Molecular biology of cell division.

Different domains of the SV40 A gene have different functions, such as viral DNA replication, cell DNA replication, and stimulation of cellular RNA synthesis. The sequences in the SV40 A gene that are critical for the induction of cell DNA synthesis lie on the map between nucleotide 4360 and nucleotide 4001, a stretch of 360 nucleotides coding for 120 of the 708 amino acids of the large T antigen. The sequences critical for stimulation of rRNA synthesis lie on the map further downstream, between nucleotides 3827 and 3506, thus indicating that the signals for growth in size and for cell DNA replication can be dissociated. Methylation of the SV40 A gene at multiple ECoRI* sites has no effect on its expression. However, methylation of the HSV-TK gene at one single ECoRI site 70 base pairs upstream from the cap site inhibits its expression. The results indicate that methylation of genes affects their expression, but only when methylation occurs at specific sites.

Digital radiology using active matrix readout of amorphous selenium: radiation hardness of cadmium selenide thin film transistors.

A flat-panel x-ray imaging detector using active matrix readout of amorphous selenium (a-Se) is being investigated for digital radiography and fluoroscopy. The active matrix consists of a two-dimensional array of thin film transistors (TFTs). Radiation penetrating through the a-Se layer will interact with the TFTs and it is important to ensure that radiation induced changes will not affect the operation of the x-ray imaging detector. The methodology of the present work is to investigate the effects of radiation on the characteristic curves of the TFTs using individual TFT samples made with cadmium selenide (CdSe) semiconductor. Four characteristic parameters, i.e., threshold voltage, subthreshold swing, field effect mobility, and leakage current, were examined. This choice of parameters was based on the well established radiation damage mechanisms for crystalline silicon metal-oxide-semiconductor field-effect transistors (MOSFETs), which have a similar principle of operation as CdSe TFTs. It was found that radiation had no measurable effect on the leakage current and the field effect mobility. However, radiation shifted the threshold voltage and increased the subthreshold swing. But even the estimated lifetime dose (50 Gy) of a diagnostic radiation detector will not affect the normal operation of an active matrix x-ray detector made with CdSe TFTs. The mechanisms of the effects of radiation will be discussed and compared with those for MOSFETs and hydrogenated amorphous silicon (a-Si:H) TFTs.

Digital radiology using active matrix readout of amorphous selenium: detectors with high voltage protection.

A flat-panel x-ray imaging detector is being investigated for digital radiography and fluoroscopy. The detector uses a layer of amorphous selenium (a-Se) to convert x rays to a charge image, which is then electronically read out with a two-dimensional array of thin film transistors (TFTs). In order to sensitize the a-Se layer to x rays, a high voltage (of the order of several thousand volts) is applied to its top surface. The TFTs, which are at the bottom surface of the a-Se layer, are not subjected to any high voltage under normal radiological operational conditions since the pixel potential is < 10 V. However under a fault condition where these two events occur simultaneously: (1) suspended detector scan; and (2) an x-ray exposure more than ten times higher than normal, the voltage on the TFTs could rise to a damaging value. This paper describes a method for protecting the TFTs from high voltage damage under this fault condition. It employs a dual-gate TFT structure, one gate is for scanning control and the other is connected to the pixel electrode for high voltage protection. Before the pixel potential reaches a damaging value, the protection gate turns on the TFT automatically and drains excess charge away from the pixel thus providing a safe pixel saturation potential. In this paper, the characteristic curves of dual-gate TFTs are studied both theoretically and experimentally. The pixel x-ray response for imaging detectors with high voltage protection are predicted, and it is shown that with practical TFT designs the detector can provide a safe pixel saturation potential as well as satisfy the dynamic range required for diagnostic x-ray imaging applications.

Digital radiology using active matrix readout of amorphous selenium: construction and evaluation of a prototype real-time detector.

The goal of the present work is to develop a large area, flat-panel solid-state detector for both digital radiography and fluoroscopy. The proposed detector employs a photoconductive layer of amorphous selenium (a-Se) to convert x rays into charge. The charge image formed by the a-Se layer is electronically read out in situ using a two dimensional array of thin film transistors (TFTs), or active matrix. Since the active matrix readout is capable of producing x-ray images in real-time, it can potentially be applied in both radiography and fluoroscopy. In this paper, the imaging performance of this concept is investigated using a prototype x-ray imaging detector. The designs for the active matrix, the peripheral electronic circuits, and the image acquisition system are described. Measurements of x-ray imaging properties of the prototype detector, i.e., x-ray sensitivity, presampling modulation transfer function (MTF), and noise power spectrum (NPS), were performed, and from which the spatial frequency dependent detective quantum efficiency (DQE) of the prototype was derived. The experimental results are in agreement with the results of our theoretical analysis. The factors affecting the imaging performance and methods of improvement in the future are discussed.

Broadening our understanding of clinical quality: from attribution error to situated cognition.

The tendency to overestimate the influence of personal characteristics on outcomes, and to underestimate the influence of situational factors, is known as the fundamental attribution error. We argue that medical-education researchers and policy makers may be guilty of this error in their quest to understand clinical quality. We suggest that to truly understand clinical quality, they must examine situational factors, which often have a strong influence on the quality of clinical encounters.

A curricular divide: basic pharmacology vs. clinical pharmacology.

Two current surveys of graduating medical students and/or recently graduated physicians from the United Kingdom raise significant concerns about the quality and quantity of clinical pharmacology instruction. Compared with the relative abundance of information about the instruction of basic pharmacology in US medical schools, little information exists about similar curricular content of clinical pharmacology. Here, we highlight this lack of information and encourage clinical pharmacology educators to address this curricular divide.

Effect of methylation on expression of microinjected genes.

The cloned genes for the simian virus 40 large tumor antigen and for herpes simplex virus (HSV) thymidine kinase (TK) were methylated with EcoRI methylase. The genes were microinjected into the nuclei of TK-deficient (tk-) cells, and expression of the genes was determined by immunofluorescence staining for the simian virus 40 large tumor antigen and by [3H]thymidine incorporation followed by autoradiography for HSV TK. We found that methylation of the simian virus 40 gene, under EcoRI or EcoRI* conditions, resulting in methylation at sites within the gene and in the surrounding sequences, has no effect on expression of the large tumor antigen when the gene is manually microinjected into mammalian nuclei. However, methylation of the HSV tk gene at the two EcoRI sites markedly reduces or abolishes the expression of this gene. One of the EcoRI sites of HSV tk is approximately 1.1 kilobases downstream from the 3' end of the gene and is believed to have no regulatory function in the expression of the tk gene. The other EcoRI site is 79 base pairs upstream from the 5' end of the gene and has considerable homology to the regulatory sequence proposed by [Benoist C., O'Hare, K., Breathnach, R., & Chambon, P. (1980) Nucleic Acids Res. 8, 127-142]. Our results are direct proof that methylation can alter gene expression and also that the effect depends strictly on the sites that are methylated.

Flat-panel digital radiology with amorphous selenium and active-matrix readout.

Future imaging techniques will be capable of high image quality, fast acquisition, compactness, and versatile operation. A flat-panel imager that is expected to achieve these goals is under development. It consists of a thin layer of amorphous selenium that converts x rays directly to an electric charge and a thin-film electronic circuit, or active matrix, to read out the electronic signal directly to a computer host. The advantages of amorphous selenium include high resolution and low noise without loss of signal strength. The advantages of the active matrix are real-time readout, flexible design parameters, and compactness of the readout structures. A prototype of this imager has been built and operated, and initial images (of an x-ray test bar pattern and a hand phantom) have been acquired. Although the prototype was built to test scientific principles and many possibilities for optimization remain, the images already possess the quality necessary for many radiographic procedures. A large range of current and new applications exist for this imager.

Microinjection of RNA polymerase II corrects the temperature-sensitive defect of tsAF8 cells.

tsAF8 cells are a temperature-sensitive (ts) mutant of BHK cells that arrest in the G1 phase of the cell cycle at the non-permissive temperature of 40.6 degrees C. Previous reports had suggested that the temperature-sensitivity of these cells was based on a defect in either the synthesis, assembly or turnover of RNA polymerase II. We now show that the direct microinjection of purified RNA polymerase II into nuclei of tsAF8 cells corrects the ts defect and allows these cells to enter the S phase of the cell cycle.