An open-label study of repeated use of diazepam rectal gel (Diastat) for episodes of acute breakthrough seizures and clusters: safety, efficacy, and tolerance. North American Diastat Group.

Purpose: To assess safety of diazepam rectal gel (DZPRG) for control of acute seizures in epilepsy patients and to evaluate tolerance with repeated use of DZPRG at intervals of > or =5 days.

Methods: Subjects were persons with epilepsy, age 2 years or older, with seizure clusters or prolonged seizures. Onset of a treatable episode was defined; caregivers were trained to administer DZPRG and to monitor respiration, seizures, and adverse effects (AEs).

Molecular control of cell cycle progression in primary human hematopoietic stem cells: methods to increase levels of retroviral-mediated transduction.

Pluripotent hematopoietic stem cells (HSC) are the ideal targets for gene transfer because they can repopulate a sublethally irradiated recipient, giving rise to all lineages of blood cells. Thus, introduction of a corrected gene into HSC (stem cell gene therapy) should ensure persistent transmission of the gene. To date, the most efficient mode of gene delivery is via Moloney murine leukemia virus (MoMuLV)-based retroviral vectors which stably integrate into the genome of the target cell.

Multiple factors determine the selection of the ectodomain cleavage site of human cell surface macrophage colony-stimulating factor.

Human cell surface macrophage colony-stimulating factor (CSF-1256, M-CSF alpha) is converted to a soluble growth factor by a regulated proteolytic cleavage process at amino acid residues 157-159. We have previously shown that multiple factors specified by the juxtamembrane region determine the cleavage efficiency [Deng, P., Rettenmier, C.

Differential interaction of Crkl with Cbl or C3G, Hef-1, and gamma subunit immunoreceptor tyrosine-based activation motif in signaling of myeloid high affinity Fc receptor for IgG (Fc gamma RI).

Cbl-Crkl and Crkl-C3G interactions have been implicated in T cell and B cell receptor signaling and in the regulation of the small GTPase, Rap1. Recent evidence suggests that Rap1 plays a prominent role in the regulation of immunoreceptor tyrosine-based activation motif (ITAM) signaling. To gain insight into the role of Crkl in myeloid ITAM signaling, we investigated Cbl-Crkl and Crkl-C3G interactions following Fc gamma RI aggregation in U937IF cells.

The molecular basis of keloid and hypertrophic scar formation.

Excess scar formation secondary to traumatic or surgical injuries can have devastating consequences, ranging from body disfigurement to organ dysfunction. Hypertrophic scars and keloids are skin fibrotic conditions that can be caused by minor insults to skin, such as acne or ear piercing, or by severe injuries such as burns. Differences between keloids, hypertrophic scars and normal scars include distinct scar appearance, histologic morphology and cellular function in response to growth factors.

Safety of hospitalized ventilator-dependent children outside of the intensive care unit.

Objective: Hospitalization of clinically stable ventilator-dependent children in an intensive care unit (ICU) remains the standard in most pediatric centers. The aim of this study was to determine whether chronically ventilator-dependent children could be hospitalized safely in a non-ICU setting.

Methods: All ventilator-dependent children who were hospitalized on the pediatric wards at Childrens Hospital Los Angeles from December 1992 through June 1996 were reviewed retrospectively (N = 63) and compared with the general pediatric ward population hospitalized during the same period.

Saccharomyces cerevisiae Cdc6 stimulates Abf1 DNA binding activity.

In budding yeast Saccharomyces cerevisiae, an ARS binding factor 1 (Abf1) binds to the sequence-specific DNA element involved in DNA replication and transcription. We describe in this study how yeast Cdc6 protein stimulates Abf1 protein DNA binding activities. The Abf1 binding activity was reduced approximately 20-fold in a cdc6-1 mutant than in the wild-type strain.

Protein tyrosine phosphatase inhibitors in Fc gamma RI-induced myeloid oxidant signaling.

Fc-receptor stimulation in myeloid cells results in increased oxygen consumption, termed the respiratory burst, which is coupled to a rapid and transient increase in tyrosine phosphorylation of cellular proteins. In a previous paper in this journal we showed that the protein tyrosine phosphatase (PTPase) inhibitors sodium orthovanadate and phenylarsine oxide (PAO) block the Fc gamma RI-induced respiratory burst in interferon-gamma-differentiated U937 cells (U937IF) while augmenting the Fc gamma RI-induced tyrosine phosphorylation of cellular proteins. Herein we examine the effects of PTPase inhibitors on specific molecules involved in Fc gamma RI signaling.

Cyclin D1 antisense RNA destabilizes pRb and retards lung cancer cell growth.

To investigate the role of cyclin D1 in the regulation of lung cancer cell growth, we created five stably transfected cell lines carrying a cyclin D1 antisense construct. The transfected cells exhibited a marked decrease in the rate of cell growth, in contrast to the original lines (A549 and NCI-H441). The expression of several cell cycle-regulating proteins, including cyclin A, the cyclin-dependent kinases (cdk) 2 and cdk4, in addition to cyclin D1 itself, was markedly decreased.

The effects of bronchodilators on spontaneous ventilation and oxygen consumption in rhesus monkeys.

The effects of breathing normal saline, salmeterol, fenoterol, ipratropium bromide, or formoterol, and of i.v. infusion of theophylline on oxygen consumption (VO2), carbon dioxide production (VCO2), minute ventilation (VE), heart and respiratory rates, and end-tidal carbon dioxide tension (P(ET)CO2) have been defined in 10 anesthetized, intubated rhesus monkeys (mean age 7.

Recurrence patterns and anaplastic change in a long-term study of pilocytic astrocytomas.

Thirty-six cases of pilocytic astrocytomas treated at the Childrens Hospital of Los Angeles from 1984 through 1995 were reviewed. The mean age at initial presentation was 8 years (range 15 months to 14 years). These patients were followed for an average of 5.

Suitability of bone marrow from HIV-1-infected donors for retrovirus-mediated gene transfer.

Bone marrow samples from 21 human immunodeficiency virus type 1 (HIV-1)-infected subjects were evaluated for their suitability for retrovirus-mediated gene transduction with anti-HIV-1 genes. The percentages of CD34+ cells that could be isolated from the mononuclear fraction of bone marrow samples were determined. Fifteen of the 21 marrow samples had normal percentages of CD34+ cells isolated by immunomagnetic methods.

Severe bronchospasm during cardiopulmonary bypass.

Purpose: To describe the rare problem of severe bronchospasm occurring during cardiopulmonary bypass in a six-year-old-child.

Clinical Features: Severe bronchospasm became apparent on attempting to resume controlled ventilation prior to weaning from cardiopulmonary bypass. The patient had a previous history of asthma but was asymptomatic preoperatively.

Molecular genetics of small round cell tumors.

The small round cell tumors of children and young adults constitute part of a group of undifferentiated tumors, the precise diagnosis of which is often a challenge for the pathologist because of their uniform morphological appearance. Diagnostic cytogenetic analysis and identification of specific chromosomal abnormalities have been especially useful for the classification of some of these tumors. The cloning of the genes and molecular characterization of the associated genetic anomalies have led to the discovery of the mechanisms involved in their neoplastic transformation and identified a variety of tumor-specific molecular genetic markers.

Dynamics of TGF-beta 3 peptide activity during rat alveolar epithelial cell proliferative recovery from acute hyperoxia.

Hyperoxia causes a reproducible pattern of lung injury and recovery, characterized by proliferation of type II alveolar epithelial cells (AEC2) during the recovery phase. We measured TGF-beta peptide production by AEC2 and macrophages from lungs of adult male rats exposed to 100% oxygen for 48 h and then allowed to recover for up to 72 h in room air. TGF-beta peptide activity levels were measured using the PAI-1 promoter-luciferase mink lung epithelial cell assay and characterized with peptide specific inhibitory antibodies.

Obstructive sleep apnea in infants and young children.

The obstructive sleep apnea syndrome (OSAS), is a common cause of morbidity during childhood. Childhood OSAS usually stems from adenotonsillar hypertrophy. OSAS in infants is usually related to craniofacial anomalies.

Engineering, expression and renaturation of targeted TGF-beta fusion proteins.

This study reports the expression, purification, and renaturation of biologically active Transforming Growth Factor-beta 1 (TGF-beta 1) fusion proteins from Escherichia coli (E. coli). A prokaryotic expression vector was engineered to produce tripartite fusion proteins consisting of (i) a purification tag, (ii) a protease-sensitive linker/collagen binding domain, and (iii) a cDNA sequence encoding the active fragment of human TGF-beta 1.

Perioperative management with prolonged intravenous insulin infusion versus subcutaneous insulin in children with type I diabetes mellitus.

Our objective was to retrospectively evaluate glycemic excursion and insulin dosage in the perioperative period in children and adolescents with type I diabetes mellitus receiving prolonged intravenous insulin infusion for 2-3 days compared to conventional subcutaneous insulin treatment. A retrospective review of surgical admissions at the Children's Hospital of Los Angeles in patients with type I diabetes mellitus was conducted for the 3-year period from July 1989 to June 1992, to evaluate two treatment protocols used during that period. For the nine admissions in group 1, patients received 0.

Effects of overnight supplemental oxygen in obstructive sleep apnea in children.

Supplemental oxygen during sleep may be useful as a temporary palliative treatment in children with obstructive sleep apnea syndrome (OSAS) associated with significant hypoxemia. However, supplemental O2 may also blunt hypoxic ventilatory drive and worsen ventilation. To assess the safety of the use of supplemental O2 in children with OSAS, we studied 16 children ages 2-8 (mean: 4.

Arousal and cardiorespiratory responses to hypoxia in Prader-Willi syndrome.

Ventilatory responses to peripheral chemoreceptor stimuli are absent in patients with Prader-Willi syndrome (PWS) during wakefulness. Because arousal from sleep after rapidly developing hypoxia may require intact peripheral chemoreceptor function, we hypothesized that blunted hypoxic arousal responses during sleep Stage 3/4 would be present in PWS. Thirteen patients with PWS (mean age, 23.

Intratracheal pulmonary ventilation versus conventional mechanical ventilation in a rabbit model of surfactant deficiency.

Intratracheal pulmonary ventilation (ITPV) enhances the clearance of CO2 from dead space and lungs by a bias flow of gas administered in the distal trachea. ITPV flow is continuously administered through a separate catheter placed within an endotracheal tube (ETT). After exiting from catheter's tip in the distal trachea, the flow of gas is redirected outward away from the lungs.

Hypercapnic arousal responses in Prader-Willi syndrome.

Study Objective: Prader-Willi syndrome (PWS) is characterized by a number of abnormalities of hypothalamic function, such as hyperphagia, short stature, temperature instability, hypogonadotropic hypogonadism, and neurosecretory growth hormone deficiency. Patients with PWS are reported to have sleep-disordered breathing and have blunted hypercapnic ventilatory responses secondary to abnormal peripheral chemoreceptor function. Thus, we hypothesized that hypercapnic arousal responses would be abnormal in PWS.

Ventilatory response to consecutive short hypercapnic challenges in children with obstructive sleep apnea.

In healthy adults, a ventilatory pattern characterized by progressively increased tidal volume (VT), and decreased respiratory rate (RR) accompany repeated short hypercapnic ventilatory challenges, while minute ventilation (VE) remains constant. We hypothesized that the peculiar ventilatory pattern seen in adults would be blunted in children with obstructive sleep apnea syndrome (OSAS) who undergo comparable intermittent or chronic alveolar PCO2 elevation. We measured ventilatory responses to five challenges of 2-min duration (C1-C5) with 5% CO2 in O2, separated by 5-min room-air breathing intervals (R1-R4), in nine children with OSAS and in eight age-, sex-, and body mass index-matched controls.

Lipopolysaccharide stimulates the tyrosine phosphorylation of mitogen-activated protein kinases p44, p42, and p41 in vascular endothelial cells in a soluble CD14-dependent manner. Role of protein tyrosine phosphorylation in lipopolysaccharide-induced stimulation of endothelial cells.

Vascular endothelial cell (EC) injury or activation by LPS plays a critical role in the pathogenesis of Gram-negative meningitis and endotoxic shock. EC do not express membrane CD14, but respond to LPS in a soluble CD14-dependent manner. The signal transduction mechanisms involved in LPS-induced EC responses are largely unknown.

Engraftment of gene-modified umbilical cord blood cells in neonates with adenosine deaminase deficiency.

Haematopoietic stem cells in umbilical cord blood are an attractive target for gene therapy of inborn errors of metabolism. Three neonates with severe combined immunodeficiency were treated by retroviral-mediated transduction of the CD34+ cells from their umbilical cord blood with a normal human adenosine deaminase complementary DNA followed by autologous transplantation. The continued presence and expression of the introduced gene in leukocytes from bone marrow and peripheral blood for 18 months demonstrates that umbilical cord blood cells may be genetically modified with retroviral vectors and engrafted in neonates for gene therapy.