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Biomedical subjects

M Serrano

Publications and source records attributed to M Serrano.

At least 55 records · Page 3Linked to original sources
PubMed1998

Crystal structure of the complex of the cyclin D-dependent kinase Cdk6 bound to the cell-cycle inhibitor p19INK4d.

The crystal structure of the cyclin D-dependent kinase Cdk6 bound to the p19 INK4d protein has been determined at 1.9 A resolution. The results provide the first structural information for a cyclin D-dependent protein kinase and show how the INK4 family of CDK inhibitors bind. The structure indicates that the conformational changes induced by p19INK4d inhibit both productive binding of ATP and the cyclin-induced rearrangement of the kinase from an inactive to an active conformation. The structure also shows how binding of an INK4 inhibitor would prevent binding of p27Kip1, resulting in its redistribution to other CDKs. Identification of the critical residues involved in the interaction explains how mutations in Cdk4 and p16INK4a result in loss of kinase inhibition and cancer.

Adenosine Triphosphate↗
PubMed1998

Auditory event-related potentials to semantic priming during sleep.

The present study uses the N400 component of event-related potentials (ERPs) as a processing marker of single spoken words presented during sleep. Thirteen healthy volunteers participated in the study. The auditory ERPs were registered in response to a semantic priming paradigm made up of pairs of words (50% related, 50% unrelated) presented in the waking state and during sleep stages II, III-IV and REM. The amplitude, latency and scalp distribution parameters of the negativity observed during stage II and the REM stage were contrasted with the results obtained in the waking state. The 'N400-like' effect elicited in these stages of sleep showed a mean amplitude for pairs of unrelated words significantly greater than for related pairs and an increment of latency. These results suggest that during these sleep stages a semantic priming effect is maintained actively although the lexical processing time increases.

Adult↗
PubMed1998

A phase I clinical trial of prolonged infusion of hydroxyurea in combination with hyperfractionated, accelerated, external radiation therapy in patients with advanced squamous cell cancer of the head and neck.

BACKGROUND: Preclinical data suggested that sustained inhibition of the anabolic enzyme, ribonucleotide reductase (RR), by hydroxyurea (HU) may be critical for the anticancer effects of the drug. A phase I trial of continuous infusion HU with concomitant hyperfractionated, accelerated radiation therapy (CHU-CHRT) was initiated to determine the maximum tolerated dose (MTD) and dose limiting toxicities (DLT) of HU in patients with locally advanced squamous cell carcinoma (SCC) of the head and neck. METHODS: Patients were required to have histologically-documented and radiographically-staged locally advanced SCC of the hypopharynx (AJC stages II, III or IV), oropharynx (AJC stage IV), or oral cavity (AJC stage IV) not amenable to reasonable surgical resection. Eligible patients had adequate bone marrow, hepatic, and renal function and had to give informed consent. Concomitant, hyperfractionated, accelerated radiation therapy (CHRT) consisted of 1.2 Gy BID (6 hour minimum interfraction interval) on weekdays and 1.2 Gy delivered daily on the weekends to a total tumor dose of 74.4 Gy. Continuous infusion hydroxyurea (CHU) was administered at 0.25-0.375 mg/m2/min as a continuous intravenous infusion daily for 5 days with weekends days off for the duration of the radiation therapy. The dose of HU was increased by 0.125 mg/m2/min between dose levels until DLT was reached in 2/6 patients. If the primary had a complete clinical response and biopsies were negative, planned neck dissections were performed. RESULTS: Fifteen patients were enrolled and are evaluable. The initial dose level, 0.25 mg/m2/min was tolerated by 3/3 patients. At 0.375 mg/m2/min, 3/6 patients experienced grade 3-4 infections, with one patient having a non-fatal, subendocardial infarction. At 0.313 mg/m2/min, no patient experienced DLT. CONCLUSION: The MTD for CHU-CHRT was 0.313 mg/m2/min. The toxicities were primarily mucosal and a phase II study is in progress.

Adult↗
PubMed1998

Inhibition of tumour necrosis factor and reversal of endotoxin-induced shock by U-83836E, a 'second generation' lazaroid in rats.

1. Antioxidants can exert protective effects in endotoxic shock by either a reduction of the oxidant damage or attenuation of Tumour Necrosis Factor (TNF-alpha) production. 2. Lazaroids are a family of compounds that inhibit lipid peroxidation. Besides, they can also reduce TNF-alpha. U-83836E is a new lazaroid lacking the glucocorticoid ring. 3. Aim of our study was to investigate the effect of U-83836E on TNF-alpha production either in vivo or in vitro. Endotoxic shock was produced in male rats by a single intravenous (i.v.) injection of 20 mg kg(-1) of S. enteritidis lipopolysaccharide (LPS). LPS administration reduced survival rate (0% survival, 72 h after endotoxin administration), decreased mean arterial blood pressure, increased serum and macrophage TNF-alpha and enhanced plasma malonylaldehyde (MAL) levels. Furthermore aortic rings from shocked rats showed a marked hyporeactivity to phenylephrine (PE 1 nM-10 microM). 4. Treatment with U-83836E (7.5, 15 and 30 mg kg(-1), i.v.) 5 min after endotoxin challenge significantly protected against LPS induced lethality (90% survival rate and 80% survival rate 24 h and 72 h after LPS injection respectively, following the highest dose of the drug), reduced hypotension, blunted plasma MAL, decreased serum and macrophage TNF-alpha and restored the hyporeactivity of aortic rings to control values. In vitro LPS stimulation (50 microg ml(-1) for 4 h) significantly increased cytokine production in macrophages (Mphi) harvested from untreated normal rats. Pretreatment with pertussis toxin (PT; 0.1, 1 and 10 ng ml(-1) 4 h before LPS) significantly increased TNF-alpha production. PT effects on these LPS responses were correlated with a PT mediated ADP ribosylation of a 41 kDa protein. U-83836E (50 microM) reduced, in a dose dependent manner, LPS induced TNF-alpha production and inhibited the PT effects on cytokine production and on ADP ribosylation of the protein. 5. Our data suggest that lazaroids may affect the early events associated with LPS receptor mediated activation of a G protein in LPS induced TNF-alpha production. These molecular events may explain, at least in part, the in vivo inhibition of cytokine production and reversal of endotoxic shock.

Animals↗
PubMed1998

Prevention of aluminium exposure through dialysis fluids. Analysis of changes in the last 8 years.

Despite extensive measures to control aluminium exposure, chronic and acute episodes of aluminium intoxication still occur. The objective of this study was to analyse the changes in the aluminium content of dialysis fluid and the effect on serum aluminium in different dialysis centres in Spain in the last 8 years. For this purpose, the aluminium content in dialysis fluid and serum samples (N=5609) from 17 dialysis centres was analysed for >8 years (from the last quarter of 1988 to 1996). In that period of time, the percentage of dialysis fluid samples with acceptable concentrations of aluminium (<2 microg/l) increased from 0% in 1988 to 80% in 1996. The percentage of dialysis fluid samples with high aluminium levels (>6 microg/l) ranged between 37.5% in 1988 and 2.3% in 1996. The improvement in the quality of the dialysis fluid resulted in lower values of serum aluminium. The percentage of serum samples with low aluminium (<20 microg/l) increased from 16.5% in 1988 to 54.2% in 1966. The mean serum aluminium correlated with the mean dialysis fluid aluminium (r=0.55, P<0.001). A higher correlation was found when the aluminium in dialysis fluid ranged between 4 and 10 microg/l (r=0.802, P<0.001), and no correlation was found when the aluminium in dialysis fluid was <4 microg/l. Even taking into account that the dialysis fluid is not the only source of aluminium for dialysis patients, our study clearly demonstrated a close relationship with the serum aluminium content. Therefore, we must emphasize the necessity for controlling the aluminium content in dialysis fluid more often than is done at present.

Aluminum↗
PubMed1998

Increased Na,K,Cl cotransporter and Na, K-ATPase activity of vascular tissue in two-kidney Goldblatt hypertension.

The properties of the Na/K pump and Na,K,Cl cotransporter were studied in vascular tissue of two-kidney Goldblatt hypertensive rats. These transport systems were measured as ouabain-sensitive and bumetanide-sensitive 86Rb/K uptake in aortic rings, left ventricular muscle and soleus skeletal muscle fibers of control and hypertensive Sprague-Dawley rats. A dramatic increment in Na/K pump activity was observed in intact aortic rings from the hypertensive group. The same was true for the Na,K,Cl cotransporter. The transport parameters related to the left ventricular muscle and soleus skeletal muscle were not significantly altered in the hypertensive rats. Measurements of the catalytic isoforms of the Na+, K(+)-ATPase in the aortic rings indicated that both isoforms (alpha 1 and alpha 2) were elevated in the same proportion in the hypertensive rats. The results also indicate that the endothelium plays an important role in both transport systems: in the absence of endothelium, a much lower 86Rb/K uptake was observed than in intact aortic rings, either from control or hypertensive vascular tissue. Nevertheless, when the 86Rb/K transport activity was measured in denuded aortic rings, a significantly higher ouabain and bumetanide sensitive 86Rb/K uptake was also observed in the hypertensive rats. These data also show no alteration in the endothelium of the hypertensive rats as compared to control animals. The presence of endothelium had a more striking effect on the alpha 2 catalytic isoform activity than on the alpha 1 isoform. We conclude that there is a significant increment in the Na/K pump and Na,K,Cl cotransporter in two kidney-Goldblatt hypertension, that is specific for vascular smooth muscle.

Animals↗
PubMed1997

The tumor suppressor protein p16INK4a.

The tumor suppressor protein p16INK4a (inhibitor of CDK4) is one of the most direct links between cell-cycle control and cancer. The p16INK4a gene is frequently inactivated in human tumors, and inheritance of mutant alleles results in susceptibility to several types of cancer. p16INK4a is part of a cell-cycle regulatory pathway that converges in the tumor suppressor protein Rb. The mechanisms that regulate p16INK4a are starting to be characterized.

Alleles↗
PubMed1997

17Beta-oestradiol reduces cardiac leukocyte accumulation in myocardial ischaemia reperfusion injury in rat.

We investigated whether oestrogens modulate the phenomenon of leukocyte accumulation during ischaemia and reperfusion of the myocardium. Anaesthetized rats were subjected to total occlusion (1 h) of the left main coronary artery followed by 1 h reperfusion. Sham myocardial ischaemia-reperfusion rats (Sham) were used as controls. Myocardial necrosis, myocardial myeloperoxidase activity, serum creatinine phosphokinase activity, serum and macrophages tumour necrosis factor (TNF-alpha) and the myocardial staining of intercellular adhesion molecule-1 (ICAM-1) were evaluated. Myocardial ischaemia plus reperfusion in untreated rats produced marked myocardial necrosis, increased serum creatinine phosphokinase activity (348 +/- 38 U/ml) and cardiac myeloperoxidase activity, a marker of polymorphonuclear leukocyte accumulation, both in the area at risk and in the necrotic area (MPO 9 +/- 1.1 mU/g tissue and 8.2 +/- 1 mU/g tissue, respectively), and induced a marked increase in the macrophage (156 +/- 14 U/ml at the end of reperfusion) and serum (344 +/- 12 U/ml, at the end of reperfusion) levels of TNF-alpha. Finally, myocardial ischaemia-reperfusion injury increased ICAM-1 staining in the myocardium. Administration of 17beta-oestradiol (5, 10 and 20 microg/kg, i.m. 5 min after induction of myocardial ischaemia-reperfusion injury), lowered myocardial necrosis and myeloperoxidase activity in the area at risk and in the necrotic area, reduced serum and macrophages TNF-alpha (20 +/- 3 U/ml and 9 +/- 3 U/ml, respectively) and decreased serum creatinine phosphokinase activity (67 +/- 3 U/ml). Oestrogen treatment also blunted the increased staining of ICAM-1 in the injured myocardium. Finally, 17beta-oestradiol added in vitro to peritoneal macrophages collected from untreated rats subjected to myocardial ischaemia-reperfusion injury, significantly reduced TNF-alpha production. Our results suggest that 17beta-oestradiol, by inhibiting TNF-alpha production, limits the deleterious ICAM-1-mediated binding of leukocytes to injured myocardium and protects against myocardial ischaemia-reperfusion injury.

Animals↗
PubMed1997

Inactivation of the cyclin-dependent kinase inhibitor p15INK4b by deletion and de novo methylation with independence of p16INK4a alterations in murine primary T-cell lymphomas.

A wide panel of murine induced T-cell lymphomas have been analysed for p16INK4a or p15INK4b alterations. Only one gamma-radiation-induced lymphoma showed p16INK4a homozygous deletion and no other intragenic mutations were found in these INK4 genes. However, de novo methylation of the 5' CpG islands of the murine p15INK4b and p16INK4a genes was found to be highly frequent. While p16INK4a hypermethylation was found in 36% of the neutron-radiation-induced lymphomas and 15% of the gamma-radiation-induced lymphomas, de novo methylation of p15INK4b occurs in 88% and 42% of these tumors respectively, correlating with deficient expression of the corresponding mRNA and allelic losses in the p15INK4b and p16INK4a chromosome location. These data represent, to our knowledge, the first report on the significant involvement of hypermethylation of these INK4 genes in murine primary tumors. Moreover, they show the importance of allelic losses and CpG island methylation of p15INK4b gene inactivation and support a tumor suppressor role for p15INK4b in T-cell lymphomas independent of p16INK4a.

Animals↗
PubMed1997

Oncogenic ras provokes premature cell senescence associated with accumulation of p53 and p16INK4a.

Oncogenic ras can transform most immortal rodent cells to a tumorigenic state. However, transformation of primary cells by ras requires either a cooperating oncogene or the inactivation of tumor suppressors such as p53 or p16. Here we show that expression of oncogenic ras in primary human or rodent cells results in a permanent G1 arrest. The arrest induced by ras is accompanied by accumulation of p53 and p16, and is phenotypically indistinguishable from cellular senescence. Inactivation of either p53 or p16 prevents ras-induced arrest in rodent cells, and E1A achieves a similar effect in human cells. These observations suggest that the onset of cellular senescence does not simply reflect the accumulation of cell divisions, but can be prematurely activated in response to an oncogenic stimulus. Negation of ras-induced senescence may be relevant during multistep tumorigenesis.

Animals↗
PubMed1997

Gliomatosis cerebri mimicking a metastatic breast cancer: fatal outcome.

Gliomatosis cerebri (GC) is defined by the World Health Organization as a neoplasm of the glial cells. It is extremely rare, and there exists only 160 documented cases since 1897. There is no known treatment and the median survival rate is one year. The association of extracranial and CNS tumors is unusual, only three cases of breast cancer have been associated with gliomas and meningiomas but none with GC. Below we describe a case of breast cancer associated with GC and review the anatomoclinical and radiological manifestations of Gliomatosis Cerebri.

Aged↗
PubMed1997

Prognostic value from DNA quantification by static cytometry in breast cancer.

OBJECTIVE: To determine if DNA quantification on fine needle aspiration (FNA) has predictive value in breast cancer. STUDY DESIGN: Forty-nine patients with breast cancer were selected for this study because of their common characteristics. The smears were studied using image cytometry. RESULTS: With the type of histogram and value of entropy, two large groups, with high and low degrees of malignancy, were obtained. Survival was utilized as a variable of interest. Kaplan-Meier survival curves for both groups were formulated, and the results were supported with statistical data. CONCLUSION: The survival differences between both groups were statistically significant (P < .001), thus demonstrating the predictive value of DNA quantification.

Biopsy, Needle↗
PubMed1997

[Age as source of variation in various parameters of 'delta sleep'].

INTRODUCTION: The clinic usefulness of a diagnostic test is in relationship to the precision with which measures the studied phenomenon. The lack of precision involve the reliability upon causing confounded results of the normal and diseased populations. OBJECTIVE: Since the sleep varies in function of the age, to find sleep parameters that fit better to the changes that the aging produces in the sleep. MATERIAL AND METHODS: Spectral analysis through the Fast Fourier Transformation of the ambulatory EEG of 28 healthy subjects. RESULTS: Maximum value of power (maximum depth) in a frequency of ended in a point of the sleep goes losing in a way specifies and systematical with the age. CONCLUSIONS: The variance accounted by this parameter is of the 87%, what, being tried to a phenomenon so variable as the sleep, supposes a interesting starting point to be applied to some pathologies in those which is presumed that the slow sleep (to which is attributed a paper in the cerebral restoration) is decreased.

Adolescent↗
PubMed1996

Activation of replication origins in phi29-related phages requires the recognition of initiation proteins to specific nucleoprotein complexes.

Protein p6 of Bacillus subtilis phage phi29 activates the initiation of viral DNA replication by forming a multimeric nucleoprotein complex at the origins of replication, located at both ends of the linear genome. This activation requires a precise positioning of the protein p6 array with respect to the initiation site. To investigate this activation mechanism, we have purified the phi29 protein p6 counterparts from the related phages Nf and GA-1 and analyzed the formation of complexes with DNA. In the homologous protein p6-DNA complexes the phi29 and Nf protein arrays showed an identical positioning, different than that of the GA-1 protein array. In contrast, in the heterologous complexes the protein showed a different arrangement except in the case of the Nf protein-phi29 DNA complex. We have also purified the proteins involved in the initiation of replication (terminal protein and DNA polymerase) from phages Nf and GA-1 and measured the ability of the different p6 proteins to activate homologous and heterologous replication origins. The results obtained indicate that the activation requires not only the formation of a specific nucleoprotein complex but also its specific recognition by the proteins involved in the initiation of DNA replication.

Bacillus Phages↗
PubMed1996

Role of the INK4a locus in tumor suppression and cell mortality.

The cell cycle inhibitor p16INK4a is inactivated in many human tumors and in families with hereditary melanoma and pancreatic cancer. Tumor-associated alterations in the INK4a locus may also affect the overlapping gene encoding p19ARF and the adjacent gene encoding p15I1NK4b, both negative regulators of cell proliferation. We report the phenotype of mice carrying a targeted deletion of the INK4a locus that eliminates both p16INK4a and p19ARF. The mice are viable but develop spontaneous tumors at an early age and are highly sensitive to carcinogenic treatments. INK4a-deficient primary fibroblasts proliferate rapidly and have a high colony-formation efficiency. In contrast with normal cells, the introduction of activated Ha-ras into INK4a-deficient fibroblasts can result in neoplastic transformation. These findings directly demonstrate that the INK4a locus functions to suppress neoplastic growth.

Alleles↗
PubMed1996

Analysis of p16INK4a and its interaction with CDK4.

The interaction between cyclin-dependent kinase 4 (CDK4) and its inhibitor p16INK4a (p16) was studied by random mutagenesis and yeast two-hybrid system. The gene encoding p16 was mutagenized randomly and the amino acid changes that affect the binding of p16 to CDK4 were identified. Several amino acid residues were shown to be important for the binding and many of these changes occur at residues conserved in all known human p16 family proteins Most of the mutant p16 proteins that failed to bind to CDK4 contained multiple amino acid changes, and these alterations were observed throughout the entire gene with no apparent mutational patterns or hot spots. Some of the mutations that moderately reduced the binding activity severely affected the kinase-inhibitory activity of p16.

Amino Acid Sequence↗