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Biomedical subjects

J Domínguez

Publications and source records attributed to J Domínguez.

At least 19 recordsLinked to original sources

Monoclonal antibodies specific for porcine monocytes/macrophages: macrophage heterogeneity in the pig evidenced by the expression of surface antigens.

Macrophages are widely distributed in most tissues of the body, where they play important roles in host defense and repair of tissue damage. In this report we describe the production and characterization of a panel of six monoclonal antibodies (mAb) against porcine macrophages and their use for phenotyping tissue macrophages. All mAbs were produced by immunizing mice with porcine alveolar macrophages. Three of them (2A10/11, 3B11/11 and 3F7/11) react mainly with macrophages and, at a lower extent, blood monocytes, whereas the others (1E12/11, 2C12/10 and 4E9/11) also recognize granulocytes. Antigens recognized by these antibodies could be characterized by Western blot and/or immunoprecipitation, with the exception of that one recognized by 2C12/10. By their behavior in SDS-PAGE under reducing and nonreducing conditions, all seem to be single polypeptides, whose apparent molecular weight under reducing conditions are: 1E12/11 and 3B11/11 larger than 204 kDa; 2A10/11, 150 kDa; 4E9/11, 125-170 kDa; and 3F7/11, 135 kDa. Immunohistochemical analyses of both lymphoid and non-lymphoid organs using these mAbs reveal important antigenic heterogeneity among tissue macrophages. These mAbs are, therefore, useful tools for the study of porcine macrophage maturation and differentiation and for determining their heterogeneity both in normal and pathological conditions.

Animals

Direct detection of Mycobacterium tuberculosis complex in nonrespiratory specimens by Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test.

The Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test (AMTDT) was adapted for the detection of Mycobacterium tuberculosis complex in 224 nonrespiratory specimens from 188 patients. The sensitivity and specificity of the AMTDT for such specimens, after resolution of discrepant results, were 85.7 and 100%, respectively. Pretreatment of nonrespiratory specimens with sodium dodecyl (lauryl) sulfate is mandatory to obtain consistent and reproducible AMTDT results. The use of 500 microliters of decontaminated specimen improves the sensitivity of the test. Because the AMTDT detects stable rRNA from noncultivable bacilli, it is not useful for monitoring patients receiving treatment.

Bacterial Typing Techniques

Diagnostic and therapeutic thoracoscopy in esophageal cancer.

The decision before operation whether an esophagus affected by cancer can be resected is difficult. In order to determine if the tumor can be removed or not, patients must undergo surgery. This high risk intervention is often carried out only to prescribe palliative treatment. We used a thoracoscopic technique for the diagnosis and resection of esophageal cancer with the aim of improving the high morbidity rate associated with esophageal open surgery. A right thoracoscopy was performed in seven patients and only four underwent resection. The entire esophagus was mobilized thoracoscopically and the intervention was completed through the abdomen and the neck. In three patients in whom resection was not considered possible, the thoracoscopic procedure was done with excellent tolerance, and treated by palliative methods. Thoracoscopy is a very valuable procedure for the diagnosis of resectability as well as for the mobilization of the esophagus prior to resection.

Endoscopes

The role of the nodI and nodJ genes in the transport of Nod metabolites in Rhizobium etli.

A kinetic analysis of secretion of lipo-chitin oligosaccharides (LCO) produced by Rhizobium etli (Re) wild-type (wt) strain and derivatives carrying disrupted nodI or nodJ genes was performed. LCO were detected in the growth media of the wt strain as early as 1 h after nod gene induction. In contrast, strains carrying nodI or nodJ mutations secreted less LCO, and accumulated LCO metabolites intracellularly after 4 h of induction. These Re mutants presented a delayed nodulation phenotype and a reduction in the maximum number of nodules formed in Phaseolus vulgaris roots.

ATP-Binding Cassette Transporters

Monoclonal antibody 2F4/11 recognizes the alpha chain of a porcine beta 2 integrin involved in adhesion and complement mediated phagocytosis.

The characterization of a new mAb, named 2F4/11, specific for porcine myelomonocytic cells is described. This mAb immunoprecipitates a non-covalently linked heterodimer of 155,000/95,000, which is expressed by granulocytes, monocytes and tissue macrophages but not by lymphocytes, erythrocytes or platelets. Immunoblot analysis localizes the 2F4/11 epitope on the largest subunit of the heterodimer. Mab 2F4/11 is able to block phagocytosis of complement-opsonized zymosan particles by PMN granulocytes and alveolar macrophages, as well as adherence to plastic surfaces of PMA-activated PMN. Together, these results suggest that mAb 2F4/11 recognizes the CD11b or alpha chain of the porcine complement type 3 receptor (CR3).

Animals

[The degree of compliance with the radiology requests from primary care].

OBJECTIVE: To study the level of compliance with the radiological tests requested from primary care. DESIGN: A crossover, descriptive study with systematic randomised distribution. SETTING: Catchment area of a specialist centre covering eight urban health districts in Zaragoza. MATERIAL: 413 X-ray requests out of those requested between January and March, 1995. MEASUREMENTS AND RESULTS: The filling in of the following sections was evaluated: patient's basic data, referral from, clinical/investigatory data, suspected diagnosis and its agreement with the radiology report, the investigation requested, date, signature and legibility of the request. Compliance was: complete name, 100%; telephone, 37%; address, 50%; social security number, 99%; referral from, 71%; symptomatology, 92%; clinical investigation, 35%; suspected diagnosis, 56%; date, 92%; signature, 86%; good legibility in 13% of cases and poor in 75%. Distribution by gender: men 44%, women 56%, a significant difference. Average age 47.2, women 49.9 and men 43.8, also a significant difference. Simple osteoarticular x-ray was the commonest request (43.3%). CONCLUSIONS: 1) There was poor compliance in the telephone and address sections. 2) Clinical data (symptoms, investigations) and the suspected diagnosis were also insufficient. 3) Date and signature did not reach the desirable 100% compliance. 4) Deficient legibility, with very few requests not causing problems.

Adult

Brain oedema in patients with intracranial meningioma. Correlation between clinical, radiological, and histological factors and the presence and intensity of oedema.

The authors analysed the correlation between different clinical, radiological, and pathological variables and the presence and intensity of brain oedema associated to intracranial meningioma in 400 consecutive patients studied by computerized tomography (CT). The following factors did not show significant correlation with brain oedema development: the age and sex of the patient, the occurrence of focal deficits, the presence of skull changes (endostosis, exostosis, osteolysis), the occurrence of tumour calcification, the density of the tumour on plain CT scan, the presence of a cystic component, the pathological subtype of meningioma (both conventional and non-conventional), and the presence of histological features of tumour aggressiveness, such as an increased vascularization, high cellularity, high mitotic index, pleomorphism, necrosis, and brain infiltration. Factors showing a statistically significant correlation with the presence and intensity of brain oedema at the bivariate analysis were: the presence of symptoms (p < 0.001), the duration of the clinical history (p < 0.05), the location and size of the tumour (p < 0.001), the type (heterogeneous vs homogeneous), and intensity of tumour contrast enhancement (p < 0.001), the presence of irregular tumour margins (p < 0.001), and the existence of focal low density intratumoural areas (p < 0.001). The multivariate analysis using only clinical parameters showed that the group of variables with the highest power for predicting the presence of brain oedema (concordance level of 76.8%) included: the presence of symptoms, the occurrence of seizures (focal or generalized), the presence of an intracranial hypertension syndrome, and the age of the patient. The multivariate analysis using only anatomico-radiological parameters showed that the model which included the size of the tumour, the intensity of contrast enhancement, the tumour margins, and meningioma location, predicted the presence of brain oedema in 80.8% of the cases. Though the results of the present study do not definitively support any of the major physiopathological theories proposed to explain brain oedema formation in patients with intracranial meningioma, some findings could favour the so-called hydrodynamic theory.

Adolescent

Irreversible tetraplegia after tracheal resection.

The case of a 21-year-old woman without previous cervical pathology in whom irreversible tetraplegia developed after operation for tracheal stenosis is reported. After tracheal resection the neck was kept in extreme flexion and after extubation she was moved to a sitting position. The different causal agents that could produce the neurologic damage remain unclear, although we think that the combination of relative arterial hypotension secondary to the sitting position and disturbed autorregulation, caused by extreme neck flexion, could result in ischemic spinal cord injury.

Adult

Evaluation of the disinfectant effect of Solprogel against human immunodeficiency virus type 1 (HIV-1).

The antiviral activities of sodium dichloroisocyanurate (NaDCC) and a commercial product (Solprogel 2%) against human immunodeficiency virus type 1 (HIV-1) were investigated using a quantitative suspension test method. Solprogel is a compound that contains NaDCC and a biodegradable polymer of acrylic acid. Viral suspensions were prepared containing 3.2 x 10(6) tissue culture infective dose 50 (TCID50) in culture media. Syncytium formation in the MT-2 line and HIV antigen p24 on the supernatant of the cultures were used to determine viral titre. Results indicate that satisfactory disinfection (1000-fold reduction in 5 min) can be achieved using NaDCC and Solprogel at concentrations of 100 and 120 ppm available chlorine, respectively.

Antiviral Agents

Synthesis and activity of some quinolone derivatives against Plasmodium falciparum in vitro.

Quinolones 7a-i and 8a-i were synthesized and tested in vitro as antimalarial against Plasmodium falciparum in chloroquine resistant strain. The above compounds were inactive at concentrations of 1.0 x 10(-4) M except the quinolone 3-amino-9-(2,4-dichlorophenyl)-1H-pyrazolo[3,4-b]-4-quinolone 7h which showed IC50 of 5.06 microM given the most interesting results.

Animals

Synthesis of pyridopyrimidone derivatives and their activity against P. falciparum in vitro.

Pyridopyrimidone derivatives 4a-d and 5a- were synthesized as potential antimalarial agents on the basis of the pharmacological properties of existing quinolone analogues such as ciprofloxacine IC50 39.8 microM. Meanwhile among these new compounds, only the 3-amino-7-methyl-1(H)pyrazolo[3,4-b]-pyrido [1,2: 1'2']pyrimido-4-ona 4b, produces significant antimalarial activity IC50 0.42 microM against P. falciparum in vitro. The remaining compounds were effective as antimalarial agents leading from IC50 1.0 microM to IC50 4.47 microM.

Animals

Isolation, chemical structures and biological activity of the lipo-chitin oligosaccharide nodulation signals from Rhizobium etli.

Rhizobium etli is a microsymbiont of plants of the genus Phaseolus. Using mass spectrometry we have identified the lipo-chitin oligosaccharides (LCOs) that are produced by R. etli strain CE3. They are N-acetylglucosamine pentasaccharides of which the non-reducing residue is N-methylated and N-acylated with cis-vaccenic acid (C18:1) or stearic acid (C18:0) and carries a carbamoyl group at C4. The reducing residue is substituted at the C6 position with O-acetylfucose. Analysis of their biological activity on the host plant Phaseolus vulgaris shows that these LCOs can elicit the formation of nodule primordia which develop to the stage where vascular bundles are formed. The formation of complete nodule structures, including an organized vascular tissue, is never observed. Considering the very close resemblance of the R. etli LCO structures to those of R. loti (I. M. López-Lara, J. D. J. van den Berg, J. E. Thomas Oates, J. Glushka, B. J. J. Lugtenberg, H. P. Spaink, Mol Microbiol 15: 627-638, 1995) we tested the ability of R. etli strains to nodulate various Lotus species and of R. loti to nodulate P. vulgaris. The results show that R. etli is indeed able to nodulate Lotus plants. However, several Lotus species are only nodulated when an additional flavonoid independent transcription activator (FITA) nodD gene is provided. Phaseolus plants can also be nodulated by R. loti bacteria, but only when the bacteria contain a FITA nodD gene. Apparently, the type of nod gene inducers secreted by the plants is the major basis for the separation of Phaseolus and Lotus into different cross inoculation groups.

Acetylglucosamine

[Neoplastic obstructive jaundice: palliative treatment with self-expandable metallic prosthesis].

PURPOSE: Study of the effectiveness and morbidity of palliative treatment of malignant obstructive jaundice with metallic biliary endoprosthesis compared to surgical palliation. DESIGN: Retrospective review. PATIENTS: 35 patients with non-resectable neoplasms causing jaundice were treated with percutaneous stent (pancreatic carcinoma, n = 11; cholangiocarcinoma, n = 11; gallbladder carcinoma, n = 4; extrahepatic metastases of various malignancies, n = 8). CONTROL GROUP: 23 patients with malignant jaundice treated with palliative surgery. RESULTS: Most frequent complications were cholangitis and stent obstruction. The mean hospital stay after the stent placement was 6.8 days, longer in patients with complications (p = 0.035). Recurrence of jaundice was seen in 22.9% of the patients and the rate of readmission was 42.9%. The mean survival was 163.33 days (range 19-522). Reduction in serum bilirubin after BE was significant (215 vs. 82 mmol/l, p < 0.001). CONCLUSIONS: Comparing to our previous experience with surgical palliative treatment, there was no significant difference neither in morbidity-mortality, nor recurrence or readmission. Patients with pancreatic cancer and cholangiocarcinoma benefit from a shorter hospital stay.

Aged

Interaction between noradrenergic and histamine-containing neurons in the rat vas deferens.

A decussation of histamine (HA)-containing nervous pathways, at the level of the sympathetic ganglionic clusters of the rat vas deferens, was formerly described in this laboratory. This type of neuronal distribution is confirmed in the present study by using surgical interruptions of nervous pathways and determining changes in L-histidine decarboxylase (HD) activity, as an HA-containing neuron marker, after nerve degeneration. Decreases of HD activity follow previously found decreases of HA levels in the vas deferens submitted to the same type of surgical interruptions. An interaction between noradrenergic neurons and HA-containing neurons is suggested, because electrical stimulation of the preganglionic sympathetic nerve causes a rapid increase in HD activity in the ipsilateral vas deferens, possibly as a signal of the activation of HA-containing neurons, whose pathways come from the opposite ganglion. This activation appears to be indirect, presumably through a sensory interneuron, which upon being excited by the enhanced sympathetic discharge crosses over to stimulate the contralateral ganglionic HA-containing neuron. The sympathetic discharge-induced peripheral loop reflex resulting in an increase in HD activity depends upon the integrity of the noradrenergic neuron. An, as yet unknown, interaction seems to occur between the noradrenergic neurons and the HA-containing neurons in the rat vas deferens, when the sympathetic activity of the latter is enhanced.

Animals

Carboxyl terminal sequences of beta-tubulin involved in the interaction of HMW-MAPs. Studies using site-specific antibodies.

After the finding of the involvement of the C-terminal moieties of tubulin subunits in the interaction of MAPs, different studies have focused on the substructure of the binding domains for the different MAPs. Current biochemical evidence point to the role of a low-homology sequence between alpha and beta-subunits within the conserved region of the C-terminal domain of tubulin, in the binding of MAP-2 and tau. Another line of studies indicates that a site for interaction of the high molecular weight MAPs is located in the variable region defined by the glutamic-rich C-terminus of beta-tubulin. Here, we report the usefulness of idiotypic site-directed antibodies, produced by immunization with peptides from different beta-tubulin isoforms, to study both MAP-1 and MAP-2 binding sites on tubulin. On the basis of these results with site-specific antibodies along with previous structural information (Cross et al., 1991, Biochemistry 30: 4362-4366), we propose the role of consensus sequences, from the invariant beta-tubulin C-terminal domain in the binding of MAP-2 and from the variable domain in the interactions of MAP-1 and MAP-2.

Amino Acid Sequence

Monoclonal antibodies to turbot (Scophthalmus maximus) immunoglobulins: characterization and applicability in immunoassays.

Five monoclonal antibodies (mAbs) to immunoglobulins (Igs) of the turbot Scophthalmus maximus were produced and characterized. All the mAbs (denominated UR1, UR3, UR4, UR6 and UR7) are of isotype IgG1/kappa and show good anti-turbot Ig reactivity in enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Results of competitive ELISA and immunoblotting analysis indicate that these five mAbs react with at least three different epitopes on the turbot Ig H chain. Except in the case of UR1, reactivity with periodate-treated purified turbot Ig was much lower than with the untreated Ig, suggesting that carbohydrate residues are involved in epitope recognition. All the mAbs showed reactivity with sera from the closely related species Scophthalmus rhombus but not with sera from species of other flatfish genera. One of these mAbs (UR3) has been successfully applied for the detection of antibodies against Vibrio anguillarum in ELISA.

Animals