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Biomedical subjects

M Rodriguez

Publications and source records attributed to M Rodriguez.

At least 415 records · Page 23Linked to original sources

Abrogation of resistance to Theiler's virus-induced demyelination in H-2b mice deficient in beta 2-microglobulin.

Intracerebral infection of susceptible strains of mice with Theiler's virus, a picornavirus, results in central nervous system demyelination, which is similar to multiple sclerosis. Immunogenetic experiments indicate that the MHC (H-2) and, in particular, the D region that controls class I-restricted immune responses, is an important determinant to development of demyelination. We tested whether disruption of beta 2-microglobulin (beta 2-m) would abrogate resistance to demyelinating disease normally observed in H-2b mice. All (C57BI/6 x 129)F3 mice transgenic for homozygous beta 2-m gene disruption (-/-) developed chronic demyelination after Theiler's murine encephalomyelitis virus infection, whereas none of the infected littermates with normal expression of class I MHC (beta 2-m, +/+) developed demyelination. Demyelinated lesions showed class II MHC expression, macrophages, and TNF but no class I MHC expression or CD8+ T cells. No correlation was observed between development of demyelination and delayed-type hypersensitivity responses to virus Ag. Despite the presence of demyelinating lesions, none of the infected beta 2-m (-/-) mice developed neurologic deficits. Infectious virus and virus Ag persisted in the central nervous systems of infected beta 2-m (-/-) mice but not in beta 2-m (+/+) mice. These experiments support the hypothesis that a class I immune response mediated by CD8+ T cells is important in resistance to Theiler's murine encephalomyelitis virus-induced demyelination. Development of chronic neurologic deficits as observed in immunocompetent susceptible strains of mice may be dependent on the presence of class I MHC and CD8+ T cells.

Animals↗

Molecular cloning of a human beta 3-adrenergic receptor cDNA.

We report the molecular cloning of a beta 3-adrenergic receptor [beta 3-AR] cDNA from human brown adipose tissue. The cDNA-encoded protein is identical to the previously cloned beta 3-AR but with 6 additional amino acids at the C-terminus. The C-terminus is shared by the beta 3 receptors expressed in human neuroblastoma cells [SK-N-MC] [Mol. Pharmacol. 42 (1992) 964-970]. Furthermore, using a polymerase chain reaction strategy we have cloned and sequenced the beta 3-AR introns. Sequence analysis demonstrates that the human beta 3-AR gene comprises at least 3 exons and 2 introns and that the most abundant beta 3-AR transcripts encode a protein with an exon 3-derived C-terminus. Interestingly, although a similar organization has been found in rodent genes, the rat beta 3-AR transcripts encode a receptor with an exon 2-derived C-terminus.

Adipose Tissue, Brown↗

Synthesis and analgesic effects of N-[3-[(hydroxyamino) carbonyl]-1-oxo-2(R)-benzylpropyl]-L-isoleucyl-L-leucine, a new potent inhibitor of multiple neurotensin/neuromedin N degrading enzymes.

The synthesis of N-[3-[(hydroxyamino) carbonyl]-1-oxo-2(R)-benzylpropyl]-L-isoleucyl-L-leucine (JMV-390-1, 6a), a multipeptidase inhibitor based on the C-terminal sequence common to neurotensin (NT) and neuromedin N (NN), is described. This compound behaves as a full inhibitor of the major NT/NN degrading enzymes in vitro, e.g. endopeptidase 24.16, endopeptidase 24.15, endopeptidase 24.11, and leucine aminopeptidase (type IV-S), in the nanomolar range (IC50's from 30 to 60 nM). Compound 6a was found to increase endogenous recovery of NT and NN from slices of mice hypothalamus depolarized with potassium. In various assays commonly used to select analgesics, e.g. hot-plate test, tail-flick test, acetic acid-induced writhing test, in mice, compound 6a proved to be potent when intracerebroventricularly (icv) injected. The analgesic effects observed were totally (hot-plate test) or largely (tail-flick test) reversed by the opioid antagonist naltrexone. Furthermore, icv injection of compound 6a (10 micrograms/mouse) was found to significantly potentiate the hypothermic effects of NT or NN.

Analgesics↗

The rat beta 3-adrenergic receptor gene contains an intron.

We report that the rat beta 3-adrenergic receptor (beta 3-AR) gene has an intron. The intron starts with an in-frame stop codon with the result that unspliced transcripts will encode a C-terminal truncated protein. The reported protein sequences of mouse and human beta 3-AR were both deduced from genomic DNA sequences. Given the heterogeneity at the C-termini of the otherwise highly similar rat, mouse and human sequences, we discuss the intriguing possibility that the beta 3-AR gene of the latter two species also contain an intron near the extremity of the open reading frame. A beta-adrenergic receptor (beta-AR) cDNA we have cloned from rat colonic tissue which has a sequence essentially identical to that previously reported for the rat adipose beta 3-AR cDNA [(1991) J. Chem. 266, 24053], encodes the spliced version of the beta 3-AR.

Amino Acid Sequence↗

Human class I major histocompatibility complex transgene prevents virus-induced demyelination in susceptible mutant B10.D2dml mice.

Theiler's murine encephalomyelitis virus (TMEV) induces immune-mediated demyelination in susceptible strains of mice, providing an excellent model for multiple sclerosis. Class I genes within the major histocompatibility complex locus (H-2D region) play a major role in determining whether strains of mice develop chronic demyelination and TMEV persistence. B10.D2dml mice with deletion in the 3' end of Dd and the 5' end of Ld genes develop the most prominent demyelination in comparison with resistant B10.D2 mice with normal complementation of H-2D region genes. We tested whether expression of a class I human transgene (HLA-B27) would modulate virus-induced demyelination in mutant B10.D2dml mice. Transgenic B10.D2dml (HLA-B27+) mice infected with virus showed dramatic decrease in the extent of demyelination (p < 0.0001) and virus antigen expression in spinal cord compared with littermate controls without the human class I transgene. These experiments demonstrate that transgenic expression of a human class I major histocompatibility complex locus molecule can prevent demyelination induced by a virus in mutant mice.

Animals↗

Platelet adhesion/aggregation in an in vitro model of coronary artery stenosis.

Platelet adhesion/aggregation (PAA) at a site of coronary artery stenosis is believed to be a process strongly modulated by local shear rates and the functional state of neighboring endothelium. One purpose of the present work, therefore, is to describe an in vitro model for the direct imaging of such PAA. Another is to apply the model to the question as to whether the use of nonionic vs. ionic contrast media (CM) in the presence of vascular endothelium contributes to PAA at the stenosis site. Toward these ends, we utilized a special flow chamber which incorporates a monolayer of endothelial cells (ECs), a step 66% flowpath constriction at a site preadsorbed with microfibrillar collagen, and arterial shear rates. By epifluorescence microscopy and digital image analysis of video recordings, PAA was found to be greater with dysfunctional ECs (pretreated with lysine acetylsalicyclate) than with normal ECs, thereby confirming a modulatory role in PAA of functionally intact ECs. When nonionic (iohexol) or ionic (ioxaglate, diatrizoate) CM was added to the flowing blood at a concentration of 20% by non-red cell volume, PAA was inhibited in the order diatrizoate > ioxaglate > iohexol > saline control. No inhibition by any CM was seen, however, when chamber prefill culture medium containing 20% by volume CM was displaced by CM-free blood, in simulation of bolus administration of CM. In terms of inhibition of PAA during percutaneous transluminal coronary angioplasty (PTCA), therefore, our model provides a conceptual basis by which one may anticipate in flowing blood no clear benefit of ionic over nonionic CM.(ABSTRACT TRUNCATED AT 250 WORDS)

Aspirin↗

Detection of human papillomavirus L1 protein in condylomata acuminata from adults with defects in cell-mediated immunity.

Immunohistochemical assays for human papillomavirus (HPV) L1 protein, using antiserum directed against the L1 major capsid protein of bovine papillomavirus (anti-BPV serum), were performed on 101 condylomata acuminata biopsy samples from 47 men (40 of whom had intact cell mediated immunity [CMI], and 7 with conditions known to cause CMI defects), and 54 women (48 with normal CMI, and 6 with CMI defects). L1 protein was detected in 28% of all biopsies, including 20.5% of samples from patients with normal CMI and 76.9% of patients with CMI defects (P = .00002). For both males and females, L1 protein was detected significantly more often in samples from patients with CMI defects than those with normal CMI. Immunohistochemical assays of HPV 11-infected human foreskin implants grown in athymic mice were performed to optimize the conditions of the assay. Three dilutions of anti-BPV serum or preimmune rabbit serum were used. A 1:500 dilution provided readily interpretable results, while preimmune serum at this dilution did not stain to any significant degree. However, at 1:10 and 1:100 dilution, both the anti-BPV serum and preimmune serum caused an unacceptable amount of nuclear staining, making results uninterpretable. Defects in CMI may allow active viral particle assembly at an increased rate, as judged by detection of L1 protein in condylomata acuminata from these patients.

Capsid↗

ST0 or ST60.

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Electrocardiography↗

Improved exercise test accuracy using discriminant function analysis and "recovery ST slope".

The objective of the study was to optimize the accuracy of the exercise test for predicting the presence of significant angiographic coronary artery disease. A retrospective analysis of stored digital exercise electrocardiographic data on 147 men who had undergone exercise testing and cardiac catheterization was performed. With significant coronary artery disease defined as > or = 70% stenosis, 95 patients had one or more vessel(s) diseased. None were receiving digoxin, had a myocardial infarction or previous coronary artery bypass graft, or exhibited left bundle branch block, left ventricular hypertrophy, Q waves, or ST depression on their resting electrocardiogram. Analysis was performed using the authors' averaging and measurement software at rest and at each 30 seconds throughout the exercise and recovery in leads II, V2, and V5. Discriminant function analysis was used to analyze pretest variables, as well as hemodynamic and electrocardiographic changes and symptoms during exercise. A discriminant function score was developed and compared to other treadmill scores. The setting was a 1,000 bed Veterans Affairs Medical Center (Long Beach, CA). Discriminant function analysis chose age, smoking status, presenting chest pain characteristics, and lead V5 ST slope in recovery to have independent power for separating those with and without coronary artery disease. A discriminant function score using these four variables was used to form a receiver operating characteristics curve (and derive receiver operating characteristics curve areas) for comparison to other exercise test methods and scores: (discriminant function score = .81; slope 3.5 minutes into recovery in lead V5 = .73; traditional ST amplitude method = .72; ST60/HR index (amplitude of ST depression 60 ms after the J point/delta heart rate) = .66; traditional ST amplitude/HR index (traditional method/delta heart rate) = .75; Hollenberg score = .68; Hollenberg areas only = .66; and ST integral = .66. Receiver operating characteristics curve analysis revealed a trend for the discriminant function score to be superior to all other measurements and scores. Recovery ST slope in lead V5 performed as well as or better than all other electrocardiographic criteria or treadmill scores except for the authors' discriminant function score.

Cardiac Catheterization↗

Ontogeny of T-maze behavioral lateralization in rats.

Behavioral lateralization has been reported in both humans and animals. In humans, lateralization can be detected in neonates and increases to adult levels during postnatal development. Recently we reported lateralization of head and tail movements in neonatal rats. However, the postnatal ontogeny of lateralization in animals has not been previously studied. This work presents a study of rat behavioral lateralization in the T-maze test during postnatal development (from day 30 to day 60). A decrease was found in absolute (percent preferred-side choices) and population (right-left arm choices) laterality between day 30 and day 45 of postnatal life. The lateralization degree remained unchanged between days 45 and 60. Because behavioral alternation increases from day 30 to day 45, the present data suggest that animal lateralization of behavior is a phenomenon that remains throughout the subject's life span, but whose behavioral quantification could be concealed by the ontogenic increases of other phenomena such us behavioral alternation. This hypothesis could explain the high level of lateralization in neonatal rats, the low level of rat lateralization during adulthood, and the increases in lateralization induced by stress.

Aging↗

Expression of T cell receptor V beta transcripts in central nervous system of mice susceptible and resistant to Theiler's virus-induced demyelination.

We utilized the polymerase chain reaction (PCR) to examine for preferential expression of T cell receptor (TcR) V beta s in T cells infiltrating the central nervous system (CNS) of mice infected with Theiler's virus. Infection of susceptible strains of mice with Theiler's virus results in demyelinating disease similar to multiple sclerosis. At 7 days following infection, no difference was observed in TcR V beta usage in lymphocytes infiltrating the CNS between resistant (B10.K) and susceptible (B10.Q or SJL/J) mice. In susceptible mice with prominent demyelination (day 45 and 238 following infection), no preferential expression of TcR V beta s was observed in the CNS. There is strong evidence that T cells play a major role in pathogenesis of TMEV-induced demyelination. There is increasing evidence using recombinant inbred strain mice with TcR V beta deletions that T cells expressing certain TcR V beta genes may be critical in the disease process. Yet, analysis of TcR V beta expression on T cells in CNS using PCR technology did not provide a way to dissect which antigen-specific T cells play a role in disease. These results confirm that during active demyelination specific as well as non-specific T cells are recruited to the CNS. Even though the pathogenesis of TMEV-induced disease may not be identical to that in multiple sclerosis, it is unlikely that similar approaches utilizing polymerase chain reaction will provide insights to the role of T cell receptors in the pathogenesis of multiple sclerosis.

Animals↗

Neonatal lateralization of behavior and brain dopaminergic asymmetry.

Behavioral lateralization has previously been reported in adult animals. This work presents a study of behavioral laterality and spontaneous alternation behavior in 156 neonatal rats (39 litters with two males and two females per litter). The initial tail and head lateral movements (axis body-tail or head higher than 30 degrees) were recorded after neonatal rats were gently placed along a straight line drawn on a glass surface. This test was repeated 10 times. A leftward population lateralization was found for tail movement while head movement was rightward. The alternation behavior was lower (10-25%) than that previously reported for adult rats (80-90%) and than that expected if movement was made at random (50%). Males were more lateralized than females for head and tail movements. No sex differences were observed for behavioral alternation. In 2-day-old rats (10 litters with one male and one female per litter), the right brain side had a higher content in dopamine (mesencephalon) and DOPAC (proencephalon) than the left brain side. Thus, we concluded that behavioral and biochemical asymmetries in animals are conditioned by phenomena present during prenatal or early neonatal (first hours after birth) life and that spontaneous alternation behavior is not present during the earlier stages of postnatal development.

3,4-Dihydroxyphenylacetic Acid↗

Oligodendrocyte injury is an early event in lesions of multiple sclerosis.

The ultrastructural features of 11 stereotaxic brain biopsy specimens that demonstrated inflammatory primary demyelination consistent with acute multiple sclerosis were examined. Uniform widening of inner myelin lamellae (biphasic myelinopathy) and degeneration of inner glial loops ("dying-back" oligodendrogliopathy) were early pathologic abnormalities that antedated complete destruction of myelin sheaths. Perivascular inflammatory cells (lymphocytes, macrophages, and occasional plasma cells) were in intimate contact with degenerating myelin sheaths. The response of astrocytes was prominent, even in areas of minimal demyelination. Oligodendrocytes were morphologically preserved in early lesions but proliferated at the periphery of active lesions. Thinly myelinated axons indicative of central nervous system-type remyelination by oligodendrocytes were observed primarily at the edge of plaques. Disturbances of the myelinating function of oligodendrocytes--unaccompanied by death of these cells--may be among the earliest pathologic features in multiple sclerosis.

Adolescent↗

The relationship between serum calcitonin and calcium in the hemodialysis patient.

An increase in the serum calcium level is known to stimulate calcitonin secretion, but whether calcitonin contributes to calcium regulation in the dialysis patient is not known. Because we recently observed in the rat that (1) a sigmoidal calcitonin-calcium curve was present, and (2) calcitonin stimulation decreased the calcemic response to parathyroid hormone (PTH), the relationship between serum calcitonin and calcium was studied in six hemodialysis patients. To evaluate both the calcitonin-calcium and PTH-calcium relationships, hemodialysis with a low-calcium dialysate (2 mg/dL) was performed and followed 1 week later by hemodialysis with a high-calcium dialysate (8 mg/dL). The calcitonin-calcium relationship was best represented as a sigmoidal curve that was opposite in direction to the PTH-calcium curve. The range of the calcitonin-calcium curve was less than the PTH-calcium curve; as a percent of maximal, the minimal calcitonin was higher than the minimal PTH, 61% +/- 5% versus 26% +/- 4% (P < 0.001). As opposed to the PTH-calcium curve in which the basal PTH value was intermediate between the minimal and maximal PTH, the basal calcitonin value was close to the minimal calcitonin. Despite a wide range of ionized serum calcium concentrations (3.5 to 5.3 mg/dL), maximal stimulation was greater than maximal inhibition of calcitonin, 60% +/- 13% versus 3% +/- 2% (P = 0.01). Finally, a significant correlation was observed between basal and maximal calcitonin (r = 0.99, P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Progression from asymptomatic to symptomatic urinary tract infection in patients with SCI: a preliminary study.

The purpose of this prospective pilot study was to (1) evaluate the role of pyuria in predicting the progression from asymptomatic to symptomatic urinary tract infection (UTI) in spinal cord-injured patients who undergo sterile intermittent bladder catheterization and (2) evaluate the impact of treating asymptomatic UTI on this progression. Twenty hospitalized patients were randomized to either the treatment group (10 subjects) or the control group (10 subjects). Weekly urine samples were obtained for quantitation of bacterial growth and pyuria. Neither the level nor the trend of pyuria helped predict the imminent progression to symptomatic UTI. Thirty percent of patients in the treatment group developed symptomatic UTI vs 70 percent of patients in the control group; it took a significantly longer time for patients in the treatment vs control group to develop symptomatic UTI (median number of days: 72 vs 7, respectively; p < 0.003). Further analysis of the long-term impact of antibiotic treatment of asymptomatic UTI is warranted.

Adult↗

The potential for oligodendrocyte proliferation during demyelinating disease.

The potential for oligodendrocytes to proliferate in response to central nervous system injury was examined. We used intracerebral infection of Theiler's murine encephalomyelitis virus, a model for multiple sclerosis, which results in chronic demyelinating disease of SJL/J mice. Proliferating cells in spinal cord sections of adult mice were identified using simultaneous immunohistochemistry and in situ autoradiography ([3H]-thymidine incorporation). Seven different cell-specific markers were used to characterize proliferating cells as oligodendrocytes (myelin basic protein, proteolipid protein, galactocerebroside, CNPase), astrocytes (glial fibrillary acidic protein), microglia/macrophages (Griffonia simplicifolia isolectin B4) or T-lymphocytes (CD3). The average number of proliferating cells per area of spinal cord white matter was 11/mm2 in normal young adult mice compared to 61/mm2 in chronically infected mice. Most proliferating cells in normal spinal cord were not identified with these markers and were presumed to be progenitor glial cells. However, in spinal cord white matter of mice infected with Theiler's virus for approximately 4 months, 88% of proliferating cells were identified. Approximately one-third of all proliferating cells were in the oligodendrocyte lineage and expressed markers observed late in myelin differentiation. In demyelinated areas as compared to normal white matter, there was an 80- to 211-fold increase in the number of proliferating oligodendrocytes expressing myelin basic protein or proteolipid protein, respectively. The remainder of the proliferating cells in areas of demyelination were astrocytes, microglial cells and T-cells. These experiments support the hypothesis that factors within a demyelinating lesion promote the proliferation and differentiation of cells within the oligodendroglial lineage.

2',3'-Cyclic-Nucleotide Phosphodiesterases↗

Changes in malathion and pyrethroid resistance after cypermethrin selection of Culex quinquefasciatus field populations of Cuba.

Use of the organophosphorus insecticide malathion for mosquito control in Cuba, for 7 years up to 1986, selected elevated non-specific esterase and altered acetylcholinesterase (AChE) resistance mechanisms in Culex quinquefasciatus. In central Havana space-spraying of malathion was replaced by the pyrethroid cypermethrin in 1987: alternate cycles of malathion and cypermethrin were applied in some of the more rural areas of Havana district during 1987-91. Consequently, populations of Cx quinquefasciatus in the central area of Havana developed resistance to cypermethrin, but there is no evidence of pyrethroid resistance in the outlying areas. Malathion resistance levels declined significantly after 1986, measured both by bioassay and the frequency of the elevated esterase resistance mechanism, and then stabilized with no measurable decline during 1990 in any of the populations tested. These populations had less than 10% frequency of susceptible homozygotes for both the esterase and AChE resistance mechanisms, indicating that organophosphate resistance is still prevalent in Cuban Cx quinquefasciatus. These two mechanisms appear to be in linkage equilibrium, suggesting that current selection for double resistance is not strong. In the central Havana region, pirimiphos-methyl, an organophosphorus insecticide unaffected by the two common malathion resistance mechanisms, is now being used in a resistance management strategy designed to avoid pyrethroid resistance spreading.

Animals↗