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J Zamorano

Publications and source records attributed to J Zamorano.

At least 37 records · Page 2Linked to original sources
PubMed1999

The IL-4 receptor: signaling mechanisms and biologic functions.

Interleukin-4 is a multifunctional cytokine that plays a critical role in the regulation of immune responses. Its effects depend upon binding to and signaling through a receptor complex consisting of the IL-4R alpha chain and the common gamma chain (gamma c), resulting in a series of phosphorylation events mediated by receptor-associated kinases. In turn, these cause the recruitment of mediators of cell growth, of resistance to apoptosis, and of gene activation and differentiation. Here we describe our current understanding of the organization of the IL-4 receptor, of the signaling pathways that are induced as a result of receptor occupancy, and of the various mechanisms through which receptor function is modulated. We particularly emphasize the modular nature of the receptor and the specialization of different receptor regions for distinct functions, most notably the independent regulation of cell growth and gene activation.

Amino Acid Sequence↗
PubMed1998

Regulation of apoptosis by tyrosine-containing domains of IL-4R alpha: Y497 and Y713, but not the STAT6-docking tyrosines, signal protection from apoptosis.

IL-4 is a cytokine with important antiapoptotic activity. We have analyzed the role that tyrosine-containing domains within the cytoplasmic tail of IL-4R alpha play in IL-4-mediated protection from apoptosis. 32D cells expressing a wt huIL-4R alpha or one truncated at aa 557 were protected by huIL-4 from apoptosis while cells expressing a receptor truncated at aa 657 were not, suggesting that the carboxyl-terminal domain signals protection from apoptosis. However, changing Y713 within this region to phenylalanine had no effect. To analyze the contribution of tyrosine-containing domains independently, we transplanted regions of the huIL-4R alpha to a truncated form of the huIL-2R beta that could not signal protection from apoptosis. Transplantation of the huIL-4R alpha domains containing Y497 or Y713 partially prevented cell death and together signaled protection from apoptosis in response to IL-2 as well as the wt IL-2R beta. Mutation of Y497 and Y713 to phenylalanine inhibited protection. In contrast, transplantation of the domain containing the potential STAT6-docking tyrosines alone had no effect, yet it inhibited the protection mediated by the other domains. Although IL-4R alpha signals Shc and SH2-containing inositol phosphatase (SHIP) phosphorylation, we could not establish an association between their activation and protection from apoptosis. Taken together, this study suggests that the domains of the huIL-4R alpha containing Y497 and Y713 positively regulate protection from apoptosis while the domain containing the STAT6 docking sites suppresses this protection, and that additional signaling molecules other than insulin receptor substrate-1 (IRS1), Shc, or SHIP may be involved in antiapoptotic signaling.

Animals↗
PubMed1998

A role for the insulin-interleukin (IL)-4 receptor motif of the IL-4 receptor alpha-chain in regulating activation of the insulin receptor substrate 2 and signal transducer and activator of transcription 6 pathways. Analysis by mutagenesis.

The interleukin (IL)-4 receptor alpha-chain (IL-4Ralpha) contains a sequence motif (488PLVIAGNPAYRSFSD) termed the insulin IL-4 receptor motif (I4R motif). Mutation of the central Tyr497 to Phe blocks the tyrosine phosphorylation of the insulin receptor substrate 1 (IRS1) and diminishes proliferation in response to IL-4. Recent data suggest that the I4R motif encodes binding sites for several protein tyrosine binding (PTB) domain-containing proteins such as IRS1 and Shc and potentially for the Src homology 2 domain of signal transducer and activator of transcription 6 (STAT6). To analyze the function of the I4R motif in regulating IL-4 signaling, we changed conserved residues upstream and downstream of the central Tyr to Ala in the human IL-4Ralpha. We analyzed the ability of these constructs to signal the tyrosine phosphorylation of IRS2 and STAT6, the induction of DNA binding activity, and CD23 induction in response to human IL-4 (huIL-4) in transfected M12.4.1 cells. Mutagenesis of residues downstream of Tyr497, such as Arg498 or Phe500, to Ala had no effect on any of these responses, suggesting that the I4R motif may not be important for functional Src homology 2 domain interactions. However, mutagenesis of Pro488 to Ala (P488A) greatly diminished the tyrosine phosphorylation of IRS2 and abolished tyrosine phosphorylation of STAT6, induction of DNA binding activity, and CD23 induction in response to huIL-4. By contrast, a P488G mutant signaled these responses to huIL-4. Mutagenesis of hydrophobic amino acids previously shown to contact the PTB domain of IRS1, Leu489 or Ile491, to Ala had only minimal effects on responses to huIL-4. However, changing both Leu498 and Ile491 to Ala greatly diminished the tyrosine phosphorylation of IRS2 and abolished STAT6 activation. Taken together, these results indicate the important role of the I4R motif in regulating IRS docking and suggest that I4R docking to a PTB domain-containing protein regulates activation of the STAT6 pathway.

Alanine↗
PubMed1998

Regulation of cell growth by IL-2: role of STAT5 in protection from apoptosis but not in cell cycle progression.

Cytokines play an essential role in the regulation of lymphocyte survival and growth. We have analyzed the pathways activated by IE-2 that lead to protection from apoptosis and cell proliferation. IL-2 can act as a long-term growth factor in 32D cells expressing the wild-type human (hu)IL-2R beta. By contrast, cells expressing a truncated form of the huIL-2R beta, which is able to induce Bcl-2 and c-myc expression but not STAT5 activation, were not protected from apoptosis by IL-2; consequently, they could not be grown long term in the presence of IL-2. However, IL-2 promoted cell cycle progression in cells bearing the truncated huIL-2R beta with percentages of viable cells in the G0/G1, S, and G2/M phases similar to cells expressing the wild-type huIL-2R beta. Transplantation of a region from the erythropoietin receptor, which contains a docking site for STAT5 (Y343) to the truncated huIL-2R beta, restored the ability of IL-2 to signal both activation of STAT5 and protection from apoptosis. By contrast, transplantation of a region from the huIL-4R alpha containing STAT6 docking sites did not confer protection from apoptosis. These results indicate that the IL-2-induced cell cycle progression can be clearly distinguished from protection from apoptosis and that STAT5 participates in the regulation of apoptosis.

Animals↗
PubMed1998

Glucocorticoids inhibit IL-4 and mitogen-induced IL-4R alpha chain expression by different posttranscriptional mechanisms.

BACKGROUND: A high level of expression of IL-4Ralpha chain on the surface of lymphocytes has been described in certain allergic and inflammatory autoimmune diseases. Progression of these diseases are usually controlled by steroid treatment. One mechanism by which these drugs exert their antiinflammatory and immunosuppressive effects is by widely repressing or enhancing the production of cytokines and their receptors. OBJECTIVES: The effect of glucocorticoids on IL-4Ralpha chain expression has not been previously studied, and this is the aim of the present report. For this purpose, human lymphocytes were induced to express IL-4Ralpha chain by means of protein kinase C (PKC) activation with phorbol myristate acetate (PMA) or by triggering the Janus kinase-Stat pathway with IL-4 in the presence or absence of pharmacologic doses of dexamethasone. METHODS: IL-4Ralpha cell surface expression was studied by flow cytofluorometry. The levels and stability of mRNA were assessed by Northern blot analysis. The effect of dexamethasone on the IL-4Ralpha rate of transcription was determined by nuclear run-on experiments. RESULTS: Dexamethasone significantly downregulated PMA-induced IL-4Ralpha mRNA and protein levels in total peripheral blood mononuclear cells and in isolated T cells. The mechanism involved a posttranscriptional regulation of IL-4Ralpha expression because dexamethasone decreased the PMA-induced IL-4Ralpha mRNA half-life. However, we found that PMA did not influence the transcription rate of IL-4Ralpha gene, irrespective of the presence or absence of dexamethasone. This immunosuppressor also diminished the IL-4-induced IL-4Ralpha expression on the surface of isolated T and B lymphocytes but, interestingly, without modifying mRNA levels that indicates that dexamethasone downregulated IL-4-dependent IL-4Ralpha expression by acting at a translational or posttranslational level. In fact, we observed that the drug did not affect IL-4-induced IL-4Ralpha gene transcription rate nor did it shorten mRNA half-life. The effect of dexamethasone on the IL-4Ralpha was steroid specific because it was totally reversed by the glucocorticoid receptor antagonist RU486. CONCLUSION: Our results support that dexamethasone may influence the course of allergic and inflammatory diseases by downregulating the expression of IL-4Ralpha.

Cells, Cultured↗
PubMed1998

[Staphylococcus aureus bacteremia and a mass in the right atrium in a patient carrying a central port].

A male heroin abuser was admitted because of Candida endophthalmitis. A permanent central venous line was inserted. Three months later, he had positive catheter and blood cultures with Staphylococcus aureus. Transthoracic and transesophageal echocardiography and magnetic resonance revealed a right atrial mass. Treatment with antibiotics in addition to clinical and echocardiographic follow up was decided upon. Six weeks later, a transesophageal echocardiography showed a decrease in right atrial mass size and the patient was discharged asymptomatic without treatment. Three months later, he was well and a new transesophageal echocardiography showed a further decrease in atrial mass size with signs of fibrosis. This follow up supports the clinical suspicion of thrombus in the right atrium attributable to a central venous line and Staphylococcus aureus bacteremia.

Adult↗
PubMed1998

[Applications of contrast media in echocardiography].

The development of contrast media in ultrasound has been slow and sporadic, and there are no fully satisfactory agents for clinical imaging to date. Most contrast agents consist of air filled microbubbles which generate scattered echoes and enhance the ultrasound information. In this article we review the different phases of contrast echocardiography their potentials and clinical applications. First, right-sided echocontrast which is mainly used for the assessment of intracardiac shunts. Second, left-sided contrast agents with smaller and more stable microbubbles, that allow the visualization of the left ventricle after intravenous injection. With these agents, one it is now possible to study myocardial perfusion which is one of the most attractive potentials of these types of agents.

Contrast Media↗
PubMed1998

Regulation of gene expression, growth, and cell survival by IL-4: contribution of multiple signaling pathways.

Interleukin-4 is a cytokine produced by activated T cells, mast cells, and basophils that elicits many important biological responses[1] (see Tab 1). These responses range from the regulation of helper T cell differentiation[2] and the production of IgE[3] to the regulation of the adhesive properties of endothelial cells via VCAM-1[4]. In keeping with these diverse biological effects, high-affinity binding sites for IL-4 (Kd 20 to 300 pM) have been detected on many hematopoietic and non-hematopoietic cell types at levels ranging from 50 to 5000 sites per cell[5]. This review will focus on the discrete signal transduction pathways activated by the IL-4 receptor and the coordination of these individual pathways in the regulation of a final biological outcome.

Amino Acid Sequence↗
PubMed1998

[Implantable defibrillators in the management of malignant ventricular tachyarrhythmias. Experience at the Arrhythmia National Center].

BACKGROUND: Implantable defibrillators are the most effective means to prevent sudden death in patients with malignant ventricular tachyarrhythmias. The availability of this type of devices is limited in Chile, due to their high price. AIM: To report the first patients treated with implantable defibrillators in our hospital. PATIENTS AND METHODS: Nine males and one female aged 13 to 65 years old are reported. Three presented with ventricular fibrillation (presenting out of the hospital in three) and the rest had ventricular tachycardia resistant to drugs or radiofrequency ablation. RESULTS: All implants were performed using intracardiac electrodes. The generator was implanted in the pectoral region in nine and in the abdomen in one. A successful defibrillation was obtained with less than 15 J in four patients, with 20 J in three and with 24 J in three. There were no complications during the procedure. After a 12 months follow up, four patients have been treated by the implantable device. One of these subjects had a ventricular fibrillation in two occasions. One patient died of a bronchopneumonia two years after the implant. CONCLUSIONS: Implantable defibrillators are an effective therapy for the treatment of malignant ventricular arrhythmias with a high risk of sudden death.

Adolescent↗
PubMed1997

HMG-I(Y) phosphorylation status as a nuclear target regulated through insulin receptor substrate-1 and the I4R motif of the interleukin-4 receptor.

Interleukin (IL)-4 is a cytokine that regulates both the growth and differentiation of hematopoietic cells. Its ligand binding specificity and important signal transduction mechanisms are conferred by the IL-4 receptor alpha chain (IL-4Ralpha). The I4R is a tyrosine-containing motif within IL-4Ralpha that is critical for proliferative responses to IL-4. Although the I4R also contributes to gene regulation, nuclear targets directly regulated by this motif have not been described. It is shown here that the tyrosine at position 497 in the I4R is critical for regulation of the phosphorylation status of a set of nuclear proteins that includes HMG-I(Y), small non-histone chromosomal proteins involved in the control of gene expression in hematopoietic cell lines. Moreover, IL-4 is unable to induce HMG-I(Y) phosphorylation in insulin receptor substrate-1-deficient cells, and the inhibitor wortmannin completely blocks IL-4 regulation of HMG-I(Y) phosphorylation status but not activation of an IL-4 Stat protein. Taken together, these data indicate that HMG-I(Y) is a nuclear target whose phosphorylation status is regulated through the I4R motif via insulin receptor substrate proteins, independent of activation of the Stat pathway.

Androstadienes↗
PubMed1997

The IL-4-induced tyrosine phosphorylation of the insulin receptor substrate is dependent on JAK1 expression in human fibrosarcoma cells.

It has been shown that IL-4 induces the tyrosine phosphorylation of JAK1 and JAK3 in the majority of hemopoietic cell types, and JAK2 and TYK2 in several other types. However, the significance of this tyrosine phosphorylation in regulating IL-4 signaling has not been shown. To determine whether JAKs play a role in activating a signal transduction pathway different from the classical JAK/STAT pathway, we analyzed the ability of huIL-4 to stimulate the tyrosine phosphorylation of one of its major cellular substrates, the insulin receptor substrate (IRS). Using human fibrosarcoma cell lines with mutations in JAK1, JAK2, and TYK2, we found that expression of functional JAK1, but not TYK2 or JAK2, is essential for IL-4-induced tyrosine phosphorylation of IRS. We also provide evidence that the IRS pathway is independent of STAT-6, showing that JAK1 is essential for activating a STAT-independent pathway.

Fibrosarcoma↗
PubMed1997

Requirement of a second signal via protein kinase C or protein kinase A for maximal expression of CD40 ligand. Involvement of transcriptional and posttranscriptional mechanisms.

High levels of CD40 ligand (CD40L) protein expression are induced on native T cells by increasing the intracellular Ca2+ concentration. In the present study we have shown that ionomycin induces CD40L gene transcription leading to mRNA accumulation which translates to high levels of protein expression. Conversely, agents which increase the intracellular levels of cyclic AMP (cAMP), such as prostaglandin E2 (PGE2) or dibutyryl cyclic AMP (dbcAMP), were unable to induce CD40L expression on T lymphocytes. Cell activation by phorbol 12-myristate 13-acetate (PMA) treatment had a slight effect on increasing CD40L mRNA and protein levels. However, PMA and dbcAMP synergized with ionomycin to significantly increase and to prolong the CD40L expression. Nuclear run-on assays revealed that PMA, but not dbcAMP, increased threefold the CD40L gene transcription rate induced by ionomycin. This effect was independent of de novo protein synthesis. In addition, at a posttranscriptional level, both reagents synergized with the Ca2+ ionophore to prolong the CD40L mRNA half-life by a mechanism which was also independent of de novo protein synthesis. Moreover, when transcription was blocked with actinomycin D, an increment of the CD40L transcript levels induced by PMA or dbcAMP on ionomycin-treated cells was observed in the presence of cycloheximide. This probably means that newly synthesized protein may contribute to the CD40L mRNA destabilization. In summary, these data show that PMA and dbcAMP synergized with ionomycin to increase the CD40L mRNA and protein levels. The up-regulatory effect of PMA was accomplished at a transcriptional and posttranscriptional level, whereas dbcAMP exerted its synergistic effect exclusively at a posttranscriptional level.

Bucladesine↗
PubMed1997

[Detection of ventricular asynchrony using tissue Doppler echocardiography--a new method for assessing left ventricular wall motion].

UNLABELLED: The asynchrony of the heart in patients with coronary artery disease can be detected by digitized cine- and radionuclidventriculography. Both methods require time-consuming offline analysis. The aim of the current study was the assessment of the clinical value of the recently developed tissue Doppler echocardiography (TDE) to detect myocardial asynchrony. In the current study, 21 healthy subjects (age 49 +/- 14 y) and 22 patients with known coronary artery disease (20 with > 70% luminal narrowing of the LAD, 4 with a history of CABG, age 58 +/- 12 y) were included. In the apical 4-chamber-view, midseptal and midlateral LV segments were analyzed by 2-D and M-Mode-TDE. Evaluation was possible in 20 healthy subjects (95%) and 20 CAD patients (91%). During isovolumic relaxation time (IVRT) healthy subjects showed slow synchronous outward motion of the septum and the free wall with homogenous color coding (blue/green) and low negative tissue velocities followed by rapid symmetrical outward motion during rapid filling (RF) and atrial contraction (AC) phase (high negative velocities). During diatasis (DI) almost no wall motion could be detected. In 17 (85%) of 20 CAD patients, myocardial asynchrony during IVRT was detected; while the septum was moving inward (red coding with low positive velocities), the free wall was moving outward (blue green coding with low negative/velocities). After opening of the mitral valve, all CAD patients showed rapid, symmetrical outward motion of both the septum and the free wall with homogenous color coding and high negative tissue velocities. CONCLUSION: Tissue Doppler echocardiography detects ventricular asynchrony online. In patients with significant LAD stenosis, a pathological septal movement is observed during isovolumic relaxation time. Determinants of the etiology could be chronic hypoperfusion or ischemia ("hibernating myocardium").

Adult↗
PubMed1997

Non-invasive assessment of cardiac physiology by tissue Doppler echocardiography. A comparison with invasive haemodynamics.

BACKGROUND: Tissue Doppler echocardiography reveals characteristic patterns of myocardial velocities within systole and diastole which are not well understood. AIM: The purpose of this study was to determine the relationship of myocardial velocity patterns, as assessed by tissue Doppler echocardiography, to the contraction and relaxation phases of the cardiac cycle, as determined during cardiac catheterization. METHODS: Recordings of left ventricular/aortic and left ventricular/pulmonary wedge pressures were obtained simultaneously with apical tissue Doppler echocardiographic images of the left ventricle. A total of 210 cardiac cycles from 22 patients (mean age 58 years, 18 male) undergoing cardiac catheterization were analysed. The time intervals of the different phases of the cardiac cycle were measured from the pressure tracings. These time intervals were correlated to the interfaces of colour myocardial velocity patterns obtained by M-mode tissue Doppler echocardiography. RESULTS: There was a good correlation between the time intervals assessed haemodynamically and those based on the different velocity interfaces obtained with M-mode tissue Doppler echocardiography. Comparable time intervals (from the R wave) obtained by pressure recordings and tissue Doppler echocardiography were, respectively: isovolumic contraction (70 +/- 14 vs 67 +/- 9 ms, r = 0.79); rapid ejection (206 +/- 54 vs 202 +/- 49 ms; r = 0.95); late ejection (357 +/- 36 vs 346 +/- 42 ms. r = 0.93); isovolumic relaxation (405 +/- 43 vs 409 +/- 56 ms; r = 0.95); rapid filling (514 +/- 67 vs 523 +/- 64 ms, r = 0.91); diastasis (697 +/- 153 vs 709 +/- 146 ms, r = 0.98); atrial contraction (890 +/- 128 vs 899 +/- 132 ms, r = 0.96). CONCLUSION: Tissue Doppler echocardiography has the potential to accurately measure the different phases of the cardiac cycle which until now could only be determined invasively. It may provide a sensitive method for the assessment of changes in both cardiac contraction and relaxation in different clinical settings.

Atrial Fibrillation↗
PubMed1997

[Seroprevalence of cytomegalovirus and Toxoplasma gondii in healthy subjects under 30 years old in Santiago, Chile].

BACKGROUND: Infections by Cytomegalovirus and Toxoplasma gondii are endemic in Chile and only a low proportion of infected individuals have clinical manifestations. AIM: To study the prevalence of infection by Cytomegalovirus and Toxoplasma gondii in Chile. SUBJECTS AND METHODS: The prevalence of IgG antibodies against Cytomegalovirus and Toxoplasma gondii were studied in 560 subjects under 30 years old, using an ELISA technique. Age, socioeconomic level, breast feeding, assistance to nurseries and number of family members were considered as risk factors for these infections. RESULTS: Infection by Cytomegalovirus had a global prevalence of 60%. It showed an epidemiological pattern of late acquisition in high socioeconomic levels and a pattern of early infection in medium and low socioeconomic levels. Eighty to 90% of sera were positive for the infection in adult subjects of the three socioeconomic levels. There was a positive correlation between the duration of breast feeding and the frequency of Cytomegalovirus infection. Infection by Toxoplasma gondii had a global prevalence of 24.6%. The rates of susceptible individuals were 80 and 50% in high and medium-low socioeconomic levels respectively. CONCLUSIONS: The knowledge about the frequency of these infections in high risk populations such as women during their reproductive years and immunodepressed individuals, will allow the implementation of preventive measures.

Adolescent↗