GTP binding by class II transactivator: role in nuclear import.

Class II transactivator (CIITA) is a global transcriptional coactivator of human leukocyte antigen-D (HLA-D) genes. CIITA contains motifs similar to guanosine triphosphate (GTP)-binding proteins. This report shows that CIITA binds GTP, and mutations in these motifs decrease its GTP-binding and transactivation activity. Substitution of these motifs with analogous sequences from Ras restores CIITA function. CIITA exhibits little GTPase activity, yet mutations in CIITA that confer GTPase activity reduce transcriptional activity. GTP binding by CIITA correlates with nuclear import. Thus, unlike other GTP-binding proteins, CIITA is involved in transcriptional activation that uses GTP binding to facilitate its own nuclear import.

Transcriptional scaffold: CIITA interacts with NF-Y, RFX, and CREB to cause stereospecific regulation of the class II major histocompatibility complex promoter.

Scaffold molecules interact with multiple effectors to elicit specific signal transduction pathways. CIITA, a non-DNA-binding regulator of class II major histocompatibility complex (MHC) gene transcription, may serve as a transcriptional scaffold. Regulation of the class II MHC promoter by CIITA requires strict spatial-helical arrangements of the X and Y promoter elements. The X element binds RFX (RFX5/RFXANK-RFXB/RFXAP) and CREB, while Y binds NF-Y/CBF (NF-YA, NF-YB, and NF-YC). CIITA interacts with all three. In vivo analysis using both N-terminal and C-terminal deletion constructs identified critical domains of CIITA that are required for interaction with NF-YB, NF-YC, RFX5, RFXANK/RFXB, and CREB. We propose that binding of NF-Y/CBF, RFX, and CREB by CIITA results in a macromolecular complex which allows transcription factors to interact with the class II MHC promoter in a spatially and helically constrained fashion.

Synthesis of "cactus" top-decorated aligned carbon nanotubes and their third-order nonlinear optical properties.

We report a new morphology of "cactus" top-decorated aligned carbon nanotubes grown by the PECVD method using pure C2H2 gas. Unlike most previous reports, no additional carrier gas is used for pretreatment. Carbon nanotubes can still grow and maintain the tubular structure underneath the "cactus" tops. It is proposed that the H atoms produced by the dissociation of C2H2 activate the catalyst nanoparticles. Scanning electron microscopy (SEM) shows that the top "cactus" morphology is composed of a large quantity of small nanosheets. Transmission electron microscopy (TEM) reveals the amorphous carbon nature of these "cactus" structures. The formation of these "cactus" structures is possibly due to covalent absorption and reconstruction of carbon atoms on the broken graphite layers of nanotubes produced by the strong ion bombardment under plasma. The third-order optical nonlinearities and nonlinear dynamics are also investigated. The third-order nonlinear susceptibility magnitude /chi(3)/ is found to be 2.2 x 10(-11) esu, and the relaxation process takes place in about 1.8 ps.

Fabrication of size-tunable gold nanoparticles array with nanosphere lithography, reactive ion etching, and thermal annealing.

Two-dimensional ordered arrays of gold (Au) nanoparticles were fabricated using two different variants of the nanosphere lithography technique. First, ordered arrays of polystyrene nanospheres on Si substrate were used as deposition masks through which gold films were deposited by electron beam evaporation. After the removal of the nanospheres, an array of triangular Au nanodisks was left on the Si substrate. After thermal annealing at increasing temperature, systematic shape transition of the nanostructures from original triangular Au nanodisks to rounded nanoparticles was observed. This approach allows us to systematically vary the size and morphology of the particles. In the second and novel technique, we made use of reactive ion etching to simultaneously reduce the dimension of the masking nanospheres and create arrays of nanopores on the substrate prior to the deposition of the Au films. These samples were subsequently annealed, which resulted in size-tunable and ordered Au nanoparticle arrays with the nanoparticles nested in the nanopores of the templated substrate. With the nanoparticles anchored in the nanopores, the substrate could be useful as a template for growth of other nanomaterials.

A pharmacological study of cefaclor in the newborn infant.

The absorption and excretion of cefaclor were studied in 10 newborn infants. A mean peak serum concentration of 7.7 microgram/ml was achieved at 1 hour after an oral dose of 7.5 mg/kg. It is concluded that cefaclor is a well absorbed and tolerated cephalosporin for use in newborn infants.

Primary T-cell-rich B-cell lymphoma in the kidney presenting with acute renal failure and a second malignancy.

Infiltration of the kidney is commonly found in lymphoma, but acute renal failure arising from bilateral renal infiltration is uncommon. Primary renal lymphoma may occur and is usually of B-cell lineage. It is rare for patients with lymphoma to develop acute renal failure as their initial clinical presentation. Recently, an association between primary renal lymphoma and a second primary malignancy has been reported. We describe the first case of a renal T-cell-rich B-cell lymphoma presenting as acute renal failure, which was associated with a second primary pulmonary malignancy.

Prognostic utility of visual evoked potentials in term asphyxiated neonates.

The prognosis for term infants with birth asphyxia is variable and does not correlate well with the acute clinical state. Diagnostic tools such as electroencephalography or imaging techniques have not been satisfactory predictors of outcome. Visual evoked potentials (VEPs) have changed acutely in birth asphyxia and may provide information for long-term prognosis. We studied VEPs serially in 25 term infants with documented birth asphyxia. The VEPs were classified into three categories: normal, mildly abnormal, or severely abnormal, and then compared to each infant's acute clinical state and outcome at age six months. Eight of the nine infants with normal or mildly abnormal VEPs were normal when examined subsequently. All the patients with severely abnormal VEPs died, or suffered severe neurologic sequelae. The VEPs demonstrated good correlation with neurodevelopmental outcomes in infants with birth asphyxia and may be useful prognostically.

Concomitant fracture of the coracoid and acromion after direct shoulder trauma.

Fractures of the acromial process or coracoid process of the scapula are rare. We present a combined fracture of the coracoid and acromion after direct trauma to the shoulder. An anteroposterior radiography with the central x-ray beam angled 25 degrees cephalad or an axillary lateral radiograph may be needed to detect these fractures.

DREAM2 challenge.

In the Dialogue for Reverse Engineering Assessments and Methods Conference (DREAM2) BCL6 target identification challenge, we were given a list of 200 genes and tasked to identify which ones are the true targets of BCL6 using an independent panel of gene-expression data. Initial efforts using conventional motif-scanning approaches to find BCL6 binding sites in the promoters of the 200 genes as a means of identifying BCL6 true targets proved unsuccessful. Instead, we performed a large-scale comparative study of multiple expression data under different conditions. Specifically, we employed a supervised learning approach that learns and models the expression patterns under different conditions and controls from a training collection of known BCL6 targets and randomly chosen decoys. Genes in the given list whose expression matches well with that of the training set of known BCL6 targets are more likely to be BCL6 targets. Using this approach, we are able to identify BCL6 targets with high accuracy, making us joint best performers of the challenge.

Viperin (cig5), an IFN-inducible antiviral protein directly induced by human cytomegalovirus.

Little is known about the mechanism by which IFNs inhibit human cytomegalovirus (HCMV) replication. Indeed, infection of fibroblasts with HCMV initiates the expression of a subset of type I IFN-inducible genes whose role in the infectious process is unclear. We describe here the identification of a cytoplasmic antiviral protein that is induced by IFNs, by HCMV infection, and by the HCMV envelope protein, glycoprotein B (gB). Stable expression of the protein in fibroblasts inhibits productive HCMV infection, down-regulating several HCMV structural proteins (gB, pp28, and pp65) known to be indispensable for viral assembly and maturation. We have named the protein viperin (for virus inhibitory protein, endoplasmic reticulum-associated, interferon-inducible). HCMV infection causes the redistribution of the induced viperin from its normal endoplasmic reticulum association, first to the Golgi apparatus and then to cytoplasmic vacuoles containing gB and pp28. Expression before HCMV infection reduces viperin redistribution from the endoplasmic reticulum to the Golgi apparatus and prevents vacuolar localization, perhaps reflecting the mechanism used by HCMV to evade the antiviral function.

Improvement in auditory and visual evoked potentials in jaundiced preterm infants after exchange transfusion.

Two preterm infants with peak serum bilirubin concentrations of 270 mumol/l and 200 mumol/l, respectively showed improvement in the wave peak latencies of the auditory and visual evoked potentials after exchange transfusion. The implications of this observation and the use of evoked potential recording in neonatal jaundice are discussed.

Activation and transdominant suppression of MHC class II and HLA-DMB promoters by a series of C-terminal class II transactivator deletion mutants.

The class II transactivator (CIITA) is a highly specific transcription factor that activates only genes known to be involved in the class II MHC processing pathway, including class II MHC, invariant chain, and HLA-DMA/B genes. In this work, we show the requirement of a new region in CIITA that is critical for its function. Deletion mutants lacking varying length of the C terminus show that the C-terminal 41 amino acids of CIITA are indispensable for the activation of both the conventional DRA and nonconventional DMB promoters. This region contains a highly charged stretch of amino acids that is homologous to a yeast transcription factor, repression activator protein-1 (RAP1)-interacting factor 1. Mutants lacking the C terminus were tested in a transdominant-negative assay to examine their capacity to block the wild-type CIITA function. Two of these deletion mutants suppressed the activity of endogenously expressed wild-type CIITA to activate both DRA and DMB promoters. It may be possible to utilize these mutants to modulate MHC class II and DM gene expression.

Importance of acidic, proline/serine/threonine-rich, and GTP-binding regions in the major histocompatibility complex class II transactivator: generation of transdominant-negative mutants.

The class II transactivator (CIITA) is a master transcription regulator of gene products involved in the exogenous antigen presentation pathway, including major histocompatibility complex (MHC) class II, invariant chain, and DM. An extensive analysis of the putative functional domains of CIITA is undertaken here to explore the action of CIITA. Antibodies to CIITA protein were produced to verify that these mutant proteins are expressed. Both acidic and proline/serine/threonine-rich domains are essential for class II MHC promoter activation. In addition, three guanine nucleotide-binding motifs are essential for CIITA activity. Of these mutants, two exhibited strong transdominant-negative functions. These two mutants provide a plausible approach to manipulate MHC class II expression and immune responses.

Neonatal typhoid fever.

Three infants of Pakistani immigrant mothers developed typhoid fever in the neonatal period. All three survived, but two became chronic excretors of Salmonella typhi. The risk of an outbreak of typhoid fever in a maternity unit or special care baby unit is emphasized.

Light transmission of phototherapy eyeshields.

Twelve types of phototherapy eyeshields showed peak light transmission of less than 0.1%, and none transmitted greater than 0.04% light in the 460 nm spectral region. Commercial eyeshields offered no advantage over locally made ones. The choice of shield may be less important than how it is secured over the infant's eyes.

Development of visual evoked potentials in neonates. A study using light emitting diode goggles.

We used a signal averager with light emitting diode goggles as the photostimulator to study the development of the visual evoked potentials in 40 normal neonates of between 23 and 42 weeks' gestation. All except two infants of less than 24 weeks' gestation had replicable visual evoked potentials. A negative peak of latency (mean (SD), 308 (21) msec) was present in all infants, but the development of the primary positive peak depended on maturity. Only infants of 37 weeks or more had a consistent positive peak of latency (mean (SD), 220 (22) msec). The practical simplicity and reliability of this technique has distinct advantages over previous conventional recording systems. Neonatal visual evoked potentials are shown to change with maturity.

The DMB promoter: delineation, in vivo footprint, trans-activation, and trans-dominant suppression.

The HLA-DM loci encode the heterodimeric unconventional class II MHC molecules that are coexpressed with conventional class II MHC molecules. DM molecules are essential for the proper formation and function of conventional class II MHC molecules. This report characterizes the DMB promoter both by in vivo footprint and by in vitro functional analysis and reveals a promoter structure similar to that of conventional class II MHC genes. DR-negative mutant cell lines selectively defective in the transcription factor or class II trans-activator (CIITA) were used to reveal a requirement for both these factors in DMB promoter activation. Complementation of defective cell lines with the appropriate transcription factor reconstituted DMB promoter activation. Further analysis with CIITA identified several mutant forms of CIITA that are trans-dominant-negative mutants, i.e., they suppressed DMB promoter activation by transfected and endogenous CIITA. These mutants may be used in abiological setting to down-regulate the function of DM in Ag processing.

A defect in the nuclear translocation of CIITA causes a form of type II bare lymphocyte syndrome.

The severe immunodeficiency type II bare lymphocyte syndrome (BLS) lacks class II MHC gene transcription. One defect from a complementation group A type II BLS patient is a 24 aa deletion in the MHC class II transactivator (CIITA). We show here that the molecular defect present in this protein is a failure of CIITA to undergo nuclear translocation. This defect was mapped to a position-dependent, novel nuclear localization sequence that cannot be functionally replaced by a classical NLS. Fusion of this 5 aa motif to an unrelated protein leads to nuclear translocation. Furthermore, this motif is not critical for transactivation function. This is a description of a genetic disease resulting from a novel defect in the subcellular localization of a transcriptional coactivator.

Concentrated autologous plasma protein: a biochemically neutral solder for tissue welding.

BACKGROUND AND OBJECTIVE: Xenographic or allographic serum protein solders used for laser welding may have immunologic and/or pathogenic complications. The objective of these studies was to develop a safe, autologous solder. STUDY DESIGN/MATERIALS AND METHODS: Five methods of preparing concentrated autologous plasma protein solder (CAPPS) were evaluated. Next, the CAPPS was evaluated via (1) thermal denaturation studies using differential scanning calorimetry, (2) tissue welding studies to characterize both acute and healing properties. RESULTS: The optimal concentration method to produce CAPPS rapidly was a dialysis method using chemical (osmotic) forces. The CAPPS showed similar denaturation profiles to serum albumin (SA) solders. Acutely, CAPPS provided comparable breaking strengths to SA solders. At 7 days, there was no significant difference in breaking strength or histology between 50% human SA solder and CAPPS (using a porcine skin model). CONCLUSIONS: These studies demonstrate that the CAPPS system provides acceptable acute and chronic properties for laser welding.