Effect of high-frequency oscillatory ventilation on intracranial pressure in a child with herpes encephalitis and acute respiratory distress syndrome.
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Child, Preschool↗
Biomedical subjects
Y S Hahn
Publications and source records attributed to Y S Hahn.
Risk factors of intracranial pressure monitoring in children with fiberoptic devices: a critical review.
BACKGROUND: Intracranial pressure (ICP) monitoring has become standard in the management of severe head injuries. A variety of monitoring techniques and devices are available, each with advantages and disadvantages; however there have been few studies in the pediatric population. METHODS: To study the risk factors, efficacy, and complication rate of fiberoptic ICP monitoring we studied 98 consecutive children with severe head injuries over a 2-year period. The average patient age was 9 years and most had an initial Childrens Coma Score (CCS) of 8 or less. The monitoring devices were placed in frontal parenchyma of all children and no prophylactic antibiotics were used. All fiberoptic catheter tips were cultured upon removal. RESULTS: The average duration of ICP monitoring was 7 days; the usual range was 3-15 days, with the exception of one patient who underwent monitoring for 40 days. No complications occurred during insertion of the ICP monitors. Catheter tip cultures were positive for Staphylococcus epidermidis in 7% of the children, but none developed clinical features of CNS infection. The hospital location of placement or duration of ICP monitoring did not affect the rate of catheter tips with positive cultures. There was a 13% mechanical failure rate of the fiberoptic device. CONCLUSIONS: We conclude that fiberoptic ICP monitoring is safe and effective; however, there is a relatively low mechanical failure rate. Infection in uncommon despite prolonged use and there is little risk of complications associated with placement of this monitor.
Adolescent↗
Formation of native hepatitis C virus glycoprotein complexes.
The hepatitis C virus (HCV) glycoproteins (E1 and E2) interact to form a heterodimeric complex, which has been proposed as a functional subunit of the HCV virion envelope. As examined in cell culture transient-expression assays, the formation of properly folded, noncovalently associated E1E2 complexes is a slow and inefficient process. Due to lack of appropriate immunological reagents, it has been difficult to distinguish between glycoprotein molecules that undergo productive folding and assembly from those which follow a nonproductive pathway leading to misfolding and aggregation. Here we report the isolation and characterization of a conformation-sensitive E2-reactive monoclonal antibody (H2). The H2 monoclonal antibody selectively recognizes slowly maturing E1E2 heterodimers which are noncovalently linked, protease resistant, and no longer associated with the endoplasmic reticulum chaperone calnexin. This complex probably represents the native prebudding form of the HCV glycoprotein heterodimer. Besides providing a novel reagent for basic studies on HCV virion assembly and entry, this monoclonal antibody should be useful for optimizing production and isolation of native HCV glycoprotein complexes for serodiagnostic and vaccine applications.
Animals↗
The requirement for proteasome activity class I major histocompatibility complex antigen presentation is dictated by the length of preprocessed antigen.
Accumulating evidence has implicated the proteasome in the processing of protein along the major histocompatibility complex (MHC) class I presentation pathway. The availability of potent proteasome inhibitors provides an opportunity to examine the role of proteasome function in antigen presentation by MHC class I molecules to CD8+ cytotoxic T lymphocytes (CTLs). We have investigated the processing and presenting of antigenic epitopes from influenza hemagglutinin in target cells treated with the inhibitor of proteasome activity MG132. In the absence of proteasome activity, the processing and presentation of the full-length hemagglutinin was abolished, suggesting the requirement for proteasome function in the processing and presentation of the hemagglutinin glycoprotein. Epitope-containing translation products as short as 21 amino acids when expressed in target cells required proteasome activity for processing and presentation of the hemagglutin epitope to CTLs. However, when endogenous peptides of 17 amino acids or shorter were expressed in target cells, the processing and presentation of epitopes contained in these peptides were insensitive to the proteasome inhibitor. Our results support the hypothesis that proteasome activity is required for the generation of peptides presented by MHC class I molecules and that the requirement for proteasome activity is dependent on the size of the translation product expressed in the target cell. The implications of these findings are discussed.
Amino Acid Sequence↗
Endogenous presentation of a nascent antigenic epitope to CD8+ CTL is more efficient than exogenous presentation.
Cytotoxic T lymphocytes (CTL) recognize short antigenic peptides in association with class I MHC molecules at the cell surface. Newly synthesized viral polypeptides are processed in the cytoplasm and the fragments of antigen are transported into the endoplasmic reticulum (ER) via a peptide transporter where they complex with nascent class I molecules. The peptide-MHC complex is transported to the cell surface and presented to CTL. Sequence analysis of endogenously expressed, MHC-associated self or viral antigens indicates that the naturally processed peptides bound to class I MHC molecules are in general 9 +/- 1 residues long. Peptides bound to specific class I MHC molecules have in common allele-specific motifs of conserved residues. The motif for the class I Kd molecules has been shown to be nine or 10 residues with the sequence X-Tyr-(X)6-I/L or X-Tyr-(X)7-I/L. The Tyr residue at the second position and the I/L residue at the ninth position are allele-specific anchor residues which appear to be required for binding of the peptide to Kd. To examine the stringency of the requirement for Tyr at the second position, we have performed saturation mutagenesis of a minigene encoding the class I Kd-restricted influenza HA210-219 site at the Tyr residue 211. A series of 10 mutants was tested for effects on target-cell sensitization. Most amino acid substitutions for the Tyr residue resulted in a loss of endogenous peptide recognition by HA210-219 reactive CTL, consistent with the critical role of the Tyr at the second position for interaction with Kd molecules. One mutant gene-product encoding a His substitution for the Tyr residue was recognized by CTL. However, the corresponding synthetic peptide containing a His substitution at the dominant anchor position bound only weakly to Kd, and target cells treated with the peptide were poorly recognized by CTL. The endogenous His-containing peptide was also less stably associated with class I MHC Kd molecules at the cell surface than the wild-type Tyr peptide. These data indicate that endogenous antigenic peptides may bind newly-synthesized class I MHC molecules in the ER more efficiently than fully formed class I molecules at the cell surface and that endogenous peptides may dissociate from class I MHC molecules at different rates. The implication of these findings for CTL recognition and epitope mapping are discussed.
Amino Acid Sequence↗
Regulation of antigen processing and presentation to class I MHC restricted CD8+ T lymphocytes.
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Amino Acid Sequence↗
Neurocysticercosis: report of unusual pediatric cases.
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Brain Diseases↗
Open myelomeningocele.
This article defines open myelomeningocele and describes its development in the embryo. The anatomic features of the neural placode, pia-arachnoid, and dorsal versus ventral roots are well described. The surgical technique and prenatal and postnatal care are detailed. Anomalies associated with open myelomeningocele are also described. The concept of a team approach to treating children with myelomeningocele and their families is addressed.
Female↗
Use of the distal double-slit valve system in children with hydrocephalus.
Shunts with distal slit valves have been in use for almost three decades for children with hydrocephalus. Drawbacks of this device include shunt obstruction, infection and overdrainage. One-piece shunts have been used widely for ventriculoperitoneal insertion because they are easy to place, do not need manipulation of connecting parts during surgery, and rarely disconnect. Disadvantages of these shunts, however, include a relatively higher risk of distal obstruction than with the open-ended peritoneal tube and a tendency for overdrainage. With these disadvantages in mind, we hypothesized that using double-distal-slit valves could increase the survival rate of the shunt. For the past 2 years we have been using a one-piece shunt with double-distal-slit valves and an elliptical reservoir under a strict protocol. The study showed the following (1) Among the children operated on, 30.3% required shunt revisions (0.9 revisions per patient). Children with intraventricular hemorrhage-posthermorrhagic hydrocephalus had a three times higher risk of shunt revision than children with posttraumatic hydrocephalus (55.4% vs 18.8%). (2) The so-called slit-ventricle syndrome was observed in only 4 children (8.5%), who subsequently required surgical attention. Most of the children with other forms of the overdrainage phenomenon continued to do well without clinical problems. (3) Of the 155 children, 7 (4.5%) experienced infections after the initial shunt installation. (4) Of the 136 shunt revisions, 90.4% (123 procedures) were related to obstruction. Distal obstruction (47.1%) was more frequent than proximal obstruction (36.6%). (5) In 8 children, the shunt was still working through the proximal set of valves when the distal set of valves was completely obstructed.(ABSTRACT TRUNCATED AT 250 WORDS)
Adolescent↗
CD8+ T cell recognition of an endogenously processed epitope is regulated primarily by residues within the epitope.
Cytotoxic T lymphocytes (CTL) recognize short antigenic peptides associated with cell surface class I major histocompatibility complex (MHC) molecules. This association presumably occurs between newly synthesized class I MHC molecules and peptide fragments in a pre-Golgi compartment. Little is known about the factors that regulate the formation of these antigenic peptide fragments within the cell. To examine the role of residues within a core epitope and in the flanking sequences for the generation and presentation of the newly synthesized peptide fragment recognized by CD8+ CTL, we have mutagenized the coding sequence for the CTL epitope spanning residues 202-221 in the influenza A/Japan/57 hemagglutinin (HA). In this study over 60 substitution mutations in the epitope were tested for their effects on target cell sensitization using a cytoplasmic viral expression system. The HA202-221 site contains two overlapping subsites defined by CTL clones 11-1 and 40-2. Mutations in HA residues 204-213 or residues 210-219 often abolished target cell lysis by CTL clones 11-1 and 40-2, respectively. Although residues outside the core epitope did not usually affect the ability to be lysed by CTL clones, substitution of a Gly residue for Val-214 abolished lysis by clone 11-1. These data suggest that residues within a site that affect MHC binding and T cell receptor recognition appear to play the predominant role in dictating the formation of the antigenic complex recognized by CD8+ CTL, and therefore the antigenicity of the protein antigen presented to CD8+ T cells. Most alterations in residues flanking the endogenously expressed epitope do not appreciably affect the generation and recognition of the site.
Amino Acid Sequence↗
Infectious Sindbis virus transient expression vectors for studying antigen processing and presentation.
Sindbis virus (SIN) is a small positive-strand enveloped RNA virus that infects a broad range of vertebrate and insect cells. A SIN vector (called dsSIN), designed for transient expression of heterologous RNAs and proteins, was engineered by inserting a second subgenomic mRNA promoter sequence into a nonessential region of the SIN genome. By using this vector, dsSIN recombinants have been constructed that express either bacterial chloramphenicol acetyltransferase, a truncated form of the influenza hemagglutinin (HA), or mini-genes encoding two distinct immunodominant cytotoxic T lymphocyte (CTL) HA epitopes. Infection of murine cell lines with these recombinants resulted in the expression of approximately 10(6)-10(7) chloramphenicol acetyltransferase polypeptides per cell and efficient sensitization of target cells for lysis by appropriate major histocompatibility complex-restricted HA-specific CTL clones in vitro. In addition, priming of an influenza-specific T-cell response was observed after immunizing mice with dsSIN recombinants expressing either a truncated form of HA or the immunodominant influenza CTL epitopes. This SIN expression system allows the generation of high-titered recombinant virus stocks in a matter of days and should facilitate mapping and mutational analysis of class I major histocompatibility complex-restricted T-cell epitopes expressed via the endogenous pathway of antigen processing and presentation.
Amino Acid Sequence↗
Management of the child with severe brain injury.
The pathophysiology and clinical management of acute brain injury in infancy and childhood are presented using acute traumatic brain injury as a model. The principles of stabilization, transport, and intensive care management are critically reviewed.
Brain Injuries↗
Presentation of viral antigen to class I major histocompatibility complex-restricted cytotoxic T lymphocyte. Recognition of an immunodominant influenza hemagglutinin site by cytotoxic T lymphocyte is independent of the position of the site in the hemagglutinin translation product.
Class I major histocompatibility complex (MHC) restricted T lymphocytes preferentially recognize fragments of polypeptides processed through a nonendosomal presentation pathway. At present the intracellular compartment(s) in which polypeptide fragmentation occurs and factors which influence the formation of an antigenic epitope are not well understood. To assess the role of residues flanking an antigenic site in the generation of the antigenic moiety recognized by class I MHC restricted T lymphocytes we have moved the coding sequence for an immunodominant H-2Kd restricted site on the influenza A/JAPAN/57 hemagglutinin (residues 202-221) by site-directed mutagenesis to six different positions along the coding sequence of the hemagglutinin gene. We have found that all six classes of mutants are recognized by MHC class I restricted T cells as efficiently as the wild type hemagglutinin gene product. Thus neither N-terminal to C-terminal position within the translation product nor sequences flanking the antigenic site influence processing.
Amino Acid Sequence↗
Traumatic intracavernous aneurysm in children: massive epistaxis without ophthalmic signs.
Traumatic intracavernous aneurysms are a distinctly uncommon lesion in children. They usually present with compressive symptoms of the III, IV, V, or VI cranial nerves rather than epistaxis. Epistaxis is rare and usually minor and brief in duration. A child with severe head injury and basal skull fracture, who did not have any preceding symptoms or signs of ophthalmoplegia or exophthalmus, developed massive epistaxis as a leading symptom of the ruptured intracavernous aneurysm. Since there are no reported pediatric cases with fatal epistaxis as a presenting sign, we would like to share our experience.
Adolescent↗
Expression of the structural proteins of dengue 2 virus and yellow fever virus by recombinant vaccinia viruses.
Vaccinia virus recombinants were constructed which contained cDNA sequences encoding the structural region of dengue 2 virus (PR159/S1 strain) or yellow fever virus (17D strain). The flavivirus cDNA sequences were expressed under the control of the vaccinia 7.5k early/late promotor. Cultured cells infected with these recombinants expressed immunologically reactive flavivirus structural proteins, precursor prM and E. These proteins appeared to be cleaved and glycosylated properly since they comigrated with the authentic proteins from dengue 2 virus- and yellow fever virus-infected cells. Mice immunized with the dengue/vaccinia recombinant showed a dengue-specific immune response that included low levels of neutralizing antibodies. Immunization of mice with the yellow fever/vaccinia recombinant was less effective at inducing an immune response to yellow fever virus and in only some of the mice were low titers of neutralizing antibodies produced.
Amino Acid Sequence↗
Synthesis and processing of the nonstructural polyproteins of several temperature-sensitive mutants of Sindbis virus.
We have examined the synthesis and processing of nonstructural polyproteins by several temperature-sensitive mutants of Sindbis virus, representing the four known RNA-minus complementation groups. Four mutants that possess mutations in the C-terminal domain of nonstructural protein nsP2 all demonstrated aberrant processing patterns when cells infected with these mutants were shifted from a permissive (30 degrees) to a nonpermissive (40 degrees) temperature. Mutants ts17, ts18, and ts24 showed severe defects in processing of nonstructural polyproteins at 40 degrees, whereas ts7 showed only a minor defect. In each case, cleavage of the bond between nsP2 and nsP3 was greatly reduced whereas cleavage between nsP1 and nsP2 occurred almost normally, giving rise to a set of polyprotein precursors not seen in wild-type-infected cells at this stage of infection. The nsP1 produced by these mutants was unstable and only small amounts could be detected in infected cells at the nonpermissive temperature. Submolar quantities of nsP2 were also present. We suggest that nsP1 and nsP2 may function as a complex and that free nsP1, and possibly nsP2, is degraded. Cleavage between nsP3 and nsP4 appeared to be normal in the mutants except in the case of ts17, where upon shift to 40 degrees P34 was unstable and nsP4 accumulated. We propose that the change in the P34/nsP4 ratio upon shift is responsible for the previously observed temperature sensitivity of subgenomic 26 S RNA synthesis in ts17 and for the failure of the mutant to regulate minus strand synthesis at 40 degrees. Other mutations tested, including ts21, which is found in the N-terminal half of nsP2, ts11, which has a mutation in nsP1, and ts6, which has a mutation in nsP4, all demonstrated nonstructural polyprotein processing indistinguishable from that in wild-type-infected cells. These results support our conclusion, based upon deletion mapping studies, that the C-terminal domain of nsP2 contains the nonstructural proteinase activity.
Animals↗
Mapping of RNA- temperature-sensitive mutants of Sindbis virus: complementation group F mutants have lesions in nsP4.
Temperature-sensitive (ts) mutants of Sindbis virus belonging to complementation group F, ts6, ts110, and ts118, are defective in RNA synthesis at the nonpermissive temperature. cDNA clones of these group F mutants, as well as of ts+ revertants, have been constructed. To assign the ts phenotype to a specific region in the viral genome, restriction fragments from the mutant cDNA clones were used to replace the corresponding regions of the full-length clone Toto1101 of Sindbis virus. These hybrid plasmids were transcribed in vitro by SP6 RNA polymerase to produce infectious transcripts, and the virus recovered was tested for temperature sensitivity. After the ts lesion of each mutant was mapped to a specific region of 400 to 800 nucleotides by this approach, this region of the cDNA clones of both the ts mutant and ts+ revertants was sequenced in order to determine the precise nucleotide change and amino acid substitution responsible for each mutation. Rescued mutants, which have a uniform background except for one or two defined changes, were examined for viral RNA synthesis and complementation to show that the phenotypes observed were the result of the mutations mapped. ts6 and ts110 had a single base substitution in nsP4, resulting in replacement of Gly by Glu at position 153 or position 324, respectively. It is of interest that nsP4 contains the Gly-Asp-Asp motif characteristic of a number of viral replicases, and this, together with the fact that all RNA synthesis in ts6-infected cells and, to a lesser extent, in ts110-infected cells shut off when the cells were shifted from a permissive to a nonpermissive temperature, suggests that nsP4 is the virus polymerase. ts118 was a double mutant. It contained a single base substitution in nsP2, resulting in replacement of Val by Ala at position 425 that resulted in the formation of minute plaques, but not in a reduction in the plaque number at the nonpermissive condition. The second change, a substitution of Gln by Arg in ts118 at residue 93 in nsP4, had little apparent phenotype on its own, but in combination with the change in nsP2 led to a ts phenotype. Thus, in each case the mutation responsible for the temperature sensitivity of the three known complementation group F mutants lay in nsP4. In addition, the result with ts118 suggests that nsP2 and nsP4 may interact with each other in a complex.
Amino Acid Sequence↗
Mapping of RNA- temperature-sensitive mutants of Sindbis virus: assignment of complementation groups A, B, and G to nonstructural proteins.
Four complementation groups of temperature-sensitive (ts) mutants of Sindbis virus that fail to make RNA at the nonpermissive temperature are known, and we have previously shown that group F mutants have defects in nsP4. Here we map representatives of groups A, B, and G. Restriction fragments from a full-length clone of Sindbis virus, Toto1101, were replaced with the corresponding fragments from the various mutants. These hybrid plasmids were transcribed in vitro by SP6 RNA polymerase to produce infectious RNA transcripts, and the virus recovered was tested for temperature sensitivity. After each lesion was mapped to a specific region, cDNA clones of both mutants and revertants were sequenced in order to determine the precise nucleotide change responsible for each mutation. Synthesis of viral RNA and complementation by rescued mutants were also examined in order to study the phenotype of each mutation in a uniform genetic background. The single mutant of group B, ts11, had a defect in nsP1 (Ala-348 to Thr). All of the group A and group G mutants examined had lesions in nsP2 (Ala-517 to Thr in ts17, Cys-304 to Tyr in ts21, and Gly-736 to Ser in ts24 for three group A mutants, and Phe-509 to Leu in ts18 and Asp-522 to Asn in ts7 for two group G mutants). In addition, ts7 had a change in nsP3 (Phe-312 to Ser) which also rendered the virus temperature sensitive and RNA-. Thus, changes in any of the four nonstructural proteins can lead to failure to synthesize RNA at a nonpermissive temperature, indicating that all four are involved in RNA synthesis. From the results presented here and from previous results, several of the activities of the nonstructural proteins can be deduced. It appears that nsP1 may be involved in the initiation of minus-strand RNA synthesis. nsP2 appears to be involved in the initiation of 26S RNA synthesis, and in addition it appears to be a protease that cleaves the nonstructural polyprotein precursors. It may also be involved in shutoff of minus-strand RNA synthesis. nsP4 appears to function as the viral polymerase or elongation factor. The functions of nsP3 are as yet unresolved.
Amino Acid Sequence↗