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Biomedical subjects

M E Rodriguez

Publications and source records attributed to M E Rodriguez.

At least 19 recordsLinked to original sources

The calcimimetic R-568 increases vitamin D receptor expression in rat parathyroid glands.

We previously demonstrated that extracellular calcium regulates vitamin D receptor (VDR) expression by parathyroid cells. Since the calcimimetic R-568 potentiates the effects of calcium on the calcium-sensing receptor, it was hypothesized that administration of R-568 may result in increased VDR expression in parathyroid tissue. In vitro studies of the effect of R-568 on VDR mRNA and protein were conducted in cultures of whole rat parathyroid glands and human hyperplastic parathyroid glands. In vivo studies in Wistar rats examined the effect of R-568 and calcitriol alone and in combination. Incubation of rat parathyroid glands in vitro with R-568 (0.001-1 microM) resulted in a dose-dependent decrease in parathyroid hormone (PTH) secretion and an increase in VDR expression (mean +/- SE). Incubation in 1 mM calcium + 0.001 microM R-568 elicited an increase in VDR mRNA (306 +/- 46%) similar to the maximum increase detected with 1.5 mM calcium (330 +/- 42%). In vivo, VDR mRNA was increased after administration of R-568 (168 +/- 9%, P < 0.001 vs. control) or calcitriol (198 +/- 16%, P < 0.001 vs. control). Treatment with R-568 also increased VDR protein in normal rat parathyroid glands and in human parathyroid glands with diffuse, but not nodular, hyperplasia. In conclusion, the present study shows that the calcimimetic R-568 exerts a stimulatory effect on VDR expression in the parathyroid glands of study models and provides additional evidence for the use of calcimimetics in the treatment of secondary hyperparathyroidism.

Aniline Compounds↗

Cyclin D3 expression in primary Ta/T1 bladder cancer.

Cyclin D3 deregulation has recently been reported in bladder cancer but its prognostic significance remains uncertain. A cohort of 159 patients with stage Ta or T1 primary bladder tumours was investigated to determine the significance of cyclin D3 expression in association with other G1-S phase regulators of the cell cycle (p53, p21Waf1, p27kip1, cyclin D1), including tumour proliferation (ki67-MIB1); its association with conventional clinicopathological parameters; and the relationship between cyclin D3 and loss of heterozygosity (LOH) at the 9p21 (p16INK4a locus) chromosome region. The end point of the study was progression-free survival. Cyclin D3, other G1-S phase regulators, and tumour proliferation were investigated by immunohistochemistry and measured by the grid-counting method. To validate the immunohistochemical expression, cyclin D3 was additionally assessed by western blotting in selected cases. LOH at the 9p21 chromosome region (marker D9S171) was assessed in 125 cases using an AB Prism 310 genetic analyser and a set of microsatellite fluorescence-labelled primers. Cyclin D3 overexpression was related to larger tumour size (>5 cm; p < 0.0001) and high tumour proliferation (>10%; p = 0.025). Mean cyclin D3 expression increased with 2004 WHO grading categories in stage Ta (p = 0.035, ANOVA) and stage T1 (p = 0.047, t test) tumours. Cyclin D3 was not related to other clinicopathological parameters, G1-S phase modulators, or 9p21 LOH. Cox's multivariate analysis selected cyclin D3 as an independent predictor of progression-free survival (p = 0.0012, relative risk (RR) = 5.2366) together with tumour size (p = 0.0115, RR = 4.4442) and cyclin D1 (p = 0.0065, RR = 3.3023). Cyclin D3 expression had the highest risk ratio. Our results suggest that expression of cyclin D3 is relevant to the progression-free survival of patients with Ta/T1 bladder carcinomas.

Adult↗

Fc receptor-mediated immunity against Bordetella pertussis.

The relevance of specific Abs for the induction of cellular effector functions against Bordetella pertussis was studied. IgG-opsonized B. pertussis was efficiently phagocytosed by human polymorphonuclear leukocytes (PMN). This process was mediated by the PMN IgG receptors, FcgammaRIIa (CD32) and FcgammaRIIIb (CD16), working synergistically. Furthermore, these FcgammaR triggered efficient PMN respiratory burst activity and mediated transfer of B. pertussis to lysosomal compartments, ultimately resulting in reduced bacterial viability. Bacteria opsonized with IgA triggered similar PMN activation via FcalphaR (CD89). Simultaneous engagement of FcalphaRI and FcgammaR by B. pertussis resulted in increased phagocytosis rates, compared with responses induced by either isotype alone. These data provide new insights into host immune mechanisms against B. pertussis and document a crucial role for Ig-FcR interactions in immunity to this human pathogen.

Antibodies, Bacterial↗

Effects of Ca(OH)(2) treatments ("overliming") on the composition and toxicity of bagasse hemicellulose hydrolysates.

Hemicellulose syrups from dilute sulfuric acid hydrolysates of hemicellulose contain inhibitors that prevent efficient fermentation by yeast or bacteria. It is well known that the toxicity of these hydrolysate syrups can be ameliorated by optimized "overliming" with Ca(OH)(2). We have investigated the optimization of overliming treatments for sugar cane bagasse hydrolysates (primarily pentose sugars) using recombinant Escherichia coli LY01 as the biocatalyst. A comparison of composition before and after optimal overliming revealed a substantial reduction in furfural, hydroxymethylfurfural, and three unidentified high-performance liquid chromatography (HPLC) peaks. Organic acids (acetic, formic, levulinic) were not affected. Similar changes have been reported after overliming of spruce hemicellulose hydrolysates (Larsson et al., 1999). Our studies further demonstrated that the extent of furan reduction correlated with increasing fermentability. However, furan reduction was not the sole cause for reduced toxicity. After optimal overliming, bagasse hydrolysate was rapidly and efficiently fermented (>90% yield) by LY01. During these studies, titration, and conductivity were found to be in excellent agreement as methods to estimate sulfuric acid content. Titration was also found to provide an estimate of total organic acids in hydrolysate, which agreed well with the sum of acetic, levulinic, and formic acids obtained by HPLC. Titration of acids, measurement of pH before and after treatment, and furan analyses are proposed as relatively simple methods to monitor the reproducibility of hydrolysate preparations and the effectiveness of overliming treatments.

Biotechnology↗

Pneumococcal capsular polysaccharide-specific IgA triggers efficient neutrophil effector functions via FcalphaRI (CD89).

Specific anti-capsular polysaccharide IgG is believed to be important for protection against infection by Streptococcus pneumoniae. Significant IgA responses have been observed after vaccination with pneumococcal vaccines, but the role of this isotype in anti-pneumococcal host defense is unclear. Here, it is shown that purified serum IgA specific for pneumococcal capsular polysaccharides can initiate efficient cellular effector functions, such as phagocytosis, via interaction with the myeloid IgA receptor, FcalphaRI (CD89). The efficiency of FcalphaR-triggered granulocyte effector functions was comparable to that of FcgammaRIIa (CD32), as shown in experiments with bispecific antibodies. These results support a role for polysaccharide-specific IgA in antipneumococcal cellular effector function and suggest that FcalphaRI represents an important leukocyte receptor for immunity against S. pneumoniae.

Antibodies, Bacterial↗

Release of outer membrane vesicles from Bordetella pertussis.

The aim of the study reported here was to investigate the production of Bordetella pertussis outer membrane vesicles (OMVs). Numerous vesicles released from cells grown in Stainer-Scholte liquid medium were observed. The formation of similar vesicle-like structures could also be artificially induced by sonication of concentrated bacterial suspensions. Immunoblot analysis showed that OMVs contain adenylate cyclase-hemolysin (AC-Hly), among other polypeptides, as well as the lipopolysaccharide (LPS). Experiments carried out employing purified AC-Hly and OMVs isolated from B. pertussis AC-Hly- showed that AC-Hly is an integral component of the vesicles. OMVs reported here contain several protective immunogens and might be considered a possible basic material for the development of acellular pertussis vaccines.

Adenylate Cyclase Toxin↗

Crucial role of FcgammaRIIa (CD32) in assessment of functional anti-Streptococcus pneumoniae antibody activity in human sera.

Immunoglobulin G (IgG)-mediated phagocytosis by polymorphonuclear leukocytes (PMNL) constitutes the main defense against Streptococcus pneumoniae. Two leukocyte IgG receptors, FcgammaRIIa and FcgammaRIIIb, are constitutively expressed on PMNL. Blocking experiments showed FcgammaRIIa is crucial for opsonophagocytosis of serum-opsonized S. pneumoniae. The biallelic, genetically determined FcgammaRIIa polymorphism (FcgammaRIIa-R131 vs. IIa-H131) determines the capacity of IgG2-mediated phagocytosis via this receptor. Comparative studies with PMNL from donors either homozygous for FcgammaRIIa-R131 or IIa-H131 showed the latter had higher phagocytic capacity. These results were confirmed in FcgammaRIIa-R131- and FcgammaRIIa-H131-transfected IIA1.6 cells. The performance of FcgammaRIIa-transfected cells in S. pneumoniae phagocytosis was validated using sera from adults and children. Serum-induced phagocytic activity depended mainly on anti-pneumococcal IgG2 antibodies. Results obtained with PMNL and IIA1.6 cells showed high correlation (r=0.94; P<.001), and support that FcgammaRIIa transfectants are a good alternative to PMNL as effector cells in opsonophagocytosis assays.

Antibodies, Bacterial↗

Immunogenicity of Streptococcus pneumoniae type 6B and 14 polysaccharide-tetanus toxoid conjugates and the effect of uncoupled polysaccharide on the antigen-specific immune response.

The immunogenicity of two types of Streptococcus pneumoniae capsular polysaccharide-tetanus toxoid conjugates (PS6BTT and PS14TT) was evaluated in mice. Both conjugates induced high titres of high avidity type-specific anti-PS IgG, which include all IgG isotypes except IgG2a. Repeated immunization resulted in booster responses in both cases. The antibodies induced exhibited opsonic activity, as measured in an in vitro opsonophagocytosis assay, using the mouse macrophage cell line RAW-264. Furthermore, the influence of spiking PS6BTT with free PS6B of either 1000 kDa (native) or 37 kDa was investigated. The results indicate that not only the amount but also the molecular weight of the free PS6B present in the conjugate vaccine affect the anti-PS6B immune response. Large amounts of free PS6B of both molecular weights decrease each anti-PS6B IgG isotype response. However, unlike admixture of the low molecular weight PS6B, addition of the high molecular weight PS6B leads to a rather persistent state of unresponsiveness.

Animals↗

Enantioselective local disposition of semotiadil (R-enantiomer) and levosemotiadil (S-enantiomer) in perfused rat liver.

The enantioselective local disposition of semotiadil (R-enantiomer) and levosemotiadil (S-enantiomer) in rat liver was investigated in the single-pass perfusion system containing 1% bovine serum albumin (BSA). After an instantaneous injection of semotiadil, levosemotiadil, or Evans Blue (a marker of BSA), each outflow time profile from the liver was analyzed by a two-compartment dispersion model. The recovery ratio, FH (1.88 +/- 0.28%), of semotiadil was significantly smaller than that (8.99 +/- 1.40%) of levosemotiadil. The mean transit time, fH (0.146 +/- 0.014 min) of semotiadil was significantly smaller than that (0.191 +/- 0.012 min) of levosemotiadil. The biliary excretion kinetics of these enantiomers was also evaluated by moment analysis. The parent compound (semotiadil or levosemotiadil) was not detected in bile, but four metabolites generated from each parent enantiomer were found in the bile. A portion (16.5 +/- 1.2%) of the drug eliminated by the liver was recovered as R-metabolites in the bile within 1 hr after an injection of semotiadil, whereas 11.2 +/- 1.6% was recovered as S-metabolites in the bile within 1 hr after an injection of levosemotiadil. This excreted percentage into the bile was significantly different between R- and S-metabolites. The mean biliary excretion time MRTe (19.1 +/- 2.2 min) of total R-metabolites was significantly larger than that (14.8 +/- 1.1 min) of total S-metabolites. In conclusion, stereo-selectivity was suggested both at the hepatic elimination of the parent compound and at the biliary excretion of the metabolites.

Animals↗

Guanine, pyrazolo[3,4-d]pyrimidine, and triazolo[4,5-d]pyrimidine (8-azaguanine) phosphonate acyclic derivatives as inhibitors of purine nucleoside phosphorylase.

Phosphonate acyclic derivates of guanines, pyrazolo[3,4-d]pyrimidines, and triazolo[4,5-d]-pyrimidines (8-azaguanines) are inhibitors of the enzyme purine nucleoside phosphorylase (PNPase) with Ki' values ranging from 0.05 to 1.6 microM. These compounds are enzymatically stable congeners of the potent PNPase inhibitor acyclovir diphosphate (53).

Azaguanine↗

Effect of hydromechanical stress on cellular antigens of Bordetella pertussis.

Cells of Bordetella pertussis grown in a bioreactor under stirring conditions were studied to investigate the effect of shear stress on cellular-bound filamentous haemagglutinin (FHA). FHA attached to the bacterial surface, unlike extracellular FHA, was not affected at the shear levels tested. Moreover, no other cellular immunogen involved in the whole-cell protective activity seemed to be affected by hydromechanical forces.

Animals↗

Quantitation of adenylate cyclase of Bordetella pertussis by enzyme linked immunosorbent assay.

Bordetella pertussis produces extracytoplasmic adenylate cyclase toxin (AC) which has received considerable attention as a potential vaccine candidate. Great interest from laboratories involved in production, purification and quality control of acellular pertussis vaccine is focused on finding an appropriate technique for rapid and accurate quantitation of AC antigen. In this paper a competitive ELISA is proposed. A polystyrene microplate coated with purified AC was incubated with the sample to be tested plus anti-AC serum. The bound anti-AC antibodies were measured by sequential reaction with alkaline phosphatase-labelled anti-mouse IgG and p-nitrophenylphosphate. This method showed high specificity, with the 50% inhibition corresponding to 4 micrograms/ml of AC. It also proved to be useful to assess the presence of AC in culture supernatants, with high reproducibility.

Adenylyl Cyclases↗

Use of cyclodextrin as an agent to induce excretion of Bordetella pertussis antigens.

This paper attempts to provide an explanation for the effect of cyclodextrin on the yield of Bordetella pertussis soluble antigens. It was demonstrated that the addition of cyclodextrin to the synthetic Stainer-Scholte liquid medium enhances the level of the intracellular form of adenylate cyclase (200 kDa) in the supernate. In addition to this effect, it has been reported that cyclodextrin also enhances the levels of two other extracellular proteins, pertussis toxin and filamentous hemagglutinin. As these antigens are structurally different, it seems that the effect of cyclodextrin is not specific. With the use of different buffer systems of well-known action on outer membrane stability it was possible to determine a relationship between the presence of cyclodextrin, destabilisation of the outer membrane and the release of proteins. It was determined that the cyclodextrin did not modify the fluidity of B. pertussis cells but produced a change of outer membrane permeability.

Adenylate Cyclase Toxin↗

Reflex myoclonus in olivopontocerebellar atrophy.

The presence of reflex myoclonus in response to touching and pin-pricking the wrist or stretching the fingers and to photic stimulation was assessed in 24 patients with a presumed diagnosis of olivopontocerebellar atrophy (OPCA) and in 30 age matched control subjects. Reflex myoclonus to soma-esthetic stimulation was found in 23 patients and in none of the controls. Photic myoclonus was present in 12 patients and in none of the controls. Electrophysiological study of the reflex myoclonus showed enhanced (> 10 microV) somatosensory evoked potentials and an associated reflex electromyographic discharge (C-wave) in 15 patients. These findings indicate that reflex myoclonus is common in OPCA and probably of cortical origin.

Atrophy↗

[The movement from the humoral theory of disease to the birth of the modern clinic in Mexico].

This article refers to the time when the "Real y Pontificia Universidad de México" was closed and the Establecimiento de Ciencias Médicas (Medical Sciences Establishment) was created in 1833. Due to this historical event, it was done firstly as an epistemological approach in the history of medicine in Mexico; this is to say, disease concept changed. Instead of the Humoral Theory, the Biological Model regarding the lesion was developed. This model supports that disease is an anatomical or functional disturbance in the human body. The research, theoretically, is based on the philosophy of Gaston Bachelard.

Education↗

An enzyme immunoassay for determining total thyroxine in human serum using an ultramicroanalytical system.

A simple, rapid competitive ultramicroElisa assay has been developed for the measurement of total thyroxine (T4) using only 10 microliters of serum. Our novel UME is based on fluorescence measurements of the hydrolytic product of 4-methyl-umbelliferyl-beta-D-galactopyranoside. T4-beta-Galactosidase conjugates and monoclonal antibodies were immobilized on polyvinyl plates, with sodium salicylate used as a blocking agent for thyroxine binding protein. The analytical steps were carried out using a semiautomated batch-assay system entitled "SUMA" (system for ultramicroanalysis). The T4 assay was completed in 2 h, with a measuring range of 24-386 nmol/L. The intra-assay coefficient of variation (CV) was 4.6-6.9% and the inter-assay C.V. 9.0-12.4% depending on the T4 concentrations. Percentage recovery ranged from 99.2-111%. Regression analysis showed a good correlation with an established radioimmunoassay (n = 121, r = 0.946, P less than 0.01).

Antibodies, Monoclonal↗

Neuroleptic malignant syndrome treated with subcutaneous lisuride infusion.

A schizophrenic patient developed a characteristic clinical picture of neuroleptic malignant syndrome (NMS) while admitted to the hospital during an exacerbation of his psychiatric symptoms. Oral treatment of the NMS with bromocriptine (7.5 mg/day) or levodopa/carbidopa (125/12.5 mg) provoked intense vomiting in spite of domperidone (60 mg/day), which led to their discontinuation. In view of the deterioration of the symptoms, treatment was begun with lisuride (1-2 mg/24 h) subcutaneously. An obvious improvement was shown in 24 h, but levodopa/carbidopa (125/12.5 mg t.d.s. orally) had to be added later to achieve complete resolution of the NMS. During the recovery phase, while being treated with subcutaneous lisuride infusion and levodopa (p.o.), the patient presented with confusion, agitation, and hallucination. Lisuride infusion was stopped and levodopa was continued until complete resolution of the NMS. This case indicates that parenteral administration of lisuride or other dopamine agents such as levodopa (i.v.) or apomorphine (s.c.) may be considered an effective and practical way of treating NMS, particularly when the patient's condition makes it difficult or impossible to use other dopaminergic drugs by the oral route.

Adult↗