Identification of a BRCA1-associated kinase with potential biological relevance.

A biochemical approach was used to identify proteins which interact with human BRCA1. Through this work, a kinase activity which co-purifies with BRCA1 has been identified. This kinase activity, which phosphorylates BRCA1 in vitro, was originally identified in Sf9 insect cells but is also present in cells of human origin including breast and ovarian carcinoma cell lines.

Effect of antihypertensive treatment with doxazosin on insulin sensitivity and fibrinolytic parameters.

The effects of the selective alpha-1-adrenoceptor antagonist doxazosin on metabolic and fibrinolytic parameters were studied in hypertensive patients with various degrees of fasting plasma insulin levels (Group A: 22.5 +/- 3 microU/ml, Group B: 8.1 +/- 1.

Alignment of conduits for the nascent polypeptide chain in the ribosome-Sec61 complex.

An oligomer of the Sec61 trimeric complex is thought to form the protein-conducting channel for protein transport across the endoplasmic reticulum. A purified yeast Sec61 complex bound to monomeric yeast ribosomes as an oligomer in a saturable fashion. Cryo-electron microscopy of the ribosome-Sec61 complex and a three-dimensional reconstruction showed that the Sec61 oligomer is attached to the large ribosomal subunit by a single connection.

Fast visual evoked potential input into human area V5.

Studies of the human visual cortex have demonstrated that an area for motion processing (V5) is located in the lateral occipito-temporal cortex. To study the timing of arrival of signals in V5 we recorded multi-channel visual evoked potentials (VEPs) to checkerboard stimuli. We then applied dipole source analysis which was computed on a grand average of 10 subjects, and on five individual subjects, respectively.

Large-conductance cation channels in the envelope of nuclei from rat cerebral cortex.

Eucaryotic nuclei are surrounded by a double-membrane system enclosing a central cisterna which is continuous with the endoplasmic reticulum and serves as a calcium store for intracellular signaling. The envelope regulates protein and nucleic acid traffic between the nucleus and the cytoplasm via nuclear pores. These protein tunnels cross through both nuclear membranes and are permeable for large molecules.

Isolation of DNAs that selectively bind a baculovirus produced mutant p53 (Ala 143) protein but not an RRL or WGL produced mutant p53 protein.

A PCR-based technique was used to generate a large pool of random sequence double-stranded DNAs. Four DNA sequences that selectively bound in vitro to a mutant p53 143A protein, synthesized in baculovirus infected cells, were characterized. The four DNA sequences all approximated the known consensus sequence for wild-type p53.

Inverse localization of electric dipole current sources in finite element models of the human head.

The paper describes finite element related procedures for inverse localization of multiple sources in realistically shaped head models. Dipole sources are modeled by placing proper monopole sources on neighboring nodes. Lead field operators are established for dipole sources.

The influence of skull-conductivity misspecification on inverse source localization in realistically shaped finite element head models.

The electric conductivities of different tissues are important parameters of the head model and their precise knowledge appears to be a prerequisite for the localization of electric sources within the brain. To estimate the error in source localization due to errors in assumed conductivity values, parameter variations on skull conductivities are examined. The skull conductivity was varied in a wide range and, in a second part of this paper, the effect of a nonhomogeneous skull conductivity was examined.

Subcellular localization and analysis of apparent 180-kDa and 220-kDa proteins of the breast cancer susceptibility gene, BRCA1.

The breast cancer susceptibility gene BRCA1 encodes an 1863-amino acid protein that acts as a tumor suppressor. The biochemical function of BRCA1 is unknown, and there are conflicting results describing its subcellular location. We have identified a 220-kDa protein, which is reactive with three antibodies raised against the amino- and carboxyl-terminal regions of BRCA1.

Serum albumin is a specific inhibitor of apoptosis in human endothelial cells.

Excess blood vessels are removed by apoptosis of endothelial cells, however, the signals responsible for this have not been defined. Apoptosis of cultured human umbilical vein endothelial cells is induced by deprivation of serum or adhesion. In this paper, apoptosis in human umbilical vein and microvascular endothelium was induced by deprivation of serum and or adhesion.

The ACE-inhibitor lisinopril affects plasma insulin levels but not fibrinolytic parameters.

There is evidence that ACE-inhibitors exert beneficial effects on endogenous fibrinolysis in patients with previous myocardial infarction. It is still unknown if this effect is restricted to this patient group only and by which mechanisms ACE-inhibitors exhibit the profibrinolytic effects. One possible explanation might be the positive influence of ACE-inhibitors on insulin metabolism by decreasing plasma insulin which in turn could decrease PAI-1, a major regulator of the fibrinolytic system.

Bradykinin induces rise of free calcium in nuclei of neuroblastoma x glioma hybrid NG 108-15 cells.

Confocal fluorescence microscopy was used to study the bradykinin-induced calcium signals in the neuroblastoma x glioma cell line NG 108-15. We found that bradykinin induced a rise in free calcium, not only in the cytoplasm but also in the nucleus. The nuclear and cytosolic calcium concentrations were not significantly different and rose to about 1.

Origin of P16 median nerve SEP component identified by dipole source analysis--subthalamic or within the thalamo-cortical radiation?

Following median nerve stimulation, several monophasic peaks were recorded at the scalp in the 15-18 ms time range. Source analysis, using three different methods, modelled a source near the centre of the head with an orientation towards the activated hemisphere and a peak activity at 16 ms post stimulus. Magnetic recordings detected no signal in this time range, which confirmed a subcortical location of the source.

Fibrinogen, t-PA, and PAI-1 plasma levels in patients with pulmonary hypertension.

We measured fibrinogen levels as well as the fibrinolytic parameters tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor 1 (PAI-1) in plasma samples obtained at basal conditions and after stimulating the fibrinolytic system by venous occlusion (VO). Samples were taken from patients with primary pulmonary hypertension (PPH), with secondary thromboembolic pulmonary hypertension (SPHTH), with secondary pulmonary hypertension due to congenital heart disease with Eisenmenger's reaction (SPHCD), and from healthy control individuals (CON). Fibrinogen levels were not significantly different between the groups with PPH and SPHTH or between SPHCD and CON.

Hepatocyte growth factor stimulates expression of plasminogen activator inhibitor type 1 and tissue factor in HepG2 cells.

HGF is a powerful mitogen for both rat and human hepatocytes, epithelial cells and endothelial cells in vitro, and is angiogenic in vivo. It has considerable homology with plasminogen and has been shown to upregulate urokinase-type plasminogen activator (u-PA) in endothelial cells as well as u-PA and its receptor in kidney epithelial cells. In this study, we report that human recombinant HGF stimulates expression of plasminogen activator inhibitor type 1 (PAI-1) and tissue factor (TF) in the human hepatoma cell line HepG2.

Differential nuclear localization of protein kinase C isoforms in neuroblastoma x glioma hybrid cells.

The protein kinase C (PKC) alpha, beta and epsilon isoforms have distinct nuclear localizations in neuroblastoma x glioma hybrid cells NG 108-15. We found by immunoblotting that PKC alpha, beta II, delta and epsilon are the predominant isoforms in these cells. In contrast to other neuronal cell lines, none of these isoforms is down-regulated during differentiation.

Improved expression of a hybrid Streptomyces clavuligerus cefE gene in Penicillium chrysogenum.

A hybrid cefE gene, encoding penicillin N expandase, was constructed by fusing the promoter sequences, Pcp, and terminator sequences, Pct from the Penicillium chrysogenum pcbC gene to the open reading frame (orf), cefEorf, from the Streptomyces clavuligerus cefE gene. The resulting hybrid gene, Pcp/cefE'orf/Pct, differed from a previously reported hybrid cefE gene contained on plasmid pPS65. The latter gene, Pcp/cefE'orf/Sct, contained the Pcp sequences fused to the S.

Potency Testing of Erysipelas Vaccines in Mice. ELISA vs. Challenge.

Vaccines are widely used for the prophylaxis of swine erysipelas. According to national pharmacopoiea requirements, potency of these biologicals has to be tested in a mouse challenge model. Many animals are needed and the test causes much suffering.

Effects of papaverine and prostaglandin E1 on corpus cavernosum smooth muscle of arteriogenically and diabetically impotent men.

Relaxant effect of prostaglandin E1 (PGE1) and papaverine (PAP) were measured in strips of corpus cavernosum smooth muscle taken from a healthy control group of men (A; n = 5), from arteriogenically impotent men (B; n = 6) and from additionally diabetic impotent men (C; n = 5) with venous leakage. Maximal relaxant effect was achieved with PAP at a mean of 10(-4) mol/l and PGE1 at a mean of 5.8 x 10(-6) mol/l.

The constitutive and stress inducible forms of hsp 70 exhibit functional similarities and interact with one another in an ATP-dependent fashion.

Mammalian cells constitutively express a cytosolic and nuclear form of heat shock protein (hsp) 70, referred to here as hsp 73. In response to heat shock or other metabolic insults, increased expression of another cytosolic and nuclear form of hsp 70, hsp 72, is observed. The constitutively expressed hsp 73, and stress-inducible hsp 72, are highly related proteins.

Influence of cardiac output on peak t-PA plasma levels in patients receiving thrombolytic therapy with recombinant tissue-type plasminogen activator--correlation with patency rate.

We studied 35 consecutive patients with short onset of myocardial infarction who underwent thrombolytic therapy with rt-PA at a standard dosage regimen of 100 mg rt-PA total (10 mg given as a bolus followed by 50 mg, 20 mg and 20 mg per hour for 3 hours). These patients were monitored for t-PA antigen and t-PA activity and PAI-1 activity plasma levels during rt-PA infusion. Success or failure of thrombolytic therapy was evaluated by non-invasive criteria (early plasma creatine kinase peaks, early peak plasma myoglobin values, and electrocardiographic criteria) as well as by means of coronary angiography at the fourth day after thrombolytic treatment.

The role of type-1 plasminogen activator inhibitor in failure of thrombolytic therapy with recombinant tissue plasminogen activator.

We investigated the possible role of type-1 plasminogen activator inhibitor (PAI-1) on success or failure of thrombolytic therapy with recombinant tissue plasminogen activator (rt-PA) in 10 responders and 10 non-responders with acute myocardial infarction and early initiation of therapy within 2 h of onset using the common infusion scheme (100 mg rt-PA over 3 h). We determined plasma levels of t-PA (activity and antigen) as well as PAI-1 (activity and antigen) in samples obtained before, during and after thrombolytic treatment and compared the course of each of those parameters between responders and non-responders to therapy. Success or failure of treatment was determined by a combination of noninvasive methods and proven by coronary angiography within 5 days of initiation of thrombolysis.

Nuclear substrates of protein kinase C.

Starting from the finding that, for neuronal cells, the nuclear-membrane-associated protein kinase C (PKC) is the so-called 'membrane inserted', constitutively active form, we attempted to identify substrates of this nuclear PKC. For this purpose, nuclear membranes and other subcellular fractions were prepared from bovine brain, and in-vitro phosphorylation was performed. Several nuclear membrane proteins were found, the phosphorylation of which was inhibited by specific PKC inhibitors and effectively catalyzed by added PKC.

Nitric oxide mediates neurogenic relaxation induced in rabbit cavernous smooth muscle by electric field stimulation.

We investigated the relaxant effect of electric field stimulation (EFS) on rabbit cavernous smooth muscle strips in vitro precontracted by phenylephrine. Effects of EFS were monitored alone, and following muscarinic receptor blockade, and inhibition of nitric oxide (NO) formation by L-N-monomethylarginine (L-NMMA) or by L-N-nitroarginine (L-NOARG). Atropine only slightly reduced the relaxant effect of EFS to 89.