RNA movement between the nucleus and the cytoplasm

The past year has seen significant advances in our understanding of the mechanism of RNA movement between the nucleus and the cytoplasm. The emerging view is that proteins bind to and escort RNAs to their proper subcellular location. The discovery of peptide signals that target nuclear export and the identification of novel protein mediators of RNA export are examples of significant recent discoveries.     

Prenatal diagnosis: update addresses technological advances

This is the second in a series of articles provided to the readers of PENNSYLVANIA MEDICINE as an update from researchers and clinicians in this cutting-edge medical field. The series is partially sponsored through a grant from the Pennsylvania Department of Health to the University of Pittsburgh Department of Human Genetics.     

An in vitro model of traumatic neuronal injury: loading rate-dependent changes in acute cytosolic calcium and lactate dehydrogenase release

We developed a new in vitro model of neuronal injury using NT2-N cells to examine the effects of hydrodynamic loading rate on intraneuronal calcium dynamics and lactate dehydrogenase (LDH) release. Our apparatus consisted of a parallel disk viscometer which induced fluid shear stress with well-defined magnitudes and loading rates to cultured cells. We found that the deformation response of the cells was dependent on the severity of the insult, with increased cellular strains generated for higher shear stresses at a constant loading rate. Peak intracellular free calcium concentration correlated with strain, suggesting that mechanical deformation may regulate calcium response. Slowly applied fluid shear stress elicited no response, whereas high loading rates resulted in peak calcium increases 2.9 to 3.6 times baseline values as injury severity was increased. LDH release measured within 5 min after the insult correlated with loading rate. In addition, LDH release continued to increase out to 24 h following high loading rate conditions, demonstrating that the application of fluid shear stress led to prolonged cell damage. The acute response in NT2-N cells subjected to an insult with the CSID is dependent on the loading rate, and these results suggest that initial membrane deformation may trigger subsequent events.     

Interfacial reactions in the squeeze-cast (SAFFIL+C)/SAE 329 Al composite

Interfacial reactions in the squeeze-cast SAE 329 Al alloy reinforced with SAFFIL and carbon fibers were investigated using scanning and transmission electron microscopy and secondary ion mass spectroscopy. The SiO₂ layer added as a binder for the preform of SAFFIL fiber appeared to prevent excessive reaction between the fiber and the molten Al alloy during casting. In the as-cast composite, the reaction between the SiO₂ layer and the Al alloy is considered to produce MgO crystals in the vicinity of SAFFIL fiber. With subsequent heat treatment of the composite to the T6 condition, small crystals of MgO and MgAl₂O₄ were found to form at the surface of SAFFIL fiber as a result of interfacial reaction between the SiO₂ layer and the Al alloy. Even at the T6 condition, significant reactions between the Al alloy and SAFFIL fiber itself were not observed, indicating the effective role of the binder layer in suppressing the degradation of SAFFIL fiber by reaction with the Al alloy.     

Molecular follow-up of disease progression and interferon therapy in chronic myelocytic leukemia

We previously reported that the abl promoter (Pa) undergoes de novo DNA methylation in the course of chronic myelocytic leukemia (CML). The clinical implications of this finding are the subject of the present study in which samples of CML patients, including a group treated with interferon alpha (IFNalpha) were surveyed. The methylation status of the abl promoter was monitored by polymerase chain reaction (PCR) amplification of the Pa region after digestion with several site-methylation sensitive restriction enzymes. Some 74% of the DNA samples from blood and marrow drawn in the chronic phase were nonmethylated, similar to control samples from non-CML patients. The remaining 26% were partially methylated in the abl Pa region. The latter samples were derived from patients who were indistinguishable from the others on the basis of clinical presentation. Methylated samples were mostly derived from patients known to have a disease of longer duration (26 months v 7.5 months, P = .01). Samples of 30 IFNalpha-treated patients were sequentially analyzed in the course of treatment. Fifteen patients with no evidence of Pa methylation before treatment remained methylation-free. The remainder, who displayed Pa methylation before treatment, reverted to the methylation-free status. The outcome is attributed to IFNalpha therapy, as the Pa methylation status was not reversed in any of the patients treated with hydroxyurea. Methylation of the abl promoter indicates a disease of long-standing, most likely associated with a higher probability of imminent blastic transformation. It appears to predict the outcome of IFNalpha therapy far better than the cytogenetic response.     

Insulin and insulin-like growth factor-1 regulate tau phosphorylation in cultured human neurons

Hyperphosphorylated tau is the major component of paired helical filaments in neurofibrillary lesions associated with Alzheimer's disease. Hyperphosphorylation reduces the affinity of tau for microtubules and is thought to be a critical event in the pathogenesis of this disease. Recently, glycogen-synthase kinase-3 has been shown to phosphorylate tau in vitro and in non-neuronal cells transfected with tau. The activity of glycogen-synthase kinase-3 can be down-regulated in response to insulin or insulin-like growth factor-1 through the activation of the phosphatidylinositol 3-kinase pathway. We therefore hypothesize that insulin or insulin-like growth factor-1 may affect tau phosphorylation through the inhibition of glycogen-synthase kinase-3 in neurons. Using cultured human neuronal NT2N cells, we demonstrate that glycogen-synthase kinase-3 phosphorylates tau and reduces its affinity for microtubules and that insulin and insulin-like growth factor-1 stimulation reduces tau phosphorylation and promotes tau binding to microtubules. We further demonstrate that these effects of insulin and insulin-like growth factor-1 are mediated through the inhibition of glycogen-synthase kinase-3 via the phosphatidylinositol 3-kinase/protein kinase B signaling pathway.     

A novel method for elimination of line-current harmonics insingle-stage PFC switching regulators

This paper studies a particular single-stage power-factor-correction (PFC) switching regulator employing a discontinuous-conduction-mode (DCM) boost-input cell and a continuous-current-mode (CCM) forward output cell. Although this single-stage PFC regulator can provide a reasonably high power factor when its PFC stage is operating in discontinuous mode, substantial reduction in line-current harmonics is possible by applying a suitable frequency-modulation scheme. This paper derives a frequency-modulation scheme and proposes a practical implementation using a simple translinear analog circuit. A quantitative analysis on the total harmonic distortion (THD) of the line current when the circuit is subject to a limited range of frequency variations is presented along with some considerations for practical design. Experimental data obtained from a prototype confirms the effectiveness of the proposed frequency-modulation scheme. The proposed analog translinear circuit allows custom integrated circuit implementation, making it a viable low-cost solution to the elimination of line-current harmonics in switching regulators     

An epidemiologic, clinical, and therapeutic study of childhood Guillain-Barré syndrome in Kuwait: is it related to the oral polio vaccine?

We studied Guillain-Barré syndrome, affecting children 12 years old or less, throughout Kuwait, in the period between January 1, 1992, and March 31, 1997. Nineteen children had the diagnostic criteria of Guillain-Barré syndrome, with an overall annual incidence rate of 0.95/100,000 population at risk. Female patients outnumbered male patients with a sex ratio of 1.4:1. There was a clustering of cases in winter and spring and in the year 1996. The disease symptoms were relatively severe in our patients because only 16% (3 of 19) of them were able to walk at the height of their illness, whereas the rest were bed or chair bound or needed assisted ventilation. Two patients had the electrodiagnostic features of axonal neuropathy and both had residual deficits on follow-up, whereas the rest recovered fully. All the patients received intravenous immunoglobulin. The mean time to walk unaided was 23.5 days (range, 2-84 days) after intravenous immunoglobulin and excluding the two patients with axonal neuropathy, and full recovery was achieved in a mean time of 103 days (range, 30-300 days). Contrary to previous studies, we found no correlation between oral polio vaccine administration and Guillain-Barré syndrome in 2 successive years (1995 and 1996) during a nationwide campaign targeting children less than 5 years old.     

Micromachined piezoelectric force sensors based on PZT thin films

A micromachined lead zirconate titanate (PZT) force sensor for scanning force microscope (SFM) is conceptualized by its piezoelectricity. The fabrication procedure is interpreted, and mechanical characteristics of the micromachined PZT force sensors with various lengths are studied in this paper. A compact SFM is constructed by using the piezoelectric PZT sensor. A very clear image is taken by this SFM. The current study of the micromachined PZT force sensor can be considered as a breakthrough of design of SFM as well as a good example of integrated piezoelectric microdevices