Although DISC1 has been implicated in many psychiatric disorders, including schizophrenia, bipolar disorder, schizoaffective disorder and major depression, its biological role in these disorders is unclear. To better understand this gene and its role in psychiatric disease, we conducted transcriptional profiling and genome-wide association analysis in 1 232 pedigreed Mexican American individuals for whom we have neuroanatomic images, neurocognitive assessments and neuropsychiatric diagnoses. SOLAR was used to determine heritability, identify gene expression patterns and perform association analyses on 188 quantitative brain-related phenotypes. We found that the DISC1 transcript is highly heritable (h2=0.50; p=1.97 × 10−22), and that gene expression is strongly cis-regulated (cis-LOD=3.89) but is also influenced by trans-effects. We identified several DISC1 polymorphisms that were associated with cortical gray-matter thickness within the parietal, temporal and frontal lobes. Associated regions affiliated with memory included the entorhinal cortex (rs821639, p=4.11 × 10−5; rs2356606, p=4.71 × 10−4), cingulate cortex (rs16856322, p=2.88 × 10−4) and parahippocampal gyrus (rs821639, p=4.95 × 10−4); those affiliated with executive and other cognitive processing included the transverse temporal gyrus (rs9661837, p=5.21 × 10−4; rs17773946, p=6.23 × 10−4), anterior cingulate cortex (rs2487453, p=; 4.79 × 10−4; rs3738401, p= 5.43 × 10−4) and medial orbitofrontal cortex (rs9661837; p=7.40 × 10−4). Cognitive measures of working memory (rs2793094, p=3.38 × 10−4), as well as lifetime history of depression (rs4658966, p=4.33 × 10−4; rs12137417, p=4.93 × 10−4) and panic (rs12137417, p=7.41 × 10−4) were associated with DISC1 sequence variation. DISC1 has well-defined genetic regulation and clearly influences important phenotypes related to psychiatric disease.
DISC1; association; neuroanatomical; neurocognitive; endophenotype; cis-regulation; depression; panic; memory; learning
Background and Purpose: We hypothesized that the P-selectin (SELP) gene, localized to a region on chromosome 1q24, pleiotropically contributes to increased blood pressure and cerebral atrophy. We tested this hypothesis by performing genetic correlation analyses for 13 mRNA gene expression measures from P-selectin and 11 other genes located in 1q24 region and three magnetic resonance imaging derived indices of cerebral integrity. Methods: The subject pool consisted of 369 (219F; aged 28–85, average = 47.1 ± 12.7 years) normally aging, community-dwelling members of large extended Mexican-American families. Genetic correlation analysis decomposed phenotypic correlation coefficients into genetic and environmental components among 13 leukocyte-based mRNA gene expressions and three whole-brain and regional measurements of cerebral integrity: cortical gray matter thickness, fractional anisotropy of cerebral white matter, and the volume of hyperintensive WM lesions. Results: From the 13 gene expressions, significant phenotypic correlations were only found for the P- and L-selectin expression levels. Increases in P-selectin expression levels tracked with decline in cerebral integrity while the opposite trend was observed for L-selectin expression. The correlations for the P-selectin expression were driven by shared genetic factors, while the correlations with L-selectin expression were due to shared environmental effects. Conclusion: This study demonstrated that P-selectin expression shared a significant variance with measurements of cerebral integrity and posits elevated P-selectin expression levels as a potential risk factor of hypertension-related cerebral atrophy.
cerebral atrophy; genetics; DTI; aging; hypertension; gene expression; P-selectin
This report describes a case of well differentiated fetal adenocarcinoma of the lung in a 29 year old female smoker. The histological pattern and immunohistochemical profile were consistent with well differentiated fetal adenocarcinoma and the patient made an uneventful postoperative recovery with no recurrence after 18 months. This neoplasm is a rare lung tumour that is composed of glycogen rich neoplastic glands and tubules that resembles fetal lung at 10 to 15 weeks of gestation. It is important to identify this rare variant of adenocarcinoma because it is a low grade malignancy with low associated mortality.
fetal adenocarcinoma; lung
Allosteric DNA oligonucleotides are potentially useful diagnostic reagents. Here we develop a model system for the study of allosteric interactions in DNAs. A DNA that binds either Cibacron blue or cholic acid was isolated and partially characterized. Isolation was performed using a multi-stage SELEX. First, short oligos that bind either Cibacron blue or cholic acid were enriched from random oligonucleotide pools. Then, members of the two pools were fused to form longer oligos, which were then selected for theability to bind Cibacron blue columns and elute with cholic acid. One resulting isolate (A22) was studied. Dye- and cholate-binding functions can be separated on sequences from the 5'- and 3'-regions, respectively. Ligand-column affinity assays indicate that each domain binds only its respective ligand. However, the full-length A22 will bind either dye or cholate columns and elute with the other ligand, as if binding by the ligands is mutually exclusive. Furthermore, S1 nuclease protection assays show that Cibacron blue causes a structural change in A22 and that cholic acid inhibits this change. This system will be useful for elucidating mechanisms of allosteric interactions in synthetic DNAs.
Malignant hyperthermia (MH) is an autosomal dominant genetic condition that presents in susceptible people undergoing general anaesthesia. The clinical disorder is a major cause of anaesthetic morbidity and mortality. The UK Malignant Hyperthermia Group has performed genetic linkage analysis on 20 large, well defined malignant hyperthermia families, using hypervariable markers on chromosome 19q13.1, including the candidate MH gene RYR1, the gene coding for the skeletal muscle ryanodine receptor protein. The results were analysed using LINKAGE to perform two point and multipoint lod scores, then HOMOG to calculate levels of heterogeneity. The results clearly showed genetic heterogeneity between MH families; nine of the families gave results entirely consistent with linkage to the region around RYR1 while the same region was clearly excluded in three families. In the remaining eight MHS families there were single recombinant events between RYR1 and MH susceptibility. HOMOG analysis was of little added benefit in determining the likelihood of linkage to RYR1 in these families. This confirmation of the presence of heterogeneity in the UK MH population, along with the possibility of the presence of two MH genes in some pedigrees, indicates that it would be premature and potentially dangerous to offer diagnosis of MH by DNA based methods at this time.
The role of the negative-stranded virus accessory C proteins is difficult to assess because they appear sometimes as nonessential and thereby of no function. On the other hand, when a function is found, as in the case of Sendai virus, it represents an enigma, in that the C proteins inhibit replication under conditions where the infection follows an exponential course. Furthermore, this inhibitory function is exerted differentially: in contrast to the replication of internal deletion defective interfering (DI) RNAs, that of copy-back DI RNAs appears to escape inhibition, under certain experimental conditions (in vivo assay). In a reexamination of the C effect by the reverse genetics approach, it was found that copy-back RNA replication is inhibited by C in vivo as well, under conditions where the ratio of C to copy-back template is increased. This effect can be reversed by an increase in P but not L protein. The "rule of six" was differentially observed in the presence or absence of C. Finally, a difference in the ability of the replicating complex to tolerate promoter modifications in RNA synthesis initiation was shown to occur in the presence or the absence of C as well. We propose that C acts by increasing the selectivity of the replicating complex for the promoter cis-acting elements governing its activity. The inhibitory effect of C becomes the price to pay for this increased selectivity.
Others have recently shown that the UUU phenylalanine codon is highly frameshift-prone in the 3'(rightward) direction at pyrimidine 3'contexts. Here, several approaches are used to analyze frameshifting at such sites. The four permutations of the UUU/C (phenylalanine) and CGG/U (arginine) codon pairs were examined because they vary greatly in their expected frameshifting tendencies. Furthermore, these synonymous sites allow direct tests of the idea that codon usage can control frameshifting. Frameshifting was measured for these dicodons embedded within each of two broader contexts: the Escherichia coli prfB (RF2 gene) programmed frameshift site and a 'normal' message site. The principal difference between these contexts is that the programmed frameshift contains a purine-rich sequence upstream of the slippery site that can base pair with the 3'end of 16 S rRNA (the anti-Shine-Dalgarno) to enhance frameshifting. In both contexts frameshift frequencies are highest if the slippery tRNAPhe is capable of stable base pairing in the shifted reading frame. This requirement is less stringent in the RF2 context, as if the Shine-Dalgarno interaction can help stabilize a quasi-stable rephased tRNA:message complex. It was previously shown that frameshifting in RF2 occurs more frequently if the codon 3'to the slippery site is read by a rare tRNA. Consistent with that earlier work, in the RF2 context frameshifting occurs substantially more frequently if the arginine codon is CGG, which is read by a rare tRNA. In contrast, in the 'normal' context frameshifting is only slightly greater at CGG than at CGU. It is suggested that the Shine-Dalgarno-like interaction elevates frameshifting specifically during the pause prior to translation of the second codon, which makes frameshifting exquisitely sensitive to the rate of translation of that codon. In both contexts frameshifting increases in a mutant strain that fails to modify tRNA base A37, which is 3'of the anticodon. Thus, those base modifications may limit frameshifting at UUU codons. Finally, statistical analyses show that UUU Ynn dicodons are extremely rare in E.coli genes that have highly biased codon usage.
Many paramyxoviruses express small basic C proteins, from an alternate, overlapping open reading frame of the P gene mRNA, which were previously found to inhibit mRNA synthesis. During recent experiments in which infectious Sendai virus (SeV) was recovered from cDNA via the initial expression of the viral N, P, and L genes from plasmids, the abrogation of C protein expression from the plasmid P gene was found to be necessary for virus recovery. We have investigated the effect of C coexpression on the amplification of an internally deleted defective interfering (DI) genome directly in the transfected cell, for which, in contrast to virus recovery experiments, genome amplification is independent of mRNA synthesis carried out by the SeV polymerase. We find that C protein coexpression also strongly inhibits the amplification of this DI genome but has little or no effect on that of a copy-back DI genome (DI-H4). We have also characterized the C protein from a mutant SeV and found that (i) it had lost most of its inhibitory activity on internally deleted DI genome amplification and (ii) its coexpression no longer prevented the recovery of SeV from DNA. However, consistent with the insensitivity of copy-back DI genomes to C protein inhibition, C coexpression did not prevent the recovery of copy-back nondefective viruses from DNA. The inhibitory effects of C coexpression thus appear to be promoter specific.
Because of the enormity of the HIV-AIDS epidemic and the urgency for preventing transmission, HIV prevention programs are a high priority for careful and timely evaluations. Information on program effectiveness and efficiency is needed for decision-making about future HIV prevention priorities. General characteristics of successful HIV prevention programs, programs empirically evaluated and found to change (or not change) high-risk behaviors or in need of further empirical study, and economic evaluations of certain programs are described and summarized with attention limited to programs that have a behavioral basis. HIV prevention programs have an impact on averting or reducing risk behaviors, particularly when they are delivered with sufficient resources, intensity, and cultural competency and are based on a firm foundation of behavioral and social science theory and past research. Economic evaluations have found that some of these behaviorally based programs yield net economic benefits to society, and others are likely cost-effective (even if not cost-saving) relative to other health programs. Still, specific improvements should be made in certain HIV prevention programs.
Abstract: Optimal conditions for cryopreserving of populations of root lesion nematode (Pratylenchus spp.) were determined. Nematode survival was achieved using glycerol pre-treatments in the range of 14-17% (w/w). Increasing duration of the incubation in glycerol (up to 5 days) before immersion in liquid nitrogen significantly influenced nematode survival The highest mean survival for P. thornei was 76% after incubation in glycerol for 5 days. Nematodes were able to reproduce and infect carrot disc cultures after cryopreservation. This technique has valuable applications for long-term germplasm storage and maintenance of genetic lines.
cryopreservation; lesion nematode; Pratylenchus
Two domains involved in RNA synthesis have recently been found within the N-terminal 77 amino acids of the Sendai virus P protein. One domain is required for RNA synthesis per se and has properties in common with the transactivation domains of cellular transcription factors. The second domain is thought to be specifically required for the nascent chain assembly step in genome replication. We have further mapped this second domain by the construction of chimeric and deleted P proteins to amino acids 33 to 41 of P and by examining the abilities of these P proteins to support DI genome replication in vivo. Using glycerol gradient sedimentation, we have shown that this domain is required to form a stable complex with unassembled NP (P-NP0) and to prevent NP from assembling illegitimately, i.e., independently of the concurrent assembly of a nascent viral genome. Since the P-NP0 complex represents the functional form of unassembled NP which is delivered to the nascent chain during genome replication, and since amino acids 33 to 41 are not required for the stable interaction of P with the assembled NP of the nucleocapsid, this chaperone function of P is not required for mRNA synthesis or the RNA synthesis step of genome replication.
As the epidemic of human immunodeficiency virus (HIV) infection and acquired immunodeficiency syndrome (AIDS) has evolved over the past 10 years, the Centers for Disease Control (CDC) has been at the forefront of the scientific efforts that have characterized HIV-AIDS research. Because of CDC's central role in these efforts, the medical and public health communities have come to depend on the agency for prompt reporting of new developments related to the epidemiology of HIV infection and AIDS and for advice on risk management, prevention, and control. CDC disseminates this information through epidemiologic updates and prevention guidelines published in the periodical, Morbidity and Mortality Weekly Report, through articles in scientific journals and summary tabulations of AIDS case data and HIV seroprevalence data, and through interviews and presentations at scientific meetings. These formal information dissemination activities are supplemented with training and support efforts directed at health care providers, health department and laboratory personnel, educators, and centralized HIV-AIDS information resources. As questions are answered, controversies resolved, and new research applications explored, CDC will continue to provide the medical and public health communities with the most recent epidemiologic information and recommendations developed to help direct efforts in HIV prevention and risk reduction.
The paramyxovirus nucleocapsid proteins (NPs) are relatively well conserved, except for the C-terminal 20% (or ca. 100 amino acids), referred to as the tail. We have examined whether this hypervariable tail is required for genome synthesis, both in vitro, where synthesis is predominantly from the input templates, and in vivo, where multiple rounds of amplification occur. In these viruses, genome synthesis and assembly of the nascent chain are coupled. We find that the tail is required in vivo but not in vitro. Closer examination of the in vivo system showed that the tailless NP could encapsidate the genome chain but that amplification did not occur. We interpret these results as indicating that the tail is not required for RNA assembly but is required for the template to function in RNA synthesis. Relatively small deletions within the conserved N-terminal 80% of the protein, on the other hand, rendered the protein nonfunctional in either system. The possible functions of the tail in RNA synthesis are discussed.
Application method had an appreciable effect on the efficacy of Heterorhabditis sp. (isolate T390) in reducing the numbers of Otiorhynchus sulcatus infesting field-grown strawberries. Results were related to nematode placement. In Trial 1, the mean weevil mortality was 36% for trickle-irrigated Steinernema sp. (isolate NC513) at a dose of 100,000 nematodes per plant, whereas the same dose of Heterorhabditis sp. (isolate T390) resulted in mortality of 65% and 86% for trickle-irrigated and surface-sprayed nematodes, respectively. Mortality rate (y) was inversely related to initial weevil population size (x) by y = 4.96x-0.957 and y = 4.71x-0.558 for trickle-irrigated Steinernema sp. (isolate NC513) and Heterorhabditis sp. (isolate T390), respectively. In Trial 2, using 100,000 Heterorhabditis sp. (isolate T390) per plant, mean weevil mortalities were 61%, 63%, and 79% for single-injection, irrigation, and multiple-injection techniques, respectively.
application method; biological control; Fragaria; Heterorhabditis; nematode; Otiorhynchus sulcatus; pest density; Steinernema; strawberry
We present evidence that the formation of NP-P and P-L protein complexes is essential for replication of the genome of Sendai defective interfering (DI-H) virus in vitro, using extracts of cells expressing these viral proteins from plasmids. Optimal replication of DI-H nucleocapsid RNA required extracts of cells transfected with critical amounts and ratios of each of the plasmids and was three- to fivefold better than replication with a control extract prepared from a natural virus infection. Extracts in which NP and P proteins were coexpressed supported replication of the genome of purified DI-H virus which contained endogenous polymerase proteins, but extracts in which NP and P were expressed separately and then mixed were inactive. Similarly, the P and L proteins must be coexpressed for biological activity. The replication data thus suggest that two protein complexes, NP-P and P-L, are required for nucleocapsid RNA replication and that these complexes must form during or soon after synthesis of the proteins. Biochemical evidence in support of the formation of each complex includes coimmunoprecipitation of both proteins of each complex with an antibody specific for one component and cosedimentation of the subunits of each complex. We propose that the P-L complex serves as the RNA polymerase and NP-P is required for encapsidation of newly synthesized RNA.
Subcutaneous emphysema is an iatrogenic complication by which air is introduced into the tissues either during or immediately after surgery. A case is presented that demonstrates the complication, following the removal of third molars, believed to be due to violation of the maxillary sinus, an underinflated cuff of a nasotracheal tube, and coughing on extubation.
Infective-stage juveniles of Steinernema and Heterorhabditis spp. were cryopreserved using two-stage incubation in glycerol and 70% methanol before storage in cryotubes in liquid nitrogen. Optimal glycerol concentrations and incubation times for survival were determined for different species, but acceptable survival of all species and isolates of entomopathogenic nematodes can be obtained using 15% (w/w) glycerol and incubation for 48 hours. Mean survival was 69% for isolates of Steinernema and 68% for isolates of Heterorhabditis (n = 84). The maximum survival recorded was 97% for S. feltiae K254 stored in liquid nitrogen for 12 months.
cryopreservation; Heterorhabditis; nematode; Steinerneraa
During 1987-89, the Centers for Disease Control (CDC), in collaboration with State and local health departments, other Federal agencies, blood collection agencies, and medical research institutions, implemented a national sentinel surveillance system for human immunodeficiency virus (HIV) infection. This ongoing surveillance system, known as the CDC family of HIV seroprevalence surveys, uses standardized survey and HIV serologic testing procedures in a group of sentinel populations from geographically diverse metropolitan areas, States, and Territories of the United States. As of September 1989, sentinel surveillance for HIV infection was being conducted in 41 States, Puerto Rico, and 39 metropolitan areas, including the District of Columbia. Information from this system complements AIDS surveillance data to assist health officials to direct resources and develop strategies for HIV prevention and health-care programs.
Numbers of Steinernema sp. (CB2B) and S. carpocapsae (Agriotos) exponentially declined after application into a clay loam soil. Over a 35-day sampling period, Steinernema sp. (CB2B) was more persistent than S. carpocapsae (Agriotos). The presence or absence of the second-stage cuticle on the third-stage juveniles (J3) at the time of application did not alter the rate of population decline of Steinernema sp. (CB2B). Nearly all J3 of Steinernema sp. (CB2B) and S. carpocapsae (Agriotos) lost their cuticle within 24 hours of being in soil. Centrifugal flotation recovered the greatest number of nematodes, with a lower variance than either the live bait or Baermann funnel techniques. A strong positive linear relationship was evident between numbers of nematodes present in the soil and the numbers that established in a bait insect. Approximately 40% of Steinernema sp. (CB2B) and 30% of the S. carpocapsae (Agriotos) present in the soil established in Galleria mellonella larvae. The extraction techniques had different efficiencies and gave different relative estimates of persistence for the two species. Persistence and infectivity was best measured using a combination of live bait and flotation techniques.
entomopathogenic nematode; extraction technique; Galleria; infectivity; nematode; persistence; Steinernema carpocapsae; Steinernema sp.
Parainfluenza virus type 1 (PIV1) and Sendai virus (SEN) are very closely related, but the PIV1 P/C gene does not contain the ACG codon which initiates the SEN C' protein. Nevertheless, a protein corresponding to the PIV1 C' protein was observed both in vivo and in vitro. The initiation site of this protein maps upstream of the PIV1 C protein AUG in a region that does not contain an AUG codon. We have used site-directed mutagenesis to demonstrate that the PIV1 C' protein initiates from a GUG codon, four codons upstream of where the ACG is found in SEN. Remarkably, this GUG appears to initiate in vivo almost as frequently as AUG in the same context. However, whereas GUG permits downstream expression of the P and C proteins, AUG in this context does not. The conservation of an upstream non-AUG initiation codon for C' among PIV1 and SEN suggests that it is important for virus replication, even though some paramyxoviruses express only the C protein and others have no C open reading frame at all.
The nucleotide sequence of the P gene of human parainfluenza virus type 1 (PIV1) was determined from cloned cDNA copies of the mRNA. By analogy with the gene organization of Sendai virus, two open reading frames in the mRNA sense of the gene were identified as coding sequences for the P protein (568 amino acids with an estimated molecular weight of 64,655) and the C protein (204 amino acids with an estimated molecular weight of 24,108). Comparison of the deduced amino acid sequences of the P and C proteins of PIV1 with those of Sendai virus showed a high degree of homology. However, a sequence for the cysteine-rich V protein, which was considered a common feature of other paramyxoviruses, was interrupted by the presence of multiple stop codons. The sequence analysis of three P-gene-specific cDNA clones generated from genomic RNA by polymerase chain reaction and one additional clone generated from mRNA confirmed that the coding sequence for the cysteine-rich region is silent in the PIV1 gene and thus is not translated into protein. Two potential editing sites with the consensus sequence 3'UUYUCCC were found in the PIV1 P gene at positions 564 to 570 and 1430 to 1436. However, examination of the PIV1 mRNA population by a primer extension method indicated that neither of these sites is utilized. These results indicate that the PIV1 P gene has a coding strategy different from those of other paramyxovirus P genes.
Two forms of the Sendai virus P/C mRNA have been predicted: one an exact copy of the viral genome, and the other with a single G insertion within a run of three G's. We directly cloned the mRNA or portions of it containing the insertion site and screened the resulting colonies with oligonucleotides that could distinguish the presence of three or four G's at this position. We found that 31% of the mRNAs did in fact contain the predicted insertion, whereas the viral genomes contained no heterogeneity at this position. A smaller fraction (7%) of the mRNA contained two to eight G's inserted at this position. The insertions also took place during RNA synthesis in vitro with purified virions but were not detected when the mRNA was expressed in vivo via a vaccinia virus recombinant. When the Sendai virus- and vaccinia virus-derived P/C mRNAs were coexpressed in the same cells under conditions in which each could be distinguished, those from the Sendai genome were altered as before, but those from the vaccinia virus genome remained unaltered. The activity that alters the mRNA is therefore likely to be coded for by the virus and cannot function in trans.