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Biomedical subjects

M Rivera

Publications and source records attributed to M Rivera.

At least 91 records · Page 5Linked to original sources

MRI visualization of small structures using improved surface coils.

In this paper we present the spatial resolution enhancement and noise reduction level achieved with an optimized inductively coupled surface coil specifically designed for our experiments. The technique of designing and implementing customized coils for magnetic resonance imaging of very small structures is described. We have designed a low cost prototype of an inductively coupled circular surface coil, tuned for 1H magnetic resonance imaging at 200 MHz. The coil is mounted on a customized teflon support. The inductive coupling used in this coil improves the signal-to-noise ratio by reducing various loss mechanisms (specially the dielectric losses). Test images have been acquired to determine the evolution of induced articular lesions in a rabbit animal model, as well as brain tumors in rats. The images show high spatial resolution, excellent B1 field homogeneity and no "hot spots". Comparing these images with those acquired with conventional coils, one finds better spatial resolution and signal-to-noise ratio, as well as larger field of view with less intense illumination artifact. The methodology can be used in any application that requires high quality imaging of small structures.

Animals↗

A role for CREB binding protein and p300 transcriptional coactivators in Ets-1 transactivation functions.

The Ets-1 transcription factor plays a critical role in cell growth and development, but the means by which it activates transcription are still unclear (J. C. Bories, D. M. Willerford, D. Grevin, L. Davidson, A. Camus, P. Martin, D. Stehelin, F. W. Alt, and J. C. Borles, Nature 377:635-638, 1995; N. Muthusamy, K. Barton, and J. M. Leiden, Nature 377:639-642, 1995). Here we show that Ets-1 binds the transcriptional coactivators CREB binding protein (CBP) and the related p300 protein (together referred to as CBP/p300) and that this interaction is required for specific Ets-1 transactivation functions. The Ets-1- and c-Myb-dependent aminopeptidase N (CD13/APN) promoter and an Ets-1-dependent artificial promoter were repressed by adenovirus E1A, a CBP/p300-specific inhibitor. Furthermore, Ets-1 activity was potentiated by CBP and p300 overexpression. The transactivation function of Ets-1 correlated with its ability to bind an N-terminal cysteine- and histidine-rich region spanning CBP residues 313 to 452. Ets-1 also bound a second cysteine- and histidine-rich region of CBP, between residues 1449 and 1892. Both Ets-1 and CBP/p300 formed a stable immunoprecipitable nuclear complex, independent of DNA binding. This Ets-1-CBP/p300 immunocomplex possessed histone acetyltransferase activity, consistent with previous findings that CBP/p300 is associated with such enzyme activity. Our results indicate that CBP/p300 may mediate antagonistic and synergistic interactions between Ets-1 and other transcription factors that use CBP/p300 as a coactivator, including c-Myb and AP-1.

Acetyltransferases↗

Organochlorine compounds (DDE and PCB) in plasma and breast cyst fluid of women with benign breast disease.

The organochlorines, dichloro-diphenyl-trichloroethane and polychlorinated biphenyl (PCB) are pervasive environmental contaminants. Results from previous studies have been conflicting regarding the relationship between the internal dose of these organochlorine residues and breast cancer risk. To determine whether these compounds are present in breast cyst fluids and whether cyst fluid and plasma concentrations are correlated, we analyzed organochlorines in paired cyst fluid and plasma samples from 24 subjects using gas chromatography and electron capture detection. All but one of the women had a history of multiple cysts, suggesting that they were at elevated risk for future breast cancer. DDE (a metabolite of dichloro-diphenyl-trichloroethane) was present in 22 of the cyst samples and PCB was detected in 19 of the cyst samples. Organochlorine levels were more concentrated in the plasma than in breast cyst fluids. Levels of DDE in plasma were significantly correlated with those in cyst fluid (r = 0.73; P < 0.001); in contrast to PCB levels in cyst and plasma (r = 0.37; P = 0.12). Congener specific analysis of the PCBs showed that some individual congeners were preferentially excluded from or concentrated in the cyst fluid. To our knowledge, this study is the first to demonstrate that PCB and DDE are present in cyst fluids and thus in contact with the ductal epithelium of the breast. These results support the use of plasma DDE as a proxy for DDE in the target tissue in research on the role of environmental factors in breast cancer.

Adult↗

Chronic morphine augments G(beta)(gamma)/Gs(alpha) stimulation of adenylyl cyclase: relevance to opioid tolerance.

In the current study, we investigated the neurochemical basis for the previously reported predominance of stimulatory mu-opioid signaling in guinea pig longitudinal muscle/myenteric plexus (LMMP) preparations after chronic in vivo morphine exposure. As expected, recombinant Gsalpha (rGsalpha) dose-dependently stimulated adenylyl cyclase (AC) activity in LMMP membranes obtained from opioid naive as well as tolerant LMMP tissue. However, the magnitude of the increase was significantly greater in the latter than in the former. The Gbetagamma blocking peptide QEHA (50 microM) essentially abolished stimulation by rGsalpha in LMMP membranes obtained from both opioid naive and tolerant animals. Interestingly, after partial blockade by lower QEHA concentrations, the incremental AC stimulation by rGsalpha in tolerant LMMP membranes was no longer observed, indicating augmented Gbetagamma stimulatory responsiveness. Concomitant changes in the content of AC isoform protein are consistent with these biochemical observations. After chronic systemic morphine, AC protein is augmented significantly (56%). This increment is most likely to be composed of AC isoforms that are stimulated by Gbetagamma. This is the first demonstration in a complex mammalian tissue that persistent activation of opioid receptors results in augmented Gbetagamma/Gsalpha AC stimulatory interactiveness. The relevance of such changes to the manifestation of opioid tolerance is discussed.

Adenylyl Cyclases↗

Assessment of the Massachusetts Medicaid managed Behavioral Health Program: year three.

This evaluation of the third year of the Massachusetts Medicaid managed Mental Health/Substance Abuse Program showed that overall utilization increased slightly and expenditures were nearly the same in FY1994 compared to FY1993; however, they were lower for disabled members. Providers believed that access to care, utilization, and quality of care were the same or better than a year earlier and that the clinical review process was improved. Client severity was higher. Aftercare planning improved but gaps in services persisted. Integration of care improved. Administrative and management problems continued. Lessons for similar, more recent initiatives are discussed.

Adult↗

Nandrolone decanoate reduces serum lipoprotein(a) concentrations in hemodialysis patients.

We have studied the changes in the lipid profile of 14 chronic hemodialysis patients receiving a 6-month cycle of nandrolone decanoate as treatment for anemia. Nandrolone decanoate was administered in a weekly intramuscular dose of 200 mg and resulted in an increase in the hemoglobin concentration (baseline, 7.9 +/- 0.9 g/dL; month 6, 10.8 +/- 1.7 g/dL; P < 0.001, ANOVA) and also produced relevant modifications in the lipid concentrations. The most significant finding was a decrease in the concentration of lipoprotein(a) [Lp(a)]: baseline, 19.8 mg/dL (median), month 2, 10.6 mg/dL; month 4, 8.7 mg/dL; and month 6, 7.1 mg/dL (P < 0.001, Friedman). Other lipid changes induced by nandrolone decanoate were an increase in the concentrations of apolipoprotein B (P < 0.02, ANOVA) and triglyceride (P = NS, ANOVA) and a decrease of high-density lipoprotein (HDL) cholesterol (P < 0.001, ANOVA) and apolipoprotein A-I (P = NS, ANOVA). The decrease in HDL cholesterol was at the expense of the HDL2 cholesterol subfraction, whereas HDL3 remained unchanged. These lipid modifications were reversible; 4 months after nandrolone decanoate withdrawal, the lipid concentrations were similar to the basal values. The changes in Lp(a) levels did not correlate with those of hemoglobin or the other lipid parameters, suggesting that the underlying mechanisms are unrelated. Our findings could be clinically relevant if confirmed by further studies.

Aged↗

Trypanosoma evansi in capybara from Venezuela.

During the slaughtering season of February and March 1991, 559 capybaras (Hydrochoerus hydrochaeris) were tested for Trypanosoma evansi in two areas in Venezuela: El Frio Ranch and El Cedral Ranch. Blood and serum samples were evaluated for T. evansi. Forty-eight (9%) of 559 capybaras had T. evansi using the microcentrifugation technique. Further, 279 (50%) of the 559 capybaras had antibodies against T. evansi immunofluorescence test in both ranches. Thus, capybaras may be important in the epizootiology of T. evansi in this enzootic area.

Animals↗

Vaccine evaluation studies of replication-defective SIVsmB7.

Non-infectious virus-like particles of SIVsmB7 that expresses env and gag gene products but are defective in pol and vpx/vpr were assessed for their ability to induce protective immunity against infection with pathogenic SIVsmE660 in rhesus macaques. Animals were immunized in three groups: group A was primed with cell-associated SIVsmB7 and boosted with cell-free SIVsmB7; group B was primed with cell-free SIVsmB7 and boosted with cell-free SIVsmB7 conjugated to iron oxide microbeads; group C was primed with cell-free SIVsmB7 mixed with Titer Max adjuvant and boosted with cell-free SIVsmB7 mixed with SAF-M adjuvant followed by secondary boosting with cell-free SIVsmB7 conjugated to microbeads. Animals were challenged intravenously with 20 animal infectious doses of SIVsmE660 grown in rhesus peripheral blood mononuclear cells 3 weeks after final boosting. All animals became infected as evidenced by quantitative virus cultivation. Sera from immunized animals contained low-titer antibodies by ELISA and low or undetectable neutralizing antibodies on the day of challenge but strong anamnestic antibody responses were observed following challenge. Interestingly, 2 of 3 animals in group A showed evidence of transient viremia and more stable CD4 counts following challenge as compared to the other immunized animals and to non-immunized controls. Thus, immunization with cell-associated SIVsmB7 did not provide sterilizing immunity against challenge with a highly pathogenic SIV strain but might have caused virus clearance later in infection.

Animals↗

13C NMR spectroscopic and X-ray crystallographic study of the role played by mitochondrial cytochrome b5 heme propionates in the electrostatic binding to cytochrome c.

The role played by the outer mitochondrial membrane (OM) cytochrome b5 heme propionate groups in the electrostatic binding between OM cytochrome b5 and horse heart cytochrome c was investigated by 13C NMR spectroscopy and X-ray crystallography. To achieve these aims, 13C-labeled heme OM cytochrome b5 was expressed in Escherichia coli as previously described [Rivera M., Walker, F.A. (1995) Anal. Biochem. 230, 295-302]. Assignment of the resonances arising from the heme propionate carbons in ferricytochrome b5 was carried out by a combination of one- and two-dimensional NMR experiments. Titrations of [13C]heme-labeled OM cytochrome b5 with horse heart cytochrome c were carried out in order to monitor the resonances arising from the heme propionate carbonyl carbons in OM cytochrome b5. The results from these titrations clearly show that only the heme propionate located on the exposed heme edge in OM cytochrome b5 participates in the electrostatic stabilization of the complex between OM cytochrome b5 and horse heart cytochrome c. Similar experiments carried out monitoring 13C resonances arising from several other heme substituents demonstrated that the stoichiometry of the complex is 1:1. A conditional binding constant, K which equals 3.8 x 10(4) +/- 1.4 x 10(4) at mu = 0.02 M, was obtained for the formation of the complex by fitting the binding curves obtained experimentally to a model based on this stoichiometry. The X-ray crystal structure of rat liver OM cytochrome b5 solved to 2.7 A resolution shows that the structures of bovine liver microsomal cytochrome b5 and rat liver OM cytochrome b5 are almost identical when compared at medium resolution. The similarity between the two structures, combined with the findings that only the heme propionate located on the exposed heme edge of OM cytochrome b5 participates in the electrostatic binding to cytochrome c and that the stability of this complex is similar to that measured for the association between microsomal cytochrome b5 and cytochrome c, clearly indicates that the site of interaction on OM cytochrome b5 is almost identical to the one elucidated for microsomal cytochrome b5. It is therefore possible to conclude that the large body of information gathered by many investigators for the nonphysiological interaction between microsomal cytochrome b5 and cytochrome c (recently reviewed) [Mauk, A. G. Mauk, M. R., Moore, G. R., & Northrup, S. H. (1995) Bioenerg. Biomembr. 27, 311-330] has indeed biological as well as pedagogical validity.

Animals↗

Electrochemical measurement of second-order electron transfer rate constants for the reaction between cytochrome b5 and cytochrome c.

The second-order electron transfer reaction between cytochrome b5 and cytochrome c has been studied by cyclic voltammetry utilizing a gold electrode modified with beta-mercaptopropionate. When cyclic voltammetry is performed on a solution containing a mixture of cytochrome b5, cytochrome c and polylysine, cytochrome b5 undergoes reversible electrochemistry at the electrode surface while cytochrome c discriminates against the electrode surface. The selectivity of the modified electrode for negatively charged proteins makes it possible to selectively reduce a protein possessing a net negative charge and a relatively low reduction potential (outer mitochondrial membrane cytochrome b5, Eo = -102 mV; microsomal cytochrome b5, Eo = 3 mV) in the presence of another protein possessing a net positive charge and a relatively high reduction potential (cytochrome c, Eo = 265 mV). The electrochemical reduction of ferricytochrome b5 at the electrode surface is followed by a second-order electron transfer reaction between ferrocytochrome b5 and ferricytochrome c that yields ferricytochrome b5 and ferrocytochrome c. This fast homogeneous electron transfer reaction which is preceded by a heterogeneous electron transfer reaction results in a characteristic cyclic voltammogram containing a pre-peak to the reduction current. The second-order rate constant for the homogeneous reaction was obtained by invoking the above reaction scheme for digital simulation of a cyclic voltammogram which was subsequently fitted to the experimental data. Second-order rate constants obtained with this method are 2.9 x 10(8) and 8.9 x 10(8) for the homogeneous electron transfer reactions between rat liver outer mitochondrial membrane (OM) ferrocytochrome b5 and beef liver microsomal ferrocytochrome b5, with horse heart ferricytochrome c, respectively. These values are in good agreement with second-order rate constants obtained for the same protein systems by flash photolysis. [Meyer, T. E., Rivera, M., Walker, F. A., Mauk, M. R., Mauk, A. G., Cusanovich, M. A., & Tollin, G. (1993) Biochemistry 32, 622-627].

3-Mercaptopropionic Acid↗

Relevance of phosphorylation state to opioid responsiveness in opiate naive and tolerant/dependent tissue.

This laboratory previously reported that the mu-selective opiate receptor agonist, sufentanil, produces a naloxone-reversible, concentration-dependent facilitation or inhibition of the stimulated formation of cAMP in the myenteric plexus. Chronic in vivo exposure to morphine results not only in the loss of inhibitory opioid responsiveness but in the reversal of inhibition to enhancement. The present study demonstrates, in tolerant/dependent as well as opiate naive tissue, that the state of phosphorylation is a critical determinant of the balance between positive and negative opioid modulation of stimulated cAMP formation. In vitro treatment of chronic morphine-treated preparations with inhibitors of protein kinases, abolishes the previously observed reversal of opioid inhibition to enhancement and restores sufentanil inhibitory responsiveness. The established kinase-type selectivity profile of the inhibitors employed suggests the involvement of protein kinase C (PKC) in the tolerant-associated reversal from opioid inhibition to enhancement of cAMP formation. Conversely, treatment of opiate naive tissue with the protein phosphatase inhibitor okadaic acid or a phorbol ester activator of protein kinase C, phorbol 12-myristate 13-acetate (PMA), not only attenuates sufentanil inhibition of evoked cAMP formation but reverses it to a facilitation (as occurs following chronic in vivo morphine exposure). This effect of PMA is abolished by the PKC-selective inhibitor chelerythrine. Moreover, the longitudinal muscle myenteric plexus content of PKC alpha and PKC beta is substantially elevated following chronic morphine treatment. These results underscore the relevance of opioid bimodality to the manifestation of tolerance/dependence and suggest that augmented phosphorylation (mediated at least in part via PKC) is a critical determinant of some of the sequelae of chronic morphine exposure.

Animals↗

The experiences of ataques de nervios: towards an anthropology of emotions in Puerto Rico.

Ataques de nervios are an idiom of distress used by Puerto Ricans and other Latinos to express dislocations in the social world of the family. This paper contributes to the growing study of the "anthropology of the emotions". Through detailed interviews with 121 people in Puerto Rico, 78 of whom had had an ataque de nervios, we are developing a thick description of both the prototypical models for ataques de nervios and the varied individual experiences of ataques. The interview used in this study is a version of the Explanatory Model Interview Catalogue specifically adapted for use in a community study of ataques de nervios in Puerto Rico. Responses to questions on the experience of ataque de nervios were analyzed using a team of reviewers who represented differing knowledge and experience with Puerto Rican culture and mental health practice. The experience of ataques de nervios involves a loss of control in several important domains of experience: emotional expressions, bodily sensations, action dimensions and alterations in consciousness. That loss of control is closely linked to important social contexts relating to major life problems and the experience of suffering.

Adolescent↗

A concept-based retrieval system for thoracic radiology.

Current digital information systems in radiology are insufficient to accommodate the retrieval needs of academicians. Significant efforts are required in retrieving clinical cases for teaching and research. We describe a prototype system that supports intelligent case retrieval based on a combined specification of patient demographics, radiologic findings, and pathologic diagnoses. The documents for these cases can be distributed among multiple heterogeneous data bases. The system features automatic indexing of radiology and pathology reports, a comprehensive lexicon for thoracic radiology, an interface to a hospital information system, radiology information system, and picture archiving and communication systems, and a graphical user interface for query formulation and results visualization. The prototype system was developed within the domain of thoracic radiology involving patients with lung cancer.

Abstracting and Indexing↗