A targetable HB-EGF-CITED4 axis controls oncogenesis in lung cancer.

Aberrant epidermal growth factor (EGF) receptor (EGFR) signaling contributes to neoplastic initiation and progression in lung. Mutated EGFR has become as an important therapeutic target in lung cancer, whereas targeted treatment is not available for wild-type EGFR or its ligands. In this study, we found that heparin-binding (HB)-EGF, a member of the EGF family, was highly expressed in a subset of lung cancer, proliferation of which was dependent on HB-EGF signaling. Silencing of HB-EGF with RNA interference inhibited cell cycle progression in lung cancer cells. We observed that, upon HB-EGF induction, CITED4 was induced through a signal transducer and activator of transcription 3 (STAT3)-dependent pathway, regulating cell proliferation. CITED4 interacted with MYC and potentiated MYC-mediated transactivation of the CCND1 promoter, leading to cell cycle progression. Correlation analysis revealed that HB-EGF and CITED4 were significantly positively associated in primary lung tumors, and expression of HB-EGF predicted a poor survival outcome in patients. In vitro and in vivo experiments revealed that pharmacological inhibition of HB-EGF with CRM197 significantly attenuated tumor cell growth. Thus, CITED4 functions as a molecular switch in HB-EGF-induced growth control, and HB-EGF provides a novel therapeutic target for lung cancer intervention.

Designer germanium quantum dot phototransistor for near infrared optical detection and amplification.

We demonstrated a unique CMOS approach for the production of a high-performance germanium (Ge) quantum dot (QD) metal-oxide-semiconductor phototransistor. In the darkness, low off-state leakage (Ioff ∼ 0.27 pA µm(-2)), a high on-off current ratio (Ion/Ioff ∼ 10(6)), and good switching behaviors (subthreshold swing of 175 mV/dec) were measured on our Ge-QD phototransistor at 300 K, indicating good hetero-interfacial quality of the Ge-on-Si. Illumination makes a significant enhancement in the drain current of Ge QD phototransistors when biased at both the on- and off-states, which is a great benefit from Ge QD-mediated photoconductive and photovoltaic effects. The measured photocurrent-to-dark-current ratio (Iphoto/Idark) and the photoresponsivities from the Ge QD phototransistor are as high as 4.1 × 10(6) and 1.7 A W(-1), respectively, under an incident power of 0.9 mW at 850 nm illumination. A superior external quantum efficiency of 240% and a very fast temporal response time of 1.4 ns suggest that our Ge QD MOS phototransistor offers great promise as optical switches and transducers for Si-based optical interconnects.

Human Factors in the Large: Experiences from Denmark, Finland and Canada in Moving Towards Regional and National Evaluations of Health Information System Usability. Contribution of the IMIA Human Factors Working Group.

OBJECTIVES: The objective of this paper is to explore approaches to understanding the usability of health information systems at regional and national levels. METHODS: Several different methods are discussed in case studies from Denmark, Finland and Canada. They range from small scale qualitative studies involving usability testing of systems to larger scale national level questionnaire studies aimed at assessing the use and usability of health information systems by entire groups of health professionals. RESULTS: It was found that regional and national usability studies can complement smaller scale usability studies, and that they are needed in order to understand larger trends regarding system usability. Despite adoption of EHRs, many health professionals rate the usability of the systems as low. A range of usability issues have been noted when data is collected on a large scale through use of widely distributed questionnaires and websites designed to monitor user perceptions of usability. CONCLUSION: As health information systems are deployed on a widespread basis, studies that examine systems used regionally or nationally are required. In addition, collection of large scale data on the usability of specific IT products is needed in order to complement smaller scale studies of specific systems.

Size tunable Ge quantum dots for near-ultraviolet to near-infrared photosensing with high figures of merit.

We report a unique approach for the inclusion of size-tunable (7-50 nm), spherical Ge quantum dots (QDs) into gate stacks of metal-oxide-semiconductor (MOS) diodes, through selective oxidation of SiGe layers over the buffer layer of Si3N4 deposited over the Si substrate. In this complementary MOS (CMOS)-compatible approach, we successfully realized high performance nm scale Ge-QD MOS photodetectors with high figures of merit of low dark current density (1.5 × 10(-3) mA cm(-2)), superior photo-current-to-dark current ratio (13 500), high photoresponsivity (2.2 A W(-1)), and fast response time (5 ns), which are ready for direct integration with Si CMOS electronic circuits. Most importantly, the detection wavelength of the Ge QDs is tunable from near infrared to near ultraviolet by reducing the QD size from 50 to 7 nm as well as the optimal photoresponsivity is tailored by the Ge QD size and the effective thickness of gate dielectrics.

Moving mobile: using an open-sourced framework to enable a web-based health application on touch devices.

Computer devices using touch-enabled technology are becoming more prevalent today. The application of a touch screen high definition surgical monitor could allow not only high definition video from an endoscopic camera to be displayed, but also the display and interaction with relevant patient and health related data. However, this technology has not been quickly embraced by all health care organizations. Although traditional keyboard or mouse-based software programs may function flawlessly on a touch-based device, many are not practical due to the usage of small buttons, fonts and very complex menu systems. This paper describes an approach taken to overcome these problems. A real case study was used to demonstrate the novelty and efficiency of the proposed method.

A proof of concept for assessing emergency room use with primary care data and natural language processing.

OBJECTIVE: The objective of this study was to undertake a proof of concept that demonstrated the use of primary care data and natural language processing and term extraction to assess emergency room use. The study extracted biopsychosocial concepts from primary care free text and related them to inappropriate emergency room use through the use of odds ratios. METHODS: De-identified free text notes were extracted from a primary care clinic in Guelph, Ontario and analyzed with a software toolkit that incorporated General Architecture for Text Engineering (GATE) and MetaMap components for natural language processing and term extraction. RESULTS: Over 10 million concepts were extracted from 13,836 patient records. Codes found in at least 1% percent of the sample were regressed against inappropriate emergency room use. 77 codes fell within the realm of biopsychosocial, were very statistically significant (p < 0.001) and had an OR > 2.0. Thematically, these codes involved mental health and pain related concepts. CONCLUSIONS: Analyzed thematically, mental health issues and pain are important themes; we have concluded that pain and mental health problems are primary drivers for inappropriate emergency room use. Age and sex were not significant. This proof of concept demonstrates the feasibly of combining natural language processing and primary care data to analyze a system use question. As a first work it supports further research and could be applied to investigate other, more complex problems.

Analyzing primary care data to characterize inappropriate emergency room use.

Primary care data represents the complete biopsychosocial profile of the patient and has recently become available for secondary analysis in Ontario. This study involved extracting de-identified primary care data and analyzing it with natural language processing to extract UMLS codes. These codes were used to statistically relate inappropriate emergency room use to biopsychosocial concepts with logistic regression. The concepts of pain and mental health were statistically significant. This technique demonstrates a creative use of primary care data. It could be used to analyze system use in other health care settings.

Application of the Apriori algorithm for adverse drug reaction detection.

The objective of this research is to assess the suitability of the Apriori association analysis algorithm for the detection of adverse drug reactions (ADR) in health care data. The Apriori algorithm is used to perform association analysis on the characteristics of patients, the drugs they are taking, their primary diagnosis, co-morbid conditions, and the ADRs or adverse events (AE) they experience. This analysis produces association rules that indicate what combinations of medications and patient characteristics lead to ADRs. A simple data set is used to demonstrate the feasibility and effectiveness of the algorithm.

Free anterolateral thigh flap for extremity reconstruction: clinical experience and functional assessment of donor site.

From August of 1995 through July of 1998, 38 free anterolateral thigh flaps were transferred to reconstruct soft-tissue defects. The overall success rate was 97 percent. Among 38 anterolateral thigh flaps, four were elevated as cutaneous flaps based on the septocutaneous perforators. The other 34 were harvested as myocutaneous flaps including a cuff of vastus lateralis muscle (15 to 40 cm3), either because of bulk requirements (33 cases) or because of the absence of a septocutaneous perforator (one case). However, vastus lateralis muscle is the largest compartment of the quadriceps, which is the prime extensor of the knee. Losing a portion of the vastus lateralis muscle may affect knee stability. Objective functional assessments of the donor sites were performed at least 6 months postoperatively in 20 patients who had a cuff of vastus lateralis muscle incorporated as part of the myocutaneous flap; assessments were made using a kinetic communicator machine. The isometric power test of the ratios of quadriceps muscle at 30 and 60 degrees of flexion between donor and normal thighs revealed no significant difference (p > 0.05). The isokinetic peak torque ratio of the quadriceps and hamstring muscles, including concentric and eccentric contraction tests, showed no significant difference (p > 0.05), except the concentric contraction test of the quadriceps muscle, which revealed mild weakness of the donor thigh (p < 0.05). In summary, the functional impairment of the donor thighs was minimal after free anterolateral thigh myocutaneous flap transfer.

Static wrist position associated with least median nerve compression: sonographic evaluation.

OBJECTIVE: To determine the wrist angle that produces the least compression to the median nerve and to evaluate the usefulness of sonography in determining the optimal position. DESIGN: Seventeen wrists of 17 healthy volunteers received dynamic, high-frequency (8 MHz), high-resolution sonography with the wrist splinted at various positions: 15 degrees of flexion, neutral position, and 15 degrees and 30 degrees of extension. The morphologic changes of the median nerve were evaluated with the wrist positioned at various angles. RESULTS: The neutral position caused significantly lower compression of the median nerve than it did in the other positions. However, in some cases, the lowest pressure was found when the wrist was fixed in 15 degrees of flexion or 15 degrees of extension. Because median nerve compression may decrease the anteroposterior diameter, increase the transverse diameter, and decrease the cross-sectional area, greater anteroposterior diameter, lower flattening ratio (transverse diameter/anteroposterior diameter), and greater cross-sectional area were considered to indicate lower median nerve compression. CONCLUSIONS: Neutral position of the wrist is the best position with the least median nerve compression in most individuals. However, the optimal position may vary from person to person. Sonographic examination can help to determine the splint position that results in the lowest median nerve compression.

Nickel compounds are novel inhibitors of histone H4 acetylation.

Environmental factors influence carcinogenesis by interfering with a variety of cellular targets. Carcinogenic nickel compounds, although generally inactive in most gene mutation assays, induce chromosomal damage in heterochromatic regions and cause silencing of reporter genes when they are located near telomere or heterochromatin in either yeast or mammalian cells. We studied the effects of nickel on the lysine acetylation status of the NH2-terminal region of histone H4. At nontoxic levels, nickel decreased the levels of histone H4 acetylation in vivo in both yeast and mammalian cells, affecting only lysine 12 in mammalian cells and all of the four lysine residues in yeast. In yeast, lysine 12 and 16 were more greatly affected than lysine 5 and 8. Interestingly, a histidine Ni2+ anchoring site is found at position 18 from the NH2-terminal tail of H4. Nickel was also found to inhibit the acetylation of H4 in vitro using purified recombinant histone acetyltransferase. To our knowledge, this is the first agent shown to decrease histone H4 acetylation at nontoxic levels.

Gcn4 activator targets Gcn5 histone acetyltransferase to specific promoters independently of transcription.

Histone acetylation correlates well with transcriptional activity, and histone acetyltransferases (HATs) selectively regulate subsets of target genes by mechanisms that remain unclear. Here, we provide in vivo evidence that the yeast transcriptional activator Gcn4 recruits Gcn5 HAT complexes to selective promoters positioned in natural or ectopic locations, thereby creating local domains of histone H3 hyperacetylation and subsequent transcriptional activation. A significant portion of the Gcn4-targeted histone acetylation by Gcn5 is independent of transcriptional activity. These observations provide strong evidence for promoter-selective, targeted histone acetylation by Gcn5 that facilitates transcription in a causal fashion. In addition, Gcn5 also functions in an untargeted manner to acetylate H3 on a genome-wide scale.

In vivo cross-linking and immunoprecipitation for studying dynamic Protein:DNA associations in a chromatin environment.

Chromatin structure plays important roles in regulating many DNA-templated processes, such as transcription, replication, and recombination. Considerable progress has recently been made in the identification of large, multisubunit complexes dedicated to these nuclear processes, all of which occur on nucleosomal templates. Mapping specific genomic loci relative to the position of selectively modified or unique histone variants or nonhistone components provides valuable insights into how these proteins (and their modifications) function in their normal chromatin context. Here we describe a versatile and high-resolution method which involves two basic steps: (1) in vivo formaldehyde cross-linking of intact cells followed by (2) selective immunoprecipitation of protein-DNA complexes with specific antibodies. This method allows for detailed analyses of protein-DNA interactions in a native chromatin environment. Recently, this technique has been successfully employed to map the boundaries of specifically modified (e.g., acetylated) histones along target genes, to define the cell cycle-regulated assembly of origin-dependent replication and centromere-specific complexes with remarkable precision, and to map the in vivo position of reasonably rare transcription factors on cognate DNA sites. Thus, the basic chromatin immunoprecipitation technique is remarkably versatile and has now been used in a wide range of cell types, including budding yeast, fly, and human cells. As such, it seems likely that many more studies, centered around chromatin structure and protein-DNA interactions in its native setting, will benefit from this technique. In this article, a brief review of the history of this powerful approach and a discussion of the basic method are provided. Procedures for protein recovery as well as limitations and extensions of the method are also presented.

Cell cycle-regulated histone acetylation required for expression of the yeast HO gene.

Expression of the yeast HO gene in late G1 of the cell cycle requires the SWI/SNF chromatin remodeling complex, the Gcn5p histone acetyltransferase, and two different sequence-specific transcriptional activators, Swi5p and Swi4p/Swi6p. We have used chromatin immunoprecipitation assays to investigate the role of each of these trans-acting factors in establishing a cell cycle-regulated domain of histone acetylation surrounding the HO upstream regulatory region. We detect a approximately 1-kb domain of H3 and H4 acetylation that is established in mid-G1, prior to and independent of HO transcription, which then declines with kinetics similar to inactivation of HO. This cell cycle burst of histone acetylation requires Gcn5p, SWI/SNF, and the Swi5p activator, but occurs in the absence of the Swi4p activator. We also find that inactivation of the Sin3p/Rpd3p deacetylase complex leads to a high level of acetylation at the HO locus throughout the cell cycle. We propose a sequential model for activation of HO in which the Swi5p-dependent recruitment of the Gcn5p acetyltransferase requires chromatin remodeling events by the SWI/SNF complex.

Catalytic mechanism and function of invariant glutamic acid 173 from the histone acetyltransferase GCN5 transcriptional coactivator.

Within chromatin, reversible acetylation of core histones is critical for transcriptional activation of eukaryotic target genes. The recent identification of intrinsic histone acetyltransferase (HAT) catalytic activity from a number of transcriptional co-activators (including yeast GCN5, p300/CBP, P/CAF, and TAFII250), has underscored the importance of protein acetylation in transcriptional control. The GCN5 family is the prototype for a diverse group of at least four distinct human HATs families. Although there is now a clear link between in vivo HAT catalytic activity and gene activation, little is known about the molecular mechanisms of histone acetylation. Herein, we report the first detailed biochemical study that probes the catalytic mechanism and the function of invariant glutamic acid 173 within the GCN5 family of HATs. Our results suggest that the HAT reaction involves the formation of a ternary complex (histones, acetyl-CoA, and enzyme) where the epsilon-amino group of histone lysine residues directly attacks the bound acetyl-CoA. The acetylation reaction requires deprotonation of the epsilon-amino group prior to nucleophilic attack. Employing site-directed mutagenesis, chemical modification, steady-state, and pH-dependent rate analysis, it is demonstrated that glutamic acid 173 is an essential catalytic residue, acting as a general base catalyst by deprotonating the histone substrate.

Roles of histone acetyltransferases and deacetylases in gene regulation.

Acetylation of internal lysine residues of core histone N-terminal domains has been found correlatively associated with transcriptional activation in eukaryotes for more than three decades. Recent discoveries showing that several transcriptional regulators possess intrinsic histone acetyltransferase (HAT) and deacetylase (HDAC) activities strongly suggest that histone acetylation and deacetylation each plays a causative role in regulating transcription. Intriguingly, several HATs have been shown an ability to acetylate nonhistone protein substrates (e.g., transcription factors) in vitro as well, suggesting the possibility that internal lysine acetylation of multiple proteins exists as a rapid and reversible regulatory mechanism much like protein phosphorylation. This article reviews recent developments in histone acetylation and transcriptional regulation. We also discuss several important, yet unanswered, questions.

Histone acetyltransferase activity of yeast Gcn5p is required for the activation of target genes in vivo.

Gcn5p is a transcriptional coactivator required for correct expression of various genes in yeast. Several transcriptional regulators, including Gcn5p, possess intrinsic histone acetyltransferase (HAT) activity in vitro. However, whether the HAT activity of any of these proteins is required for gene activation remains unclear. Here, we demonstrate that the HAT activity of Gcn5p is critical for transcriptional activation of target genes in vivo. Core histones are hyperacetylated in cells overproducing functional Gcn5p, and promoters of Gcn5p-regulated genes are associated with hyperacetylated histones upon activation by low-copy Gcn5p. Point mutations within the Gcn5p catalytic domain abolish both promoter-directed histone acetylation and Gcn5p-mediated transcriptional activation. These data provide the first in vivo evidence that promoter-specific histone acetylation, catalyzed by functional Gcn5p, plays a critical role in gene activation.

Multiple phosphorylated forms of the Saccharomyces cerevisiae Mcm1 protein include an isoform induced in response to high salt concentrations.

The Saccharomyces cerevisiae Mcm1 protein is an essential multifunctional transcription factor which is highly homologous to human serum response factor. Mcm1 protein acts on a large number of distinctly regulated genes: haploid cell-type-specific genes, G2-cell-cycle-regulated genes, pheromone-induced genes, arginine metabolic genes, and genes important for cell wall and cell membrane function. We show here that Mcm1 protein is phosphorylated in vivo. Several (more than eight) isoforms of Mcm1 protein, resolved by isoelectric focusing, are present in vivo; two major phosphorylation sites lie in the N-terminal 17 amino acids immediately adjacent to the conserved MADS box DNA-binding domain. The implications of multiple species of Mcm1, particularly the notion that a unique Mcm1 isoform could be required for regulation of a specific set of Mcm1's target genes, are discussed. We also show here that Mcm1 plays an important role in the response to stress caused by NaCl. G. Yu, R. J. Deschenes, and J. S. Fassler (J. Biol. Chem. 270:8739-8743, 1995) showed that Mcm1 function is affected by mutations in the SLN1 gene, a signal transduction component implicated in the response to osmotic stress. We find that mcm1 mutations can confer either reduced or enhanced survival on high-salt medium; deletion of the N terminus or mutation in the primary phosphorylation site results in impaired growth on high-salt medium. Furthermore, Mcm1 protein is a target of a signal transduction system responsive to osmotic stress: a new isoform of Mcm1 is induced by NaCl or KCl; this result establishes that Mcm1 itself is regulated.

Transcription-linked acetylation by Gcn5p of histones H3 and H4 at specific lysines.

The yeast transcriptional adaptor, Gcn5p, is a catalytic subunit of a nuclear (type A) histone acetyltransferase linking histone acetylation to gene activation. Here we report that Gcn5p acetylates histones H3 and H4 non-randomly at specific lysines in the amino-terminal domains. Lysine 14 of H3 and lysines 8 and 16 of H4 are highly preferred acetylation sites for Gcn5p. We also demonstrate that lysine 9 is the preferred position of acetylation in newly synthesized yeast H3 in vivo. This finding, along with the fact that lysines 5 and 12 in H4 are predominant acetylation sites during chromatin assembly of many organisms, indicates that Gcn5p acetylates a distinct set of lysines that do not overlap with those sites characteristically used by type B histone acetyltransferases for histone deposition and chromatin assembly.