Duration, response, and location: the influence of upcoming 32% sucrose on rats' licking or lever pressing for 1% liquid sucrose

The present study investigated whether rats' rates of licking or pressing a lever for 1% liquid sucrose delivered by a continuous reinforcement schedule would decrease (contrast) or increase (induction) when the upcoming period would allow access to 32% sucrose and whether such changes would be influenced by how long each substance was available. In Experiment 1, different groups of rats licked a spout or pressed a lever for 1% sucrose in the first half of the session and, in different conditions, for 1% or 32% sucrose in the second half. Across conditions, halves of the session were 3, 6, 12, or 24 min long. Upcoming 32% sucrose significantly decreased rates of licking at each duration whereas it increased rates of lever pressing except when access duration was 3 min. Experiment 2 replicated Experiment 1 with the exception that rats that licked did so from the same spout in both halves of the session and rats that pressed a lever collected the sucrose reinforcers in the different halves at different locations. In these procedures, upcoming 32% sucrose significantly increased rates of licking. Significant, but small, increases in rates of lever pressing were still observed. The present results suggest that continuous reinforcement or duration of access to sucrose are not primary determinants of whether contrast or induction is observed. Rather, they suggest that the type of behavior (licking versus pressing a lever) and the location at which the substances are collected and consumed play a large role in which effect occurs.     

Models for reserpine effects on normal and brain-operated animals on a continuous reinforcement bar pressing schedule

Mathematical models are presented for the relationship between changes in the bar pressing rate of albino rats on a continuous reinforcement schedule and alterations of the brain amines norepinephrine and serotonin induced by electrolytic lesions in the hypothalamus and by the drug, reserpine. The discussion utilizes central concepts from some of the more prominent hypotheses on the relation of the amines to behavior and for the potentiation effects of the drug by the lesion (A. Carlsson, 1964. “Functional Significance of Drug-induced Changes in Brain Monoamine Levels.” InBiogenic Amines, eds. H. E. Himwich and W. A. Himwich, New York: Elsevier, 9–27; B. B. Brodie and E. Costa, 1962. “Some Current Views on Brain Monoamines.”Psychopharm. Service Centre Bull.,2, 1–25.) Laboratory data (F. Grabarits and J. Harvey, 1966. “The Effects of Reserpine on Behavior and on Brain Concentrations of Serotonin and Norepinephrine in Control Rats and Rats with Hypothalamic Lesions.”J. Pharm.,153, 403–411) are interpreted, and suggestions for further experimental and theoretical work are made.     

Three tests of 'anticipatory responding' as an account for induction produced by upcoming food-pellet reinforcement

Previous research has demonstrated that rats' rates of lever pressing for low-concentration liquid-sucrose reinforcers are increased when food-pellet, rather than sucrose, reinforcement will be upcoming in the same session (i.e. induction). The present experiments were designed to determine whether this induction was the product of 'anticipatory responses' for the upcoming food pellets being added to the responses being made for the currently available sucrose reinforcement. Experiment 1 tested this idea by summing sucrose-reinforced responding and 'anticipatory responding' from different conditions and comparing the sum to responding from a third condition in which subjects responded for sucrose when food-pellet reinforcement was upcoming. The comparison yielded similar response rates. Experiment 2 employed a blackout, of different durations in different conditions, to delay the upcoming food-pellet reinforcement. Consistent with the anticipatory-responding account, the delay decreased the size of the induction. However, results from the blackouts were not entirely consistent with the anticipatory-responding explanation. Experiment 3 provided, in some conditions, sucrose and food-pellet reinforcement in the first and second halves of the session, respectively, for responding on separate levers. These conditions separated 'anticipatory responses' for the food pellets from responses for the sucrose reinforcers. However, induction in responding for sucrose was still present. Together, these experiments demonstrate that, although anticipatory responses may contribute to induction in some instances, they are not solely responsible for the effect.     

Bombesin effects on responding maintained by a fixed interval schedule of food reinforcement

The present experiment examined rats' responding maintained by a fixed interval two-min schedule of food reinforcement following IP injections of bombesin (4, 6, 16, 32 μg/kg). The results showed that bombesin's effects were rate dependent where the responses per minute emitted during the early portion of the fixed interval were reduced, but responding during the latter portion was unaffected. Bombesin did not reduce overall session responses per minute, pause after reinforcement, or amount of water consumed in the test chamber. The results are in accord with prior research examining the effects of bombesin and cholecystokinin on operant behavior. Together, the data challenge the notion that bombesin affects food-motivated behavior generally; rather, the results indicate that bombesin's effect may interact with the demands required of the animal for reinforcement.     

Highly efficient reinforcement of poly-L-lactide materials by polymer blending of a thermotropic liquid crystalline polymer

A novel polymer blend system consisting of poly(l-lactide) (PLLA) and a thermotropic liquid crystalline polymer (LCP: an aromatic polyester comprising poly(4-hydroxybenzoate) sequences) was investigated in the presence and absence of a polycabodiimide (PCD). Scanning electron micrographs of the injection-molded polymer blends revealed the formation of fibrous structure of LCP in the PLLA matrix, supporting the efficient toughening. In particular, the LCP fibrils became semimicrometer in diameter in the presence of PCD with which both PLLA and LCP had reacted during the melt blending to form their block and graft copolymers working as compatibilizer. The blend specimens containing LCP in 20-30 wt % were found to hold high dynamic storage-moduli (E') at high temperature. In addition, the E' value of the specimens containing 30 wt % of LCP reached 10.7 GPa at room temperature, being significantly higher than that of PLLA.     

Identifying sucrose as a signal for nitrate uptake by wheat roots

Some sugars supplied directly to roots can stimulate nitrate uptake by wheat (Triticum aestivum L.) roots. To identify a signaling molecule, we compared the response of net nitrate influx to sugar supply. A method with a high time resolution (minutes) enabled to make a comparison. A signaling sugar should cause a faster and greater response than other compounds. Among nine sugars and mannitol tested, sucrose alone caused an immediate active stimulation of net nitrate influx. Glucose, fructose, and raffinose caused weak responses with a lag. Other carbohydrates had no effect. Sucrose behaves as a specific signal for nitrate uptake, which has long been supposed but not supported experimentally.     

Regioselective O-glucosylation by sucrose phosphorylase: a promising route for functional diversification of a range of 1,2-propanediols

1,2-Propanediol and 3-aryloxy/alkyloxy derivatives thereof are bulk commodities produced directly from glycerol. Glycosylation is a promising route for their functional diversification into useful fine chemicals. Regioselective glucosylation of the secondary hydroxyl in different 1,2-propanediols was achieved by a sucrose phosphorylase-catalyzed transfer reaction where sucrose is the substrate and 2-O-α-d-glucopyranosyl products are exclusively obtained. Systematic investigation for optimization of the biocatalytic synthesis included prevention of sucrose hydrolysis, which occurs in the process as a side reaction of the phosphorylase. In addition to ‘nonproductive’ depletion of donor substrate, the hydrolysis also resulted in formation of maltose and kojibiose (up to 45%) due to secondary enzymatic glucosylation of the glucose thus produced. Using 3-ethoxy-1,2-propanediol as the acceptor substrate (1.0 M), the desired transfer product was obtained in about 65% yield when employing a moderate (1.5-fold) excess of sucrose donor. Loss of the glucosyl substrate to ‘glucobiose’ by-products was minimal (<7.5%) under these conditions. The reactivity of other acceptors decreased in the order, 3-methoxy-1,2-propanediol > 1,2-propanediol > 3-allyloxy-1,2-propanediol > 3-(o-methoxyphenoxy)-1,2-propanediol > 3-tert-butoxy-1,2-propanediol. Glucosylated 1,2-propanediols were not detectably hydrolyzed by sucrose phosphorylase so that their synthesis by transglucosylation occurred simply under quasi-equilibrium reaction conditions.     

A randomised controlled trial of a self-management education program for osteoarthritis of the knee delivered by health care professionals

Introduction

Our aim in the present study was to determine whether a disease-specific self-management program for primary care patients with osteoarthritis (OA) of the knee (the Osteoarthritis of the Knee Self-Management Program (OAK)) implemented by health care professionals would achieve and maintain clinically meaningful improvements in health-related outcomes compared with a control group.

Methods

Medical practitioners referred 146 primary care patients with OA of the knee. Volunteers with coexistent inflammatory joint disease or serious comorbidities were excluded. Randomisation was to either a control group or the OAK group. The OAK group completed a 6-week self-management program. The control group had a 6-month waiting period before entering the OAK program. Assessments were taken at baseline, 8 weeks and 6 months. The primary outcomes were the results measured using the Western Ontario and McMaster Universities Arthritis Index (WOMAC) Pain and Function subscales on the Short Form 36 version 1 questionnaire (SF-36) Secondary outcomes were Visual Analogue Scale (VAS) pain, Timed Up &; Go Test (TUG), knee range of motion and quadriceps and hamstring strength-isometric contraction. Responses to treatment (responders) and minimal clinically important improvements (MCIIs) were determined.

Results

In the OAK group, VAS pain improved from baseline to week 8 from mean (SEM) 5.21 (0.30) to 3.65 (0.29) (P ≤ 0.001). During this period, improvements in the OAK group compared with the control group and responses to treatment were demonstrated according to the following outcomes: WOMAC Pain, Physical Function and Total dimensions, as well as SF-36 Physical Function, Role Physical, Body Pain, Vitality and Social Functioning domains. In addition, from baseline to week 8, the proportion of MCIIs was greater among the OAK group than the control group for all outcomes. For the period between baseline and month 6, WOMAC Pain, Physical Function and Total dimensions significantly improved in the OAK group compared to the control group, as did the SF-36 Physical Function, Role Physical, Body Pain, Vitality and Social Functioning domains, as well as hamstring strength in both legs. During the same period, the TUG Test, range of motion extension and left-knee flexion improved compared with the control group, although these improvements had little clinical relevance.

Conclusions

We recorded statistically significant improvements compared with a control group with regard to pain, quality of life and function for participants in the OAK program on the basis of WOMAC and SF-36 measures taken 8 weeks and 6 months from baseline.     

Mapping QTLs for sucrose content,yield and quality in a sugar beet population fingerprinted by EST-related markers

Seventy five expressed sequence tags (ESTs) that are associated with functions in carbohydrate and nitrogen metabolism were genotyped in 108 plants of an F2 population of sugar beet ( Beta vulgaris L.) segregating for sugar quality and yield parameters. Supplemented by known RFLP and AFLP markers, the resulting map spans 446 cM of the 758-Mbp genome of sugar beet. F3 test-cross plants were analysed for corrected sugar yield, beet yield, ion balance and the content of sugar, amino nitrogen, potassium and sodium in six locations. Twenty one significant quantitative trait loci (QTLs) were detected using the composite interval mapping approach. Expressed genes flanking the QTLs were identified in all cases. Correlations between QTLs and potential candidate genes are discussed.     

Optimization of reaction variables for sucrose monoester production using lipase in a solvent free system by Taguchi's method

Taguchi's method was attempted successfully to optimize the reaction variables and their ranges for sucrose monoester production. Optimal conditions determined by Taguchi's method were 35°C, 200rpm, capric acid (40mmol) and sucrose (3mmol), 24h, 0.5g lipase, 0.72g Ba(OH)2 ·8H2O, 0.72g Ba(OH)2·1H2O. A product yield of 21% was obtained. © Rapid Science Ltd. 1998     

Development and validation of a sensitive liquid chromatography/mass spectrometry method for quantitation of flavopiridol in plasma enables accurate estimation of pharmacokinetic parameters with a clinically active dosing schedule

A high-performance liquid chromatographic assay with tandem mass spectrometric detection was developed and validated for quantitation of the broad spectrum kinase inhibitor, flavopiridol, in human plasma. Sample preparation conditions included liquid-liquid extraction in acetonitrile (ACN), drying, and reconstitution in 20/80 water/ACN. Flavopiridol and the internal standard (IS), genistein, were separated by reversed phase chromatography using a C-18 column and a gradient of water with 25 mM ammonium formate and ACN. Electrospray ionization and detection of flavopiridol and genistein were accomplished with single reaction monitoring of m/z 402.09>341.02 and 271.09>152.90, respectively in positive-ion mode [M+H](+) on a triple quadrupole mass spectrometer. Recovery was greater than 90% throughout the linear range of 3-1000 nM. Replicate sample analysis indicated within- and between-run accuracy and precision to be less than 13% throughout the linear range. This method has the lowest lower limit of quantitation (LLOQ) reported to date for flavopiridol, and it allows for more accurate determination of terminal phase concentrations and improved pharmacokinetic parameter estimation in patients receiving an active dosing schedule of flavopiridol.     

Mannosylated lipoarabinomannans inhibit IL-12 production by human dendritic cells: evidence for a negative signal delivered through the mannose receptor

IL-12 is a key cytokine in directing the development of type 1 Th cells, which are critical to eradicate intracellular pathogens such as Mycobacterium tuberculosis. Here, we report that mannose-capped lipoarabinomannans (ManLAMs) from Mycobacterium bovis bacillus Calmette-Guérin and Mycobacterium tuberculosis inhibited, in a dose-dependent manner, the LPS-induced IL-12 production by human dendritic cells. The inhibitory activity was abolished by the loss of the mannose caps or the GPI acyl residues. Mannan, which is a ligand for the mannose receptor (MR) as well as an mAb specific for the MR, also inhibited the LPS-induced IL-12 production by dendritic cells. Our results indicate that ManLAMs may act as virulence factors that contribute to the persistence of M. bovis bacillus Calmette-Guérin and M. tuberculosis within phagocytic cells by suppressing IL-12 responses. Our data also suggest that engagement of the MR by ManLAMs delivers a negative signal that interferes with the LPS-induced positive signals delivered by the Toll-like receptors.     

Urine sample preparation of tricyclic antidepressants by means of a supported liquid membrane technique for high-performance liquid chromatographic analysis

Supported liquid membrane (SLM) technique for sample work-up and enrichment was used for determination of tricyclic antidepressant drugs in urine by high-performance liquid chromatography (HPLC) with UV detection. The studied antidepressant drugs were amitriptyline, opipramol, noxiptyline and additionally diethazine was used as possible internal standard. Alkaline phosphoric buffer with urine sample, as the donor solution, was passed over the liquid membrane into which investigated substances were extracted. On the other side of the membrane, analyzed compounds were trapped due to creating non-extractable form in acidic acceptor solution. Enriched and cleaned up drugs were then injected into a HPLC system with ultraviolet detection to analyze of their concentration in acceptor solution. Optimum extraction efficiency was determined by changing acceptor and donor solutions pH, application of different flow rates of donor solution and by using different solvents in the membrane. Also, donor solution volume, extraction time and concentration of analytes were varied to check the linearity of extraction process. The highest extraction efficiency: 43% for opipramol, 56% for noxiptyline, 43% for amitriptyline and 42% for diethazine (R.S.D. values were <6% and n=3) was achieved when 0.05 M phosphate buffer pH 4.0 and 9.5 were used as donor and acceptor solutions, respectively, n-undecane with 5% tri-n-octylphosphine oxide (TOPO) was used as liquid membrane. Limit of quantification (LOQ) for tricyclic antidepressants after enrichment of 100ml of urine sample was about 1 ng/ml.     

Analysis of a G protein-coupled receptor for neurotensin by liquid chromatography-electrospray ionization-mass spectrometry

The type 1 neurotensin receptor (NTS1) belongs to the G protein-coupled receptor (GPCR) family. GPCRs are involved in important physiological processes, but for many GPCRs ligand binding sites and other structural features have yet to be elucidated. Comprehensive analyses by mass spectrometry (MS) could address such issues, but they are complicated by the hydrophobic nature of the receptors. Recombinant NTS1 must be purified in the presence of detergents to maintain solubility and functionality of the receptor, to allow testing of ligand, or to allow G protein interaction. However, detergents are detrimental to MS analyses. Hence, steps need to be taken to substitute the detergents with MS-compatible polar/organic solvents. Here we report the characterization of NTS1 by electrospray ionization (ESI)-MS with emphasis on methods to transfer intact NTS1 or its proteolytic peptides into compatible solvents by protein precipitation and liquid chromatography (LC) prior to ESI-MS analyses. Molecular mass measurement of intact recombinant NTS1 was performed using a mixture of chloroform/methanol/aqueous trifluoroacetic acid as the mobile phase for size exclusion chromatography-ESI-MS analysis. In a separate experiment, NTS1 was digested with a combination of cyanogen bromide and trypsin and/or chymotrypsin. Subsequent reversed phase LC-ESI-tandem MS analysis resulted in greater than 80% sequence coverage of the NTS1 protein, including all seven transmembrane domains. This work represents the first comprehensive analysis of recombinant NTS1 using MS.     

Multiple steady states in a continuous stirred tank reactor: an experimental case study for hydrolysis of sucrose by invertase

The purpose of this work was to validate experimentally that multiple steady states may be achieved in a continuous stirred tank reactor (CSTR) during hydrolysis of sucrose by invertase. Experiments were done with four initial sucrose concentrations (0.1, 0.175, 0.584 and 1 M) to study their effect on residual sucrose and reaction rate at steady state. Two different steady states (S=0.7 M, r=9×10⁻⁴ mol/l min and S=0.135 M, r=1.54×10⁻³ mol/l min) were found depending on initial concentration of sucrose in the reactor. Two stable steady states were possible in a CSTR using invertase for the hydrolysis of sucrose. A third possible steady state can be derived theoretically, but it should be a metastable condition because any small disturbance in the system will result in transitory states stabilizing at sugar concentrations of either 0.135 or 0.7 M.     

Adaptive response for chromosomal inversions in pKZ1 mouse prostate induced by low doses of X radiation delivered after a high dose

Adaptive responses are induced by stress such as X radiation and result in a lower than expected biological response. Two-dose adaptive response experiments typically involve a low priming dose followed by a subsequent high radiation dose. Here, we used a sensitive in vivo chromosomal inversion assay to demonstrate for the first time an adaptive response when a low dose (0.01-1 mGy) was given several hours after a high 1000-mGy radiation dose. The adaptive responses in this study were of similar magnitude to the two-dose adaptive responses previously observed in this test system when the low dose was given first. A chromosomal inversion adaptive response was also induced by two 1000-mGy doses and when a 1-mGy dose was preceded or followed by a dose of 0.01 mGy, but not by two 4000-mGy doses. This is also the first example of an adaptive response when both doses are low. Our data agree with previous reports of an on-off mechanism of adaptive response. The induction of an adaptive response by a low dose after a high damaging dose provides evidence that the mechanisms underlying radiation adaptive responses are not due to prevention of damage induced by the high dose but to modulation of the cellular response to this damage.     

Microanalysis for MDR1 ATPase by high-performance liquid chromatography with a titanium dioxide column

MDR1 is clinically important because it is involved in multidrug resistance of cancer cells and affects the pharmacokinetics of various drugs. Because MDR1 harnesses adenosine 5'-triphosphate (ATP) hydrolysis for transporting drugs, examining the effect on ATPase activity is imperative for understanding the interactions between drugs and MDR1. However, conventional assay systems for ATPase activity are not sensitive enough for screening drugs using purified MDR1. Here we report a novel method to measure ATPase activity of MDR1 using high-performance liquid chromatography equipped with a titanium dioxide column. The amount of adenosine 5'-diphosphate (ADP) produced by the ATPase reaction was determined within 2 min with a titanium dioxide column (4.6 mm ID x 100 mm). The relationship between ADP amount and chromatogram peak area was linear from 5 pmol to 10 nmol. This method made it possible to reduce the amount of purified MDR1 required for a reaction to 0.5 ng, about 1/20th of the conventional colorimetric inorganic phosphate detection assay. This method is sensitive enough to detect any subtle changes in ATPase activity of MDR1 induced by drugs and can be applied to measure ATPase activity of any protein.