Coding properties of reticulocyte lysine transfer RNA's in hemoglobin synthesis.

Two isoacceptor transfer RNA's for lysine were found in rabbit reticulocytes. The codon recognition properties of these isoacceptors were studied in hemoglobin synthesis in a cell-free systemn. The two isoacceptors transferred lysine into different sites in hemoglobin, but showed no preference for one chain over the other. Codon cross recognition was less than 4 percent.

Publisher Correction: Transcriptomic and neurochemical analysis of the stellate ganglia in mice highlights sex differences.

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Transcriptomic and neurochemical analysis of the stellate ganglia in mice highlights sex differences.

The stellate ganglia are the predominant source of sympathetic innervation to the heart. Remodeling of the nerves projecting to the heart has been observed in several cardiovascular diseases, however studies of adult stellate ganglia are limited. A profile of the baseline transcriptomic and neurochemical characteristics of the stellate ganglia in adult C57Bl6j mice, a common model for the study of cardiovascular diseases, may aid future investigations. We have generated a dataset of baseline measurements of mouse stellate ganglia using RNAseq, HPLC and mass spectrometry. Expression differences between male and female mice were identified. These differences included physiologically important genes for growth factors, receptors and ion channels. While the neurochemical profiles of male and female stellate ganglia were not different, minor differences in neurotransmitter content were identified in heart tissue.

Targeting protein tyrosine phosphatase σ after myocardial infarction restores cardiac sympathetic innervation and prevents arrhythmias.

Millions of people suffer a myocardial infarction (MI) every year, and those who survive have increased risk of arrhythmias and sudden cardiac death. Recent clinical studies have identified sympathetic denervation as a predictor of increased arrhythmia susceptibility. Chondroitin sulfate proteoglycans present in the cardiac scar after MI prevent sympathetic reinnervation by binding the neuronal protein tyrosine phosphatase receptor σ (PTPσ). Here we show that the absence of PTPσ, or pharmacologic modulation of PTPσ by the novel intracellular sigma peptide (ISP) beginning 3 days after injury, restores sympathetic innervation to the scar and markedly reduces arrhythmia susceptibility. Using optical mapping we observe increased dispersion of action potential duration, supersensitivity to ß-adrenergic receptor stimulation and Ca(2+) mishandling following MI. Sympathetic reinnervation prevents these changes and renders hearts remarkably resistant to induced arrhythmias.

Plasma 5-S-cysteinyldopa correlates with tumor size in melanoma-bearing mice.

A sensitive assay method employing high performance liquid chromatography with electrochemical detection (HPLC-ED) was used to compare 5-S-cysteinyldopa (CD) levels in plasma to tumor size in a murine melanoma model system. Plasma CD levels correlated with the sizes of primary tumor masses in mice, and the presence of metastatic tumors did not significantly affect the relationship. Elevated plasma CD levels appear to be directly related to tumor pigmentation: mice who had nonpigmented tumors induced by injections of amelanotic melanoma cells (NP) did not have elevated plasma CD levels. These studies indicate that plasma CD levels may serve as a marker for pigmented malignant melanomas and may be useful in following patients who are at high risk for these tumors.

Time course of tolerance to apomorphine in parkinsonism.

We sought to determine if tolerance developed to the antiparkinsonian effects of apomorphine and, if so, what temporal factors influenced its development. Seven patients with parkinsonism and motor fluctuations received short (6-hour) and long (22- to 31-hour) apomorphine infusions. Tolerance was evaluated by comparison of the responses to test doses of apomorphine that were administered before and after each infusion. The responses to the test doses that followed either infusion were reduced by 35% after the short infusion and by 68% after the long infusion, although plasma apomorphine levels were similar to or higher than levels achieved with preinfusion test doses. The duration of improvement in parkinsonism after discontinuation of the long infusion was briefer than that after the short infusion. We conclude that tolerance to apomorphine occurs in parkinsonism, and the loss of response is greater after longer periods of drug administration.

Studies of effects of kainic acid lesions in the dorsal lateral geniculate nucleus of rat.

Kainic acid has been described as a highly specific neurotoxin that when injected locally into the nervous system destroys neuronal perikarya but spares axons of passage and terminals in the vicinity of the injection site. The effects of injection of this agent into the rat dorsal lateral geniculate nucleus on geniculocortical and corticofugal pathways have been examined. Neuronal perikarya were absent from injected geniculates, and products of neuronal degeneration were observed in the external stratum of the optic radiation and in layer IV of striate cortex. Furthermore, no visually driven units could be found in physiological recordings from ipsilateral visual cortex, and no orthograde axonal transport of radioactivity to cortex was detectable by autoradiography. These observations are consistent with the complete destruction of the geniculocortical pathway. On the other hand orthograde axonal transport appeared normal in corticogeniculate and corticotectal neurons on the injected side. In addition to silver grains appearing over the geniculate and superior colliculus label was observed over corticofugal axons in the internal stratum of the optic radiation. It is noteworthy that axons of the corticofugal pathways are clearly segregated from geniculocortical axons over a part of their course through white matter. Corticogeniculate neurons on the injected side were able to transport D-aspartate but not nuclear yellow retrogradely to cell bodies in layer VI. These results are consistent with the notion that cortical neurons terminating in and passing through the kainate lesion site survive but suggests, however, that corticogeniculate neurons may be functionally altered by these lesions.

Developmental time course of acidic and basic fibroblast growth factors' expression in distinct cellular populations of the rat central nervous system.

Acidic and basic fibroblast growth factors (aFGF and bFGF, respectively) are expressed in high levels in adult central nervous system (CNS). We report the time course of developmental appearance and distribution of these factors and of two FGF receptors, FGFR-1 and FGFR-2, in the CNS of rats ranging in age from embryonic day 16 to adult. Immunohistochemical analysis showed that sensory neurons in the midbrain were the first cells to contain detectable aFGF immunoreactivity at embryonic day 18. The next cell group to contain aFGF were motor neurons, which were found to be aFGF-positive at the day of birth. A number of other subcortical neuronal populations were observed to contain aFGF immunoreactivity after postnatal day 7. Adult levels and distribution patterns of aFGF were reached in all CNS areas by postnatal day 28. Basic FGF immunoreactivity was observed at postnatal day 0 in neurons in the CA2 subfield of hippocampus. Astrocytes contained detectable bFGF immunoreactivity, starting at postnatal day 7. Adult levels and patterns of distribution of bFGF were reached in all CNS areas by postnatal day 28. These immunohistochemical observations were confirmed by using bioassay and Western blot techniques. FGFR-1 and FGFR-2 mRNA were expressed in significant levels in all CNS areas at all time points analyzed. The observation that aFGF and bFGF appear in specific and distinct cellular populations at relatively late developmental times suggests that these FGFs may be involved in specific mechanisms of CNS maturation, maintenance, and repair.

Characterization and distribution of basic fibroblast growth factor-containing cells in the rat hippocampus.

Basic fibroblast growth factor (bFGF), a member of the heparin-binding growth factor family, is present in relatively high levels in the brain where it may play an important role in the maintenance, repair, and reorganization of the tissue. Although bFGF is associated mainly with astrocytes throughout most of the central nervous system (CNS), a narrow but prominent band of pyramidal neurons, which coincides with the CA2 subregion of Ammon's horn in the hippocampus, stains intensely for bFGF. In order to gain an understanding of which cells express bFGF and whether or not BFGF is a good marker for CA2 neurons, we have used a mouse monoclonal antibody directed against recombinant human bFGF to characterize the distribution and localization of bFGF expression in the hippocampus. We find that about one-quarter of the neurons in CA2 are bFGF positive, and they appear smaller and have more irregular-shaped nuclei than their unstained counterparts. In addition, all glial fibrilary acidic protein (GFAP)-positive astrocytes in the hippocampus stain for bFGF, and the distribution of these astrocytes is heterogeneous in the hippocampus. Finally, in both astrocytes and CA2 pyramidal neurons, bFGF immunoreactivity is localized primarily in the nucleus and to a lesser extent in the cytoplasm and processes of stained cells.

Effect of long-term therapy on the pharmacodynamics of levodopa. Relation to on-off phenomenon.

To determine how the response to levodopa is altered by long-term therapy, we examined the dose response to 2-hour infusions of levodopa in three groups of parkinsonian patients: those who were previously untreated, those who exhibited stable responses, and those who exhibited fluctuating responses to levodopa therapy, using tapping speed as an index of bradykinesia. The baseline tapping speed was greater in the patients with stable responses than in the untreated patients, probably representing a "long-duration response" to levodopa therapy. A "short-duration response," indicated by an increase in tapping speed lasting hours, was observed in most patients in all groups. The onset of the short-duration effect was more rapid and the incremental increase in tapping speed was twice as large in the patients with fluctuating responses compared with the untreated patients and patients with stable responses. The duration of the short-duration effect was greatest in the untreated group but did not differ between the groups with stable and fluctuating responses. Dyskinesia was not observed in any of the de novo patients but was observed in three of 12 patients with stable responses and eight of nine patients with fluctuating responses to levodopa therapy. Dyskinesia appeared before or with the antiparkinsonian effects in patients with stable responses, giving no indication of a higher threshold for dyskinesia in these patients compared with those with fluctuating responses. The plasma half-life clearance, volume of distribution, and maximum plasma concentrations of levodopa did not differ among groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Levodopa pharmacokinetics and pharmacodynamics in fluctuating parkinsonian patients.

We investigated levodopa intravenous infusion rates, plasma concentrations, and clinical responses in 23 parkinsonian patients with a fluctuating response to the drug. There was a linear relationship between rate of levodopa infusion and plasma concentration when carbidopa was coadministered. Minimum-therapeutically effective plasma concentrations of levodopa ranged from 3 to 12 nmol/ml. The duration, but not the magnitude, of clinical response was linearly related to plasma concentrations. The clinical response lagged behind the rise and fall of plasma levodopa. There was no evidence of idiosyncratic pharmacokinetic or pharmacodynamic responses to levodopa to explain the fluctuating response.

Effect of brief levodopa holidays on the short-duration response to levodopa: evidence for tolerance to the antiparkinsonian effects.

To determine whether tolerance to the antiparkinsonian actions of levodopa develops during longterm levodopa therapy, we compared the response to 2-hour levodopa infusions before and after 2- to 4-day levodopa holidays using tapping and walking speeds and tremor/dyskinesia scores as measures of response in 17 parkinsonian patients with a fluctuating response to levodopa. As expected, motor function deteriorated during the levodopa holiday, but the maximum motor tapping and walking speeds and dyskinesia scores produced by the levodopa infusion before the holiday were the same as those produced by the infusion after the holiday. Because the baseline motor function was lower after the holiday, the increment in tapping and walking speeds (ie, the difference between the baseline and the maximum response) was larger with the postholiday infusion (p < 0.01). The postholiday infusion produced a longer response than did the preholiday infusion as measured by tapping score (p = 0.047), walking speed (p = 0.02), and tremor or dyskinesia scores (p = 0.02). The prolongation of the response was greater in patients receiving larger daily doses of levodopa (r = 0.55; p = 0.03). These changes in the duration of response suggest that progressive shortening of the response to levodopa during long-term therapy is partially caused by development of tolerance to levodopa and not just by loss of dopamine storage sites. Tolerance to levodopa should be considered in establishing oral dosing regimens and in developing new strategies for drug delivery.

Clinical and biochemical studies with controlled-release levodopa/carbidopa.

In five patients with parkinsonism, the optimal dosage of a controlled-release levodopa/carbidopa preparation (CR-3) was three times higher than the dosage of Sinemet and produced higher plasma levodopa concentrations, but did not reduce the fluctuations in plasma levodopa or clinical response. Plasma levodopa concentrations were higher and clinical responses better before the first dose of the day with CR-3. CR-3 treatment benefited two patients, reducing the severity of off periods and off dystonia. Two patients were worse on CR-3 despite higher plasma levodopa levels than those adequate for clinical response to Sinemet or levodopa infusions. CR-3 could benefit a few severely affected patients, but it is necessary to understand the factors that affect absorption of levodopa from sustained-release preparations, as well as the consequences of prolonged elevation of plasma levodopa levels.

Long-duration response to levodopa.

We measured the long-duration response to levodopa by changes in tapping rate in 16 patients with Parkinson's disease undergoing 3 to 5 days of levodopa withdrawal. "Off" tapping rates deteriorated 22% over the course of this holiday, the decline beginning 24 hours after levodopa withdrawal. Deterioration was more pronounced in the more affected hand in asymmetrically affected patients. A 2-hour infusion of levodopa after the levodopa holiday did not restore the long-duration response, although it produced a greater short-duration response than before the holiday. These observations indicate that the long-duration response is an important component of the therapeutic benefit of levodopa and is separable from the short-duration response.

The effect of exercise on levodopa absorption.

We studied the effect of exercise using cycle ergometry on levodopa absorption in ten patients with Parkinson's disease. Oral levodopa was administered during exercise and at rest on separate days. Exercise delayed levodopa absorption in five patients, increased it in three, and did not influence it in two. We conclude that exercise can either increase or decrease levodopa absorption.

Peripheral pharmacokinetics of levodopa in untreated, stable, and fluctuating parkinsonian patients.

We compared the pharmacokinetics of oral and IV infusions of levodopa in untreated, stable, and fluctuating patients pretreated with carbidopa. After oral dosing, mean peak plasma levodopa levels, time to peak level concentration after oral levodopa, and area under the curve were similar in all groups. Absorption was proportional to dose. Plasma elimination half-lives, clearance rates, and volumes of distribution after levodopa infusions were similar in all groups. 3-O-Methyldopa levels were similar in stable and fluctuating subjects. We conclude that the peripheral pharmacokinetics of levodopa do not differ between untreated, stable, and fluctuating patients, and that altered peripheral kinetics of the drug are unlikely to explain the development of the fluctuating state.

Does an inhibitory action of levodopa contribute to motor fluctuations?

The possibility that levodopa can both acutely improve as well as worsen motor function was investigated in eight fluctuating parkinsonian patients by administering 2-hour infusions of placebo and of levodopa at suprathreshold, threshold, and subthreshold rates when the subjects had been without medication overnight. Plasma levodopa and clinical status were monitored for an hour before, during, and for 3 hours after the infusions. Placebo infusions were associated with a mild deterioration in motor status during the period of monitoring. Subthreshold infusions were associated with greater motor deterioration than were placebo infusions in five of six subjects, but this deterioration did not occur in any specific temporal relation to the infusion. Threshold infusion rates in three subjects produced a transient improvement in motor function followed by deterioration to below the scores during the hour preceding the infusion and then returned to baseline. Suprathreshold rates in seven of the eight subjects were associated with motor improvement for 1/2 to 3 1/2 hours followed by worsening to below baseline for 30 to 90 minutes and then spontaneous improvement to baseline function. These observations suggest that levodopa causes an acute deterioration in motor function following drug-induced improvement. Thus, the nadir of motor function in levodopa-treated patients may not be simply loss of dopaminergic stimulation and return to the untreated state, but may represent an inhibitory effect of levodopa.

Amount and distribution of dietary protein affects clinical response to levodopa in Parkinson's disease.

Reducing dietary protein improves the effectiveness of levodopa (LD) but the most effective distribution of a low-protein diet (0.8 g/kg) is unclear. We compared a 1.6 g/kg protein diet, a 0.8 g/kg diet with protein evenly distributed between meals, and a 0.8 g/kg diet with protein restricted to the evening meal in 5 parkinsonian patients with motor fluctuations. We monitored clinical response, plasma LD, and plasma large amino acids (LNAAs) hourly throughout the day. Mean "on" times were 51% (1.6 g/kg diet), 67% (0.8 g/kg evenly distributed), and 77% (0.8 g/kg restricted). Hourly averages of plasma LD did not differ between the diets. The mean plasma LNAAs were 732 nmol/ml (1.6 g/kg diet), 640 (0.8 g/kg distributed), and 542 (0.8 g/kg restricted), and the diurnal pattern reflected the distribution of protein intake. In conclusion, the amount and distribution of dietary protein affect clinical response to LD. These effects are not related to LD absorption but are explained by the variation in plasma LNAAs.

Time interval between repeated injections conditions the duration of motor improvement to apomorphine in Parkinson's disease.

Behavioral hyposensitivity to repeated apomorphine administration occurs in fluctuating parkinsonian patients. To determine to what extent the interval between doses influences the response, we administered equal paired apomorphine injections to 10 fluctuating parkinsonian patients. Subjects received two apomorphine injections at 2-hour and at 4-hour intervals on different days after a 10- to 12-hour overnight period without levodopa. Following apomorphine doses at 2-hour intervals, the duration of response was reduced by 40% (61 versus 42 minutes, p less than 0.001) but was of equal duration when the doses were given at 4-hour intervals. These findings indicate that the interval between doses is a critical determinant of motor response. We postulate a time-dependent period of partial hyposensitivity to pulsatile DA stimulation.

The effect of L-dopa infusions with and without phenylalanine challenges in parkinsonian patients: plasma and ventricular CSF L-dopa levels and clinical responses.

We monitored the motor response and plasma and ventricular CSF (CSFv) concentrations of L-dopa during IV infusions of L-dopa in two patients with advanced Parkinson's disease. Concentrations of L-dopa in CSFv mirrored, but lagged behind, those in plasma. In the fasting state, the duration, but not the magnitude, of the motor response was greater with increasing plasma and CSFv levels of L-dopa. During IV infusions of L-dopa following oral administration of phenylalanine, a large neutral amino acid that shares a transport system into the brain with L-dopa, the duration of the motor response was markedly attenuated despite undiminished CSFv levels of L-dopa. These observations suggest that either L-dopa entry into CSFv and the brain are differentially affected by phenylalanine or that phenylalanine affects other steps in the motor response. These observations demonstrate that, except in the fasting state, L-dopa in CSFv is not a reliable predictor of motor response.