their own behavior were significantly associated with the offenders not admitting responsibility for their crimes. Further, independently described histories of pathological lying were associated with criminals' blaming their convictions on a faculty criminal-justice system, while histories of failure to accept responsibility for their behavior were associated with blaming someone else for their index crimes.


Postmortem stability of lung surfactant phospholipids. The postmortem stability of the main phospholipids of lung surfactant-phosphatidyl choline (PC), phosphatidyl ethanolamine (PE), phosphatidyl inositol (PI), phosphatidyl serine (PS) and sphingomyelin (S) in three different deaths; one caused by fresh-water drowning, one by salt-water drowning, and one from a sodium-pentobarbital overdose has been studied. The drug overdose was considered the control because there was no surfactant involvement. The results show the stability of these kinds of lipids in the first 24 h, with a progressive decrease from 48 h on until 96 h, with a significant correlation to the time of P less than 0.01 in most cases.


Postmortem angiography of catheter-induced pulmonary artery perforation. A case of pulmonary artery perforation by the placement of a balloon-tipped, flow-directed (Swan-Ganz) catheter, as demonstrated postmortem by angiography and confirmed by conventional autopsy method, is reported. Angiography is an effective adjunctive modality in the postmortem diagnosis and localization of pulmonary artery perforation. In cases of suspected catheter-induced pulmonary artery perforation and death, postmortem angiography may prove useful to the forensic pathologist.


Toxicologic findings in the USS Iowa disaster. The toxicologic results from the 47 victims of the explosion on the USS Iowa are presented. Good correlation between carboxyhemoglobin saturations and cause of death was found. There were no correlations between blood cyanide concentrations and causes of death. Volatile analysis suggested postmortem ethanol production rather than antemortem ethanol ingestion. No drugs except nicotine were detected in any of the victims.


The elimination rate of mouth alcohol: mathematical modeling and implications in breath alcohol analysis. Mouth alcohol, if present in high enough concentrations, can falsely bias the accurate measurement of end-expiratory breath alcohol. Mouth alcohol will be eliminated over time, however, and can be modeled with a single term decaying exponential of the form: B0e-kt + C. It is important, however, to determine the model and its parameters when alcohol is already present within the biologic system. Using three individuals as their own controls, mouth alcohol was administered both before and after alcohol consumption followed by breath alcohol analysis performed at approximately 0.5 min intervals. The results showed that both model parameters (B0 and k) are effected and that the asymptotic value (C) is reached much sooner when alcohol already exists in the end-expiratory breath. Considering only three individuals were involved, the forensic-science importance appears to be that, as the end-expiratory breath alcohol concentration increases, the time necessary for the mouth alcohol to decrease to unbiased levels is decreased. Fifteen min of observation time prior to breath alcohol analysis appears to be more than adequate at forensically relevant concentrations.


A simple technique for age estimation in adult corpses: the two criteria dental method. A method for age determination of adults from single rooted teeth is presented. It is based on the measurement of two dental features: periodontosis height times 100/root height (P) and transparency of the root height times 100/root height (T). These measurements are made on the labial surface of the entire tooth without section and do not require special equipment or training. The application of multiple regression analysis to a working sample of 306 teeth of known age, sex and race provided the following equation: Age (years) = 0.18 x P + 0.42 x T + 25.53. The mean error between the actual and estimated age was +/- 10 years on the working sample and +/- 8.4 years on a control sample made of 45 forensic science cases. Upper incisors showed a better precision than the other single rooted teeth and accuracy was not sex related. A comparison of the Gustafson and Lamendin methods on a control sample of 39 teeth resulted in an advantage of the latter considering the mean error on the estimation (14.2 +/- 3.4 years for Gustafson versus 8.9 +/- 2.2 for Lamendin). The Lamendin method can be practical interest for any forensic pathologist or dentist as it is fast, easy to use, and reasonably accurate except for cases of individuals under age 40 where other methods must be preferred.


Human osteology: key to the sequence of events in a postmortem shooting. Forensic anthropologic examination of human skeletal remains found when a field was plowed provides evidence of both perimortem trauma, suggesting cause of death, and of subsequent shooting of the disarticulated skeleton. The case exemplifies the application of the specialized skills and knowledge of the physical anthropologist to determination of the postmortem sequence of events.


Rare electrocution due to powerline contact in a hot-air balloon: comparison with fatalities from blunt trauma. Powerline contact by hot-air balloons is one of the most frequent concurrences in balloon accidents resulting in injury or death. Injuries and deaths are usually a result of blunt trauma from falls. In this report, we describe the aircraft, the circumstances of the accidents and the autopsy data in two powerline contact accidents involving three deaths, one from electrocution and two, from blunt trauma sustained in falls. Appropriate pilot behavior is briefly discussed.


Sudden death due to intravascular hemolysis after bladder irrigation with distilled water. A 45-year-old white man was hospitalized with gross hematuria, one month after cystoscopy and biopsy for the same complaint. The biopsy revealed cystitis glandularis. One day after admission, he developed seizures and died within hours. Autopsy, laboratory tests, and further questioning of the hospital staff showed that he died of acute hyponatremia and massive intravascular hemolysis after irrigating the bladder with sterile water. Two deep bladder ulcers with exposed veins served as the portals of entry. Until now, this fatal complication had been described only during transurethral surgery. Both a careful autopsy and hospital investigation is necessary to differentiate in-hospital natural death from iatrogenic fatality.


Fatal virus-associated hemophagocytic syndrome in a young adult producing nontraumatic splenic rupture. A 24-year-old man with no previous medical history was admitted to a local hospital with pancytopenia after a recent "viral illness." During his hospitalization, he developed sudden abdominal distension and hypotension. Surgical exploration of his abdomen revealed a ruptured spleen. The spleen was removed, but the patient did not survive the operation. We investigated this unexpected and unexplained hospital death for any traumatic or iatrogenic injury. The cause of death after review of the clinical history, autopsy, and microscopic sections was virus-associated hemophagocytic syndrome (VAHS). VAHS consists of a generalized histiocytic proliferation and marked hemophagocytosis associated with a systemic viral infection. Clinically it presents as pancytopenia and organomegaly. This recently described entity is often confused with malignant histiocytosis. This is the first case report of VAHS producing nontraumatic splenic rupture, thus adding to the differential diagnosis of spontaneous splenic rupture and sudden natural death.


Fatal angioedema associated with captopril. A markedly hypertensive, 70-year-old, black man had been on captopril for 2 years when he rapidly developed obstructive angioedema. The initial sign of difficulty in understanding his speech progressed to severe laryngeal and glossal edema over a 3 1/2 h period. His airway became obstructed less than a minute after arrival at the emergency room. Oral intubation was unsuccessful, and a difficult tracheostomy was too late to save the patient. The death was reported to the medical examiner because of its sudden and unusual nature. The risk of angioedema while on angiotensin converting enzyme inhibitor therapy has been noted previously in the clinical literature. Because of the sudden onset and possible confusion with an allergic reaction, this entity is brought to the attention of the forensic medical community.


Spontaneous clostridial myonecrosis.

Spontaneous clostridial myonecrosis. Spontaneous, nontraumatic clostridial myonecrosis is a rare infection with an insidious onset and usually fatal outcome. Spontaneous clostridial myonecrosis has a frequent association with colon carcinoma, leukemia, diabetes mellitus, and drug-induced immunosuppression. We present the case of a 73-year-old diabetic man who died of spontaneous Clostridium septicum myonecrosis, who had presented with fulminant gangrene of the right thigh. Clostridium septicum was cultured from the quadriceps muscle postmortem. At autopsy, in addition to the gangrene, there was a Duke's A adenocarcinoma of the cecum, which had not been diagnosed during life. When spontaneous nontraumatic clostridial myonecrosis is diagnosed at autopsy, investigation should include through exam and the obtaining of past medical history in order to elucidate predisposing factors.


Cardiovascular physiology. Effects of microgravity. Experiments during spaceflight and its groundbase analog, bedrest, provide consistent data which demonstrate that numerous changes in cardiovascular function occur as part of the physiological adaptation process to the microgravity environment. These include elevated heart rate and venous compliance, lowered blood volume, central venous pressure and stroke volume, and attenuated autonomic reflex functions. Although most of these adaptations are not functionally apparent during microgravity exposure, they manifest themselves during the return to the gravitational challenge of earth's terrestrial environment as orthostatic hypotension and instability, a condition which could compromise safety, health and productivity. Development and application of effective and efficient countermeasures such as saline "loading," intermittent venous pooling, pharmacological treatments, and exercise have become primary emphases of the space life sciences research effort with only limited success. Successful development of countermeasures will require knowledge of the physiological mechanisms underlying cardiovascular adaptation to microgravity which can be obtained only through controlled, parallel groundbased research to complement carefully designed flight experiments. Continued research will provide benefits for both space and clinical applications as well as enhance the basic understanding of cardiovascular homeostasis in humans.


Skeletal muscle responses to unloading with special reference to man. The limited space flight data suggest that exposure to microgravity decreases muscle strength in humans and muscle mass in lower mammals. Several earth-based models have been used to address the effect of unloading on the human neuromuscular system due to the limited access of biological research to long-term space flight. Bedrest eliminates body weight bearing of both lower limbs. Unilateral lower limb suspension (ULLS), where all ambulatory activity is performed on crutches with an elevated sole on the shoe of one foot, has recently been used to unload one lower limb. The results from studies using these two models support their efficacy. The decrease in strength of m. quadriceps femoris, for example, after four to six weeks of bedrest, ULLS or space flight is 20 to 25%. The results from the earth-based studies show that this response can be attributed in part to a decrease in the cross-sectional area of the KE which reflects muscle fiber atrophy. The results from the ground based studies also support the limited flight data and show that reductions in strength are larger in lower than upper limbs and in extensor than flexor muscle groups. They also raise issue with the generally held concept that postural muscle is most affected by unweighting. Slow-twitch fibers in lower limb muscles of mixed fiber type composition and muscle composed mainly of slow-twitch fibers do not preferentially atrophy after bedrest or ULLS. Taken together, the data suggest that unloading causes remarkable adaptations in the neuromuscular system of humans. It should be appreciated, however, that this area of research is in its infancy.


Biomedical engineering. A means to add new dimension to medicine and research. Biomedical engineering is an evolving science that seeks to insert technically oriented and trained personnel to assist medical professionals in solving technological problems in the pursuit of innovations in the delivery of health care. Consequently, engineering solutions are brought to bear on problems that previously were outside the training of physicians and beyond the understanding or appreciation of the conventionally educated electrical or mechanical engineers. This physician/scientist/engineer team has a capability to extend medicine and research far beyond the capability of a single entity operating alone. How biomedical engineering has added a new dimension to medical science at the Kennedy Space Center is described.


Environmental monitoring and research at the John F. Kennedy Space Center. The Biomedical Operations and Research Office at the NASA John F. Kennedy Space Center has been supporting environmental monitoring and research since the mid-1970s. Program elements include monitoring of baseline conditions to document natural variability in the ecosystem, assessments of operations and construction of new facilities, and ecological research focusing on wildlife habitat associations. Information management is centered around development of a computerized geographic information system that incorporates remote sensing and digital image processing technologies along with traditional relational data base management capabilities. The proactive program is one in which the initiative is to anticipate potential environmental concerns before they occur and, by utilizing in-house expertise, develop impact minimization or mitigation strategies to reduce environmental risk.


Kennedy Space Center environmental health program. The Kennedy Space Center's environmental health organization is responsible for programs which assure its employees a healthful workplace under diverse and varied working conditions. These programs encompass the disciplines of industrial hygiene, radiation protection (health physics), and environmental sanitation/pollution control. Activities range from the routine, such as normal office work, to the highly specialized, such as the processing of highly toxic and hazardous materials.


Emergency medical operations at Kennedy Space Center in support of space shuttle. The unique environment of the Kennedy Space Center includes a wide variety of industrial processes culminating in launch and spaceflight. Many are potentially hazardous to the work force and the astronauts. Technology, planning, training, and quality control are utilized to prevent contingencies and expedite response should a contingency occur.


Health services at the Kennedy Space Center. Comprehensive occupational health services are provided to approximately 17,000 workers at the Kennedy Space Center and an additional 6000 on Cape Canaveral Air Force Station. These areas cover about 120,000 acres encompassing part of the Merritt Island Wild Life Refuge and wetlands which are the habitat of numerous endangered and protected species of wildlife. The services provided at the Kennedy Space Center optimally assure a safe and healthy working environment for the employees engaged in the preparation and launching of this country's Space Shuttle and other important space exploration programs.


Role of atrial natriuretic factor in salt and water homeostasis. Studies over the past 10 years suggest that the atrial natriuretic factor (ANF) plays an important role in salt and water homeostasis. Responding to atrial stretch, the atria releases ANF into the circulation. The several actions of this hormone tend to increase renal NaCl excretion resulting in reduced blood volume and blood pressure. ANF increases the glomerular filtration rate and reduces sodium chloride reabsorption in the distal nephron. It also inhibits secretion of aldosterone from the adrenal cortex. Therefore actions of ANF appear to be opposed to the renin-angiotensin-aldosterone system. Drugs that alter ANF metabolism may constitute a new mechanism of treatment for hypertension and heart failure.


Chemical properties of the divalent cation binding site on potassium channels. The actions of divalent cations on voltage-gated ion channels suggest that these cations bind to specific sites and directly influence gating kinetics. We have examined some chemical properties of the external divalent cation binding sites on neuronal potassium channels. Patch clamp techniques were used to measure the electrophysiological properties of these channels and Zn ions were used to probe the divalent cation binding site. The channel activation kinetics were greatly (three- to fourfold) slowed by low (2-5 mM) concentrations of Zn; deactivation kinetics were only slightly affected. These effects of Zn were inhibited by low solution pH in a manner consistent with competition between Zn and H ions for a single site. The apparent inhibitory pK for this site was near 7.2. Treatment of the neurons with specific amino acid reagents implicated amino, but no histidyl or sulfhydryl, residues in divalent cation binding.


Divalent cation block and competition between divalent and monovalent cations in the large-conductance K+ channel from Chara australis. The patch-clamp technique is used to investigate divalent ion block of the large-conductance K+ channel from Chara australis. Block by Ba2+, Ca2+, Mg2+, and Pt(NH3)4(2+) from the vacuolar and cytoplasmic sides is used to probe the structure of, and ion interactions within, the pore. Five divalent ion binding sites are detected. Vacuolar Ca2+ reduces channel conductance by binding to a site located 7% along the membrane potential difference (site 1, delta = 0.07; from the vacuolar side); it also causes channel closures with mean a duration of approximately 0.1-1 ms by binding at a deeper site (site 2, delta = 0.3). Ca2+ can exit from site 2 into both the vacuolar and cytoplasmic solutions. Cytoplasmic Ca2+ reduces conductance by binding at two sites (site 3, delta = -0.21; site 4, delta = -0.6; from the cytoplasmic side) and causes closures with a mean duration of 10-100 ms by binding to site 5 (delta = -0.7). The deep sites exhibit stronger ion specificity than the superficial sites. Cytoplasmic Ca2+ binds sequentially to sites 3-5 and Ca2+ at site 5 can be locked into the pore by a second Ca2+ at site 3 or 4. Ca2+ block is alleviated by increasing [K+] on the same side of the channel. Further, Ca2+ occupancy of the deep sites (2, 4, and 5) is reduced by K+, Rb+, NH4+, and Na+ on the opposite side of the pore. Their relative efficacy correlates with their relative permeability in the channel. While some Ca2+ and K+ sites compete for ions, Ca2+ and K+ can simultaneously occupy the channel. Ca2+ binding at site 1 only partially blocks channel conduction. The results suggest the presence of four K+ binding sites on the channel protein. One cytoplasmic facing site has an equilibrium affinity of 10 mM (site 6, delta = -0.3) and one vacuolar site (site 7, delta less than 0.2) has low affinity (greater than 500 mM). Divalent ion block of the Chara channel shows many similarities to that of the maxi-K channel from rat skeletal muscle.


The human erythrocyte anion transport protein, band 3. Characterization of exofacial alkaline titratable groups involved in anion binding/translocation. Chloride self-exchange across the human erythrocyte membrane at alkaline extracellular pH (pHO) and constant neutral intracellular pH (pH(i)) can be described by an exofacial deprotonatable reciprocating anion binding site model. The conversion of the transport system from the neutral to the alkaline state is related to deprotonation of a positively charged ionic strength- and substrate-sensitive group. In the absence of substrate ions ([ClO] = 0) the group has a pK of approximately 9.4 at constant high ionic strength (equivalent to approximately 150 mM KCl) and a pK of approximately 8.7 at approximately zero ionic strength. The alkaline ping-pong system (examined at constant high ionic strength) demonstrates outward recruitment of the binding sites with an asymmetry factor of approximately 0.2, as compared with the inward recruitment of the transport system at neutral pHO with an asymmetry factor of approximately 10. The intrinsic half-saturation constant for chloride binding, with [Cli] = [Clo], increased from approximately 30 mM at neutral to approximately 110 mM at alkaline pHO. The maximal transport rate was a factor of approximately 1.7 higher at alkaline pHO. This increase explains the stimulation of anion transport, the "modifier hump," observed at alkaline pHO. The translocation of anions at alkaline pHO was inhibited by deprotonation of another substrate-sensitive group with an intrinsic pK of approximately 11.3. This group together with the group with a pK of approximately 9.4 appear to form the essential part of the exofacial anion binding site. The effect of extracellular iodide inhibition on chloride transport as a function of pHO could, moreover, be simulated if three extracellular iodide binding constants were included in the model: namely, a competitive intrinsic iodide binding constant of approximately 1 mM in the neutral state, a self-inhibitor binding constant of approximately 120 mM in the neutral state, and a competitive intrinsic binding constant of approximately 38 mM in the alkaline state.


Lactic acid secretion by human neutrophils. Evidence for an H+ + lactate- cotransport system. The pathway by which L-lactate (Lac) crosses the plasma membrane of isolated human neutrophils was investigated. The influx of [14C]Lac from a 2 mM Lac, 145 mM Cl-, 5.6 mM glucose medium was approximately 1.5 meq/liter of cell water.min and was sensitive to the organomercurial agent mersalyl (apparent Ki approximately 20 microM), to alpha-cyano-4-hydroxycinnamate (CHC), the classical inhibitor of monocarboxylate transport in mitochondria, and to UK-5099 (apparent Ki approximately 40 microM), a more potent analogue of CHC. Transport was also strongly blocked (greater than 80%) by 1 mM of either 3,5-diiodosalicylic acid, MK-473 (an indanyloxyacetate derivative), or diphenyl-amine-2-carboxylate, and by 0.4 mM pentachlorophenol, but not by 1 mM ethacrynic acid, furosemide, or the disulfonic stilbenes SITS or H2DIDS. One-way [14C]Lac efflux from steady-state cells amounted to approximately 6 meq/liter.min and was likewise affected by the agents listed above. Influx, which was membrane potential insensitive and Na+ independent, displayed a strong pH dependence: extracellular acidification enhanced uptake while alkalinization inhibited the process (pK' approximately 5.7 at 2 mM external Lac). The rate of [14C]Lac influx was a saturable function of external Lac, the Km being approximately 7 mM. Steady-state cells exhibited an intracellular Lac content of approximately 5 mM and secreted lactic acid into the bathing medium a a rate of approximately 4 meq/liter.min. Secretion was completely suppressed by 1 mM mersalyl which inactivates the carrier, leading to an internal accumulation of Lac. That the Lac carrier truly mediates an H+ + Lac- cotransport (or formally equivalent Lac-/OH- exchange) was documented by pH-stat techniques wherein an alkalinization of poorly buffered medium could be detected upon the addition of Lac; these pH changes were sensitive to mersalyl. Thus, the Lac carrier of neutrophils possesses several features in common with other monocarboxylate transport systems in erythrocytes and epithelia.


Catabolism in mollicutes.

Catabolism in mollicutes. The small genome size of mollicutes, and particularly mycoplasmas and ureaplasmas, precludes their possession of the extensive range of metabolic activities present in most other bacterial groups. Demonstrated catabolic activities appear primarily to be associated with energy generation, rather than the provision of substrates for synthetic pathways, and anabolism is largely dependent upon extracellular sources of amino acids, nucleic acid precursors and lipids. However, the pathways of energy generation in mollicutes are diverse and specialized, and may in vivo be dependent upon the presence of a single amino acid (arginine) or urea. Even in those species that utilize carbohydrates the range of substrates is restricted, and while Ac. laidlawii has both EMP and PP pathways and is able to oxidize pyruvate to acetate plus CO2, many mycoplasmas possess only a part of these activities. Such specialization and the infrequent demonstration of inducible enzyme activity in mollicutes implies adaptation to specific habitats in host species, and suggests that differences in the catabolic activities of mollicute strains may be significant in terms of their ecology and pathogenicity. The demonstrated energy-generating pathways of mollicutes produce low ATP yields. Thus, mollicute growth will generate relatively large quantities of metabolic end-products and may deplete host tissues of substrates. Arginine depletion may be of particular importance in pathogenesis and the close physical association between mollicutes and host cells will enhance the potential significance of NH4+ production from the hydrolysis of arginine and urea, and of H2O2 and superoxide formation during carbohydrate metabolism. In addition, lipid and protein catabolism may be associated with virulence where extracellular or membrane-bound enzyme activities exist. Membrane-bound DNAase and RNAase activities have also been demonstrated in mycoplasmas and Ac. laidlawii (Pollack et al., 1965) and U. urealyticum (Romano & La Licata, 1978). Many aspects of mollicute catabolism, including energy conservation in some groups, is poorly understood. Also, while substantial catabolic diversity has been demonstrated within mollicutes and new species are continually being isolated, metabolism has been studied in relatively few species, and even in these only single strains or small groups of strains have been used. In this review, therefore, an attempt to avoid generalizations concerning mollicute behaviour has been made. The lack of much basic knowledge concerning mollicute metabolism has also necessitated the widespread use of 'may be' and other equally vague terms.(ABSTRACT TRUNCATED AT 400 WORDS)


Cell-bound peptidase activities of Treponema denticola ATCC 33520 in continuous culture. The oral spirochaete Treponema denticola ATCC 33520 was grown at a mean generation time of 10 h in anaerobic continuous culture in a serum- and carbohydrate-free medium at pH 7.0. The extracellular proteolytic activities of this spirochaete were then investigated by incubating washed cells with 68 2-naphthylamide derivatives of the Extended API System. Chymotrypsin-like, trypsin-like, elastase-like and iminopeptidase activities were demonstrated. The phenylalanine peptidase or chymotrypsin-like activity of T. denticola ATCC 33520, estimated with N-succinyl-L-phenylalanyl-L-leucyl-L-phenylalanine-thiobenzyl ester (SPLP) had a pH optimum at pH 8.5, a specific activity of 36.6 nmol min-1 (mg dry wt)-1 and was inhibited only slightly by HgCl2. The trypsin-like activity, estimated with benzoyl-DL-arginine-7-amido-4-methylcoumarin (BAMC), had a pH optimum at pH9, and a specific activity of 0.3 nmol min-1 (mg dry wt)-1; inhibition by HgCl2 indicated the involvement of active thiol groups. The activity should preferably be termed arginine peptidase activity, according to the carboxy-terminal amino acid of the test substrate. The extracellular proline peptidase activity, estimated with L-proline-7-amido-4-methylcoumarin. HBr (PRAMC), had an activity of 1.5 nmol min-1 (mg dry wt)-1, an optimum at pH 8.5 and the properties of a thiol protease. The main cell-bound and extracellular active peptidase activities of fast-growing cells of T. denticola ATCC 33520 are phenylalanine peptidase, proline peptidase, arginine peptidase and an oligopeptide-dependent alanine peptidase activity.(ABSTRACT TRUNCATED AT 250 WORDS)


Cloning and physical mapping of the Staphylococcus aureus rplL, rpoB and rpoC genes, encoding ribosomal protein L7/L12 and RNA polymerase subunits beta and beta'. Using segments of the Escherichia coli rpoB and rpoC genes as heterologous probes, we have identified and cloned an 8.3 kb PstI fragment from the Staphylococcus aureus genome containing the rpoB and rpoC genes, which respectively encode the beta and beta' subunits of DNA-directed RNA polymerase. This region is almost certainly equivalent to the rif locus, located near to fus at interval 12/13 on the S. aureus linkage map. Limited DNA sequencing revealed the gene order rpoB-rpoC (transcribed from left to right) and identified the rplL gene, encoding ribosomal protein L7/L12, upstream of rpoB. This and other evidence suggests that the rpoBC genes of S. aureus form part of a large gene cluster encoding components of the transcription and translation apparatus which is well-conserved in other eubacteria.


Micromonospora RNA polymerase activity changes during stationary phase. RNA polymerase was isolated from Micromonospora echinospora and from Streptomyces lividans. In vitro transcription of a DNA fragment containing multiple tandem promoters from Micromonospora followed the pattern of expression observed previously for in vivo studies. RNA polymerase was prepared from cultures of Micromonospora that were harvested during the growing phase and during the stationary phase. Promoters that were utilized in Micromonospora only during the stationary phase were utilized in vitro only when RNA polymerase was purified from a stationary-phase culture, and not when RNA polymerase was purified from growing cells.


Isolation and characterization of a repeated sequence (RPS1) of Candida albicans. A repeated sequence, named RPS1, approximately 2 kb in size, is found mainly in chromosome 6, the second most variable chromosome among the eight chromosomes of Candida albicans. Most of the RPS1 units of chromosome 6 seem to be located within a single region of about 100 kb in strain FC18. In both strains FC18 and NUM812, a part of RPS1 is apparently tandemly repeated. A unit of RPS1 has been cloned and sequenced. It consists of 2114 bp and has a GC content of 40 mol%. The repeat unit contains smaller repeats of about 80-170 bp which are called REP1, REP2, REP3, REP4 and REP5; REP2 is duplicated. The small repeats are classified into two groups by their homology. One comprises REP1, REP2 and REP5, and the other REP3 and REP4. They are termed the REP1 and REP3 families, respectively. The two families both contain a common 29 bp sequence, called COM29. The dispersed repetitive sequence RPS1 may be involved in chromosomal rearrangements and may in part explain chromosome polymorphism in C. albicans. The origin of RPS1 was not determined.


Reduced azole susceptibility of oral isolates of Candida albicans from HIV-positive patients and a derivative exhibiting colony morphology variation. Approximately 50% (15/28) of a selection of oral isolates of Candida albicans from separate individuals infected with the human immunodeficiency virus (HIV) exhibited low susceptibility to ketoconazole as determined by hyphal elongation assessment. Nine of these isolates exhibited colony morphology variation or switching at 37 degrees C, of which six expressed low ketoconazole susceptibility. To determine whether colony morphology variation could give rise to derivatives with reduced azole susceptibility, several high-frequency switching variants of three HIV-patient isolates were recovered and assessed. All but one of the variants expressed similar azole susceptibility profiles to their respective parental strains. However, the C. albicans derivative 132ACR expressed significantly reduced susceptibility to ketoconazole in comparison to its parental strain 132A. In whole cells, on the basis of total growth the switched derivative 132ACR was markedly less susceptible than its parental isolate 132A to ketoconazole at 10 microM. A much smaller difference was observed with fluconazole at 10 microM, with the switched derivative 132ACR exhibiting a threefold lower susceptibility compared with the parental isolate 132A. The incorporation of [14C]acetate in control and azole-treated cells of both organisms was higher for the parental strain. When cell lysates of strain 132A and its derivative 132ACR were incubated with [14C]mevalonic acid and ketoconazole, the IC50 for 14C-label incorporation into C-4 demethyl sterols was fivefold higher for lysates of the switched derivative 132ACR compared with those of the parental strain 132A. With fluconazole the IC50 value for the derivative 132ACR was 25-fold higher than for strain 132A. The 14-sterol demethylase of the switched derivative 132ACR was possibly less sensitive to azole inhibition than that of the enzyme of strain 132A. These studies indicated that colony morphology variation in vitro can generate derivatives with stable, reduced azole susceptibility without prior exposure to azoles.


Biological activities and chemical composition of a cytotoxin of Klebsiella oxytoca. A low-molecular-mass cytotoxin produced by Klebsiella oxytoca isolated previously from patients with antibiotic-associated haemorrhagic enterocolitis was purified, and its biological and chemical properties were elucidated. The toxin inhibited the syntheses of DNA and RNA by HEp-2 cells dose-dependently, whereas protein synthesis was only slightly inhibited, as measured by the incorporation of radioactive precursors. When synchronously cultured HEp-2 cells were examined in the presence of cytotoxin, inhibition of DNA synthesis occurred promptly within 5 h, but cell-rounding, the earliest visible morphological change, was not observed until 6 h after exposure. The intracellular levels of ATP decreased with an approximately similar time course. These results suggest that cytotoxicity toward HEp-2 cells is primarily due to the inhibitory effect of the cytotoxin on nucleic acid synthesis, possibly on DNA synthesis. Cell rounding and cell death were induced even in the absence of the cytotoxin after incubation with the cytotoxin for 6 h. The cytotoxin was heat-labile, cytotoxic activity decreasing to 50% of the initial level on heating at 70 degrees C for 20 min. Plasmids were extracted from three strains of K. oxytoca producing the cytotoxin and analysed by agarose gel electrophoresis. Two strains possessed plasmids of different sizes, but one strain possessed no plasmid, indicating that the cytotoxin is probably chromosomally encoded. Analysis by NMR and FAB-mass-spectrometry revealed that the molecular mass of the cytotoxin should be 217.1062 Da (exact mass), its molecular formula being C8H15O4N3.


The effect of AMP on the NAD-dependent glutamate dehydrogenase during activation and morphogenesis in the cellular slime moulds. In extracts from vegetative Dictyostelium discoideum V12 the basal NAD-dependent glutamate dehydrogenase (NAD-GDH) activity was low, but it increased on standing at 4 degrees C. When 0.1 mM-AMP was included in the assay mix, enzyme activity was stimulated nearly 30-fold. As the extract was allowed to age, the enzyme rapidly lost its ability to be stimulated by AMP. The response of NAD-GDH to AMP was also dependent on the stage of morphogenesis. The ratios of NAD-GDH activity assayed with and without AMP (+AMP/-AMP ratios) in freshly prepared extracts from cells at 0, 4, 8 and 12 h of development were similar, but declined later in morphogenesis. The +AMP/-AMP ratio decreased sharply during activation at 4 degrees C in extracts from cells at 0, 4, 16 and 20 h of development. By contrast, extracts from cells starved for 8 and 12 h remained more responsive to AMP throughout activation. Analysis of Western blots showed that vegetative NAD-GDH did not undergo any detectable proteolytic cleavage during 96 h of activation at 4 degrees C. Also, no change in molecular mass appeared to take place within the cells until culmination (20-24 h), when some breakdown products appeared. Activation of NAD-GDH also occurred in D. discoideum strains NC4 and AX3, and in D. mucoroides. In addition, the enzyme from these four strains was stimulated by AMP and the +AMP/-AMP ratio declined with similar kinetics during activation. The enzyme from Polysphondylium violaceum was not activated on standing, but it was stimulated by AMP. The effect of activation of NAD-GDH is discussed in relation to a postulated catabolic role for this enzyme.


Long-chain alcohol and aldehyde dehydrogenase activities in Acinetobacter calcoaceticus strain HO1-N. Three alcohol dehydrogenases have been identified in Acinetobacter calcoaceticus sp. strain HO1-N: an NAD(+)-dependent enzyme and two NADP(+)-dependent enzymes. One of the NADP(+)-dependent alcohol dehydrogenases was partially purified and was specific for long-chain substrates. With tetradecanol as substrate an apparent Km value of 5.2 microM was calculated. This enzyme has a pI of 4.5 and a molecular mass of 144 kDa. All three alcohol dehydrogenases were constitutively expressed. Three aldehyde dehydrogenases were also identified: an NAD(+)-dependent enzyme, an NADP(+)-dependent enzyme and one which was nucleotide independent. The NAD(+)-dependent enzyme represented only 2% of the total activity and was not studied further. The NADP(+)-dependent enzyme was strongly induced by growth of cells on alkanes and was associated with hydrocarbon vesicles. With tetradecanal as substrate an apparent Km value of 0.2 microM was calculated. The nucleotide-independent aldehyde dehydrogenase could use either Würster's Blue or phenazine methosulphate (PMS) as an artificial electron acceptor. This enzyme represents approximately 80% of the total long-chain aldehyde oxidizing activity within the cell when the enzymes were induced by growing the cells on hexadecane. It is particulate but can be solubilized using Triton X-100. The enzyme has an apparent Km of 0.36 mM for decanal.


Purification and amino acid sequence of a bacteriocin produced by Pediococcus acidilactici. A bacteriocin produced by Pediococcus acidilactici has been purified to homogeneity by a rapid and simple four-step purification procedure which includes ammonium sulphate precipitation, chromatography with a cation-exchanger and Octyl Sepharose, and reverse-phase chromatography. The purification resulted in an approximately 80,000-fold increase in the specific activity and about a 6-fold increase in the total activity. The amino acid composition and sequencing data indicated that the bacteriocin contained 43-44 amino acid residues. The predicted M(r) and isolectric point of the bacteriocin are about 4600 and 8.6, respectively. Comparing the amino acid sequence of this bacteriocin with the sequences of leucocin A-UAL 187, sakacin P and curvacin A (bacteriocins produced by Leuconostoc gelidum, Lactobacillus sake and Lactobacillus curvatus, respectively) revealed that all four bacteriocins had in their N-terminal region the sequence Tyr-Gly-Asn-Gly-Val-Xaa-Cys, indicating that this concensus sequence is of fundamental importance for this group of bacteriocins. The bacteriocin from P. acidilactici and sakacin P were very similar, having at least 25 common amino acid residues. The sequence similarity was greatest in the N-terminal half of the molecules--17 of the first 19 residues were common--indicating the fundamental importance of this region. Leucocin A-UAL 187 and curvacin A had, respectively, at least 16 and 13 amino acid residues in common with the bacteriocin from P. acidilactici.


Defective interfering L RNA segments of tomato spotted wilt virus retain both virus genome termini and have extensive internal deletions. Defective interfering (DI) RNA molecules derived from the genomic L RNA segment of tomato spotted wilt virus (TSWV) were generated during sequential passage of the virus at high multiplicity. Characterization of DI RNAs from four distinct isolates by Northern blot analysis and sequence determination revealed that both the 5' and 3' genomic termini were retained in these molecules. Each DI RNA contained a single internal deletion of approximately 60% to 80% of the L RNA segment. All DI RNAs studied maintain an open reading frame (ORF) which suggests that these defective molecules should be translatable by ribosomes. Detection of only defective molecules with ORFs indicates either that association with ribosomes or translation is a prerequisite for the selection and maintenance of replicating DI RNAs, or that the truncated proteins produced play a role in their selection or replication. Analysis of the junction sites in the DI RNAs showed that short nucleotide sequences are repeated, one at the release and another at the reinitiation point on the L RNA. One of these is lost during the generation of the DI molecules. The presence of repeated sequences at the junction sites seems to be unique for tospovirus DI L RNAs; they have not been described for other DI systems of either positive- or negative-strand RNA viruses. A model for TSWV DI RNA generation is proposed in which the viral polymerase can 'jump' across the internal sequences from one secondary structure to another containing the repeated sequences, during the replication of the viral complementary L RNA segment.


Cross-reacting and heterospecific monoclonal antibodies produced against arabis mosaic nepovirus. Monoclonal antibodies (MAbs) were produced against arabis mosaic nepovirus (AMV). A hybridoma screening procedure was applied which involved the testing of culture supernatants, before the hybridomas were cloned to single cell lines, for their reaction with eight nepoviruses [AMV, cherry leafroll virus (CLRV), grapevine fanleaf virus (GFLV), peach rosette mosaic virus, raspberry ringspot virus (RRSV), tobacco ringspot virus, tomato black ring virus (TBRV) and tomato ringspot virus]. In addition to AMV-specific MAbs, this screening technique has allowed the selection of two cross-reacting MAbs: one reacting with AMV and GFLV, and one reacting with AMV and RRSV. This is the first report of MAbs cross-reacting with these nepoviruses. In addition, five heterospecific MAbs (HS-MAbs) could be selected: two reacting with RRSV, two with CLRV and one with TBRV. The usefulness of the screening technique that was applied for the selection of cross-reacting MAbs and HS-MAbs, and the potential use of such antibodies are discussed.