Biomedical subjects

C Alonso

Publications and source records attributed to C Alonso.

At least 109 records · Page 6Linked to original sources

PubMed2000

Effect of supragingival plaque control on subgingival microflora and gingivo-periodontal tissues.

The aim of this study was to carry out a short term evaluation of the effect of supragingival plaque control on the subgingival microflora and gingivoperiodontal tissues in patients with moderate and severe periodontitis. A total of 92 sites from 23 patients were studied; patients ranged between 35 and 60 years of age (x 46.4 +/- 1.71). Plaque Index (PI), Gingival Index (GI), Bleeding on Probing (BP), Probing Depth (PD), and Attachment Level (AL) were measured. Subgingival microflora of the same sites was studied by dark field microscopy, and cultures in non-selective and selective media for Actinobacillus actinomycetemcomitans (Aa), Prevotella intermedia/nigrescens (Pi/n), Porphyromonas gingivalis (Pg) and Fusobacterium nucleatum (Fn). The patients were included in a supragingival plaque control program which included oral hygiene training and assessment, and professional control of supragingival plaque and calculus. After 28 days, measurement of clinical parameters and the microbiological study were repeated. The sites were divided according to baseline PD values into: Group 1 (5 mm), Group 2 (6 mm) and Group 3 (> or = 7 mm). Results revealed a significant reduction in PI scores, GI scores, PB and PD in groups 2 and 3. No significant difference in AL was observed in any of the three groups. PI scores were found to decrease significantly in group 1, and GI scores, BP and PD tended to decrease. A significant increase in coccoid cells and a significant decrease in motile rods and spirochetes, both small and total, was observed in groups 2 and 3. Group 1 showed a significant decrease in medium and total spirochetes. At the end of the study, a significant decrease in Pi/n was found in all three groups. Significant decreases were found in Pg and Fn in Groups 3 and 2 respectively. Although it holds true that supragingival plaque control reduced a potentially pathogenic subgingival microbiota, this effect was not sufficient to be compatible with healthy gingivoperiodontal tissues.

Adult↗

PubMed1999

Combined effect of platination and intercalation upon DNA binding of novel cytotoxic Pt-bis(naphthalimide) complexes.

The reaction of platinum salts with bis(naphthalimide), compound 1, yielded two Pt-bis(naphthalimide) complexes, compounds 2 and 3 which differ from each other in their leaving groups being 1,1-cyclobutane dicarboxylate or chloride, respectively. The testing of the cytotoxic activity of compounds 2 and 3 against several tumor cell lines indicated that both compounds may be endowed with important antineoplastic properties since they circumvent cisplatin resistance. At similar rates of DNA platination (r(b) = 0.025), compounds 2 and 3 unwind supercoiled pUC8 DNA by (48 +/- 2) degrees. Altogether, these data suggest (i) that the cytotoxic activity of compounds 2 and 3 may be due to a combined effect of platination and intercalation and (ii) that the bis(naphthalimide) ligand is a suitable "carrier" that favors DNA targeting by cis-Pt(II) centers.

1-Naphthylamine↗

PubMed1999

Electrocardiographic predictive factors of long-term clinical improvement with multisite biventricular pacing in advanced heart failure.

Biventricular pacing has recently been proposed for treating patients with drug refractory heart failure and intraventricular conduction delay. The purpose is to restore ventricular relaxation and contraction sequences as homogeneously as possible. The aim of this study was to determine if some factors could predict the long-term clinical effectiveness of that new treatment. This study included 26 patients, aged 66 +/- 7 years, with drug refractory heart failure and wide QRS. Patients were implanted with a biventricular pacemaker. The left ventricle was paced through a coronary sinus tributary. New York Heart Association functional class, exercise tolerance, and left ventricular (LV) ejection fraction were collected at baseline and after pacemaker implantation. Patients were divided into 2 groups: group I = responders; group II = nonresponders. QRS duration and axis at baseline and during biventricular pacing, interventricular conduction time, and LV and right ventricular lead positions were compared between the 2 groups. Group I patients (n = 19) had a mean reduction of 1.3 in functional class and an increase in peak oxygen consumption rate by a mean of 50%. The only parameter that differed between the 2 groups was the QRS duration during biventricular pacing, with a significantly shorter value in group I than in group II (154 +/- 17 vs 177 +/- 26 ms; p = 0.016). Thus, a positive response to biventricular pacing is correlated with the quality of electrical resynchronization. The optimal positions of the right and LV leads would be those that could induce the greatest shortening of QRS duration.

Aged↗

PubMed1999

BRCA1 mutation analysis in 83 Spanish breast and breast/ovarian cancer families.

A group of 83 Spanish BC/OC families were analysed for BRCA1 germ-line mutations. Analysis of the entire coding sequence was carried out by SSCP and PTT. We identified 5 frameshift mutations: 185delAG (2 times), 189insTGTC, 1241delAC, and 5537delA and 3 missense mutations in BRCA1: 330A > G, 1240C > T, and 5263G > A. The 185delAG mutation was identified in 2 families. One of them was the only Gypsy family participating in our study. All carriers shared the known founder haplotype, although they did not have any Jewish ancestry. Three frameshift mutations were found among the 14 families with 3 or more BC cases and 1 or more OC, which results in a higher percentage (21.4%) of BRCA1 mutations in this group of high risk families. Two missense mutations were found in families with 2 or 3 BC and no OC, indicating a low proportion (4.8%) of mutations in these families. There was no association between bilateral BC and a mutation carrier status. The frequency of BRCA1 involvement is lower than that of any other country hitherto reviewed. Our results highlight the influence of geographical and ethnic origin of the population studied.

Adult↗

PubMed1999

Effect of okadaic acid on O-linked N-acetylglucosamine levels in a neuroblastoma cell line.

O-Linked N-Acetylglucosamine (O-GlcNAc) is a major form of post-translational modification found in nuclear and cytoplasmic proteins. Several authors have advanced the hypothesis according to which phosphorylation and O-GlcNAc glycosylation are reciprocally related to one another [1,2]. In order to test this hypothesis we have investigated the effect of a broad spectrum phosphatase inhibitor, okadaic acid (OA), generally used to induce protein hyperphosphorylation, on the GlcNAc content of cellular glycoproteins. We demonstrate that in neuronal cells lines OA decreases the level of O-GlcNAc in both nuclear and cytoplasmic proteins with a greater effect in the nuclear fraction. This phenomenon was demonstrated by the use of three different procedures for the detection of O-GlcNAc in conjunction with a systematic treatment with PNGase F. O-Linked GlcNAc was characterized using respectively lectin staining with WGA, galactosyltransferase labeling and metabolic labeling of cultured cells with [3H]glucosamine. Although the effects on individual proteins varied, a less pronounced effect was observed on HeLa or COS cell total homogenates. When Kelly cells were treated with OA, the major observation was a decrease in O-GlcNAc content of nuclear proteins. The measurement of the UDP-GlcNAc level clearly demonstrates that the decrease on the O-GlcNAc level in the neuroblastoma cell line after treatment with okadaic acid is not a consequence of the modification of the UDP-GlcNAc pool.

Acetylglucosamine↗

PubMed1999

Preparation and characterization of novel trans-[PtCl(2)(amine)(isopropylamine)] compounds: cytotoxic activity and apoptosis induction in ras-transformed cells.

The synthesis and chemical characterization of three new transplatinum complexes of structural formula trans-[PtCl(2)(amine)(isopropylamine)] (amine = n,n-dimethylamine, propylamine, and butylamine), 1-3, are described. Cytotoxicity tests in tumor cell lines sensitive to cis-DDP (Jurkat, Hela, and Vero) and also in tumor cell lines overexpressing ras oncogenes and resistant to cis-DDP (HL-60 and Pam 212-ras) show that complexes 1 and 3 have higher cytotoxic activity than cisplatin. Moreover, these two trans-Pt(II) complexes kill Pam 212-ras cells through apoptosis induction. These results suggest that trans-PtCl(2) complexes with asymmetric aliphatic amines may be considered a new class of biologically active trans-platinum drugs.

Animals↗

PubMed1999

The L1Tc, long interspersed nucleotide element from Trypanosoma cruzi, encodes a protein with 3'-phosphatase and 3'-phosphodiesterase enzymatic activities.

The presence of a long interspersed nucleotide element, named L1Tc, which is actively transcribed in the parasite Trypanosoma cruzi, has been recently described. The open reading frame 1 of this element encodes the NL1Tc protein, which has apurinic/apyrimidinic endonuclease activity and is probably implicated in the first stage of the transposition of the element. In the present paper we show that NL1Tc effectively removes 3'-blocking groups (3'-phosphate and 3'-phosphoglycolate) from damaged DNA substrates. Thus, both 3'-phosphatase and 3'-phosphodiesterase activities are present in NL1Tc. We propose that these enzymatic activities would allow the 3'-blocking ends to function as targets for the insertion of L1Tc element, in addition to the apurinic/apyrimidinic sites previously described. The potential biological function of the NL1Tc protein has also been evidenced by its ability to repair the DNA damage induced by the methyl methanesulfonate alkylating or oxidative agents such as hydrogen peroxide and t-butyl hydroperoxide in Escherichia coli (xth and xth, nfo) mutants.

Animals↗

PubMed1999

Synthesis and characterization of complexes of p-isopropyl benzaldehyde and methyl 2-pyridyl ketone thiosemicarbazones with Zn(II) and Cd(II) metallic centers. Cytotoxic activity and induction of apoptosis in Pam-ras cells.

It is known that metallic complexes of methyl 2-pyridyl ketone thiosemicarbazone (HL1) and p-isopropyl benzaldehyde thiosemicarbazone (HL2) may have potential antitumor activity. We have prepared complexes of HL1 and HL2 with Zn(II) and Cd(II). The cytotoxic activity shown by these compounds against cell lines sensitive and resistant to cis-diamminedichloroplatinum(II) (cis-DDP) indicates that coupling of HL1 and HL2 to Zn(II) and Cd(II) centers may result in metallic complexes with important biological properties since they display IC50 values in a microM range similar to that of the antitumor drug cis-DDP. Moreover, it is interesting to note that the Zn/HL2 complex exhibits specific cytotoxic activity against Pam-ras cells (cis-DDP resistant cells which over-express the H-ras oncogene) with an in vitro therapeutic index of 3.26 versus 0.78 for cis-DDP. Treatment of Pam-ras cells with the IC50 value of the Zn/HL2 compound induces a 'DNA ladder' (fragmentation of genomic DNA in nucleosome units) indicative of apoptosis in this ras-transformed cell line. In contrast, a 'DNA smear' (non-specific fragmentation of genomic DNA) is observed in Pam 212 normal cells treated with the IC50 of this compound. The analysis by circular dichroism (CD) spectroscopy of the interaction of the Zn/HL2 compound with calf thymus DNA (CT DNA) indicates that it produces stronger alterations on the double helix conformation than cis-DDP. So, these results suggest that Zn/HL2 may be considered a potential antitumor agent.

Animals↗

PubMed1999

Synthesis and characterization of Pd(II) and Pt(II) complexes of p-isopropylbenzaldehyde N-protected thiosemicarbazones. Cytotoxic activity against ras-transformed cells.

Cytotoxicity tests in tumor cells sensitive to cis-DDP (HL-60, JURKAT, Hela and 3T3) and in tumor cells transformed by ras oncogenes and resistant to cis-DDP (Pam 212-ras) show that cyclometallated complexes 1a [Pd(p-is.TSCN-NHMe)]4, 2b [Pt(p-is.TSCN-NMe2)]4 and 4a [Pd(p-is.TSCN-NHex)]4 may be endowed with specific cytotoxic properties. In fact, these three novel metal-thiosemicarbazone compounds kill Pam 212-ras cells through apoptosis induction. These results, together with others recently published, indicate that the design and synthesis of metallated-thiosemicarbazone compounds may lead to the discovery of novel antitumor agents able to circumvent cis-DDP resistance, in particular tumor cell lines.

Antineoplastic Agents↗

PubMed1999

Structural and cytotoxic study of new Pt(II) and Pd(II) complexes with the bi-heterocyclic ligand mepirizole.

Pd(II) and Pt(II) complexes of formulae [MLCl2], where L = mepirizole, were synthesized and characterized. Two complexes were obtained and studied by different techniques: IR, 1H and 13C NMR and XPS spectroscopies and mass spectrometry (electrospray). The crystal structure of the complex cis-dichloro-4-methoxy-2-(5-methoxy-3-methyl- pyrazol-1-yl)-6-methyl-pyrimidinepalladium(II), [Pd(mep)Cl2], was studied by crystal X-ray diffraction. It consists of discrete molecules with planar geometry. Pd(II) ions are four-coordinated by two mepirizole nitrogen atoms (N1 from the pyrazole ring and N4 from the pyrimidine ring) and two chlorine atoms. The geometry of the PdN2Cl2 chromophore is a distortion of the square-planar coordination. Data from powder pattern X-ray diffraction of cis-dichloro-4-methoxy-2-(5-methoxy-3-methyl-pyrazol-1-yl)-6-methyl- pyrimidineplatinum(II), [Pt(mep)Cl2], demonstrated that the two complexes are isostructural. The cytotoxic activity of both Pd and Pt complexes was checked for six different tumor cell lines and was lower than that of cisplatin. The Pt bound to DNA was also checked and only a low percentage is able to cross the cell membrane.

Animals↗

PubMed1999

Analysis of the humoral immune response against total and recombinant antigens of Leishmania infantum: correlation with disease progression in canine experimental leishmaniasis.

Leishmaniasis by Leishmania infantum in the Mediterranean Basin constitutes an important problem in both human and veterinary medicine. Based in both the importance of canids as reservoirs for the human disease and the fact that the canine disease may be an excellent model for the human condition, the present work has been conducted to analyze clinical and immune mechanisms associated with canine experimental leishmaniasis. Six-month-old mixed-breed dogs were intravenously infected with L. infantum promastigotes and the infection course was monitored along a 343 days-period. On day 75 post-infection (p.i.), amastigotes were observed in the lymph nodes of all dogs. The analysis of the humoral response against total L. infantum antigens by both ELISA and Western blotting evidenced a correlation between the levels of IgG isotypes (IgG1 and IgG2) and disease progression. It was observed that in those animals showing either a regressive or an oligosymptomatic form of the disease, the anti-Leishmania IgG1 antibodies were undetectable whereas those animals developing active disease showed high levels of anti-Leishmania IgG1 antibodies. Additionally, the time-course of antibody production against L. infantum recombinant antigens in the experimentally infected dogs has been analyzed. The present data suggest that reactivity against the heat-shock protein 70 (HSP70) may be used as diagnostic marker of early steps of infection, and that the appearance of anti-histone antibodies is associated with progression of infection to disease status.

Animals↗

PubMed1999

Synthesis, characterization and DNA modification induced by a novel Pt(IV)-bis(monoglutarate) complex which induces apoptosis in glioma cells.

Programmed cell death or apoptosis is a mechanism for the elimination of cells that occurs not only in physiological processes but also in drug-induced tumor cell death. Thus, because cisplatin, cis-diamminechloroplatinum (II), produces important damages on the DNA inducing apoptosis in several cell lines it has become a widely used antitumor drug. However, cisplatin possesses some dose-limiting toxicities mainly nephrotoxicity. Pt(IV) complexes, such as iproplatin, ormaplatin, and JM216 are a new class of platinum complexes that exhibits less toxicity than cisplatin. Some of these complexes have shown significant antitumor activity and a low cross-resistance to cisplatin. In the present paper, we have analyzed the DNA binding mode and the cytotoxicity of a novel Pt(IV)-bis (monoglutarate) complex. The data show that this novel complex produces DNA interstrand cross-links to a higher extent and with a faster kinetics than cisplatin. Also the Pt(IV)-bis (monoglutarate) complex kills glioma cells at drug concentrations significantly lower than those of cisplatin. Interestingly, this Pt(IV) complex produces in the glioma cells characteristic features of apoptosis such as 'DNA laddering' and fragmented nuclei. Moreover, the p53 protein accumulates early in glioma cells as a result of Pt(IV)-bis (monoglutarate) treatment. These data indicate that the Pt(IV)-bis (monoglutarate) complex induces apoptosis in glioma cells through a p53-dependent pathway.

Antineoplastic Agents↗

PubMed1999

Leishmania: fine mapping of the Leishmanolysin molecule's conserved core domains involved in binding and internalization.

The Leishmanolysin molecule's role in the uptake of Leishmania parasites by the human U937 pro-myelocytic cell line was studied, using synthetic peptides representing the complete Leishmania (Viannia) guyanensis Leishmanolysin protein amino acid sequence. The particular peptides present in two protein's core domains efficiently impaired the internalization of promastigotes from four different Leishmania species and modified the kinetics of the binding of heterologous recombinant Leishmanolysin protein. The functional domains which exhibited this property represent a highly conserved portion of the sequence among different Leishmania species. The peptides' inhibitory activity correlated with their ability to bind molecules present on the surface of the human cell line. One of the two functional core domains identified involves the previously described adhesive sequence (SRYD) and the putative zinc-binding motif (HExxH). The second functional core domain includes a third histidine residue coordinated with zinc which determines the molecule's structural features. These findings indicate that the molecular interactions between Leishmanolysin's conserved domains and the macrophage surface molecules efficiently contribute to the parasite's internalization. Induction of neutralizing immune responses, which impair the early parasite-host interaction described here, may be an important alternative in designing synthetic subunit human leishmaniasis vaccines.

Amino Acid Sequence↗

PubMed1999

Empirical validation of a two-state kinetic model for the B-Z transition of double-stranded poly[d(G-m5C)].

A formula based on a first-order kinetic equation is derived to evaluate the rate constant of the B-Z transition of a synthetic double-stranded poly[d(G-m5C)] in terms of the salt concentration, the absolute temperature, and the cooperativity index. The validity of the formula was tested using circular dichroism spectroscopy after variation of the type of salt (NaCl, MgCl2), the salt concentration, and the temperature of the polynucleotide solution. A consequence of the proposed function is that in conditions of high salt there is a predictable salt threshold which determines the particular molecular mechanism of the B-Z transition. The paper also describes the way in which this threshold level is temperature dependent. A detailed comparison of our data with the experimental data found by other authors is given. The function agrees quantitatively with the experiments and explains the contrasting results found in the literature about the influence in the B-Z transition of both the temperature and the polymer size.

Circular Dichroism↗

PubMed1999

Occurrence of organochlorine insecticides, PCBs and PCB congeners in waters and sediments of the Ebro River (Spain)

Organochlorine insecticides and poychlorinated biphenyls (PCBs) were measured in water and sediment samples taken from thirteen sites along the Ebro River (Spain), the most agricultural and industrial basin in the North of the Iberian Peninsula. Concentrations of 10 insecticides, PCBs and 13 PCB congeners were determined: grouping all chemicals by structural affinity, these can be arranged in descending order as follow: PCBs > total DDT > total cyclodiene insecticides > total HCH. Higher levels of organochlorine pollutants were detected in sediments from the middle and lower parts of the stream where are located the main important inputs in Ebro River.

Journal Article↗

PubMed1999

Immunostimulatory properties of the Leishmania infantum heat shock proteins HSP70 and HSP83.

Emerging evidence indicates that the heat shock proteins (HSPs), a set of highly evolutionary conserved proteins, are playing essential roles in both normal processes of the immune system and specific immune responses. In a previous work, we demonstrated that the Leishmania infantum HSP70 possesses remarkable immunostimulatory properties. In the present work, we have extended the study to another HSP from this parasite, the HSP83. We show that this protein also has an adjuvant effect to an accompanying protein by stimulation of the humoral response when both proteins are fused and co-administered to BALBjc mice. The analysis of the IgG isotypes, IgG1 and IgG2a, indicated that the immunisations with the Leishmania HSPs, mainly the HSP70, potentiate a Thl-type response. It was found that the amino-terminal domain of the HSP70, the most evolutionary conserved region of the molecule, maintains the ability to stimulate the humoral response, whereas the carboxyl-terminal domain does not have a similar effect. Unexpectedly, we found that the L. infantum HSP70 and HSP83 recombinant proteins stimulated the proliferation of spleen cells from unprimed BALB/c mice. Remarkably, this proliferation was abolished either by thermal denaturing of the proteins or by using specific antibodies. The use of the T-cell inhibitor cyclosporin A in the splenocytes proliferation assays suggested that both T- and non-T-cells are stimulated by the Leishmania HSPs. These findings may be relevant for therapeutic and prophylactic applications.

Adjuvants, Immunologic↗

PubMed1999

A cycloplatinated compound of p-isopropylbenzaldehyde thiosemicarbazone and its chloro-bridged derivative induce apoptosis in cis-DDP resistant cells which overexpress the H-ras oncogene.

cis-Diamminedichloroplatinum(II) (cis-DDP) is a widely used antitumour drug which produces important damage on the DNA inducing apoptosis in several cell lines. We have analyzed the cytotoxic activity of novel cyclometallated complexes of p-isopropylbenzaldehyde thiosemicarbazone (p-is.TSCN) and their dimeric chloro-bridged derivatives in murine keratinocytes transformed by the H-ras oncogene which are resistant to cis-DDP (Pam-ras cells). The data show that, in contrast with cis-DDP, the tetrameric cycloplatinated complex [Pt(p-is.TSCN)]4 and its dimeric chloro-bridged derivative [Pt(microCl)(p-is.TSCN)]2 have a good in vitro therapeutic index when comparing the cytotoxicity in Pam-ras cells to normal murine keratinocytes (Pam 212 cells) since they induce cell death in Pam-ras cells at drug concentrations significantly lower than those needed to kill Pam 212 cells. At equitoxic doses (IC90), both complexes produce characteristic features of apoptosis in Pam-ras cells together with a drastic decrease in levels of H-ras protein. These effects are not observed when the cells are treated with the IC90 of the cis-DDP drug nor the p-is.TSCN ligand. Altogether, these results suggest that the platinum compounds [Pt(p-is.TSCN)]4 and [Pt(microCl)(p-is.TSCN)]2 might have potential as antitumour agents in view of their specific induction of apoptosis in cis-DDP resistant cells.

Animals↗

PubMed1999

Apoptosis induction and inhibition of H-ras overexpression by novel trans-[PtCl2(isopropylamine)(amine')] complexes.

Hitherto, it has been generally accepted as a paradigm of the biochemical pharmacology of platinum antitumor drugs that a cis configuration of the leaving groups is necessary for antitumor activity of platinum compounds. However, it has been recently observed that certain trans-platinum complexes have both in vitro and in vivo antitumor activity. We previously reported the synthesis, characterization and cytotoxic activity against ras-transformed cells of several trans-[PtCl2LL'] complexes where L and L' are asymmetric aliphatic amines (L = dimethylamine and butylamine, L' = isopropylamine). The results reported in this paper show that the compounds trans-[PtCl2(isopropylamine)(dimethylamine)] and trans-[PtCl2(isopropylamine)(butylamine)] kill Pam 212-ras cisplatin resistant cells through apoptosis induction. Moreover, Western blot data show that both compounds inhibit overexpression of H-ras oncogene in Pam 212-ras cells. Altogether, these data indicate that, in contrast with cis-DDP, the apoptotic activity of these novel trans-Pt(II) compounds in ras-transformed cells is associated with their ability to abolish ras-overexpression.

Animals↗