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Biomedical subjects

M Serrano

Publications and source records attributed to M Serrano.

At least 109 records · Page 6Linked to original sources
PubMed1994

Mutations and altered expression of p16INK4 in human cancer.

Cell cycle arrest at the G1 checkpoint allows completion of critical macromolecular events prior to S phase. Regulators of the G1 checkpoint include an inhibitor of cyclin-dependent kinase, p16INK4; two tumor-suppressor proteins, p53 and RB (the product of the retinoblastoma-susceptibility gene); and cyclin D1. Neither p16INK4 nor the RB protein was detected in 28 of 29 tumor cell lines from human lung, esophagus, liver, colon, and pancreas. The presence of p16INK4 protein is inversely correlated with detectable RB or cyclin D1 proteins and is not correlated with p53 mutations. Homozygous deletions of p16INK4 were detected in several cell lines, but intragenic mutations of this gene were unusual in either cell lines or primary tumors. Transfection of the p16INK4 cDNA expression vector into carcinoma cells inhibits their colony-forming efficiency and the p16INK4 expressing cells are selected against with continued passage in vitro. These results are consistent with the hypothesis that p16INK4 is a tumor-suppressor protein and that genetic and epigenetic abnormalities in genes controlling the G1 checkpoint can lead to both escape from senescence and cancer formation.

Base Sequence↗
PubMed1994

Phage phi 29 protein p6: a viral histone-like protein.

Phage phi 29 protein p6 is one of the most abundant viral proteins in phi 29-infected B subtilis cells, constituting about 4% of the total cellular proteins (about 3 x 10(6) copies/cell) at late infection. Electron microscopic studies showed that, in vitro, protein p6 forms heterogeneously-sized complexes all along phi 29 DNA, suggesting that protein p6 may have a role in genome packaging and organization. The low stability of the protein p6-phi 29 DNA complexes observed in vitro could reflect the dynamic nature of these complexes, to allow replication, transcription, and encapsidation of the genome. The protein p6-DNA complex consists of a DNA right-handed superhelix wrapped around a multimeric protein core. The DNA in this complex is strongly distorted and compacted. Protein p6 recognition signals have been mapped near the ends of the linear phi 29 DNA and act as nucleation sites for complex formation. Protein p6 does not recognize a specific sequence, but sequences with specific bendable properties that would favor the formation of the complex. Protein p6 represses transcription from the phi 29 C2 early promoter, and activates initiation of phi 29 DNA replication that occurs from both DNA ends. The formation of nucleoprotein complexes at the origins of replication, as well as the specific positioning of protein p6 with respect to the DNA ends are required for the activation of replication. This suggests that the proteins involved in the initiation step of phi 29 DNA replication, either directly interact with protein p6, or recognize a conformational change at a specific location in the DNA. The mechanism of activation could be the local and transient unpairing of DNA at specific sites, facilitated by the strong distortion of DNA conformation in the nucleoprotein complex.

Bacillus Phages↗
PubMed1994

DNA structure in the nucleoprotein complex that activates replication of phage phi 29.

Initiation of phage phi 29 DNA replication is activated by the viral protein p6 which forms a nucleoprotein complex at the replication origins, located at the linear genome ends. The complex consists of a DNA right-handed superhelix wrapped around a multimeric protein core. We have determined the superhelical path of the DNA in the complex, measuring the change in linking number induced by the protein, the surface-related helical repeat and the compaction of the DNA. One superhelical turn has approximately 63 bp (2.6 p6 dimers). Furthermore, we have determined that the DNA binding domain of protein p6 is located at the N-terminal region, predicted to form an amphipathic alpha-helix. We have obtained, by site-directed mutagenesis, protein p6 mutants in the polar side of the putative helix in which their DNA binding and replication activation properties were impaired or undetectable, in agreement with in vivo results.

Bacillus Phages↗
PubMed1994

[The combined treatment of anaplastic thyroid carcinoma].

Five patients with anaplastic thyroid carcinoma (ATC) were treated with chemotherapy (adriamycin 60 mg/m2 and cisplatin 90 mg/m2) followed by surgery and postoperative radiotherapy. The average survival in this series was 11.2 months. Partial response of the cervical tumoration was observed in 4 patients and of the distant metastasis, in 1 of 3 cases. One patient with mixed thyroid carcinoma (follicular and anaplastic) had complete remission of the disease. Four patients died: two due to metastatic disease and two due to local tumor growth. We conclude that this type of combined therapy is affective in patients with anaplastic thyroid carcinoma in terms of local control of the disease, avoiding mortality due to local invasion in some of these patients.

Aged↗
PubMed1993

A new regulatory motif in cell-cycle control causing specific inhibition of cyclin D/CDK4.

The division cycle of eukaryotic cells is regulated by a family of protein kinases known as the cyclin-dependent kinases (CDKs). The sequential activation of individual members of this family and their consequent phosphorylation of critical substrates promotes orderly progression through the cell cycle. The complexes formed by CDK4 and the D-type cyclins have been strongly implicated in the control of cell proliferation during the G1 phase. CDK4 exists, in part, as a multi-protein complex with a D-type cyclin, proliferating cell nuclear antigen and a protein, p21 (refs 7-9). CDK4 associates separately with a protein of M(r) 16K, particularly in cells lacking a functional retinoblastoma protein. Here we report the isolation of a human p16 complementary DNA and demonstrate that p16 binds to CDK4 and inhibits the catalytic activity of the CDK4/cyclin D enzymes. p16 seems to act in a regulatory feedback circuit with CDK4, D-type cyclins and retinoblastoma protein.

Amino Acid Sequence↗
PubMed1993

Superhelical path of the DNA in the nucleoprotein complex that activates the initiation of phage phi 29 DNA replication.

Initiation of bacteriophage phi 29 DNA replication is activated by protein p6, a viral double-stranded DNA-binding protein that forms a nucleoprotein complex at the viral replication origins. This complex consists of a DNA right-handed superhelix wrapped around a multimeric protein p6 core with protein p6 dimers regularly bound every 24 base-pairs (bp). In this paper, we have constructed a concatemer formed by direct repeats of a 24 bp sequence previously proposed to act as a signal for protein p6 binding at a phi 29 replication origin. DNase I footprinting shows that protein p6 binds to the concatemer in a similar way to the phi 29 DNA replication origins but with higher affinity, indicating that the 24 bp sequence is a recognition signal for protein p6. Furthermore, the concatemer was cloned in a plasmid and, by electron microscopy, it was shown to be the highest-affinity protein p6 binding region present in the plasmid. Based on these observations, the linking number change restrained by protein p6 has been measured in a series of plasmids containing concatemers with different numbers of 24 bp repeats; from the values obtained the linking number change restrained by a single protein p6 dimer has been estimated (delta Lkd = 0.1). In addition, when protein p6-DNA complexes fixed with glutaraldehyde were analysed by electron microscopy, it was observed that protein p6 compacts 4.2-fold the length of naked DNA. These data, together with the previously known value of the surface-related DNA helical repeat in the complex (12 bp), completely define the superhelical path of the DNA in the complex: one superhelical turn approximately involves 63 bp and 2.6 protein p6 dimers, and the DNA superhelix has a diameter of 6.6 nm and a slope of 14 degrees. The data obtained also indicate that the DNA in the protein p6-DNA complex is undertwisted (11.5 bp/turn) and strongly bent (66 degrees/12 bp). These DNA conformational changes might contribute to the activation of phi 29 DNA initiation of replication by protein p6.

Bacillus Phages↗
PubMed1993

[Yield of laboratory tests in the detection of excessive drinkers in the work place].

BACKGROUND: The identification of personnel consuming excess quantities of alcoholic beverages in the work place is necessary due to the economic, health, and laboral problems caused by alcohol. The objectivity of laboratory tests provides great credibility and these tests have replaced other methods as the anamnesis. METHODS: A transversal study was carried out of the employees of a Barcelona enterprise. During the periodic medical check ups the consumption of alcoholic beverages and laboratory blood tests were obtained from the employees seven days previously. RESULTS: All the laboratory tests presented a weak positive correlation with the consumption of alcohol. The calculation of diagnostic efficacy for weekly consumption equal to or grater than 280 g of pure alcohol in a male shows that the greatest sensitivity corresponds to mean corpuscle volume (33%) followed by gammaglutamyltranspeptidase (GGT) (16%) with high specificity being observed in both (85% and 93%, respectively). The results are better in employee consuming more than or equal to 420 g of alcohol weekly. CONCLUSIONS: The diagnostic efficacy of laboratory tests in identifying subjects with elevated consumption of alcohol in the work place has low clinical performance and the use of anamnesis on the consumption of alcoholic beverages should first be evaluated.

Alanine Transaminase↗
PubMed1993

Multimeric complexes formed by DNA-binding proteins of low sequence specificity.

Some proteins bind to double-stranded DNA with low sequence specificity, forming regular multimeric complexes that extend over large regions of DNA, strongly distorting its conformation. Formation of these complexes at particular DNA sites usually depends on the structural ability of the DNA to follow the path imposed by the protein array. These complexes are found in both prokaryotic and eukaryotic organisms and participate in processes such as DNA replication, transcription and packaging.

DNA↗
PubMed1993

[Chronic renal insufficiency and secondary hyperparathyroidism in rats. Biochemical and histological evaluation].

Chronic renal failure (CRF) in rats (surgical nephrectomy, 5/6) as well as its derived bone lesions have been studied. Eighty-five male Wistar rats were used, to which chronic renal failure was induced in 1 or 2 surgical times, the parameters of renal function in basal conditions and at different times after surgery being determined. With the method used chronic renal failure is induced with values of creatinine clearance 2/3 times lower than the initial ones (p < 0.05), which stabilize at the 7th week. On the other hand the parathyroid hormone levels (PTH) in serum triple (from 125 +/- 49 to 395 +/- 191, p < 0.05), and a decrease in the tubular phosphate reabsorption is produced (p < 0.001). In bone histology an increase in resorption and bone formation is observed as well as paratrabecular fibrosis, all of which is compatible with the histological diagnosis of hyperparathyroidism. The model of surgical renal insufficiency with ablation of 5/6 of the renal mass, reduces renal function to 1/3 of the initial values after 7 weeks, this procedure having a 20% global mortality without differences being observed between the carrying out of nephrectomies in 1 or 2 surgical times. This degree of CRF was accompanied by secondary hyperparathyroidism both at the biochemical and histological levels, findings which are of great usefulness for future experimental studies.

Animals↗
PubMed1993

Approaches for providing target values to improve usefulness of external quality assessment scheme. The Spanish experience.

The aim of this communication is to highlight the specific aspects of external quality assessment schemes that need to be discussed in a European context: target values, transferability of results and accredit of laboratories. The Spanish situation is presented here. The most reliable way to provide target values is to analyse the control samples by reference methods. However, it is not possible for the majority of national schemes and other approaches are presently used: the verification of consensus means is a practicable solution adopted in Spain. An initial network involving selected routine laboratories has been developed, to attain transferability of results. The traceability of routine calibrators from certified reference materials should be demonstrated. To accredit laboratories for licensing is a complex activity that should consider many aspects, results from the national quality assessment scheme bring one. A scoring system is being used in Spain for guidance, and the complete guidelines are under preparation.

Accreditation↗
PubMed1993

[Diffuse mediastinal lipomatosis and exogenous obesity].

A case of diffuse mediastinal lipomatosis (ML) is described, associated to exogenous obesity, infrequent entity, which is comprehended within the causes which can cause mediastinal broadening. The clinical characteristics of our observation and the general aspects of interest are reviewed for this type of mediastinal affectation.

Aged↗
PubMed1992

DNA conformational change induced by the bacteriophage phi 29 connector.

Translocation of viral DNA inwards and outwards of the capsid of double-stranded DNA bacteriophages occurs through the connector, a key viral structure that is known to interact with DNA. It is shown here that phage phi 29 connector binds both linear and circular double-stranded DNA. However, DNA-mediated protection of phi 29 connectors against Staphylococcus aureus endoprotease V8 digestion suggests that binding to linear DNA is more stable than to circular DNA. Endoprotease V8-protection assays also suggest that the length of the linear DNA required to produce a stable phi 29 connector-DNA interaction is, at least, twice longer than the phi 29 connector channel. This result is confirmed by experiments of phi 29 connector-protection of DNA against DNase I digestion. Furthermore, DNA circularization assays indicate that phi 29 connectors restrain negative supercoiling when bound to linear DNA. This DNA conformational change is not observed upon binding to circular DNA and it could reflect the existence of some left-handed DNA coiling or DNA untwisting inside of the phi 29 connector channel.

Bacillus Phages↗
PubMed1992

[Detection of hepatitis B virus DNA in serum by gene amplification in patients with chronic hepatitis B and in patients chronic hepatitis C].

BACKGROUND: The polymerase chain reaction (PCR) constitutes important methodological progress for detecting the presence of viral nucleic acids when these are found in small quantities in serum or tissues. The aim of this study was the use of PCR to detect DNA of the hepatitis B virus (HBV) in patients with chronic HBsAg positive hepatitis (HC-B) and in patients with chronic non A non B hepatitis with antibodies against the virus of hepatitis C (anti-HCV) positive (HC-C). METHODS: The DNA of the HBV was determined with PCR in the serum of 40 patients with HC-B and in 15 with HC-C. Moreover, the presence of anti-HCV was studied in the patients with HC-B. RESULTS: The presence of DNA of the HBV was detected by PCR in 69% of the HC-B patients presenting HBeAg positive and DNA of the HBV negative by simple hybridization as well as in 50% of the patients with HBeAg negative, anti-HBe positive and DNA of HBV negative by simple hybridization. In addition, DNA of HBV was detected by PCR in 27% (4/15) of the subjects with HC-C, three of whom had anti-HBc antibodies. On the other hand, 20% of the patients with HC-B had anti-HCV. Anti-HCV positivity was associated to a greater hypertransaminasemia in patients with HC-B in a non replicative phase. CONCLUSIONS: PCR is a sensitive method for detecting viral replication. Its use permits the detection of low DNA concentrations of the HBV in a low but appreciable percentage of chronic negative HBsAg hepatitis. Coinfection by the B and C viruses of hepatitis is not exceptional and explains hypertransaminasemia in some HC-B in a non replicative phase.

DNA, Viral↗