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Biomedical subjects

R Flores

Publications and source records attributed to R Flores.

At least 19 recordsLinked to original sources
PubMed1992

The strands of both polarities of a small circular RNA from carnation self-cleave in vitro through alternative double- and single-hammerhead structures.

The sequence of a circular RNA from carnation has been determined and found to consist of 275 nucleotide residues adopting a branched secondary structure of minimum free energy. Both plus and minus strands of this RNA can form the hammerhead structures proposed to mediate the in vitro self-cleavage of a number of small infectious plant RNAs and the transcript of satellite 2 DNA from the newt. Minus full- and partial-length transcripts of the carnation circular RNA including the hammerhead structure showed self-cleavage during transcription and after purification, indicating the involvement of a single-hammerhead structure in the self-cleavage reaction. In the case of the plus transcripts only a dimeric RNA, but not a monomeric one, self-cleaved efficiently during transcription and after purification, strongly supporting the implication in this process of a double-hammerhead structure theoretically more stable than the corresponding single cleavage domain. However, a plus monomeric transcript self-cleaved after purification at a slow rate in a concentration-independent reaction which most probably occurs through an intramolecular mechanism. Comparative sequence analysis has revealed that the circular RNA from carnation shares similarities with some representative members of the viroid and viroid-like satellites RNAs from plants, suggesting that it is a new member of either these two groups of small pathogenic RNAs.

Base Sequence↗
PubMed1992

Plus and minus RNAs of peach latent mosaic viroid self-cleave in vitro via hammerhead structures.

Peach latent mosaic viroid (PLMVd), the causal agent of peach latent mosaic disease, has been sequenced and found to be a circular RNA molecule of 337 nucleotide residues, which adopts a branched conformation when it is folded in the model of lowest free energy. PLMVd exhibits limited homologies with other viroids and some satellite RNAs, but it does not have any of the central conserved sequences characteristic of the subgroups of typical viroids. However, a segment of approximately one-third of the PLMVd sequence has the elements required to form in the RNAs of both polarities the hammerhead structures proposed to act in the in vitro self-cleavage of avocado sunblotch viroid (ASBVd) and some satellite RNAs. Plus and minus partial- and full-length RNA transcripts of PLMVd containing the hammerhead structures displayed self-cleavage during transcription and after purification as predicted by these structures. These data are consistent with the high stability of the PLMVd hammerhead structures, more similar to the corresponding structures of some satellite RNAs than to those of ASBVd, and indicate that the self-cleavage reactions of PLMVd are most probably mediated by single hammerhead structures. Our results support the inclusion of PLMVd in a viroid subgroup represented by ASBVd, whose members are characterized by their ability to self-cleave in vitro, and probably in vivo, through hammerhead structures. A consensus phylogenetic tree has been obtained suggesting that PLMVd, together with ASBVd, may represent an evolutionary link between viroids and viroid-like satellite RNAs.

Base Sequence↗
PubMed1992

Characterization of RNAs specific to avocado sunblotch viroid synthesized in vitro by a cell-free system from infected avocado leaves.

Analysis by molecular hybridization of the RNAs transcribed by a cell-free fraction from avocado infected with avocado sunblotch viroid (ASBV) demonstrated the presence of newly synthesized viroid-specific sequences, most of which were of the same polarity as the mature infectious viroid RNA. Treatment of the cell-free fraction with DNase reduced the total synthesis of RNA considerably, but it did not influence that of the ASBV-specific RNAs, indicating that the latter were transcribed on an RNA template. Inhibition studies with alpha-amanitin showed that the synthesis of ASBV-specific RNAs was not affected by concentrations of 1 and 200 micrograms/ml of the drug, which typically inhibit RNA polymerase II and III, respectively, from most animal and plant systems. These results suggest that either RNA polymerase I or an unidentified RNA polymerase activity resistant to alpha-amanitin, acting on an RNA template, plays a role in the replication of ASBV, whereas for the rest of the viroids studied so far it appears that RNA polymerase II is involved. Analysis by polycrylamide gel electrophoresis under partially and fully denaturing conditions of the ASBV-specific RNAs synthesized in vitro showed that they contain unit and longer than unit length viroid strands, probably associated in complexes with single- and double-stranded regions. The structural properties of these complexes are similar to those of the RNAs accumulating in vivo in viroid-infected tissues, which are the postulated replicative intermediates of the rolling-circle mechanism proposed for viroid synthesis.

Amanitins↗
PubMed1992

Effects of diurnal phase and pimozide on cholecystokinin-elicited hypoactivity in the hamster.

Locomotor activity in golden Syrian hamsters was measured following IP injections of cholecystokinin (CCK; 25 micrograms/kg) and pimozide (0.5 mg/kg), the dopamine receptor antagonist. In addition, animals were tested during either the dark or light phase of the diurnal cycle in either dark or light running wheel environments. Results indicated that CCK-elicited hypoactivity was blocked by pimozide and that the effect of CCK was evident only among animals tested during the light phase of the daily cycle. Ambient lighting conditions in the test environment did not modify the drug effects. Independently of any drug effect, locomotor activity was affected by diurnal phase and ambient lighting in the test environment. Animals were more active when tested during the dark phase than during the light phase and locomotor activity was higher under dark than under light ambient conditions. It is concluded that diurnal phase modulates CCK's effect on hamster locomotion and that CCK's effect on locomotion is mediated, in part, by dopaminergic mechanisms.

Animals↗
PubMed1992

Blocking naloxone-precipitated withdrawal in rats and hamsters.

Three experiments studied the effects of putative antagonists of opiate withdrawal in hamsters and rats. In Experiment 1, the calcium channel antagonists verapamil (20 mg/kg) and nifedipine (20 mg/kg) failed to antagonize naloxone (1 mg/kg)-precipitated withdrawal in hamsters implanted with two 75-mg morphine pellets, whereas clonidine (0.4 mg/kg), the alpha 2-adrenergic agonist, blocked most withdrawal signs. In Experiment 2, clonidine (0.4 mg/kg) and verapamil (20 mg/kg) were tested against naloxone-precipitated withdrawal in hamsters made acutely dependent by a single injection of morphine (15 mg/kg). As in Experiment 1, clonidine but not verapamil was effective. In Experiment 3, the effects of verapamil on naloxone-precipitated withdrawal were studied in morphine-pelleted rats and hamsters. In rats implanted with two morphine pellets, verapamil (20 mg/kg) reversed naloxone-precipitated withdrawal. By contrast, in hamsters implanted with either one or two morphine pellets neither of two doses of verapamil (20 and 30 mg/kg) was effective. These results are discussed in terms of species' differences in sensitivity to calcium channel blockers.

Animals↗
PubMed1992

Predicting body composition from anthropometry and bioimpedance in marginally undernourished adolescents and young adults.

Body-composition prediction equations were developed using data from a sample of 201 female and male Guatemalan ladinos (ie, people of Amerindian-European descent) aged 11-25 y. Fat-free mass (FFM) values were estimated from body density by using the two-component model and age- and sex-specific values for the density of FFM. The root-mean-square error (RMSE) of the best model predicting FFM from a set of simple anthropometric variables was 1.59 kg for females and 1.90 kg for males. The addition of more extensive anthropometry to the set of candidate predictors reduced the RMSE to 1.42 kg for females and 1.88 kg for males. The subsequent addition of a bioelectrical impedance measure (Ht2/R) further reduced the RMSE to 1.32 kg for females and 1.82 kg for males. These results suggest that for a marginally undernourished population with relatively little body fat, simple anthropometrics are as useful as more sophisticated measures for the prediction of body composition.

Adolescent↗
PubMed1992

Pear blister canker viroid is a member of the apple scar skin subgroup (apscaviroids) and also has sequence homology with viroids from other subgroups.

The sequence of pear blister canker viroid (PBCVd), the putative causal agent of pear blister canker (PBC) disease, has been determined. PBCVd consists of a single-stranded circular RNA of 315 nucleotide residues which assumes a branched conformation when it is folded in the model of lowest free energy. PBCVd has highest sequence similarity with grapevine 1B viroid (52.4%), but also contains sequences related to regions present in viroids that belong to different subgroups, suggesting that PBCVd could have developed from RNA recombination between viroids replicating in a common host plant. PBCVd contains almost the entire central sequence which is conserved in the members of the apple scar skin subgroup (apscaviroids) as well as a conserved sequence located in the left-terminal region of apscaviroids and pospiviroids (whose type member is potato spindle tuber viroid). A consensus phylogenetic tree has been obtained in which PBCVd and other viroids previously classified as apscaviroids appear closely related, allowing consideration of PBCVd as a new member of this subgroup.

Base Sequence↗
PubMed1992

Body mass index, body composition and the chronic energy deficiency classification of rural adult populations in Guatemala.

The present study tested the hypotheses that: (a) individual body composition estimates obtained with the Durnin-Womersley (D-W) equations have low validity in certain populations in developing countries; (b) there exists a poor relationship between the body mass index (BMI) and body composition estimates (fat mass (BFM) and fat-free mass (FFM)), and (c) BMI cut-off estimates (fat mass (BFM) and fat-free mass (FFM)), and (c) BMI cut-off points provide an invalid classification of chronic energy deficiency (CED) in adults. The study involved four samples of rural men and women in Guatemala, who had mean BMI of approximately 21 kg/m2. Body composition estimates were obtained by densitometry in three of the samples. Mean body fat (%) and mean FFM (kg) were: men: 11.6 (+/- 4.7) and 47.7 (+/- 4.9); and women: 21.6 (+/- 5.3) and 35.8 (+/- 3.5), respectively. The D-W equations based on various combinations of skinfold measurements consistently overestimated body fat content with low precision and validity. The BMI was more related to BFM and FFM than to fat proportion, but explained little of the variation in both body components, particularly at low BMI levels. A small number of men and women had BMI values below 18.5 kg/m2, and only one woman fell below 16 kg/m2. The power coefficients of height in the weight/height ratio which provided the strongest correlations with BFM and FFM were: BFM: women: 1.0; men: 1.5; FFM: 0.5 for both women and men. We conclude that the Quetelet index should not be recommended as a universally valid indicator to classify CED in adult groups similar to the study population.

Adipose Tissue↗
PubMed1991

Phylogeny of viroids, viroidlike satellite RNAs, and the viroidlike domain of hepatitis delta virus RNA.

We report a phylogenetic study of viroids, some plant satellite RNAs, and the viroidlike domain of human hepatitis delta virus RNA. Our results support a monophyletic origin of these RNAs and are consistent with the hypothesis that they may be "living fossils" of a precellular RNA world. Moreover, the viroidlike domain of human hepatitis delta virus RNA appears closely related to the viroidlike satellite RNAs of plants, with which it shares some structural and functional properties. On the basis of our phylogenetic analysis, we propose a taxonomic classification of these RNAs.

Hepatitis Delta Virus↗
PubMed1991

Identification of a new viroid as the putative causal agent of pear blister canker disease.

A small RNA species with the structural and functional properties characteristic of viroids has been isolated from three different pear sources each of which induced symptoms of the pear blister canker (PBC) disease when indexed in the pear indicator A 20. A close association between this RNA and PBC disease was established, since two of the three studied sources were known to be affected by this malady only, and the viroid was not detected in healthy pear tissue. Moreover, the PBC-associated viroid (PBCVd) replicated when purified preparations were inoculated into cucumber and pear plants. PBCVd behaved in denaturing polyacrylamide gels as a circular RNA with a molecular size of approximately 315 nucleotide residues. Analysis by dot blot hybridization indicated that PBCVd shares similarities in sequence with peach latent mosaic viroid and hop stunt viroid, and to a lesser extent with apple scar skin viroid.

Electrophoresis, Polyacrylamide Gel↗
PubMed1991

Oxygen uptake dynamics during high-intensity exercise in children and adults.

We hypothesized that the O2 uptake (Vo2) response to high-intensity exercise would be different in children than in adults. To test this hypothesis, 22 children (6-12 yr old) and 7 adults (27-40 yr old) performed 6 min of constant-work-rate cycle-ergometer exercise. Sixteen children performed a single test above their anaerobic threshold (AT). In a separate protocol, six children and all adults exercised at low and high intensity. Low-intensity exercise corresponded to the work rate at 80% of each subject's AT. High-intensity exercise (above the AT) was determined first by calculating the difference in work rate between the AT and the maximal Vo2 (delta). Twenty-five, 50, and 75% of this difference were added to the work rate at the subject's AT, and these work rates were referred to as 25% delta, 50% delta, and 75% delta. For exercise at 50% delta and 75% delta, Vo2 increased throughout exercise (O2 drift, linear regression slope of Vo2 as a function of time from 3 to 6 min) in all the adults, and the magnitude of the drift was correlated with increasing work rates in the above-AT range (r = 0.91, P less than 0.0001). In contrast, no O2 drift was observed in over half of the children during above-AT exercise. The O2 drifts were much higher in adults (1.76 +/- 0.63 ml O2.kg-1.min-2 at 75% delta) than in children (0.20 +/- 0.42, P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1991

[Evaluation of two methods to determine the polyphenol content in raw and cooked beans and its effect on protein digestibility].

The polyphenolic compounds present in raw and cooked, and dried, with and without the cooking broth of common white, black and red beans (Phaseolus vulgaris) were measured by the Folin-Denis method for total polyphenols, and by the protein precipitation method of Hagerman-Butler, which measures their biological activity. The polyphenol content was measured during 20 consecutive days on the same sample, using three different extracts of volume from each sample. Statistical analysis of the results by the Folin-Denis method indicated that variability among the three aliquots was different for each bean color. A non-parametric analysis, however, indicated that the average in the three levels of concentration for beans of all colors, was the same. A similar analysis of the results by the Hagerman-Butler method demonstrated that variability and average values for the three aliquots were equal for black and red beans but not for white beans. The coefficient of variation was lower for the higher aliquot of the extract. A significant correlation (r = 0.72, P less than 0.05, n = 60) was found between the two methods for all beans using the larger aliquot of the extract. The correlation was highly significant (r = 0.84, P less than 0.05) when white bean values were eliminated. The polyphenolic content varied with seed color and the thermic process reduced their content, as measured by the two methods. The losses in polyphenolics as measured by the Folin-Denis in the cooked beans dried with broth, varied from 31.4 to 36.3%, and from 39.8 to 51.1% for the cooked bean flour dried without broth. The losses by the Hagerman-Butler method were from 25.0 to 93.5% in the cooked bean flours dried with cooking broth, and from 33.3 to 95.7% when dried without the broth. The higher losses were recorded for red beans. In vivo digestibility for cooked bean flours, dried and without broth, were 73.2, 69.6 and 64.5%, and 71.9, 71.9 and 68.8% for white, red and black beans, respectively. A negative correlation (r = -0.39) and significant (p less than 0.05) was found between polyphenolic content in the diet and in vivo protein digestibility.

Analysis of Variance↗
PubMed1991

Plasma catecholamine levels and lipid mobilization induced by yohimbine in obese and non-obese women.

Oral yohimbine administration (0.2 mg/kg) induced lipid mobilization (increase in plasma non-esterified fatty acids, NEFA) in fasting non-obese women (body mass index BMI = 20.2 +/- 0.5, age 35.5 +/- 2.7 years) without significant action on plasma glucose, insulin levels, heart rate or blood pressure during the time-course of the experiment (240 min). Plasma norpinephrine (but not epinephrine) concentrations were increased (100 percent) after oral yohimbine administration. Oral administration of propranolol (40 mg, 60 min before yohimbine) reduced the lipid-mobilizing action of yohimbine (70 percent) during the 60 min following its administration and then totally suppressed its effect until the end of the experimental period (180 min). In fasting obese women (BMI = 36.4 +/- 2.1, age 37 +/- 3.6 years), yohimbine provoked an increase in plasma NEFA levels which was not markedly different from that observed in non-obese subjects. It had no significant effect on plasma glucose, insulin levels, heart rate or blood pressure. Plasma norepinephrine increased in the same proportions. The lipid-mobilizing effect of yohimbine in women is mainly attributable to the increase in synaptic norepinephrine with a resultant increment in lipolysis by beta-adrenergic agonism. In the standard fasting conditions (12 hours) the blockade of the antilipolytic fat cell alpha 2-adrenoceptors seems to be a minor component of the lipomobilizing effect of yohimbine. Morever, when compared with non-obese women, the lipomobilizing effect of yohimbine is not enhanced in obese women.

Administration, Oral↗
PubMed1989

Calcium channels that are required for secretion from intact nerve terminals of vertebrates are sensitive to omega-conotoxin and relatively insensitive to dihydropyridines. Optical studies with and without voltage-sensitive dyes.

Extrinsic absorption changes exhibited by potentiometric dyes have established the ionic basis of the action potential in synchronously activated populations of nerve terminals in the intact neurohypophyses of amphibia and mammals (Salzberg et al., 1983; Obaid et al., 1983, 1985b). Also, large and rapid changes in light scattering, measured as transparency, have been shown to follow membrane depolarization and to be intimately associated with the release of neuropeptides from the nerve terminals of the mouse neurohypophysis (Salzberg et al., 1985; Gainer et al., 1986). We report some experiments that help to define the pharmacological profile of the calcium channels present in intact neurosecretory terminals of vertebrates. For these, we used the peptide toxin omega-conotoxin GVIA (1-5 microM) and the dihydropyridine compounds Bay-K 8644 and nifedipine (2-5 microM), together with the after-hyperpolarization of the nerve terminal action potential. This undershoot depends upon the activation of a calcium-mediated potassium channel, as suggested by its sensitivity to [Ca++]o and charybdotoxin. omega-conotoxin GVIA substantially reduced the after-hyperpolarization in neurosecretory terminals of Xenopus, while neither of the dihydropyridine compounds had any effect under conditions that mimic natural stimulation. The effects of these calcium channel modifiers on the action potential recorded optically from the terminals of the Xenopus neurohypophysis were faithfully reflected in the behavior of the light-scattering changes observed in the neurohypophysis of the CD-1 mouse. omega-conotoxin GVIA (5 microM) reduced the size of the intrinsic optical signal associated with secretion by 50%, while the dihydropyridines had little effect. These observations suggest that the type of calcium channel that dominates the secretory behavior of intact vertebrate nerve terminals is at least partially blocked by omega-conotoxin GVIA and is insensitive, under normal conditions, to dihydropyridines.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed1989

Interactions between citrus exocortis and potato spindle tuber viroids in plants of Gynura aurantiaca and Lycopersicon esculentum.

Plants of Gynura aurantiaca and tomato (Lycopersicon esculentum) were inoculated with severe strains of citrus exocortis viroid (CEV) and potato spindle tuber viroid (PSTV). The progeny of both viroids was analyzed by two systems of PAGE which allowed discrimination between CEV and PSTV on the basis of their different sizes. When inoculated separately to G. aurantiaca CEV and PSTV induced severe and very mild symptoms, respectively, with CEV accumulating to a higher level than PSTV, whereas when tomato was the host, each viroid induced severe symptoms (although the reaction caused by PSTV was more intense) with PSTV reaching a higher steady-state concentration than CEV. The simultaneous inoculation of both viroids induced in G. aurantiaca the characteristic symptomatology of CEV, which was the only viroid that could be detected, whereas the response observed in tomato was that typical of PSTV, and only this viroid could be recovered. On the other hand, when G. aurantiaca plants were first inoculated with PSTV and later challenged with CEV, part of the plants displayed the characteristic severe reaction of CEV and both viroids were present in the corresponding extracts, but the other part showed the mild symptomatology induced by PSTV, and only this viroid could be isolated. However, when the protected plants were topped the new shoots exhibited severe symptoms and only CEV could be detected. These results suggest that CEV and PSTV compete for a limiting host factor needed for their replication, transport or accumulation, with CEV and PSTV having the highest affinity for the G. aurantiaca and tomato factors, respectively.

Electrophoresis, Polyacrylamide Gel↗
PubMed1989

Clinical consequences of a human non-fluorescent Y chromosome (Ynf).

A new case of ambiguous genitalia and immature tissue in the left gonad is presented. Cytogenetic findings with various techniques demonstrated that the distal two-thirds of the long arm of the Y chromosome is deleted. Q-banding showed a non-fluorescent Y; three positive bands were however noted when the DA/DAPI technique was applied. After a review of the literature, it was concluded that the non-fluorescent Y chromosome (Ynf) when inherited from generation to generation is a heteromorphism in normal males. However, in our case, where the proband's Y is lacking the fluorescent segment, a simple deletion does not appear to adequately explain the DA/DAPI positive bands. Possibly, a deletion followed by a structural rearrangement of the non-fluorescent segment had occurred de novo. The highly Y-specific DNA sequences present in the fluorescent segment are absent in these patients. The abnormal development in these cases is due to the presence of the 45,X cell line. The gene responsible for spermatogenesis has been localized to the non-fluorescent region in the long arm of the Y chromosome. Furthermore, it is concluded that two types of non-fluorescent Y chromosomes can be found in the population; one is a normal inherent heteromorphic variant, while the other appears to be an abnormality, especially in cases with azoospermia. Such distinctions should clearly be established prior to genetic counseling for patients with so called Ynf or del (Yd).

Chromosome Banding↗