Loss of Tribbles pseudokinase-3 promotes Akt-driven tumorigenesis via FOXO inactivation.

Tribbles pseudokinase-3 (TRIB3) has been proposed to act as an inhibitor of AKT although the precise molecular basis of this activity and whether the loss of TRIB3 contributes to cancer initiation and progression remain to be clarified. In this study, by using a wide array of in vitro and in vivo approaches, including a Trib3 knockout mouse, we demonstrate that TRIB3 has a tumor-suppressing role. We also find that the mechanism by which TRIB3 loss enhances tumorigenesis relies on the dysregulation of the phosphorylation of AKT by the mTORC2 complex, which leads to an enhanced phosphorylation of AKT on Ser473 and the subsequent hyperphosphorylation and inactivation of the transcription factor FOXO3. These observations support the notion that loss of TRIB3 is associated with a more aggressive phenotype in various types of tumors by enhancing the activity of the mTORC2/AKT/FOXO axis.

Tribbles: a family of kinase-like proteins with potent signalling regulatory function.

The recent identification of tribbles as regulators of signal processing systems and physiological processes, including development, together with their potential involvement in diabetes and cancer, has generated considerable interest in these proteins. Tribbles have been reported to regulate activation of a number of intracellular signalling pathways with roles extending from mitosis and cell activation to apoptosis and modulation of gene expression. The current review summarises our current understanding of interactions between tribbles and various other proteins. Since our understanding on the molecular basis of tribbles function is far from complete, we also describe a bioinformatic analysis of various segments of tribbles proteins, which has revealed a number of highly conserved peptide motifs with potentially important functional roles.

Tribbles: novel regulators of cell function; evolutionary aspects.

Identification of rate-limiting steps or components of intracellular second messenger systems holds promise to effectively interfere with these pathways under pathological conditions. The emerging literature on a recently identified family of signalling regulator proteins, called tribbles gives interesting clues for how these proteins seem to link several 'independent' signal processing systems together. Via their unique way of action, tribbles co-ordinate the activation and suppression of the various interacting signalling pathways and therefore appear to be key in determining cell fate while responding to environmental challenges. This review summarises our current understanding of tribbles function and also provides an evolutionary perspective on the various tribbles genes.

Role of calcium-activated potassium channels in the regulation of basal and agonist-elevated tones in isolated conduit arteries. Short communication.

Functional role of calcium-activated potassium (KCa) channels on the basal and agonist-elevated arterial tones was investigated in isolated rabbit aorta, porcine and canine coronary arteries as well as in human internal mammary artery. The vascular tones enhanced by contractile agents were increased further by preincubation of these conduit blood vessels with selective (charybdotoxin or iberiotoxin) or nonselective (tetraethylammonium) inhibitors of KCa channels. The basal tone (without an agonist) was increased only in the canine coronary artery. The results indicate a feed-back regulatory role of KCa channels counteracting the vasospasm of conduit arteries.

The Twisted gastrulation family of proteins, together with the IGFBP and CCN families, comprise the TIC superfamily of cysteine rich secreted factors.

AIMS: To analyse the similarities between the Twisted gastrulation (TSG) proteins known to date; in addition, to determine phylogenetic relations among the TSG proteins, and between the TSGs and other protein families--the CCN (for example, CCN2 (CTGF), CCN1 (CYR61), and CCN3 (NOV)) and IGFBP (insulin-like growth factor binding protein) families. METHODS: TBLASTN and FASTA3 were used to identify new tsg genes and relatives of the TSG family. The sequences were aligned with ClustalW. The predictions of sites for signal peptide cleavage, post-translational modifications, and putative protein domains were carried out with software available at various databases. Unrooted phylogenetic trees were calculated using the UPGMA method. RESULTS: Several tsg genes from vertebrates and invertebrates were compared. Alignment of protein sequences revealed a highly conserved family of TSG proteins present in both vertebrates and invertebrates, whereas the slightly less well conserved IGFBP and CCN proteins are apparently present only in vertebrates. The TSG proteins display strong homology among themselves and they are composed of a putative signal peptide at the N-terminus followed by a cysteine rich (CR) region, a conserved domain devoid of cysteines, a variable midregion, and a C-terminal CR region. The most striking similarity between the TSGs and the IGFBP and CCN proteins occurs in the N-terminal conserved cysteine rich domain and the characteristic 5' cysteine rich domain(s), spacer region, and 3' cysteine rich domain structure. CONCLUSION: The family of highly conserved TSG proteins, together with the IGFBP and CCN families, constitute an emerging multigene superfamily of secreted cysteine rich factors. The TSG branch of the superfamily appears to pre-date the others because it is present in all species examined, whereas the CCN and IGFBP genes are found only in vertebrates.

Comparative study on the effect of phosphorylated TCR zeta chain ITAM sequences on early activation events in Jurkat T cells.

One of the main dilemma in T cell receptor (TCR) signal transduction is whether the presence of multiple Immunoreceptor Tyrosine-based Activation Motifs (ITAMs) within the TCR signaling module serves for signal amplification or signal distribution. To contribute to answer this question, we analyzed the effect of synthetic oligopeptides representing the three bi-phosphorylated zeta chain-ITAMs on the early signaling events in permeabilized leukemia T cells. Our main observations were as follows: 1/Stimulation of the cells with the bi-phosphorylated membrane proximal and central ITAMs (zeta (1)y(p)y(p) and zeta (2)y(p)y(p), respectively) resulted in a strong phosphorylation of proteins with a similar pattern. In contrast, the membrane distal ITAM, zeta (3)y(p)y(p) had a reduced ability to promote tyrosine phosphorylation and failed to induce the phosphorylation of a number of proteins. 2/ The phospho-peptide induced tyrosine phosphorylation events were at least partially mediated by p56(lck) and Syk/ZAP70 protein tyrosine kinases as it was shown in p56(lck) and Syk/ZAP70 deficient Jurkat variants. 3/The patterns of the association of the adaptor protein, Grb2 with tyrosine phosphorylated proteins following cell stimulation with the bi-phosphorylated membrane proximal or the central ITAMs were similar, while the membrane distal ITAM was unable to induce any of these associations. Our data provide additional evidence that the three zetaITAMs differ in their capacity to induce tyrosine phosphorylation of intracellular proteins in permeabilized T cells, depending to their primary sequence. The first and second ITAM sequences of the zeta chain may have similar but not totally overlapping functions. This conclusion results from their similar but not identical abilities to induce tyrosine phosphorylation and association of Grb-2 with intracellular phosphoproteins. In contrast, the third ITAM (zeta3) may have distinct functions since this peptide fails to induce tyrosine phosphorylation of a number of proteins compared to the other two ITAMs, and it is unable to induce either new association or the increase in the amount of Grb-2 associated phosphoproteins.

The probable involvement of soluble and deposited melanins, their intermediates and the reactive oxygen side-products in human diseases and aging.

The plasma soluble melanins (PSM) form spontaneously in vitro and in vivo and their formation involves oxidative polymerization and copolymerization of dopa, catecholamines, homogentisic acid, 3-hydroxyanthranilic acid, p-aminophenol, p-phenylenediamine, and other end(ex)ogenous ortho and para polyhydroxy-, (poly)hydroxy(poly)amino- and polyamino-phenyl compounds. The build up of PSM is visible within 2-3 h after the start of incubation at 37 degrees C with 1 mg/ml of plasma. PSM also form similarly in blood and these processes cause hemolysis. The mean quantity of PSM in normal human plasma is 1.61+/-0.1 (S.D.) mg/ml (n = 20) and in normal human urine is 1.1+/-1.2 g/24 h collection (n = 8). They contribute to the yellow color of plasma and urine. Antioxidants delay the formation of PSM. The deposited melanins also form from these precursors. Reactive oxygen side products (ROSP) are generated during and after melanogenesis. Melanins in vivo are generally associated with proteins or with proteins and lipids. The PSM-protein-lipid complexes are called plasma soluble lipofuscins (PSL), because they have histochemical and fluorescence properties similar to those of solid lipofuscins. The soluble and deposited melanins (SDM) and their intermediates have similar toxic chemical reactivities. The oxidizing quinoid (they can produce partially and completely substituted conjugates) and the semiquinoid free radical intermediates are also moieties in most human melanin structures. Soluble melanins formed from dopa, or dopamine, or norepinephrine in weak alkaline solution have been shown to be toxic to human CD4+ lymphoblastic cells (MT-2) at higher than 10 microg/ml concentrations. Alkaptonuria with high levels of homogentisic acid in the plasma is a potentially fatal disease, exhibiting the toxic effects of the homogentisic acid melanin (soluble and deposited), its intermediates and the ROSP. Patients with alkaptonuria develop arthritis and often suffer from other diseases too, including cardiovascular disease (frequent cause of death) and kidney disease. Pheochromocytoma, with high levels of catecholamines in the plasma is another potentially fatal disease. The catecholamine PSM of pheochromocytoma have very light yellow or practically no colors, due to the concentrations and chemical structures. Pheochromocytomas can cause hypertension, cardiovascular disease (frequent cause of death), kidney disease, stroke, cancer, amyloid formation and can mimic many other diseases, including acute pancreatitis, carcinoid, neuroblastoma, psychiatric illness, hypercalcemia, retinal vascular lesions, and diabetes mellitus. Pheochromocytoma is potentially fatal even in patients without hypertension. Following trauma and surgery, heavily pigmented eyes are apt to experience greater inflammation than lightly pigmented eyes. In Parkinson's disease those neurons are lost first in the substantia nigra and locus ceruleus which contain the greatest amounts of neuromelanins. The antihypertensive alphamethyldopa causes Parkinson's syndrome. It forms PSM in a short time in vitro. The side effects of L-dopa (immobility episodes alternate with normal or involuntary movements; psychotic abnormalities) suggest that the SDM, their intermediates and the ROSP present naturally in vivo are involved in the cause of Parkinson's disease and Alzheimer's disease. There is a large overlap between these two diseases. (ABSTRACT TRUNCATED)

Low sideband guided-mode resonant filter.

A guided-mode resonant filter with low sideband reflections is proposed. It is shown that, for serious reduction of out-of band reflectance, the combination of waveguide-grating filter design with conventional antireflective stack design methods is not adequate. To achieve symmetrical low sideband reflectances, independent control of various layer thicknesses is necessary. At a given illumination angle with appropriate control of the waveguide thickness, a specific resonant grating filter is designed whose out-of-band reflectance on both sides of the resonant peak is well below 10(-4). Even 50 nm away from the peak, on both sides, the out-of-band reflectance remains below 10(-3). Analysis of the variation in the main manufacturing parameters indicates that such filters can be reliably produced with present-day technologies.

Contribution of kinases and the CD45 phosphatase to the generation of tyrosine phosphorylation patterns in the T-cell receptor complex zeta chain.

The zeta subunit of the T-cell receptor complex plays a crucial role in coupling the antigen binding alphabeta and gammadelta heterodimers to the downstream activation pathways. Three tandem amino acid sequence motifs containing pairs of exactly spaced Tyr-X-X-Leu/Ile sequences, designated as Immunoreceptor Tyrosine-based Activation Motifs (ITAMs), control this function. The phosphorylated forms of ITAMs serve as docking sites for several src homology 2 (SH2) domain containing signaling proteins. The composition of the assembled signaling complex and the outcome of cell activation depends on the tyrosine phosphorylation pattern of the zeta polypeptide. The mechanism that conducts the generation of various phosphorylated forms has not yet been well established. In this study we have analyzed the ability of src family tyrosine kinases and the CD45 tyrosine phosphatase in determining the phosphorylation state of the different ITAMs and the individual tyrosine residues of the TCR zeta chain. The intracellular part of the zeta chain was phosphorylated by src family tyrosine kinases, p56lck and p59fyn in vitro. Synthetic oligopeptides representing full-length or half-sized ITAMs with a single tyrosine residue were also phosphorylated by both p56lck and p59fyn. In contrast, an additional membrane proximal tyrosine residue in the human zeta chain, located outside of the ITAMs, was not phosphorylated. We also examined the activity of the CD45 phosphatase, using a panel of ITAM derivatives, in which one or both tyrosines were phosphorylated. The efficiency of ITAM dephosphorylation by CD45 was dependent on the primary sequence of the oligopeptides and the position of the phosphotyrosine residues. The in vitro data suggest that the CD45 phosphatase rather than the tyrosine kinase(s) may control the generation of specific phosphorylation patterns of the zeta chain during cell activation.

[Pacemaker syndrome without a pacemaker]. / Pacemaker szindróma pacemaker nélkül.

Circulatory consequences of cardiac arrhythmias are not always evident. Proper interpretation of the clinical symptoms in certain cases requires assessment of the patients' other hemodynamic characteristics. The authors present the case of a patient with left ventricular hypertrophy, who developed severe circulatory failure at the time of artrioventricular dyssynchrony in association with junctional rhythm. Analogy between the circulatory consequences of the junctional rhythm and ventricular pacing was documented by hemodynamic measurements. The patient was subsequently treated by implanting an atrioventricular pacemaker.

Conformational effect of phosphorylation on T cell receptor/CD3 zeta-chain sequences.

The effect of tyrosine-phosphorylation on the conformation of three tyrosine-based immunoreceptor activation motifs, zeta(69-86), zeta(106-126), and zeta(138-155), located in the T cell receptor/CD3 zeta-chain was investigated. Circular dichroism and Fourier-transform infrared spectroscopy of the nonphosphorylated and phosphorylated fragments gave evidence that phosphorylation can alter the secondary structure of the peptides. The most significant--alpha-helix to beta-sheet--conformational change was observed in the case of the zeta(138-155) peptide sequence which may be relevant to recognition by Src homology 2 (SH2) domains of signaling proteins.

Contact printing of Bragg gratings in optical fibers: rigorous diffraction analysis.

The fabrication of Bragg gratings inside the core of single-mode optical fibers has been greatly simplified by the application of contact printing [Appl. Phys. Lett. 62, 1035 (1993)]. According to this technique, the fiber is placed in proximity to an appropriate phase grating, which is illuminated with nominally monochromatic UV light. The transmitted light is redistributed into an ideally sinusoidal variation of optical intensity (more properly, irradiance), which is imprinted into the core as a result of photoinduced refractive index changes. In accordance with normal practice in this field, intensity is used instead of optical intensity throughout the text. The main advantage of such a writing method, apart from its simplicity, is that the illumination source need not be highly coherent. Rigorous analysis of this method is given with a new phase grating design.

[Defecation syncope following pacemaker implantation]. / Defecatiós syncope jelentkezése pacemaker beültetést követóen.

A case of syncope is reported, where hemodynamic responses compatible with pacemaker syndrome occurred during defecation. A 73 year old female received a VVI pacemaker for complete heart block. Subsequently she sustained repeated episodes of defecation syncope. Hemodynamic investigations revealed an abnormal blood pressure drop occurring at the overshoot phase of the Valsalva maneuver in association with alternation of sinus and pacemaker rhythm. This modified Valsalva response in the presence of contributing factors, such as hypovolemia and nitroglycerin therapy manifested as defecation syncope. The abnormal condition was treated by eliminating the contributing factors, and by programming a low pacemaker frequency, thus precluding sinus-pacemaker alterations.

Microheterogeneity of the cell surface tyrosine phosphatase, CD45RA, on T cells: phytohaemagglutinin binding and non-binding fraction of the 220 kDa isoform.

The CD45 antigen is a family of the transmembrane tyrosine phosphatases expressed on cells of haemopoietic origin. The molecules are distinguished by the different aminoacid sequence and glycosylation on the N terminus. Although all isoforms are heavily glycosylated and exert receptor like structures on the extracellular part, the role of the glycosylation in the possible receptor function and the ligand of the CD45 has not been determined yet. In this study we have examined the binding of phytohaemagglutinin (PHA) to the different isoforms and its relation to the phosphatase activity. Immunoblotting experiments revealed that the 220 kDa form of the CD45RA contained a PHA binding fraction when immunoprecipitated with CD45RA monoclonal antibody (mAb), while an isoform with identical molecular mass immunoprecipitated by anti-CD45 did not bind PHA. We concluded that the 220 kDa form was heterogeneous with respect to PHA binding. Functional data also confirmed this heterogeneity: previous extraction of the PHA binding proteins resulted in the elimination of all the phosphatase activity from CD45, while only a part of that was removed from CD45RA immunoprecipitates.

Relative fluorescence intensities of human plasma soluble melanins in normal adults.

Fluorescence spectroscopy of plasma soluble melanin derived from 3-hydroxyanthranilic acid discloses an emission (em) maximum at approximately 413 nm [excitation (ex) maximum approximately 324 nm]; plasma melanins derived from dopa, catecholamines, catechol and 3-hydroxykynurenine have em maxima at approximately 445 nm (ex maxima approximately 345 nm). All of these melanins are present in human plasma. For the present study, plasma samples were obtained from 120 healthy Caucasian, Hispanic, Mongoloid and Negroid males and females, ranging from 19 to 92 years of age, with 12 or more subjects in each racial and sex group. The average relative fluorescence intensities of all samples (diluted 1 to 50 with water) were 34.6 +/- 7.4 (standard deviation) at 413 nm and 40.8 +/- 8.2 at 445 nm. Individual intensity values in samples from women below 50 years in age were in the lower range of the values observed; at age 50 and higher no difference was seen in samples from women and men. No clear cut relationships to race or age were seen.

Effect of phytohaemagglutinin on CD45 in T cells.

In this study the effect of PHA activation on the phosphatase activity of CD45 has been investigated in human leukemic T-cell lines. It has been found that in vivo activation of the cells with PHA resulted in 2-4-fold increase in enzyme activity. Addition of PHA to the postnuclear supernatant of cell lysates also resulted in elevation of phosphatase activity. Elevation of enzyme activity resulted from an increase in the amount of antigen in the immunoprecipitates. Elevation of the quantity was not the result of a de novo protein synthesis since the presence of cycloheximide, a protein synthesis inhibitor, did not modulate the effect of PHA. The effect of PHA was specific since ConA, that also bound to the CD45 molecules, or crosslinking of the antigen by antibody did not affect CD45. Since direct binding of PHA to CD45 molecules was shown in immunoblotting analysis, we suggest that the effect of PHA is a consequence of a PHA-induced conformational change of CD45 that results in up-regulation of the analyzed CD45 epitopes.

[Continuous non-invasive blood pressure monitoring in the diagnosis of pacemaker syndrome]. / Folyamatos, noninvazív vérnyomásmérés a pacemaker szindróma diagnózisában.

The drop in blood pressure coinciding with the atrioventricular dyssynchrony plays an important role in the genesis of pacemaker syndrome. The diagnosis is often based on continuous blood pressure recording. Formerly the continuous blood pressure monitoring could only be performed by invasive methods. The authors demonstrate the feasibility of a new non invasive continuous blood pressure recorder, the Finapres 2300, by presenting illustrative case reports. The authors recommend more widespread use of non invasive haemodynamic monitoring for the diagnosis of pacemaker syndrome.