Breaking an absolute species barrier: transgenic mice expressing the mink PrP gene are susceptible to transmissible mink encephalopathy.

Transmissible mink encephalopathy (TME) is a rare disease of the North American mink, which has never been successfully transmitted to laboratory mice. We generated transgenic mice expressing the mink prion protein (PrP) and inoculated them with TME or the mouse-adapted scrapie strain 79A. TME infected mink PrP-transgenic mice on a murine PrP knockout background.

Mouse brain synaptosomes accumulate copper-67 efficiently by two distinct processes independent of cellular prion protein.

The prion protein (PrPC) is a copper-binding, cell-surface protein that plays an essential role in the etiology of transmissible spongiform encephalopathies. Atomic absorption spectroscopy studies have established that synaptosomal copper content is reduced in PrPC-deficient mice as compared with wild-type (WT) or PrP- overexpressing mice. To address the question of whether this is the result of a loss of function of PrPC in copper transport across the plasma membrane at the synapse, we have used synaptosomes incubated with 67Cu as a model system to characterize the mechanism of copper accumulation in nerve terminals.

[Family as challenge: contexts of adequate counselling].

We live in a world of competing family models of which the standard model--two adults of different sex with two children of different sex--is only one among many others. Besides this sociocultural context this article focusses on the inner contexts of familytherapeutic dialogues. Processphantasies and the role of metaphor are underlined.

Isolation, catalytic properties, and competitive inhibitors of the zinc-dependent murine glutaminyl cyclase.

Murine glutaminyl cyclase (mQC) was identified in the insulinoma cell line beta-TC 3 by determination of enzymatic activity and RT-PCR. The cloned cDNA was expressed in the secretory pathway of the methylotrophic yeast Pichia pastoris and purified after fermentation using a new three-step protocol. mQC converted a set of various substrates with very similar specificity to human QC, indicating a virtually identical catalytic competence.

Transcriptome analysis of microdissected pancreatic intraepithelial neoplastic lesions.

Pancreatic ductal adenocarcinoma (PDAC) carries the most dismal prognosis of all solid tumours. Both the late clinical presentation of patients, due to lack of early symptoms, as well as the rapid and aggressive course of the disease contribute to the extremely high mortality of this malignancy. Recently, a multistep progression model for PDAC integrating morphological, clinical and molecular evidence has been proposed.

Transcriptome analysis of human hepatic and pancreatic stellate cells: organ-specific variations of a common transcriptional phenotype.

Pancreatic stellate cells (PSCs) are thought to be the primary source of the extensive fibrotic reaction characteristic of pancreatic cancer and chronic pancreatitis in humans. PSCs share many morphological and functional characteristics with hepatic stellate cells (HSCs), whose central role in liver fibrosis is well established. However, it has remained unclear if hepatic and pancreatic stellate cells are derived from a common cell lineage and if they are completely similar or if they possess organ-specific features.

Generalized Venn diagrams: a new method of visualizing complex genetic set relations.

Motivation: Microarray experiments generate vast amounts of data. The unknown or only partially known functional context of differentially expressed genes may be assessed by querying the Gene Ontology database via GOMiner. Resulting tree representations are difficult to interpret and are not suited for visualization of this type of data.

Interleukin-12-induced interferon-gamma production by human peripheral blood T cells is regulated by mammalian target of rapamycin (mTOR).

Depending on the type of external signals, T cells can initiate multiple intracellular signaling pathways that can be broadly classified into two groups based on their sensitivity to the immunosuppressive drug cyclosporin A (CsA). Interleukin (IL)-12-mediated interferon (IFN)-gamma production by activated T cells has been shown to be CsA-insensitive. In this report, we demonstrate that the IL-12-induced CsA-resistant pathway of IFN-gamma production is sensitive to rapamycin.

aRNA-longSAGE: a new approach to generate SAGE libraries from microdissected cells.

Large-scale gene expression analyses of microdissected primary tissue are still difficult because generally only a limited amount of mRNA can be obtained from microdissected cells. The introduction of the T7-based RNA amplification technique was an important step to reduce the amount of RNA needed for such analyses. This amplification technique produces amplified antisense RNA (aRNA), which so far has precluded its direct use for serial analysis of gene expression (SAGE) library production.

KLF11 mediates a critical mechanism in TGF-beta signaling that is inactivated by Erk-MAPK in pancreatic cancer cells.

Background & Aims: Smad-regulated transcription plays a central role in transforming growth factor (TGF)-beta-induced cell growth inhibition and tumor suppression. Like the Smads, KLF11 is an early response transcription factor that mediates TGF-beta-induced growth inhibition in untransformed epithelial cells. Here, we investigated the functional implications of KLF11 in TGF-beta signaling and transcription in normal epithelial as well as pancreatic cancer cells.

Adenoviral gene transfer to the neonatal rat pulmonary circulation.

Background: Gene delivery to the pulmonary circulation has been studied in adult animals, but has not been extensively investigated in neonates.

Methods: We tested the ability of recombinant, replication-defective adenovirus to transduce the pulmonary circulation when delivered by percutaneous ventricular puncture. Five-day-old rat pups were injected with 10(7) to 10(10) particles (approximately 10(5) to 10(8) pfu) in 30 micro l total volume.

Identification of a Novel Transmembrane Protein (UKW): Association with Invasive Status of Mammary Carcinoma Cell Lines and Expression in Pancreatic Carcinoma.

A novel transmembrane-glycoprotein, referred to as UKW, was identified by expression profiling of a metastatic versus a non-metastatic pancreatic carcinoma cell line (S2-007 and S2-028). UKW is strongly expressed only in the metastatic cell line. The corresponding cDNA encodes for a protein of 374 amino acids.

LKW, a Putative Dual-specificity Kinase which is Down-regulated in Several Invasive Systems.

Using Affymetrix GeneChip® arrays, we have established the transcriptional profiles of two sublines of the human pancreatic adenocarcinoma cell lines SUIT-2, S2-007 (metastatic) and S2-028 (non-metastatic). By comparison of ESTs corresponding to differentially regulated mRNAs, we have identified the putative dual-specificity kinase LKW as down-regulated in the metastatic cell line S2-007. LKW is composed of 358 amino acids and contains catalytic domains corresponding to those of Ser/Thr- and Tyr-kinases.

Application of DNA array analyses in the management of gastrointestinal cancer patients.

Cancer is a polygenic disease arising from the accumulation of multiple genetic and epigenetic defects in the affected cells. For the majority of cancers, including many malignancies of the gastrointestinal tract, our current means of diagnosis and treatment of the tumors are grossly insufficient. DNA arrays offer the possibility to monitor the expression levels of thousands of mRNA transcripts simultaneously in a single assay, making them ideal tools to study the complex network of transcriptional changes that are associated with the malignant transformation of normal cells.

Expression profiling of the influence of RAS mutants on the TGFB1-induced phenotype of the pancreatic cancer cell line PANC-1.

Expression profiling analyses were used to elucidate the functional relevance of RAS proteins in mediating the effect of TGFB1 on the transcriptional phenotype of the pancreatic cancer cell line PANC-1. Despite the presence of one mutated KRAS2 allele in parental PANC-1 pancreatic cancer cells, RAS-dependent signal transduction remained susceptible to stimulation by EGF and TGFB1. To analyze the impact of RAS proteins on the TGFB1-induced transcriptional phenotype, we used PANC-1 cells stably transfected with a dominant negative HRAS(S17N) mutant or with a constitutively active KRAS2(G12V) mutant.

Speech recognition and dysarthria: a single subject study of two individuals with profound impairment of speech and motor control.

This study investigated the use of the speech recognition system Dragon Dictate as an augmentative method of computer access for two individuals with cerebral palsy, including severe motor dysfunction and dysarthria. Single subject design was used and measures of computer access system effectiveness and speech production were used before, during and after intervention. The users' original switch access system was compared to a combination of their switch access system and speech recognition, by counting the number of correct entries.

Claudin-4 expression decreases invasiveness and metastatic potential of pancreatic cancer.

Claudin-4 has been identified as an integral constituent of tight junctions and has been found to be highly expressed in pancreatic cancer. The aim of the present study was to elucidate the effect of claudin-4 on growth and metastatic potential in pancreatic cancer cells, as well as the regulation of claudin-4 by oncogenic pathways. Claudin-4 was stably overexpressed in SUIT-2 pancreatic cancer cells, and its effect on invasion and growth in vitro was examined by using two-chamber invasion assays, soft agar assays, and fluorescence-activated cell sorter analysis.

SERPINE2 (protease nexin I) promotes extracellular matrix production and local invasion of pancreatic tumors in vivo.

In large-scale expression profiling analyses, we have previously identified genes differentially expressed between subclones of the pancreatic cancer cell line SUIT-2. One of the genes most strongly overrepresented in the highly metastatic subclone S2-007 as compared with the rarely metastatic subclone S2-028 was the serine proteinase inhibitor SERPINE2 (protease nexin I), suggesting that this protein may play an important part in the process of metastasis. The aim of this study was to functionally characterize SERPINE2 for its potential to influence the invasive and metastatic phenotype of cancer cells in vitro and in vivo.

Effect of promoter methylation on the regulation of IFN-gamma gene during in vitro differentiation of human peripheral blood T cells into a Th2 population.

The carefully orchestrated events that result in a protective immune response are coordinated to a large extent by cytokines produced by Th1 and Th2 cell subsets. Th1 cells preferentially produce IL-2 and IFN-gamma, resulting in a cellular response that helps to eliminate infected cells. In contrast, Th2 cells produce IL-4, IL-5, IL-6, and IL-10, stimulating an Ab response that attacks extracellular pathogens, thereby preventing the cells from becoming infected.

Population genetic implications from DNA polymorphism in random human genomic sequences.

Denaturing high performance liquid chromatography (DHPLC) in combination with dye-terminator sequencing was used to survey 516 random genomic sequence tagged sites (STSs) for biallelic polymorphisms in 24 representatives of the major ethnic groups residing in the United States. Of the 301 polymorphic STSs (58.3%), 172 contained a single simple sequence polymorphism (SSP), while 78, 35, and 16 contained 2, 3, and 4-6 SSPs, respectively.

CaiT of Escherichia coli, a new transporter catalyzing L-carnitine/gamma -butyrobetaine exchange.

l-Carnitine is essential for beta-oxidation of fatty acids in mitochondria. Bacterial metabolic pathways are used for the production of this medically important compound. Here, we report the first detailed functional characterization of the caiT gene product, a putative transport protein whose function is required for l-carnitine conversion in Escherichia coli.

Use of DNA arrays/microarrays in pancreatic research.

In recent years, enormous technical advances in experimental protocols as well as robotic and bioinformatic techniques have allowed DNA array/microarray technology to emerge as the leading technology in the field of functional, disease-related genome analysis. Multiple applications exist for DNA arrays/microarrays including comparative genomic analysis to identify chromosomal imbalances (Matrix-CGH), the study of mutations and genetic polymorphisms, and the study of gene expression (expression profiling). Expression profiling is the most widely used application of DNA array/microarray technology and allows to measure gene expression of thousands of genes simultaneously.

Effect of rapamycin on the cyclosporin A-resistant CD28-mediated costimulatory pathway.

The consequences of T-cell activation depend exclusively on costimulation during antigen-T-cell receptor interaction. Interaction between the T-cell coreceptor CD28 and its ligand B7 during antigen-antigen receptor engagement results in full activation of T cells, the outcomes of which are proliferation and effector functions. The ability of CD28 to costimulate the production of interleukin-2 (IL-2) explains the importance of this costimulation.

The expression and characterization of cyclin-dependent kinase 6 during the activation of murine G0 T-cells.

Recent evidence strongly suggest that the D type cyclins with cdk4 and cdk6 form holoenzymes that regulate cell cycle events earlier in G1 than cyclin E/cdk2 complexes which functions near the G1/S transition. In human T lymphocytes cdk6 has been shown to be the initial retinoblastoma protein kinase detectable at mid G1. Following activation of splenic derived murine G0T-cells, cdk6, cyclin D2 and D3 specific mRNAs were detected early in G1 and reached maximal levels prior to or near G1/S.