Human membrane metallo-endopeptidase-like protein degrades both beta-amyloid 42 and beta-amyloid 40.

Beta-amyloid (Abeta) degrading endopeptidases are thought to protect against Alzheimer's disease (AD) and are potentially therapeutic. Of particular interest are endopeptidases that are blocked by thiorphan and phosphoramidon (T/P), as these inhibitors rapidly induce Abeta deposition in rodents. Neprilysin (NEP) is the best known target of T/P; however neprilysin knockout results in only modest Abeta increases insufficient to induce deposition. Therefore, other endopeptidases targeted by T/P must be critical for Abeta catabolism. Another candidate is the T/P sensitive membrane metallo-endopeptidase-like protein (MMEL), a close homolog of neprilysin. The endopeptidase properties of beta and gamma splice forms of human MMEL were determined in HEK293T cells transduced with the human cDNAs for the two splice forms; this showed degradation of both Abeta(42) and Abeta(40) by hMMEL-beta but not hMMEL-gamma. hMMEL-beta activity was found at the extracellular surface with no significant secreted activity. hMMEL-gamma was not expressed at the extracellular surface. Finally, it was found that hMMEL cleaves Abeta near the alpha-secretase site (producing Abeta(1-17)>>Abeta(1-16)). These data establish hMMEL as a mediator of Abeta catabolism and raise the possibility of its involvement in the etiology of AD and as a target for intervention.

Human immunodeficiency virus type 1 Vpr induces apoptosis in human neuronal cells.

Human immunodeficiency virus type 1 (HIV-1) infection of the central nervous system (CNS) causes AIDS dementia complex (ADC) in certain infected individuals. Recent studies have suggested that patients with ADC have an increased incidence of neuronal apoptosis leading to neuronal dropout. Of note, a higher level of the HIV-1 accessory protein Vpr has been detected in the cerebrospinal fluid of AIDS patients with neurological disorders. Moreover, extracellular Vpr has been shown to form ion channels, leading to cell death of cultured rat hippocampal neurons. Based on these previous findings, we first investigated the apoptotic effects of the HIV-1 Vpr protein on the human neuronal precursor NT2 cell line at a range of concentrations. These studies demonstrated that apoptosis induced by both Vpr and the envelope glycoprotein, gp120, occurred in a dose-dependent manner compared to protein treatment with HIV-1 integrase, maltose binding protein (MBP), and MBP-Vpr in the undifferentiated NT2 cells. For mature, differentiated neurons, apoptosis was also induced in a dose-dependent manner by both Vpr and gp120 at concentrations ranging from 1 to 100 ng/ml, as demonstrated by both the terminal deoxynucleotidyltransferase (Tdt)-mediated dUTP-biotin nick end labeling and Annexin V assays for apoptotic cell death. In order to clarify the intracellular pathways and molecular mechanisms involved in Vpr- and gp120-induced apoptosis in the NT2 cell line and differentiated mature human neurons, we then examined the cellular lysates for caspase-8 activity in these studies. Vpr and gp120 treatments exhibited a potent increase in activation of caspase-8 in both mature neurons and undifferentiated NT2 cells. This suggests that Vpr may be exerting selective cytotoxicity in a neuronal precursor cell line and in mature human neurons through the activation of caspase-8. These data represent a characterization of Vpr-induced apoptosis in human neuronal cells, and suggest that extracellular Vpr, along with other lentiviral proteins, may increase neuronal apoptosis in the CNS. Also, identification of the intracellular activation of caspase-8 in Vpr-induced apoptosis of human neuronal cells may lead to therapeutic approaches which can be used to combat HIV-1-induced neuronal apoptosis in AIDS patients with ADC.

Outcome of infants with birth weights less than 1000 g with respiratory distress syndrome treated with high-frequency ventilation and surfactant replacement therapy.

OBJECTIVE: To compare outcomes in premature infants with respiratory distress syndrome who received surfactant replacement therapy and were treated with either high-frequency or conventional mechanical ventilation. DESIGN: Retrospective chart review of patient series. SETTING: Tertiary academic medical center. PATIENTS: One hundred fourteen extremely low-birth-weight infants (< 1000 g) with respiratory distress syndrome treated with surfactant replacement therapy, consecutively admitted to the neonatal intensive care unit between September 1989 and August 1992. INTERVENTIONS: Treatment with either high-frequency ventilation (n = 46) or conventional mechanical ventilation (n = 68) after surfactant replacement therapy. MAIN OUTCOME MEASURES: Intraventricular hemorrhage and neurodevelopmental status. RESULTS: Infants who received high-frequency ventilation had significantly lower birth weights and were more premature than infants receiving conventional mechanical ventilation. Despite this, patients ventilated with high frequency had similar incidences of intraventricular hemorrhage and impaired neurodevelopmental outcomes when compared with the conventionally ventilated patients. As expected, the smaller and more premature infants receiving high-frequency ventilation required a longer duration of respiratory support (mechanical ventilation and nasopharyngeal continuous positive airway pressure). Additionally, multiple logistic regression analysis to control for differences in birth weight and gestational age between the two groups revealed a significant association between the combined use of high-frequency ventilation and antenatal corticosteroids and the absence of either intraventricular hemorrhage or pneumothorax. CONCLUSION: We conclude that high-frequency ventilation combined with surfactant therapy is as safe as conventional mechanical ventilation combined with surfactant therapy for treating respiratory distress syndrome in extremely low-birth-weight infants (< 1000 g) and does not increase the risk of either intraventricular hemorrhage or abnormal neurodevelopmental outcome.