Monkeypox virus detection in rodents using real-time 3'-minor groove binder TaqMan assays on the Roche LightCycler

During the summer of 2003, an outbreak of human monkeypox occurred in the Midwest region of the United States. In all, 52 rodents suspected of being infected with monkeypox virus were collected from an exotic pet dealer and from private homes. The rodents were euthanized and submitted for testing to the United States Army Medical Research Institute of Infectious Diseases by the Galesburg Animal Disease Laboratory, Illinois Department of Agriculture. The rodent tissue samples were appropriately processed and then tested by using an integrated approach involving real-time polymerase chain reaction (PCR) assays, an antigen-detection immunoassay, and virus culture. We designed and extensively tested two specific real-time PCR assays for rapidly detecting monkeypox virus DNA using the Vaccinia virus F3L and N3R genes as targets. The assays were validated against panels of orthopox viral and miscellaneous bacterial DNAs. A pan-orthopox electrochemiluminescence (ECL) assay was used to further confirm the presence of Orthopoxvirus infection of the rodents. Seven of 12 (58%) animals (seven of 52 (15%) of all animals) tested positive in both monkeypox-specific PCR assays and two additional pan-orthopox PCR assays (in at least one tissue). The ECL results showed varying degrees of agreement with PCR. One hamster and three gerbils were positive by both PCR and ECL for all tissues tested. In addition, we attempted to verify the presence of monkeypox virus by culture on multiple cell lines, by immunohistology, and by electron microscopy, with negative results. Sequencing the PCR products from the samples indicated 100% identity with monkeypox virus strain Zaire-96-I-16 (a human isolate from the Congo). These real-time PCR and ECL assays represent a significant addition to the battery of tests for the detection of various orthopoxviruses. In light of the recent monkeypox virus transmissions, early detection of the virus is crucial for both natural outbreaks and potential acts of bioterrorism.     

Family and Illness Predictors of Outcome in Pediatric Brain Tumors

Investigated the prediction of cognitive and behavioral outcomesin 63 children with heterogenous brain tumors. Hierarchicalmultiple regression analyses were used to determine how family-relatedvariables added to the prediction of children's outcome overand above illness measures. The best predictors of children'sbehavior problems and adaptive behavior were family and demographicvariables, whereas the best predictors of achievement were illnessand demographic variables. A combination of family and illnessvariables, however, was the best predictor of intellectual functioning.In addition to identifying specific predictors of cognitiveand behavioral outcome in children with brain tumors, theseresults lend initial support for the inclusion of contextualfactors such as family stress, maternal coping, number of parentsin the home, and family SES measures in studies of how diseasefactors affect outcomes in pediatric brain tumor patients.     

Changing contexts? The effects of a primary prevention program on classroom levels of peer relational and physical victimization

Whereas school‐based prevention programs often target deficits in individual children's social skills in order to limit their aggression or exposure to peer victimization, there is increasing evidence that school‐wide and classroom‐level factors can also affect the success of these programs. This short‐term longitudinal study involved 432 elementary school students from 44 classrooms in 17 urban schools. We investigated whether classroom characteristics (average levels of social competence, emotional problems, and behavioral problems) and school‐wide characteristics (proportion of children on income assistance and implementation of a peer victimization prevention program—the Walk away, Ignore, Talk, and Seek help [W.I.T.S.] program) experienced in Grade 1 influences changes in children's reports of relational and physical victimization at the end of Grade 2. Findings showed that classroom levels of emotional problems predicted increases in relational victimization (beyond individual differences in emotional and behavioral problems). Classroom levels of behavioral problems predicted reports of increases in physical victimization (beyond individual differences). Classroom levels of social competence also interacted with individual levels of emotional problems such that children with higher levels of emotional problems in classes with more socially competent children reported more relational and physical victimization. Higher school levels of poverty and lack of program involvement also predicted higher levels of physical victimization, beyond individual and classroom effects. The capacity of the W.I.T.S. program to influence classroom level characteristics and the moderating effects of school poverty on victimization were also assessed. © 2003 Wiley Periodicals, Inc. J Comm Psychol 31: 397–418, 2003.     

Measurement Issues in Emotion Research With Children and Adolescents

The need for a multimethod approach to the study of emotion in children and adolescents is crucial, as is a multilevel analysis in which emotional experience is assessed at a variety of levels (Solomon, 2002). This review highlights the critical role of emotion theory when constructing and selecting appropriate assessment tools with a focus on Functionalist theory that emphasizes the importance of contextual variables ( Barrett & Campos, 1987 ). The review begins with an examination of theoretical and pragmatic issues in emotion measurement followed by discussions of four basic methods of emotion assessment (i.e., self-report, other-report, observation, neurophysiology). Implications of emotion assessment for clinical practice and future directions for research conclude the review.     

Menstrual Cycle Phase is a Potential Confound in Psychophysiology Research

Effects of menstrual cycle phase on physiological levels and reactivity were studied in a withinsubject design with 20 women aged 18–30 years. Basal body temperature was monitored daily; follicular and luteal recording sessions occurred before and after the ovulatory temperature rise, respectively. The design consisted of task and rest periods, presented in counterbalanced order across phases. Half the subjects began during their follicular phase and half began during their luteal phase. Follicular and luteal phases were compared for differences between baseline resting levels, as well as for reactivity. The results showed menstrual phase differences in heart rate, pulse transit time, and EEG alpha, particularly during tasks, but no difference in reactivity. Habituation effects in the group that began during the follicular phase often masked luteal activation. In spite of the fact that not all statistical results achieved significance, the consistency of most trends supports the conclusion that physiological recording sessions with women might best be restricted to the follicular phase.     

Identified higher-order neurons controlling the feeding motor program of helisoma

A bilaterally symmetrical pair of neurons in the anterior region of the cerebral ganglia of the snail Helisoma trivolvis were found to have excitatory input to the feeding motor program contained in the buccal ganglia. Intracellular microelectrodes were used to stimulate and record from these cells while the motor output of the buccal ganglia was monitored with a combination of intracellular and extracellular recordings. Experimentally evoked tonic activity in an individual cerebral cell could initiate and maintain the patterned motor output from buccal ganglia, characteristic of the activity underlying buccal mass feeding movements. The rate of buccal motor output could be modified directly by varying the firing frequency of the cerebral cell. Cobaltous chloride backfills of cerebrobuccal connectives revealed that these higher-order neurons were the only large cells in the anterior portion of the ganglia to send processes into the connectives. Furthermore, they are the only cells in this region to fluoresce when processed with a sucrose-phosphate-glyoxylic acid solution, indicating that they contain an indoleamine, probably serotonin. Application of low concentrations of serotonin to isolated buccal ganglia or buccal ganglia-buccal mass preparations mimics the effects of the cerebral cells' activity on the buccal motor output, implying that serotonin is a putative transmitter for these cerebral cells.     

Infant Vocalization: An Index of Maternal Attitude Toward the Child

Brief observations of mother-stimulated infant vocalizationwere perform ed during pediatric clinic visits on 4- to 6-month-oldinfants to determine whether mothers with positive attitudestoward the pregnancy and/or the in fant would have infants whovocalized more during clinic visits. Mothers with more positiveattitudes toward the pregnancy, assessed prenatally, had infantswho showed significantly more vocalization during clinic visitsat 5 months, 6 months, and the combination of 5 and 6 months.Mothers with more positive attitudes toward the infant had infantswho showed signcantly more vocalization at 4 months and thecombination of observations at 4 months and 6 months. Resultssuggest that brief observations of vocalization during clinicvisits may be useful as a rough index of the quality of themother-child interaction.     

Haemophilus ducreyi Outer membrane determinants, including DsrA, define two clonal populations

The Haemophilus ducreyi outer membrane component DsrA (for ducreyi serum resistance A) is necessary for complete resistance to normal human serum (NHS). When DsrA expression in 19 temporally and geographically diverse clinical isolates of H. ducreyi was examined by Western blotting, 5 of the strains expressed a different immunotype of the DsrA protein (DsrA(II)) than the well-characterized prototypical strain 35000HP (DsrA(I)). The predicted DsrA proteins expressed by the DsrA(II) strains were 100% identical to each other but only 48% identical to that of strain 35000HP. In addition to the DsrA(II) protein, class II strains also expressed variant forms of other outer membrane proteins (OMPs) including NcaA (necessary for collagen adhesion A), DltA (ducreyi lectin A), Hlp (H. ducreyi lipoprotein), major OMP, and/or OmpA2 (for OMP A2) and synthesized a distinct, faster-migrating lipooligosaccharide. Based on these data, strains expressing DsrA(I) were termed class I, and those expressing DsrA(II) were termed class II. Expression of dsrA(II) from strain CIP 542 ATCC in the class I dsrA(I) mutant FX517 (35000HP background), which does not express a DsrA protein, rendered this strain resistant to 50% NHS. This demonstrates that DsrA(II) protein is also critical to serum resistance. Taken together, these results indicate that there are two clonal populations of H. ducreyi. The implications of two classes of H. ducreyi strains differing in important antigenic outer membrane components are discussed.     

Enhancement of normal neutrophil chemiluminescence by chronic granulomatous disease neutrophils

Neutrophils and other phagocytic cells from patients with chronic granulomatous disease (CGD) lack the ability to generate reactive oxygen intermediates (ROI), although other phagocytic functions appear to be intact. The effects of CGD neutrophils on the ability of normal neutrophils to produce ROI as measured by luminol-enhanced chemiluminescence (CL) were examined. Normal neutrophils (2×10⁵) had a peak CL response to phorbol myristate acetate (PMA; 20 ng/ml) of 6.5±0.9 mV, while the CL response of CGD neutrophils was completely absent. However, the addition of CGD neutrophils (8×10⁵) to normal neutrophils (2×10⁵) markedly increased the peak CL response to PMA to 11.0+-1.1 mV (P < 0.001). The peak response of normal neutrophils (2×10⁵) alone to the peptideN-formyl-methionyl-leucyl-phenylalanine (FMLP; 10^–6 M) was 9.0±1.1mV, and this increased to 22.2±3.5 mV (P < 0.001) when 8×10⁵ CGD neutrophils were added and to 18.9±3.6 mV (P<0.005) when 4×10⁵ CGD neutrophils were added. Thus, CGD neutrophils increase the release of ROI from normal cells, suggesting nonoxidative regulatory factors in ROI production.This work was supported in part by grants GI517-02 and 0008-08 from the Cystic Fibrosis Foundation and HD-09800 and AI-15332 from the National Institutes of Health.     

Perceived personal immunity to the consequences of drinking alcohol: The relationship between behavior and perception

Heavy drinkers, moderate drinkers, light drinkers, and nondrinkers were asked to rate a variety of negative health and social consequences of using alcohol. Subjects made probability ratings for fictional others who were heavy, moderate, or light drinkers or nondrinkers. Subjects also made probability ratings for themselves as hypothetical heavy, moderate, or light drinkers or nondrinkers and for themselves actually. A pattern of perceived personal immunity was found across groups. Subjects rated fictional others and themselves as hypothetical drinkers to be more likely to experience negative consequences than their actual selves. All groups of subjects (heavy, moderate, and light drinkers and abstainers) rated their actual chances of experiencing negative consequences to be approximately equal. In contrast, heavy drinkers saw the effects of drinking for other heavy drinkers as less likely than did subjects who had light or abstinent drinking patterns who rated fictional heavy drinkers. These findings suggest that individuals who drink more tend to deny the potential harm that may result from alcohol consumption. Short-term social consequences were viewed as most likely to occur. Long-term consequences were perceived as least likely to occur.This research was supported in part by grant 1-R01-AA06201 from the National Institute on Alcohol Abuse and Alcoholism.