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1.  Waist circumference is genetically correlated with incident Type 2 diabetes in Mexican-American families 
We aimed to determine the genetic and environmental correlation between various anthropometric indexes and incident Type 2 diabetes with a focus on waist circumference.
We used the data on extended Mexican-American families (808 subjects, 7617.92 person-years follow-up) from the San Antonio Family Heart Study and estimated the genetic and environmental correlations of 16 anthropometric indexes with the genetic liability of incident Type 2 diabetes. We performed bivariate trait analyses using the solar software package.
All 16 anthropometric indexes were significantly heritable (range of heritabilities 0.24–0.99). Thirteen indexes were found to have significant environmental correlation with the liability of incident Type 2 diabetes. In contrast, only anthropometric indexes consisting of waist circumference (waist circumference, waist–hip ratio and waist–height ratio) were significantly genetically correlated (genetic correlation coefficients: 0.45, 0.55 and 0.44, respectively) with the liability of incident Type 2 diabetes. We did not observe such a correlation for BMI.
Waist circumference as a predictor of future Type 2 diabetes is supported by shared genetic influences.
PMCID: PMC3849209  PMID: 23796311
2.  Localization of a major susceptibility locus influencing preterm birth 
Molecular Human Reproduction  2013;19(10):687-696.
Preterm birth (PTB) is a complex trait, but little is known regarding its major genetic determinants. The objective of this study is to localize genes that influence susceptibility to PTB in Mexican Americans (MAs), a minority population in the USA, using predominantly microfilmed birth certificate-based data obtained from the San Antonio Family Birth Weight Study. Only 1302 singleton births from 288 families with information on PTB and significant covariates were considered for genetic analysis. PTB is defined as a childbirth that occurs at <37 completed weeks of gestation, and the prevalence of PTB in this sample was 6.4%. An ∼10 cM genetic map was used to conduct a genome-wide linkage analysis using the program SOLAR. The heritability of PTB was high (h2 ± SE: 0.75 ± 0.20) and significant (P = 4.5 × 10−5), after adjusting for the significant effects of birthweight and birth order. We found significant evidence for linkage of PTB (LOD = 3.6; nominal P = 2.3 × 10−5; empirical P = 1.0 × 10−5) on chromosome 18q between markers D18S1364 and D18S541. Several other chromosomal regions (2q, 9p, 16q and 20q) were also potentially linked with PTB. A strong positional candidate gene in the 18q linked region is SERPINB2 or PAI-2, a member of the plasminogen activator system that is associated with various reproductive processes. In conclusion, to our knowledge, perhaps for the first time in MAs or US populations, we have localized a major susceptibility locus for PTB on chromosome 18q21.33-q23.
PMCID: PMC3779004  PMID: 23689979
preterm birth; family studies; susceptibility locus; variance-components linkage analysis; Mexican Americans
3.  QTL-based association analyses reveal novel genes influencing pleiotropy of Metabolic Syndrome (MetS) 
Obesity (Silver Spring, Md.)  2013;21(10):2099-2111.
Metabolic Syndrome (MetS) is a phenotype cluster predisposing to type 2 diabetes and cardiovascular diseases. In extended families of Northern European ancestry, we previously identified two significant QTLs 3q27 and 17p12 that were linked with multiple representative traits of MetS. To determine the genetic basis of these linkage signals, QTL-specific genomic and transcriptomic analyses were performed in 1,137 individuals from 85 extended families that contributed to the original linkage. We tested in SOLAR association of MetS phenotypes with QTL-specific haplotype-tagging SNPs as well as transcriptional profiles of peripheral blood mononuclear cells (PBMCs). SNPs significantly associated with phenotypes under the prior hypothesis of linkage mapped to seven genes at 3q27 and seven at 17p12. Prioritization based on biologic relevance, SNP association, and expression analyses identified two genes: insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2) at 3q27 and tumor necrosis factor receptor 13B (TNFRSF13B) at 17p12. Prioritized genes could influence cell-cell adhesion and adipocyte differentiation, insulin/glucose responsiveness, cytokine effectiveness and plasma lipids and lipoprotein densities. In summary, our results combine genomic, transcriptomic, and bioinformatic data to identify novel candidate loci for MetS.
PMCID: PMC3769476  PMID: 23418049
4.  Testing the hypothesis of accelerated cerebral white matter aging in schizophrenia and major depression 
Biological psychiatry  2012;73(5):482-491.
Elevated rate of aging-related biological and functional decline, termed accelerated aging, is reported in patients with schizophrenia (SCZ) and major depressive disorder (MDD). We used diffusion tensor imaging (DTI) derived fractional anisotropy (FA) as biomarkers of aging-related decline in white matter (WM) integrity to test the hypotheses of accelerated aging in SCZ and MDD.
The SCZ cohort was composed of 58/60 SCZ patients/controls (age=20–60years). MDD cohort was composed of 136/351 MDD patients/controls (age=20–79years). Main outcome measures were the diagnosis-by-age interaction on whole-brain-averaged WM FA values and FA values from twelve major WM tracts.
Diagnosis-by-age interaction for the whole-brain average FA was significant for the SCZ (p=0.04) but not in MDD cohort (p=0.80). Diagnosis-by-age interaction was nominally significant (p<0.05) for five WM tracts for SCZ and for none of the tracts in the MDD cohort. Tract-specific heterochronicity of the onset of age-related decline in SCZ demonstrated strong negative correlations with the age-of- peak myelination and the rates of age-related decline obtained from normative sample (r=−0.61 and −0.80, p<0.05, respectively). No such trends existed for MDD cohort.
Cerebral WM showed accelerated aging in SCZ but not in MDD, suggesting some difference in the pathophysiology underlying their WM aging changes. Tract-specific heterochronicity of WM development modulated presentation of accelerated aging in SCZ: white matter tracts that matured later in life appeared more sensitive to the pathophysiology of SCZ and demonstrated more susceptibility to disorder-related accelerated decline in FA values with age. This trend was not observed in MDD cohort.
PMCID: PMC3645491  PMID: 23200529
5.  NLRX1/NOD5 deficiency does not affect MAVS signalling 
PMCID: PMC3172102  PMID: 21617692
6.  Impact of DISC1 variation on neuroanatomical and neurocognitive phenotypes 
Molecular psychiatry  2011;16(11):1096-1104.
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.
PMCID: PMC3135724  PMID: 21483430
DISC1; association; neuroanatomical; neurocognitive; endophenotype; cis-regulation; depression; panic; memory; learning
7.  P-selectin Expression Tracks Cerebral Atrophy in Mexican-Americans 
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.
PMCID: PMC3340599  PMID: 22558002
cerebral atrophy; genetics; DTI; aging; hypertension; gene expression; P-selectin
8.  Well differentiated fetal adenocarcinoma of the lung in a 29 year old woman 
Journal of Clinical Pathology  2003;56(6):478-479.
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.
PMCID: PMC1769968  PMID: 12783979
fetal adenocarcinoma; lung
9.  An allosteric synthetic DNA. 
Nucleic Acids Research  1999;27(6):1512-1516.
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.
PMCID: PMC148346  PMID: 10037814
10.  Genetic heterogeneity and HOMOG analysis in British malignant hyperthermia families. 
Journal of Medical Genetics  1998;35(3):196-201.
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.
PMCID: PMC1051241  PMID: 9541102
11.  Inhibition of Sendai virus genome replication due to promoter-increased selectivity: a possible role for the accessory C proteins. 
Journal of Virology  1997;71(12):9588-9599.
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.
PMCID: PMC230267  PMID: 9371623
12.  Analyses of frameshifting at UUU-pyrimidine sites. 
Nucleic Acids Research  1997;25(10):2005-2011.
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.
PMCID: PMC146683  PMID: 9115369
14.  Bears, academia, and anaesthesia: a controlled study. 
BMJ : British Medical Journal  1996;313(7072):1643-1644.
PMCID: PMC2359135  PMID: 9011292
15.  The Sendai paramyxovirus accessory C proteins inhibit viral genome amplification in a promoter-specific fashion. 
Journal of Virology  1996;70(8):5067-5074.
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.
PMCID: PMC190461  PMID: 8764014
16.  An overview of the effectiveness and efficiency of HIV prevention programs. 
Public Health Reports  1995;110(2):134-146.
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.
PMCID: PMC1382092  PMID: 7630989
17.  Cryopreservation of Pratylenchys spp. 
Journal of Nematology  1995;27(4):483-485.
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.
PMCID: PMC2619634  PMID: 19277316
cryopreservation; lesion nematode; Pratylenchus
18.  An N-terminal domain of the Sendai paramyxovirus P protein acts as a chaperone for the NP protein during the nascent chain assembly step of genome replication. 
Journal of Virology  1995;69(2):849-855.
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.
PMCID: PMC188651  PMID: 7815552
19.  Communicating surveillance, epidemiologic, and laboratory information on HIV infection and AIDS. 
Public Health Reports  1991;106(6):721-726.
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.
PMCID: PMC1580332  PMID: 1659722
20.  The hypervariable C-terminal tail of the Sendai paramyxovirus nucleocapsid protein is required for template function but not for RNA encapsidation. 
Journal of Virology  1993;67(7):4358-4364.
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.
PMCID: PMC237806  PMID: 8389932
21.  Influence of Application Method and Pest Population Size on the Field Efficacy of Entomopathogenic Nematodes 
Journal of Nematology  1992;24(4S):631-636.
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.
PMCID: PMC2629875  PMID: 19283038
application method; biological control; Fragaria; Heterorhabditis; nematode; Otiorhynchus sulcatus; pest density; Steinernema; strawberry
22.  Complexes of Sendai virus NP-P and P-L proteins are required for defective interfering particle genome replication in vitro. 
Journal of Virology  1992;66(8):4901-4908.
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.
PMCID: PMC241329  PMID: 1321276
23.  Subcutaneous emphysema: a complication of surgery and anesthesia. 
Anesthesia Progress  1992;39(1-2):38-40.
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.
PMCID: PMC2148724  PMID: 8507023
24.  Routine Cryopreservation of Isolates of Steinernema and Heterorhabditis spp. 
Journal of Nematology  1992;24(2):269-270.
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.
PMCID: PMC2619270  PMID: 19282994
cryopreservation; Heterorhabditis; nematode; Steinerneraa
25.  The parainfluenza virus type 1 P/C gene uses a very efficient GUG codon to start its C' protein. 
Journal of Virology  1992;66(3):1765-1768.
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.
PMCID: PMC240931  PMID: 1310778

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