In situ observations show vertical community structure of pelagic fauna in the eastern tropical North Atlantic off Cape Verde.

Distribution patterns of fragile gelatinous fauna in the open ocean remain scarcely documented. Using epi-and mesopelagic video transects in the eastern tropical North Atlantic, which features a mild but intensifying midwater oxygen minimum zone (OMZ), we established one of the first regional observations of diversity and abundance of large gelatinous zooplankton. We quantified the day and night vertical distribution of 46 taxa in relation to environmental conditions. While distribution may be driven by multiple factors, abundance peaks of individual taxa were observed in the OMZ core, both above and below the OMZ, only above, or only below the OMZ whereas some taxa did not have an obvious distribution pattern. In the eastern eropical North Atlantic, OMZ expansion in the course of global climate change may detrimentally impact taxa that avoid low oxygen concentrations (Beroe, doliolids), but favour taxa that occur in the OMZ (Lilyopsis, phaeodarians, Cydippida, Colobonema, Haliscera conica and Halitrephes) as their habitat volume might increase. While future efforts need to focus on physiology and taxonomy of pelagic fauna in the study region, our study presents biodiversity and distribution data for the regional epi- and mesopelagic zones of Cape Verde providing a regional baseline to monitor how climate change may impact the largest habitat on the planet, the deep pelagic realm.

Bathyal feasting: post-spawning squid as a source of carbon for deep-sea benthic communities.

In many oceanic carbon budgets there is a discrepancy between the energetic requirements of deep-sea benthic communities and the supply of organic matter. This suggests that there are unidentified and unmeasured food sources reaching the seafloor. During 11 deep-sea remotely operated vehicle (ROV) surveys in the Gulf of California, the remains (squid carcasses and hatched-out egg sheets) of 64 post-brooding squid were encountered. As many as 36 remains were encountered during a single dive. To our knowledge this is one of the largest numbers of natural food falls of medium-size deep-sea nekton described to date. Various deep-sea scavengers (Ophiuroidea, Holothuroidea, Decapoda, Asteroidea, Enteropneusta) were associated with the remains. Although many of the 80 examined ROV dives did not encounter dead squids or egg sheets (n = 69), and the phenomenon may be geographically and temporally restricted, our results show that dead, sinking squid transport carbon from the water column to the seafloor in the Gulf of California. Based on food fall observations from individual dives, we estimate that annual squid carcass depositions may regionally contribute from 0.05 to 12.07 mg C m-2 d-1 to the seafloor in the areas where we observed the remains. The sinking of squid carcasses may constitute a significant but underestimated carbon vector between the water column and the seafloor worldwide, because squid populations are enormous and are regionally expanding as a result of climate change and pressure on fish stocks. In the future, standardized methods and surveys in geographical regions that have large squid populations will be important for investigating the overall contribution of squid falls to regional carbon budgets.

Morphology, ecology, and molecular biology of a new species of giant larvacean in the eastern North Pacific: Bathochordaeus mcnutti sp. nov.

Bathochordaeus mcnutti sp. nov. is described from the mesopelagic northeast Pacific Ocean (Monterey Bay, California, USA). Larvaceans in the genus Bathochordaeus are large, often abundant zooplankters found throughout much of the world ocean, but until recently it was unclear whether more than a single species of Bathochordaeus existed. Using remotely operated vehicles, we have made hundreds of in situ observations, compiled two decades of time-series data, and carefully collected enough specimens to determine that three species of Bathochordaeus occur in Monterey Bay: B. charon (Chun), B. stygius (Garstang), and B. mcnutti sp. nov. Bathochordaeus mcnutti is readily distinguished from its two congeners by the distinct blue outline visible around the periphery of its tail, and by other aspects of its morphology, ecology, and genetics. The abundance of larvaceans means they are ecologically important as particle processors. Species within the genus, Bathochordaeus, comprise the largest of described larvaceans.

Distinct patterns of DeltaFosB induction in brain by drugs of abuse.

The transcription factor DeltaFosB accumulates and persists in brain in response to chronic stimulation. This accumulation after chronic exposure to drugs of abuse has been demonstrated previously by Western blot most dramatically in striatal regions, including dorsal striatum (caudate/putamen) and nucleus accumbens. In the present study, we used immunohistochemistry to define with greater anatomical precision the induction of DeltaFosB throughout the rodent brain after chronic drug treatment. We also extended previous research involving cocaine, morphine, and nicotine to two additional drugs of abuse, ethanol and Delta(9)-tetrahydrocannabinol (Delta(9)-THC, the active ingredient in marijuana). We show here that chronic, but not acute, administration of each of four drugs of abuse, cocaine, morphine, ethanol, and Delta(9)-THC, robustly induces DeltaFosB in nucleus accumbens, although different patterns in the core vs. shell subregions of this nucleus were apparent for the different drugs. The drugs also differed in their degree of DeltaFosB induction in dorsal striatum. In addition, all four drugs induced DeltaFosB in prefrontal cortex, with the greatest effects observed with cocaine and ethanol, and all of the drugs induced DeltaFosB to a small extent in amygdala. Furthermore, all drugs induced DeltaFosB in the hippocampus, and, with the exception of ethanol, most of this induction was seen in the dentate. Lower levels of DeltaFosB induction were seen in other brain areas in response to a particular drug treatment. These findings provide further evidence that induction of DeltaFosB in nucleus accumbens is a common action of virtually all drugs of abuse and that, beyond nucleus accumbens, each drug induces DeltaFosB in a region-specific manner in brain.

Strain-specific alteration of zebrafish feeding behavior in response to aversive stimuli.

Behavioral management of risk, in which organisms must balance the requirements of obtaining food resources with the risk of predation, has been of considerable interest to ethologists for many years. Although numerous experiments have shown that animals alter their foraging behavior depending on the levels of perceived risk and demand for nutrients, few have considered the role of genetic variation in the trade-off between these variables. We performed a study of four zebrafish (Danio rerio (Hamilton, 1822)) strains to test for genetic variation in foraging behavior and whether this variation affected their response to both aversive stimuli and nutrient restriction. Zebrafish strains differed significantly in their latency to begin foraging from the surface of the water under standard laboratory conditions. Fish fed sooner when nutrients were restricted, although this was only significant in the absence of aversive stimuli. Aversive stimuli caused fish to delay feeding in a strain-specific manner. Strains varied in food intake and specific growth rate, and feeding latency was significantly correlated with food intake. Our results indicate significant genetic variation in foraging behavior and the perception of risk in zebrafish, with a pattern of strain variation consistent with behavioral adaptation to captivity.

A consolidated linkage map for rainbow trout (Oncorhynchus mykiss).

Androgenetic doubled haploid progeny produced from a cross between the Oregon State University and Arlee clonal rainbow trout (Oncorhynchus mykiss) lines, used for a previous published rainbow trout map, were used to update the map with the addition of more amplified fragment length polymorphic (AFLP) markers, microsatellites, type I and allozyme markers. We have added more than 900 markers, bringing the total number to 1359 genetic markers and the sex phenotype including 799 EcoRI AFLPs, 174 PstI AFLPs, 226 microsatellites, 72 VNTR, 38 SINE markers, 29 known genes, 12 minisatellites, five RAPDs, and four allozymes. Thirty major linkage groups were identified. Synteny of linkage groups in our map with the outcrossed microsatellite map has been established for all except one linkage group in this doubled haploid cross. Putative homeologous relationships among linkage groups, resulting from the autotetraploid nature of the salmonid genome, have been revealed based on the placement of duplicated microsatellites and type I loci.

Can coelenterates make coelenterazine? Dietary requirement for luciferin in cnidarian bioluminescence.

In the calcium-activated photoprotein aequorin, light is produced by the oxidation of coelenterazine, the luciferin used by at least seven marine phyla. However, despite extensive research on photoproteins, there has been no evidence to indicate the origin of coelenterazine within the phylum Cnidaria. Here we report that the hydromedusa Aequorea victoria is unable to produce its own coelenterazine and is dependent on a dietary supply of this luciferin for bioluminescence. Although they contain functional apophotoproteins, medusae reared on a luciferin-free diet are unable to produce light unless provided with coelenterazine from an external source. This evidence regarding the origins of luciferin in Cnidaria has implications for the evolution of bioluminescence and for the extensive use of coelenterazine among marine organisms.

Ancient and recent duplications of the rainbow trout Wilms' tumor gene.

The Wilms' tumor suppressor (WT1) gene plays an important role in the development and functioning of the genitourinary system, and mutations in this gene are associated with nephroblastoma formation in humans. Rainbow trout (Oncorhynchus mykiss) is one of the rare animal models that readily form nephroblastomas, yet trout express three distinct WT1 genes, one of which is duplicated and inherited tetrasomically. Sequence analyses suggest an ancient gene duplication in the common ancestor of bony fishes resulted in the formation of two WT1 gene families, that conserve the splicing variations of tetrapod WT1, and a second duplication event occurred in the trout lineage. The WT1 genes of one family map to linkage groups 6 and 27 in the trout genome map. Reverse transcribed polymerase chain reaction (RT-PCR) expression analysis demonstrated little difference in W

Composite interval mapping reveals a major locus influencing embryonic development rate in rainbow trout (Oncorhynchus mykiss).

Little is known about the genetics controlling the rate of embryonic development in salmonids, despite the fact that this trait plays an important role in the life history of wild and cultured stocks. We investigated the genetics of embryonic development rate by performing an analysis of quantitative trait loci (QTL) on two families of androgenetically derived doubled haploid rainbow trout produced from a hybrid of two clonal lines with divergent embryonic development rates. A total of 170 doubled haploid individuals were genotyped at 222 marker loci [219 amplified fragment length polymorphism (AFLP) markers, 2 microsatellites, and p53]. A genetic linkage analysis resulted in a map consisting of 27 linkage groups with 21 of the markers remaining unlinked at a minimum LOD of 3.0 and maximum theta of 0.40. Eight of these linkage groups were matched to published rainbow trout linkage groups. Composite interval mapping (CIM) revealed evidence for two QTL influencing time to hatch, and suggestive evidence for a third. These QTL accounted for a total of 24.6% of the variation in time to hatch. One of these QTL had a large effect on development rate, especially in one family of doubled haploids, in which it explained 25.6% of the variance in time to hatch. QTL influencing embryonic length and weight at the commencement of exogenous feeding were also identified. The QTL with the strongest effect on embryonic length (lenR13) mapped to the same position as the QTL with the strongest effect on time to hatch (tthR13), suggesting a single QTL may have a pleiotropic effect on both these traits. These results suggest that the use of clonal lines with a doubled haploid crossing design is an effective way of analyzing the genetic basis of complex traits in salmonids.

Cloning, mapping and genomic organization of a fish C-type lectin gene from homozygous clones of rainbow trout (Oncorhynchus mykiss).

Utilizing a splenic cDNA library and rapid amplification of cDNA 5' ends (5'-RACE), a C-type lectin gene was cloned from a homozygous cloned rainbow trout. The 1176 bp cDNA contains a 714 bp open reading frame from which a 238-amino-acid (aa) (27 kDa) protein was deduced. It was confirmed that this protein belongs to the C-type animal lectins, and is a type II membrane receptor. The predicted protein from this sequence contains a 48 aa cytoplasmic domain, a 20 aa transmembrane domain (TM), a 46 aa stalk region and a 124 aa carbohydrate-recognition domain (CRD). The stalk region contains a leucine-zipper, and an N-glycosylation site was also found in the CRD. Sequence alignment and phylogenetic analysis of the CRD indicate that the protein has similarity with human dendritic cell immunoreceptor (DCIR), gp120 binding C-type lectin (gp120BCL) and mammalian hepatic lectins. The N-terminus (aa 4-183) has similarity with NKG2, a group of C-type lectin receptors important in human natural killer cell function. The genomic DNA (gDNA) containing this gene was amplified and sequenced. The 4569 bp gDNA contains five exons and four introns. The first three exons encode the cytoplasmic domain, the TM and stalk region, respectively. Unlike the other type II C-type lectin receptors in which the CRD was encoded by three exons, the CRD of this lectin was encoded by two exons. A transposon Tc1-like fragment was found in intron III. Intron IV is composed of a simple repeat. Tissue-specific expression of the gene was studied by RT-PCR, and it was mainly expressed in spleen and peripheral blood leukocyte (PBL). Using AluI to digest the fragment containing exon I, intron I and exon II, an RFLP was produced between the sequences of this gene in two cloned fish, OSU 142 and Arlee (AR). Seventy-one doubled haploids (DH) of OSU X AR were screened, and the gene was mapped to linkage group XIV on the published map (Young et al., Genetics 148 (1998) 839).

Splitting on a pediatric consult liaison service.

OBJECTIVE: Although the concepts of splitting and projective identification have been useful in explaining certain group phenomena on adult psychiatric and medical wards, their application to pediatric settings has not been addressed in the literature. The authors demonstrate that early identification, staff conferencing, and family/staff conferencing can diffuse these dynamics in an academic pediatric setting. METHOD: The existing literature on splitting and projective identification is reviewed. Case vignettes are then used to illustrate the manifestations of splitting and projective identification in a pediatric setting and to demonstrate intervention strategies modified for children and their families from the adult literature. RESULTS: Splitting and projective identification can be interrupted in pediatric settings with early identification, staff conferencing, and family/staff conferencing. The cooperation of pediatric clinicians is critical in the implementation of these intervention strategies. CONCLUSIONS: The development of liaison support groups for pediatric residents and interdisciplinary treatment teams will enlist their cooperation in identifying splitting early, and in employing staff conferencing and family/staff conferencing to diffuse this group dynamic which, if left unchecked, can disrupt professional relationships and compromise the treatment of pediatric patients.

Enzyme-linked immunoassay for detection of Listeria monocytogenes in dairy products, seafoods, and meats: collaborative study.

A collaborative study was conducted to evaluate Listeria-Tek, an enzyme-linked immunosorbent assay (ELISA) for detection of Listeria monocytogenes and other Listeria spp. in foods. The present ELISA method was compared to the U.S. Food and Drug Administration culture method for detection of L. monocytogenes in dairy products and seafoods and to the U.S. Department of Agriculture Food Safety and Inspection Service method for detection of L. monocytogenes in meats. Replicate samples of 6 food types (frankfurters, roast beef, Brie cheese, 2% milk, raw shrimp, and crab meat) inoculated with L. monocytogenes and uninoculated control samples were analyzed by the collaborators. L. monocytogenes was identified in 593 samples by the ELISA method and in 574 samples using culture procedures. Identical results were obtained for 506 positive samples and 419 negative samples using the ELISA and culture methods for an overall agreement rate of 85.6%. The enzyme-linked immunoassay for detection of L. monocytogenes in dairy, seafood, and meat products has been adopted first action by AOAC INTERNATIONAL.

Elevated-temperature, colorimetric, monoclonal, enzyme-linked immunosorbent assay for rapid screening of Salmonella in foods: collaborative study.

A collaborative study was performed by 30 laboratories in 3 sets of trials to validate a modified colorimetric monoclonal enzyme-linked immunosorbent assay (ELISA) method for Salmonella detection. The modifications to the current methodology included incubation of enrichments and post-enrichments at an elevated temperature, addition of novobiocin to the M-broth post-enrichment, and elimination of the centrifugation and agitation steps. Five artificially contaminated foods (nonfat dry milk, milk chocolate, dried egg, ground black pepper, and soy flour) and 1 naturally contaminated food (raw ground turkey) were analyzed. The artificially contaminated foods were inoculated with individual Salmonella serotypes at a high (10-50 cells/25 g) and low (1-5 cells/25 g) contamination level. Results from the modified ELISA method were compared to the Bacteriological Analytical Manual (BAM)/AOAC culture method. In 2 of the food products, milk chocolate and pepper, a number of laboratories isolated Salmonella from un-inoculated control samples, thus invalidating their data. As a result, there were too few laboratories remaining with valid data, and these foods were repeated. In the completed study, there were 11 false negative results obtained by the modified ELISA method, while there were 28 false negatives produced by the BAM/AOAC procedure. There were 11 ELISA positive assays which could not be confirmed by culture methods. Statistically, there were no differences between the modified, colorimetric, monoclonal ELISA and the reference culture method in all foods except raw turkey, where the ELISA method was more productive. The colorimetric monoclonal enzyme immunoassay (Salmonella-Tek) method for detecting Salmonella in all foods has been adopted first action by AOAC INTERNATIONAL.

Comorbidity of conduct disorder and personality disorders in an incarcerated juvenile population.

OBJECTIVE: Youths with conduct disorder extract an inordinate amount of time and money from the U.S. judicial system and taxpayers, yet studies pertaining to this population have been few. This study was undertaken to examine the co-occurrence of personality disorders and conduct disorder in a group of incarcerated children and adolescents and to raise the issue of the possibility of antisocial personality disorder in persons under the age of 18 years. METHOD: One hundred incarcerated juvenile offenders aged 11-17 years were randomly selected and then interviewed with the Diagnostic Interview for Children and Adolescents--Revised and the Structured Clinical Interview for DSM-III-R Personality Disorders to establish their psychiatric diagnoses. RESULTS: Eighty-seven percent of the group met the criteria for conduct disorder. Among those diagnosed as having conduct disorder, the only comorbid personality disorder that was present with significant frequency was antisocial personality disorder. The other comorbid personality disorder diagnoses that appeared most frequently were the borderline, narcissistic, paranoid, passive-aggressive, and dependent types. Borderline personality disorder was observed more frequently in the females than in the males with conduct disorder. CONCLUSIONS: The findings suggest that by using DSM-III-R criteria for adult personality disorders, one finds a considerable number of personality disorders in a young population with conduct disorder. The findings also show that youths manifest signs of antisocial personality disorder before they are 18 years of age, raising the question of how age should be incorporated into the diagnosis of personality disorder as DSM-IV is being prepared.

Comparison of MICRO-ID Listeria method with conventional biochemical methods for identification of Listeria isolated from food and environmental samples: collaborative study.

Fourteen laboratories participated in a collaborative study to evaluate the ability of the MICRO-ID Listeria identification method to correctly identify Listeria isolated from food and environmental sources. Each collaborator received 60 isolates consisting of 51 Listeria and 9 non-Listeria cultures. All isolates were identified by conventional biochemical analyses in the principal laboratory. Cultures were checked for purity by Gram staining and examined for oxidase and catalase activities. Only Gram positive, oxidase negative, catalase positive cultures were tested with the method. Colonies from trypticase soy agar with 0.6% yeast extract were suspended in 4.6 mL physiological saline to a MacFarland No. 1 turbidity standard and used to inoculate the test strip. In addition, the hemolytic reaction of each isolate was determined by using the Christie-Atkins-Munch-Peterson (CAMP) test and by stabbing sheep blood agar. Identification of Listeria is based on the octal code obtained from the strip and the hemolytic reaction of the isolate. The MICRO-ID Listeria method agreed with conventinal biochemical identification for 98.0% of L. monocytogenes, 77.1% of L. seeligeri, 90.0% of L. ivanovii, 96.4% of L. grayi/L. murrayi, 73.9% of L. welshimeri, and 100% of L. innocua isolates. A large percentage of errors in identification of the L. seeligeri and L. ivanovii cultures was caused by inaccurate reading of the CAMP and hemolysis tests rather than errors in the test strip. The method was adopted first action by AOAC International.

Use of an Elevated Temperature and Novobiocin in a Modified Enzyme-linked Immunosorbant Assay for the Improved Recovery of Salmonella from Foods.

A modified colorimetric enzyme-linked immunosorbant assay (ELISA) for Salmonella detection was compared to the standard culture method of the Bacteriological Analytical Manual/Association of Official Analytical Chemists (BAM/AOAC) using 20 artificially contaminated foods (1,200 test samples). The modifications to the current methodology consisted of an elevated incubation temperature of 42°C for the tetrathionate selective broth and M-broth postenrichments, as well as addition of 10 µg/ml novobiocin to the M-broth. The microtiter plate as not agitated during assay incubation, and centrifugation steps were eliminated from the protocol. This modified ELISA method was at least as productive as the standard AOAC culture method for the food samples tested. No false-positive reactions were encountered. The false-negative incidence was 1.5% for the immunoassay and 5.3% by the AOAC cultural method. The incidence of agreement between the methods was 96.7%.

Clinician-directed hospital cost management for total hip arthroplasty patients.

Can patients treated with total hip arthroplasty (THA) receive high-quality inpatient care at less cost? In 1984, a group of orthopedic surgeons and nurses examined the use of resources for THA patients and changed certain clinical practices to promote more cost-effective hospital care. At the end of the two-year project, orders for complete blood counts fell 72% and mean operating room time dropped 47 minutes for the participating orthopedists. For all orthopedists in the division, average length of stay (ALOS) decreased from 13 to 11 days. By the end of the following year, when clinicians received quarterly length-of-stay (LOS) data, ALOS dropped further to 9.8 days. This significant ALOS reduction was not accompanied by an increase in hospital readmissions or nursing home placements. The ALOS reduction was also not seen in elective coronary artery bypass graft patients whose ALOS did not substantially change over the same period. Two years after the project, ALOS for THA patients remained at ten days or below. This reduction in LOS and in the use of other hospital services translated into a mean total hospital charge decrease of $2045 per THA patient.

Mechanism of action of GTP in the induction of Ca2+ release from hepatic microsomes.

The mechanism by which GTP induces Ca2+ release from Ca2(+)-preloaded rat hepatic microsomes was studied. In the same concentration range as that for Ca2+ release, GTP inhibited the initial rate of ATP-driven Ca2+ uptake. It also inhibited the formation by ATP of the phosphorylated intermediate of Ca2(+)-ATPase, which had previously been identified by us as a 97-116 kDa protein (Fleschner, C.R., et al. (1985) Biochem. J. 226, 839). Vanadate, an inhibitor of Ca2(+)-ATPase, also caused Ca2+ release in a similar fashion, but its effect was not additive to that of GTP. Although the non-metabolizable GTP analogues, GMPPNP and GTP gamma S, did not cause Ca2+ release by themselves, GTP gamma S completely and GMPPNP partially blocked the effect of GTP. Pretreatment of vesicles with either cholera or pertussis toxin did not alter the responsiveness to GTP. These results indicate that GTP inhibits microsomal Ca2(+)-ATPase, independently of the Gs and Gi proteins. Because a decrease in Ca2+ uptake results in a net increase in Ca+ release, this effect of GTP seems to account, at least partially, for the GTP-induced Ca2+ release from microsomes.