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Biomedical subjects

R J Nelson

Publications and source records attributed to R J Nelson.

At least 19 recordsLinked to original sources

The translation machinery and 70 kd heat shock protein cooperate in protein synthesis.

The function of the yeast SSB 70 kd heatshock proteins (hsp70s) was investigated by a variety of approaches. The SSB hsp70s (Ssb1/2p) are associated with translating ribosomes. This association is disrupted by puromycin, suggesting that Ssb1/2p may bind directly to the nascent polypeptide. Mutant ssb1 ssb2 strains grow slowly, contain a low number of translating ribosomes, and are hypersensitive to several inhibitors of protein synthesis. The slow growth phenotype of ssb1 ssb2 mutants is suppressed by increased copy number of a gene encoding a novel translation elongation factor 1 alpha (EF-1 alpha)-like protein. We suggest that cytosolic hsp70 aids in the passage of the nascent polypeptide chain through the ribosome in a manner analogous to the role played by organelle-localized hsp70 in the transport of proteins across membranes.

Amino Acid Sequence

Prediction of migration behavior of oligonucleotides in capillary gel electrophoresis.

The influence of the primary structure (base composition) on the electrophoretic migration properties of single-stranded oligodeoxyribonucleotides in capillary polyacrylamide gel electrophoresis was investigated using homo- and heterooligomers under denaturing and non-denaturing conditions. Homooligodeoxyribonucleotides of equal chain lengths but of different base composition showed significant differences in mobility. In addition, the migration properties of heterooligomers were found to be highly dependent on their base composition. A simple equation is presented for predicting relative migration times using denaturing and non-denaturing polyacrylamide capillary gel electrophoresis. Orange-G was used as an internal standard and as the basis of the relative migration time calculations. Examples are presented using homo- and heterooligomers in the 10-20-mer range to show the correlation of the primary structure and their predicted and observed migration rates.

Base Sequence

Striatal neuronal activity during the initiation and execution of hand movements made in response to visual and vibratory cues.

Recordings were obtained from 146 neurons in the neostriatum of rhesus monkeys while they performed wrist movements in response to visual and vibratory cues. Of these, 75 putamena nd 29 caudate neurons exhibited changes in firing rate that were temporally related to the onset of the wrist movements and that began prior to movement onset. This premovement activity (PMA) usually was directionally specific, in that the magnitude or direction of change in firing rates was different during flexion trials as compared to trials involving wrist extension. PMA onset usually preceded movement onset by more than 100 ms and in most instances preceded the average onset of task-related changes in electromyographic (EMG) activity in muscles of the wrist and forelimb. For most neurons, the changes in neuronal activity that began prior to movement were maintained during movement execution. However, approximately one-third of the neurons that exhibited PMA changed their firing rate in the opposite direction, relative to their PMA and to their baseline rate of activity, once the movement began. Several other neurons either exhibited PMA only or they altered their discharge rates during movement execution but did not exhibit PMA. These observations suggest that, despite the close temporal relationship between the onset of PMA and the onset of wrist movement, the neuronal mechanisms mediating the PMA may differ from those that occur during movement execution. The PMA onset of neostriatal neurons occurred earlier in visually cued than in vibratory cued trials. These differences were statistically significant only for flexion trials, however, in which movements were made against a load and in the same direction as the palmar vibratory stimulus. For trials involving wrist extension, PMA onsets for visually cued as compared with vibratory cued trials were not statistically different. These findings contrast with data obtained previously from somatosensory cortical neurons during performance of the same behavioral task. On average, PMA in the putamen began earlier, relative to movement onset, than it did in the somatosensory cortex. Moreover, in the somatosensory cortex, PMA onset occurred earlier in vibratory cued than in visually cued trials, irrespective of movement direction (Nelson 1988; Nelson and Douglas 1989). For putamen neurons, but not for caudate or cortical neurons, the onset of PMA also occurred significantly earlier during extension trials than flexion trials, irrespective of the modality of the "go-cue".(ABSTRACT TRUNCATED AT 400 WORDS)

Animals

Isolation and characterization of extragenic suppressors of mutations in the SSA hsp70 genes of Saccharomyces cerevisiae.

Saccharomyces cerevisiae strains that contain null alleles of two hsp70 genes, SSA1 and SSA2, are temperature sensitive for growth. In this study, extragenic suppressors of ssa1 ssa2 have been isolated. Suppression is due to mutations at nuclear loci designated EXA1, EXA2 and EXA3 for EXtragenic suppressor hsp70 subfamily A. Two of the four EXA1 alleles are dominant as is EXA3-1. The other two EXA1 alleles as well as the sole EXA2 allele are recessive. EXA1 mutations lead to accumulation of a previously uncharacterized form of hsp70. EXA2 and EXA3 mutations affect the regulation of the stress response. In exa2-1 ssa1 ssa2 strains the gene products of the remaining SSA hsp70 genes, SSA3 and SSA4 (Ssa3/4p), accumulate to higher levels. The EXA3-1 mutation results in increased accumulation of both Ssa3/4p and the hsp70s encoded by the SSB1 and SSB2 genes (Ssb1/2p), suggesting that the EXA3 gene product plays a central role in the yeast stress response. Consistent with this hypothesis, EXA3-1 is tightly linked to HSF1, the gene encoding the transcriptional regulatory protein known as "heat shock factor." All of the genes identified in this study seem to be involved in regulating the expression of SSA3 and SSA4 or the activity of their protein products.

Alleles

Photoperiod influences the critical caloric intake necessary to maintain reproduction among male deer mice (Peromyscus maniculatus).

The concept of critical day length is well established among rodents; reproductive function is maintained when day lengths are greater than some specific threshold. In addition to day length cues, seasonal breeding in deer mice can also be regulated by food availability. The caloric threshold necessary to support reproduction remains unspecified for seasonally breeding rodents. The present study examined the interaction between photoperiod and food availability on reproductive function in adult male deer mice (Peromyscus maniculatus). A critical caloric intake profile was constructed in long (16L:8D) and short (8L:16D) photoperiods; groups of deer mice in both photoperiods either received food ad libitum or 90, 80, or 70% of their individual ad libitum food intake for 10 wk. At autopsy, paired testes, epididymides, and seminal vesicles were removed and weighed. Body mass, total body fat, and total body water contents were also obtained. Short, as compared to long, day lengths inhibited the reproductive systems of male deer mice. However, food consumption interacted with photoperiod to affect reproductive function. Significant reductions in reproductive organ size as well as spermatogenic activity were observed among short-day mice after a 10% reduction in ad libitum food intake. Long-day animals required a 20% reduction in caloric intake to depress reproductive function. Body mass and total body water content were generally unaffected by either photoperiod or food consumption. Total body fat content was reduced in short- as compared to long-day mice. Individual reproductive responsiveness to short days increased as food availability decreased.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Experimental aspects of cerebrospinal hemodynamics: the relationship between blood flow velocity waveform and cerebral autoregulation.

The dynamic relationships among mean flow velocity, its pulsatile amplitude (FVa), cortical cerebral blood flow (CBF), and cerebral perfusion pressure (CPP) were studied in normal rabbits and rabbits with subarachnoid hemorrhage using 8-MHz pulsed transcranial Doppler ultrasound and hydrogen clearance under conditions of systemic hypotension and intracranial hypertension. A two-slope relationship was observed between FVa and CPP with a break point that correlated closely with the lower limit of CBF autoregulation in each animal. Below this CPP break point, FVa varied directly with CPP, and above the break point FVa varied inversely with CPP. In this experimental model, an inverse correlation between FVa and CPP indicates intact CBF autoregulation, whereas loss of that correlation implies exhaustion of autoregulatory reserve. Simultaneous recording and computation of FVa, CPP, and the correlation coefficient between FVa and CPP may be a means of monitoring CBF autoregulation in clinical practice.

Animals

Isolation of maxadilan, a potent vasodilatory peptide from the salivary glands of the sand fly Lutzomyia longipalpis.

Blood feeding by the sand fly Lutzomyia longipalpis is aided by the presence of a vasodilator in its salivary glands. This novel vasodilator has been isolated by reversed-phase high-performance liquid chromatography. Ten nanograms of the vasodilator are present in the extract of a pair of sand fly salivary glands. It has 500 times the vasodilatory activity of calcitonin gene-related peptide, previously the most potent vasodilator peptide known. This novel peptide is thus called maxadilan.

Amino Acids

Relationships between sensory responsiveness and premovement activity of quickly adapting neurons in areas 3b and 1 of monkey primary somatosensory cortex.

When monkeys make wrist movements in response to vibration of their hands, primary somatosensory (SI) cortical neurons that adapt quickly to the vibratory stimulus often exhibit two temporally separate types of activity. Initially, these neurons respond to the stimulus. They then cease discharging, only to resume firing prior to the movement. This activation, cessation and reactivation occurs even though the sensory stimulus remains on until after the movement is begun. The first change in activity is most likely related to sensory input. The second, which has been called premovement activity, may have a sensory component as well as one related to the upcoming movement. We wanted to test the hypothesis that the premovement activity exhibited when vibration is present represents both a reactivation of a neuron's vibratory response and the premovement activity that normally occurs when vibration is absent. We also wanted to determine if area 3b and 1 quickly adapting (QA) neurons show similar or different activity patterns during the initiation and execution of sensory triggered wrist movements. Four monkeys were trained to make wrist flexion and extension movements in response to vibratory stimuli delivered to the handle which the animals used to control the behavioral paradigm. Two fo the four monkeys also made similar wrist movements following visual cues. We found that the premovement activity of QA neurons located in area 1 (but not area 3b) is comprised of a sensory-related component as well as a movement-related component. The magnitude of these individual components differs in relationship to a neuron's receptive field type, the movement direction and the external force imposed on the stimulated forelimb. Premovement activity of area 3b and area 1 QA neurons occurs at the same time prior to movement, regardless of whether visual or vibratory cues are used to trigger wrist movements. This activity occurs at about the same time as others have observed elevations in the threshold for tactile perception, suggesting that premovement activity and changes in sensory responsiveness before movement may be related. These and previous findings are used to construct a model which may predict the firing patterns of SI QA neurons during behavioral tasks. These findings also suggest that areas 3b and 1 may have different roles in processing task-related somatosensory information.

Animals

Ultrasonic decalcification of the aortic annulus during aortic valve replacement.

Aortic valve replacement for calcifica aortic stenosis requires meticulous debridement of the aortic annulus to effect optimal valve seating. Since 1987, we have used ultrasonic energy to debride the aortic annulus during aortic valve replacement in 56 patients. In our experience, ultrasonic debridement of the annulus is superior to traditional methods of debridement, affords improved seating of the valve, and may allow placement of a larger valve. Our follow-up ranges from 2 to 32 months (mean follow-up, 13 +/- 9 months) with 0% incidence of paravalvular leak or valve failure. We advocate the use of ultrasonic debridement as an adjunctive tool in aortic valve replacement.

Adult

Hormonal control of gene expression: interactions between two trans-acting regulators in Drosophila.

The steroid hormone 20-hydroxyecdysone (20HE) and the Broad-Complex locus (BRC) are involved in regulating developmental changes in gene expression around the time of metamorphosis in Drosophila. We have investigated the regulatory interactions between 20HE, BRC, and a set of genes expressed in the fat body of third-instar Drosophila larvae. RNA levels for two hormone-inducible genes, Larval Serum Protein-2 and P1, accumulate to normal levels in BRC-mutant larvae. In contrast, RNA levels for the P6 gene were affected by mutations at BRC. On the basis of the results of experiments in which hormone concentrations were varied in BRC-mutant or wild-type larvae, we conclude that 20HE can both increase and decrease P6 RNA levels in the absence of BRC product(s). BRC appears to be a trans-acting modulator of the response of P6 to the hormone. We propose that BRC attenuates the repressive effect of the hormone, expanding the range of hormone concentrations that induce the gene, thus allowing P6 RNA to reach high levels during the third larval instar. The results are discussed in relation to other genes that are regulated by the same two trans-acting factors. A model is presented that refines the model of Ashburner et al. (1974, Cold Spring Harbor Symp. Quant. Biol. 38: 655-662) for the hormonal regulation of gene activity.

Age Factors

Effects of photoperiod and 6-methoxy-2-benzoxazolinone on male-induced estrus in prairie voles.

Prairie voles (Microtus ochrogaster) are characterized as facultative breeders with higher rates of reproductive activities observed during spring and summer than autumn and winter. The environmental factors regulating seasonal breeding in this species remain unspecified. Short day lengths inhibit reproductive organ development in male prairie voles in the laboratory, but these males remain fertile and capable of siring offspring; female prairie voles have been reported to be reproductively unresponsive to day length in the laboratory. The organization of estrus in this species is unusual in that females never display the cyclic changes associated with estrus; rather, female prairie voles require chemosensory stimuli associated with fertile males in order to be induced into estrus. The plant compound, 6-methoxy-2-benzoxazolinone (6-MBOA), is present in vegetatively growing grasses and sedges and acts to trigger reproduction in other rodent species exposed to short days. It was suspected that 6-MBOA present in the laboratory diet may have overridden the effects of photoperiod on female prairie voles in previous laboratory studies. In the present study, the effects of 6-MBOA and photoperiod on estrus induction were examined. Beginning at Week 0, female prairie voles were housed in long (LD 16:8) or short (LD 8:16) photoperiods for 9 weeks, then implanted subcutaneously either with an empty Silastic capsule or one packed with 6-MBOA. A special diet, devoid of 6-MBOA, was available ad lib from Week 5 to the end of the study.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Maternal diet influences reproductive development in male prairie vole offspring.

The plant compound, 6-methoxy-2-benzoxazolinone (6-MBOA), is found in the meristematic tissue of young, vegetatively growing grasses, and its ingestion stimulates the reproductive system of rodents in both short and long photoperiods. The present study demonstrates the existence of a novel mechanism whereby the presence of 6-MBOA in the diet of pregnant prairie voles (Microtus ochrogaster) enhances gonadal development in their male offspring. Female voles were fed specially milled food in which 6-MBOA was present (50 micrograms of 6-MBOA/g food) or absent (0 microgram of 6-MBOA/g food) prior to the birth of their young; all of these dams received a diet that lacked 6-MBOA/g after the pups were born. Other females were fed food without 6-MBOA throughout pregnancy, but received one of the two diets during lactation. The pups from all experimental groups were weaned at 3 weeks of age and fed thereafter one of the two diets for 3 additional weeks. Prenatal exposure to 6-MBOA enhanced male reproductive development. Maternal ingestion of 6-MBOA during lactation or postweaning intake of 6-MBOA by the young did not influence reproductive development of males. This mechanism may be involved in the rapid rate of sexual maturation observed prior to peak population densities of microtine rodents.

Animals

Sensory response enhancement and suppression of monkey primary somatosensory cortical neurons.

Vibratory stimulus-related responses were recorded from monkey primary somatosensory cortical (SI) neurons while animals performed two tasks. In the movement task, vibratory stimuli served as the go-cue for wrist flexion or extension. In the no-movement task, movements normally made in response to vibratory stimuli were extinguished. Area 3a, 3b, and 1 neurons with deep receptive fields (RFs) exhibited greater stimulus-related activity during the movement task than during the no-movement task. Area 3b neurons with cutaneous RFs were similarly enhanced during the movement task, whereas area 1 neurons with cutaneous RFs were less responsive to vibratory stimuli during the movement task. These results suggest that motor-set and/or selective attention may modulate the responsiveness of SI neurons to peripheral stimuli and that changes in sensory responsiveness in SI neurons differ as a function of their cortical location and RF type.

Animals

Nerve growth factor, nerve grafts and amniotic membrane grafts restore erectile function in rats.

In an effort to reduce complications arising from radical pelvic surgery, an improved technique for restoration of autonomic innervation has been developed. The ability of nerve growth factor (NGF) alone or in combination with interposition nerve grafts, as well as the use of fetal amniotic membrane as an alternative growth matrix to enhance regeneration of ablated cavernous nerves were investigated in rats. Rats with ablated cavernous nerve displayed little or no penile erection, either in response to direct electrical stimulation or to an estrous female rat. A step wise improvement in electrically induced erections was observed by NGF alone, nerve graft alone, and the combination of NGF and nerve graft. Restoration of sexual behavior followed the same pattern obtained with electrical stimulation. Furthermore, the use of neonatal amniotic membrane as an alternative nerve growth matrix enhanced both electrically stimulated erection and mating behavior. These results suggest that the use of NGF and appropriate grafting materials can facilitate autonomic nerve regrowth and potentially reduce the morbidity of surgically induced nerve injuries.

Amnion

Cavernous nerve grafts restore erectile function in denervated rats.

Although potency can be preserved in most men following radical prostatectomy, in some patients one or both cavernous nerves must be sacrificed in order to excise all tumor. For these patients we have considered nerve reconstruction at the time of surgery using an interposition nerve graft. This possibility has been tested in a rat model. Animals were divided into three groups. In the sham control group, a pelvic exploration was conducted without division of the cavernous nerves. In the nerve ablation group, a five mm. segment of cavernous nerve was excised. In the graft group, five mm. of cavernous nerve was excised bilaterally and replaced with an interposition graft of genito-femoral nerve. At one month intervals postoperatively animals from each group underwent mating tests to determine potency; the ratio of vaginal intromission to unsuccessful mounts (I/M ratio) was determined. Following the mating tests the animals were re-explored and attempts were made to stimulate erections electrically via the pelvic nerve. Results at two months show the I/M ratio of the rats with grafts (0.05) and the nerve ablations (0.14) are low compared with the sham operated animals (1.0). By month four the graft group (0.89) has approached the sham group (0.91) while the ablation group (0.18) remains unchanged (p less than .05). Electrical stimulation at month two produced no erections in the nerve ablated or the grafted rats and 100% erections in the sham-operated animals. At month four, 50% of the rats with grafts, 10% of the nerve ablated animals, and 100% of the intact animals produced erections upon electrical stimulation (p less than .05). These results suggest that cavernous nerve grafting in rats can be successful in restoring potency after surgical injury. Application of these techniques to man may be indicated.

Animals

The effects of hyponatraemia and subarachnoid haemorrhage on the cerebral vasomotor responses of the rabbit.

Impairment of cerebral autoregulation and development of hyponatraemia are both implicated in the pathogenesis of delayed cerebral ischaemia and infarction following subarachnoid haemorrhage (SAH) but the pathophysiology and interactions involved are not fully understood. We have studied the effects of hyponatraemia and SAH on the cerebral vasomotor responses of the rabbit. Cerebrovascular reactivity to hypercapnia and cerebral autoregulation to trimetaphan-induced hypotension were determined in normal and hyponatraemic rabbits before and 6 days after experimental SAH produced by two intracisternal injections of autologous blood. Hyponatraemia (mean plasma sodium of 119 mM) was induced gradually over 48 h by administration of Desmopressin and intraperitoneal 5% dextrose. Sham animals received normal saline. The cerebrovascular reactivity (% change +/- SD in cortical CBF/mm Hg PaCO2, measured by hydrogen clearance) of hyponatraemic (4.8 +/- 3.0%) and SAH (1.3 +/- 2.0%) animals was significantly less (p less than 0.05) than control (11.6 +/- 4.0%) and sham (8 +/- 2.0%) animals, whereas the reactivity of hyponatraemic-SAH animals was preserved (9.8 +/- 6.0%). Hyponatraemia and SAH alone each significantly impaired CBF autoregulation but their combined effects were not additive. Systemic hyponatraemia impairs normal cerebral vasomotor responses but does not augment the effects of experimental SAH in the rabbit.

Animals

The resolution of transcranial Doppler scanning: a method for in vitro evaluation.

The circle of Willis may be examined using low frequency pulsed Doppler ultrasound transmitted through the thin squamous temporal bone or 'acoustic windows' of the skull. The thickness and shape of these windows affect the characteristics of the ultrasound beam and partly determine the spatial resolution of transcranial Doppler scanning. We have studied the effects of temporal bone on the ultrasound beam and sample volume characteristics of the EME Transcan 2 MHz transducer using a stepper motor, computer-controlled ultrasound plotting system. The lateral and axial spatial dimensions of the sample volume, and a direct estimate of the lateral resolution of the system, were determined at various depths of interest using Doppler targets. The results in this initial study of a small number of specimens indicates a decrease in sample width, and hence an increase in lateral resolution, presumably due to the lensing effect of the skull cusping. The estimated lateral resolution of the system is of the order of 4 mm.

Aged

Association of hypovolemia after subarachnoid hemorrhage with computed tomographic scan evidence of raised intracranial pressure.

Hypovolemic patients are more likely to suffer delayed cerebral ischemia and infarction after a subarachnoid hemorrhage (SAH). Prompt recognition and correction of hypovolemia may improve the outcome. We have identified computed tomographic (CT) scan findings that increase the probability of a patient presenting with hypovolemia soon after an SAH. The plasma volume (PV) of 25 patients admitted within 96 hours of an SAH was measured using radioiodinated serum albumin. The normal PVs were measured in an outpatient setting 6 months later or predicted from their total body water. Nine patients (36%) were found to be hypovolemic, defined as a fall in PV exceeding 10% of the normal PV (mean fall, 18 +/- 2%). Sixteen patients were normovolemic or hypervolemic (mean PV, +9 +/- 2%). The basal cisterns were compressed or obliterated on the CT scans of all hypovolemic patients compared with 12.5% of normovolemic patients (chi-square, 14.52; P less than 0.01). The probabilities of a patient being hypovolemic if the CT scan indicated raised intracranial pressure were high: hydrocephalus, P = 0.80; compression of the basal cisterns, P = 0.82; and compression of the basal cisterns associated with intracerebral hematoma or midline shift, P = 1.00. Patients with an SAH and radiological evidence of raised intracranial pressure should be considered at particular risk for systemic hypovolemia.

Adult