ALBERT

Notes: 1 The effects of the immunosuppressants, tacrolimus (FK506) and cyclosporin A (CsA), on catecholamine (CA) release were examined in cultured bovine adrenal chromaffin cells. 2 In intact cells, FK506 (1–30 μm) inhibited CA release stimulated by acetylcholine (ACh; 100 μm), 1,1-dimethyl-4-phenyl-piperazinium (DMPP, 10 μm) or high K+ (40 mm). CsA (1–30 μm) had a little inhibitory effect on the ACh- or DMPP-stimulated CA release, whereas it enhanced the high K+-stimulated CA release. 3 In β-escin-permeabilized cells, FK506 inhibited CA release stimulated by Ca2+ (1 and 10 μm) in the presence and absence of MgATP (2 mm). CsA induced CA release under Ca2+-free condition and enhanced the Ca2+-stimulated CA release in the presence and absence of MgATP. 4 It is known that the Ca2+-dependent exocytosis involves at least two distinct steps, ATP-requiring priming stage and ATP-independent fusion step in adrenal chromaffin cells. Therefore, it is suggested that FK506 inhibits the Ca2+-dependent exocytosis probably at the fusion step whereas CsA induces CA release from bovine adrenal chromaffin cells.

Notes: 1 Prolonged bed rest or exposure to microgravity may cause several alterations in autonomic nervous system response (ANSR). 2 Hindlimb unloading (HU) rats were used as an animal model of simulated microgravity to investigate ANSR changes. The experiments were carried out to investigate the effects of simulated microgravity on the autonomic nervous response of the perfused mesenteric vascular bed (MVB), vas deferens and the colon and duodenum from 2-week HU rats. 3 In MVB preparations of HU rats, the frequency-dependent increases in perfusion pressure with perivascular nerve stimulation (PNS; 8–40 Hz) were inhibited, whereas the noradrenaline (NA) concentration-dependent (1–100 μm) perfusion pressure increases were potentiated. The latter most probably reflected up-regulation of α-adrenergic receptor function. Relaxant responses of NA-precontracted MVB to PNS (4–30 Hz) or isoprenaline were not different between control and HU preparations, while vasodilation induced by the endothelial agonist ACh was reduced. 4 Transmural stimulation (2–40 Hz) induced frequency-dependent twitches of the vas deferens which were reduced in vas deferens of HU rats, while the sensitivity to NA-induced contraction was significantly increased. 5 In the gastroenteric system of HU rat, direct contractile responses to carbachol or tachykinin as well as relaxant or contractile responses to nervous stimulation appeared unchanged both in the proximal colon rings and in duodenal longitudinal strips. 6 In conclusion, HU treatment affects peripheral tissues in which the main contractile mediators are the adrenergic ones such as resistance vessels and vas deferens, probably by reducing the release of neuromediator. This study validates NA signalling impairment as a widespread process in microgravity, which may most dramatically result in the clinical phenotype of orthostatic intolerance.

Notes: 1 This double-blind, cross-over, placebo-controlled study on six healthy male volunteers was designed to evaluate the effects of α2-adrenoceptor antagonism on cardiac parasympathetic regulation. 2 The subjects received atipamezole intravenously as a three-step infusion, which aimed at steady-state serum concentrations of 10, 30 and 90 ng ml−1 at 50-min intervals. 3 Drug effects were assessed with repeated recordings of blood pressure and electrocardiogram, in which the high-frequency (0.15–0.40 Hz) R-R interval variation is supposed to reflect cardiac parasympathetic efferent neuronal activity. 4 At the end of the three steps of the infusion, the mean (±SD) concentrations of atipamezole were 10.5 (3.9), 26.8 (5.6) and 81.3 (21.1) ng ml−1. 5 Within this concentration range, atipamezole appeared to reduce slightly the high-frequency R-R interval fluctuations, indicating a minor vagolytic effect in the heart. 6 Atipamezole increased systolic and diastolic arterial pressure, on average by 20 and 14 mmHg (maxima at the second step of the infusion), which evidently reflects an overall sympathetic augmentation.

Notes: 1 We have compared vasoconstriction responses in isolated mesenteric small arteries from mice and rats as elicited by KCl, noradrenaline and the lysosphingolipids sphingosine-1-phosphate (S1P) and sphingosylphosphorylcholine (SPC). 2 Contractile responses to KCl and noradrenaline, but not those of S1P or SPC, were significantly related to vessel diameter in both species. 3 When comparing vessels of similar diameter, contractile responses for KCl and the three agonists were much smaller in mice than in rats, e.g. 8.3 ± 0.4 vs. 14.7 ± 0.7 mn for noradrenaline. 4 Based upon the antagonist rank order of potency of prazosin (pKB 8.80) 〉 B8805-033 (pKB 7.89) 〉 yohimbine (pKB 6.18) ≈ BMY 7378 (pA2 6.03), noradrenaline responses in mice were mediated solely via α1A-adrenoceptors, similar to what repeatedly has been shown in rats. 5 The S1P3 receptor antagonist suramin (100 μm) significantly inhibited responses to S1P and SPC in rats but not in mice, and did not affect noradrenaline responses in either species. 6 We conclude that for any given diameter, mouse mesenteric arteries develop less contraction in response to various stimuli. Noradrenaline acts via α1A-adrenoceptors in both species. Responses to S1P and SPC differ between both species with regard to suramin-sensitivity indicating involvement of different receptor subtypes for lysosphingolipids in both species.

Notes: 1 A fructose (Fru)-enriched diet induces a mild increase in blood pressure associated with hyperglycaemia, hypertriglyceridaemia, and insulin resistance, resembling the human ‘syndrome X’, being an useful model to study hypertension and type 2 diabetes. 2 A sustained elevation of blood pressure is associated with cardiovascular structural modifications such as left ventricular hypertrophy and increased wall thickness:lumen diameter ratio in blood vessels. 3 Prostanoids (PR), metabolites of arachidonic acid through the cyclooxygenase pathway, include vasoactive substances synthesized and released by the vessel walls. 4 The aim of the present study was to analyse, in Fru-treated rats: (i) the morphology of mesenteric vessels and; (ii) the PR production in aorta and mesenteric vessels, in order to assess whether these parameters are related with the haemodynamic alterations observed in this experimental model. 5 Blood pressure, glycaemia and triglyceridaemia, were significantly elevated in both (4 and 22 weeks) Fru-treated groups. Meanwhile body and heart weight as well as insulinaemia were similar between experimental animals and controls. 6 The mesenteric vessels of Fru-treated rats (22 weeks) showed an increased thickness and area of the media when compared with the controls; meanwhile, the lumen diameter was similar in both groups. 7 The Fru treatment for 4 weeks did not modify PR production in aorta, whereas in the mesenteric bed it diminished prostaglandin (PG) E2 release significantly compared with the controls. However, in the group treated for 22 weeks, Fru reduced PGI2 production in the aorta, as assessed by 6-keto-PGF1α measurements. Meanwhile, in the mesenteric bed, the chronic Fru treatment decreased PGE2 release but, rather surprisingly, increased the output of PGI2 when compared with its corresponding controls. 8 In conclusion, the present study shows the existence of an alteration in the morphology of mesenteric vessels in Fru-treated rats, which could be related to an increase in peripheral resistance and the consequent mild hypertension observed in this model. However, a diminished release of vasodilator PRs, such as PGE2 in mesenteric vessels at 4 and 22 weeks and PGI2 in aorta at 22 weeks could further impair the vessel response. The increase in PGI2 observed in the chronic group in mesenteric vessels could be attributed to a compensatory mechanism.

Notes: 1 Electrical field stimulation (EFS) (0.5 ms in train of 2–32 Hz for 300 ms) in smooth muscle of pigeon oesophagus, in the presence of atropine (1 μm) and guanethidine (1 μm), elicited an inhibitory response consisting of a transient hyperpolarization (inhibitory junction potential, IJP) associated with muscle relaxation. 2 Sodium nitroprusside (SNP, 100 μm) induced hyperpolarization correlated to mechanical relaxation. 3 The nitric oxide (NO) synthase inhibitor Nω-nitro-l-arginine (from 0.1 to 100 μm) caused a concentration-dependent reduction of electromechanical response to EFS indicating a role for NO in this response. 4 Apamin (1 μm) reduced both IJP and relaxation to EFS but was without effect on the response to SNP indicating a role for purines, which are also blocked by apamin. 5 Adenosine, AMP, ADP and ATP (all from 1 μm to 1 mm) application caused transient hyperpolarization and muscular relaxation with the following order of potency: adenosine 〉 AMP 〉 ADP 〉 ATP. 6 Inhibitory responses evoked by purines are TTX (1 μm) insensitive but they were inhibited by apamin. This indicates that a purine component for the non-adrenergic non-cholinergic (NANC) response exists but the purine receptor site is not located on the neurone. 7 Overall these results suggest that NANC inhibitory response elicited by EFS presents two different components apamin-sensitive, probably purines-mediated and apamin-insensitive probably NO-mediated as apamin only partially block the response to EFS.

Notes: 1 Sibenadet (Viozan®), a dual dopamine D2/β2-adrenoceptor agonist, suppresses histamine-induced tachypnoea in the dog by activating dopamine D2 receptors. We here compare the effects of sibenadet and formoterol, a selective β2-adrenoceptor agonist, on histamine-induced tachypnoea in the rhesus monkey. 2 Anaesthetized, spontaneously breathing, rhesus monkeys were set up for measuring airways resistance, respiratory rate, blood pressure and heart rate. 3 Both sibenadet and formoterol administered by aerosol, induced inhibition of the bronchoconstrictor response to aerosolized methacholine accompanied by tachycardia. Sibenadet, but not formoterol, also reduced blood pressure. 4 Administration of histamine by inhalation induced tachypnoea which was accompanied by bronchoconstriction. Tachypnoea to histamine was suppressed by both sibenadet and formoterol at doses which manifest anti-bronchoconstrictor activity. These effects and the accompanying tachycardia but not the hypotension induced by sibenadet were abolished by pretreatment with propranolol. 5 The dopamine D2 receptor agonist, quinagolide, did not suppress tachypnoea to histamine despite inducing a fall in blood pressure indicating activation of dopamine D2 receptors. 6 Thus, both sibenadet and formoterol suppress histamine-evoked tachypnoea in the rhesus monkey. The effect arises exclusively through activation of β2-adrenoceptors and probably reflects the anti-bronchoconstrictor effects of these agents. The results reveal a fundamental difference in the role of dopamine receptors in the airways of dog and rhesus monkey.

Notes: 1 We have investigated the actions of the calcium entry blockers nifedipine, R-verapamil and S-verapamil in rat aorta, colon and vas deferens. 2 In aorta and colon, these agents produced concentration-dependent relaxations of KCl (80 mm)-induced contractions. In both tissues, the order of potency was nifedipine 〉 S-verapamil 〉 R-verapamil. However, nifedipine showed selectivity for aorta (potency ratio, colon/aorta: 4.36), S-verapamil showed no selectivity (0.62), but R-verapamil showed selectivity for colon (0.19). 3 In prostatic portions of rat vas deferens, nifedipine (10 μm) abolished the contraction to a single electrical stimulus, but R- and S-verapamil were without effect. In epididymal portions of rat vas deferens, R- and S-verapamil inhibited α1-adrenoceptor-mediated contractions to a single electrical stimulus at concentrations of 10 μm and above. 4 In conclusion, R-verapamil may prove useful as an intestinal selective calcium entry blocker in the treatment of intestinal disease with a hypermotility component, e.g. irritable bowel syndrome.

Notes: 1 The present study was carried out to pharmacologically identify the β-adrenoceptor subtype that mediates isoprenaline-elicited relaxation in the isolated guinea-pig tracheal smooth muscle, to answer the question whether it is β1- or β2-subtype? 2 Isoprenaline as well as salbutamol, a well-known β2-selective adrenoceptor agonist, produced a concentration-dependent relaxation with a pD2 value of 8.12 vs. 7.54 for salbutamol. 3 Isoprenaline-elicited relaxation was not affected by β1-selective antagonists, atenolol and CGP-20,712A, within the concentration ranges supposed to antagonize β1-subtype: atenolol, ≤10−6 m; CGP-20,712A, ≤10−8 m. 4 By contrast, the concentration–response curves for isoprenaline as well as salbutamol were shifted rightwards in a competitive fashion by atenolol at the concentrations ≥3 × 10−6 m. However, pA2 values of atenolol against isoprenaline (5.86) and salbutamol (5.71) were consistent with the value corresponding to β2- but not to β1-subtype (around 7.00), and these values were not significantly different from each other. 5 Competitive antagonism of the relaxations to isoprenaline and salbutamol were also obtained with β2-selective antagonists, butoxamine and ICI-118,551. Against isoprenaline and salbutamol, the pA2 values of butoxamine (6.51 vs. 6.81) and ICI-118,551 (8.83 vs. 8.90) were substantially identical. Thus the primary mediation of β2-receptor in the relaxations was strongly supported. 6 The present findings provide evidence that the β-adrenoceptor which mediates isoprenaline-elicited relaxation of guinea-pig tracheal smooth muscle is essentially β2- but not β1-subtype. The present study also indicates the importance of using multiple receptor antagonists with different pA2 values to pharmacologically identify the responsible receptor subtype in smooth muscle mechanical responses.

Notes: 1 The role of histamine in heat-induced cardiovascular changes is uncertain. The purpose of this study was to examine effects of histamine H-1- and H-2-antagonism on heart rate, mean arterial blood pressure (MAP), localized body temperature changes, survival times, and lethal body temperatures that occur during the exposure of anaesthetized rats to 35 GHz radio frequency radiation (RFR). 2 Forty-eight ketamine-anaesthetized Sprague–Dawley rats were exposed, in several different treatment groups (n = 8 in each), to 35 GHz RFR at a level that resulted in significant body heating and subsequent death. During irradiation, a continuous increase in heart rate and a biphasic response in blood pressure (initial increase followed by a decrease) were observed in all groups of animals. 3 An H-1-antagonist, diphenhydramine (1 mg kg−1 body wt) and an H-2-antagonist, cimetidine (5 mg kg−1), administered after sustained RFR exposure, failed to reverse the RFR-induced hypotension. High doses of the drugs (5 and 10 mg kg−1, respectively) also did not alter the response. Post-RFR survival time was significantly decreased in the high-dose drug-treated group, compared with vehicle-treated (0.9% NaCl, 50% ethanol and 50% D5W) controls. 4 In experiments in which the two drugs were administered prior to RFR exposure, MAP in animals receiving high-dose antihistamines was significantly depressed compared with that of vehicle-treated animals during the first 35 min of RFR exposure. Antihistamine pretreatment, however, did not alter the total RFR exposure time required for death to occur. 5 In summary, pharmacological blockade of H-1 and H-2 receptors is not beneficial in anaesthetized rats made hypotensive by RFR exposure. This indicates that activation of H-1 and H-2 receptors by histamine does not occur to any significant extent and does not mediate the hypotensive response developed in this model of hyperthermia.

Notes: 1 The great taxonomic and prey base diversity of colubrids (non-front-fanged snakes) suggests that their venoms may represent a ‘literal gold mine’ for scientists eager to find novel pharmacological probes (Mackessy, 2002). 2 While pharmacological characterization is lacking for most of these venoms, this is even more so with regard to activity of colubrid venoms on the mammalian autonomic nervous system. This study characterizes the activity of venom from the colubrid, Boiga dendrophila using in vitro smooth muscle preparations and the anaesthetized rat. 3 In the prostatic segment of the rat vas deferens, cumulative additions of venom (1–150 μg ml−1) induced concentration-dependent inhibition of electrically evoked (0.2 Hz, 0.3 ms, 70–100 V) twitches. The inhibitory effect of venom (100 μg ml−1) was attenuated by 8-phenyltheophylline (8-PT) (20 μm) and 8-cyclopentyl-1, 3-dipropylxanthine (20 μm) but not idazoxan (1 μm), or a combination of ranitidine (0.2 μm) and thioperamide (10 μm). The inhibitory effect of venom (100 μg ml−1) was augmented by dipyridamole (10 μm) but abolished by pretreatment with adenosine deaminase (7.5 units/100 μl) suggesting that it contains components with adenosine A1 receptor activity, most likely adenosine. 4 In isolated segments of guinea-pig ileum, venom (10–100 μg ml−1) caused concentration-dependent contractions which were inhibited by the muscarinic receptor antagonist atropine (0.1 μm) but not by the histamine receptor antagonist mepyramine (0.5 μm). 5 In the anaesthetized rat, venom (5–7.5 mg kg−1, i.v.) caused a hypotensive effect. 6 Our data suggest that the venom contains components with purinergic and muscarinic receptor activity.

Notes: 1 β-Adrenoceptor (AR) ligands have been the mainstay of cardiovascular therapy for decades, with β-AR antagonist being used for hypertension, angina and myocardial infarction and adrenaline in use for cardiopulmonary resuscitation for nearly 100 years. 2 Ischaemia of the heart through coronary artery occlusion causes cell injury and death through necrosis and apoptosis. Reperfusion of the ischaemic myocardium results in cardiac dysfunction and infarction. 3 Stimulation of α- and β-ARs in the ischaemic heart have variable and inconsistent effects depending on when the agonist is applied. This review describes the different effects of stimulation of the three established β-AR subtypes (β1-, β2- and β3-ARs) either before ischaemia (preconditioning) or during ischaemia and reperfusion of the heart (postconditioning). 4 Brief periods of ischaemia preceding a major ischaemic episode can have a protective effect against post-ischaemia–reperfusion damage, known as ischaemic preconditioning. This review considers the role of endogenous catecholamines released during preconditioning and the nature of the adrenoceptor subtypes that mediate these effects. The clinical significance of this to the use of β-AR antagonists is considered. 5 The transduction pathways and effects on apoptosis of the cardioprotective and deleterious effects of AR activation are considered. 6 This commentary reviews the literature and attempts to bring together a unified synopsis of the effects of adrenoceptor stimulation in myocardial ischaemia and the potential clinical relevance.

Notes: 1 Cupric ions (Cu2+), at concentrations above 0.03 mm, induced a progressive increase in the tonic contraction of guinea-pig ileal longitudinal muscle. Maximal contraction of 0.1 mm Cu2+ attained a level above that of the 60-mm K+-induced tonic response, within 20 min of application. The tension induced by Cu2+ persisted for more than several hours. Tetrodotoxin (3 × 10−6 m) had no effect on the contraction induced by 0.1 mm Cu2+. 2 After incubation in a Ca2+-free medium, the ileal response to 0.1 mm Cu2+ was lost. Nifedipine, a L-type Ca2+ channel blocker, dose-dependently inhibited contractions induced by Cu2+. 3 As the duration of the first application of 0.1 mm Cu2+ increased above 30 min, after washing with normal medium, the contractile response to a second application of 0.1 mm Cu2+ decreased gradually. After 150 min of the first application of 0.1 mm Cu2+, a second application of Cu2+ could not evoke any contraction. 4 After the application of 0.1 mm Cu2+ for 150 min, when muscles were washed with a medium containing 1 mm EDTA, the response to 0.1 mm Cu2+ returned to a greater extent in the normal Ca2+ medium. 5 In conclusion, Cu2+ (0.1 mm) induced a maximal ileal tension above that of the K-induced tonic response within 20 min. The ileal contraction to Cu2+ persisted for more than several hours and depended on extracellular Ca2+ concentrations. It is possible that a part of Cu2+, bound to a EDTA-inaccessible site, also has a tension inhibitory effect.

Notes: 1 Anthraquinone stimulant cathartics, such as emodin, are believed to increase the rate of contraction of ileum tissue in vitro via multiple mechanisms. The aim of this study was to probe the effects of emodin on acetylcholine (ACh)-induced contraction of the rat isolated ileum preparation. 2 Ileal sections were incubated in Tyrode's solution and responses to methacholine, ACh and emodin obtained in the absence and presence of the muscarinic antagonist atropine and the choline uptake inhibitor hemicholinium (HC-3). Depletion of endogenous ACh in the presence of HC-3 was achieved by construction of an ACh dose–response curve, using exogenous ACh, prior to re-testing the effects of emodin in the presence of HC-3. 3 Emodin caused dose-dependent tissue contraction that was abolished by inclusion of atropine (1 μm) in the buffer. Atropine (1 μm) antagonized the response caused by methacholine. 4 Incubation of tissues with HC-3 (1 and 10 μm) reduced the maximum response caused by emodin by 45% and 71% respectively, but had no effect on ACh-induced tissue contraction. 5 These data suggest that, emodin causes contraction of the ileum by triggering the release of endogenous ACh which acts on muscarinic receptors to cause contraction of the rat isolated ileum preparation.

Notes: To determine the taste components of Cheddar cheese, we fractionated one mild Cheddar cheese and one aged Cheddar cheese by water extraction, freeze-drying and gel filtration. Salty, sour and umami were the three predominant tastes present in the fractions. Neither trigeminal sensations nor astringency was perceived. We used response surface methodology to reconstruct a mild Cheddar cheese taste and an aged Cheddar cheese taste in water. Less salt and less acid were needed to simulate the taste of mild Cheddar cheese, compared with aged Cheddar cheese. Our optimized water solutions (containing sodium chloride, lactic acid and monosodium glutamate) were as similar to the real cheese samples as were the water extracts of the standard cheeses. However, neither our optimized solutions nor the water extracts matched the taste of the actual cheeses.

Notes: Judges were required to rate the total intensity of NaCl solutions using a variety of unstructured category and line scales under a ‘rank-rating’ protocol and a traditional protocol that did not allow retasting or the reviewing of scores. The various scales and protocols induced two types of scaling errors. The first type was named a different-stimulus error. This involved a judge rating a stronger stimulus as equal to or less than a weaker stimulus. The second type was named a same-stimulus error. This involved a judge giving different ratings to two stimuli of equal concentration. For all scales, judges made a higher proportion of same-stimulus errors than different-stimulus errors. ‘Rank-rating’ only reduced the proportion of different-stimulus errors. It was hypothesized that a category scale with fewer categories would induce a higher level of different-stimulus errors but lower level of same-stimulus errors. These trends were noted but not always significantly. For line scales, the equivalent hypotheses regarding line length were either weak or not supported. It would appear that increasing the length of a line scale is not always equivalent to increasing the number of categories in a category scale.

Notes: The assessor performance is a key point in a sensory evaluation. In particular, at the end of a session, a decrease of the performance can be feared. We propose to analyze this performance with various criteria: usual ones as the main product effect or the error variance; a new one measuring the perceived products variability. The performance can then be studied all along the session from two points of view: in taking into account the only products tested at a given instant (named instantaneous); in taking into account all the products tested up to a given instant (named cumulative). In the presented example, in spite of the large number of products successively tested by each assessor, the instantaneous performance of the panel shows no significant deterioration. Furthermore, when the number of products tested by each assessor increases, more significant product effects can be obtained thanks to the accumulation of the amount of data. This shows that the number of products that can be reasonably studied by one assessor during one session is generally underestimated.

Notes: Two experimental procedures recommended for the evaluation of the psychophysical characteristics of odorous compounds, olfactory matching with the 1-butanol scale and cross-modality matching with the finger span are compared. The intensity of ethyl butyrate and guaiacol solutions presented at four different concentration levels was evaluated by a panel of sixteen subjects over five repetitions using the two methods. Each stimulus was delivered to the subject from a Teflon bag through a nose-shaped glass sniffing port.The discrimination ability, repeatability, panel homogeneity and within-subject variability of the methods were assessed. Results indicate that with both methods, subjects were able to highly discriminate the presented concentration levels of the odorants. There were no great difference in repeatability and the same individual variability was observed between both methods. However, the smaller within-subject variability highlighted for the 1-butanol scale method suggested that this method is potentially more powerful than finger-span method.

Notes: People can eat a food without having a strong preference for it, and people can prefer a food without eating it. Given this seeming disconnect between attitude and behavior, which type of measure or segment can best be used to profile or identify loyal consumer segments of a food, such as soy? This research compares a usage-based method (heavy-light-nonusers) with a new attitude-based method (seeker-neutral-avoider), and finds that the attitude-based method differentiates purchase-related intentions better than the usage-based method. Implications for profiling consumer taste patterns and consumer segments are provided.

Notes: Astringent and bitter sensations are characteristic sensory qualities of black tea. Three different classes of potential astringent reference standards (two concentrations each of alum and tannic acid and three fruit juices) were evaluated in this study. The perceived astringency, bitterness and sourness of each were profiled using computerized time-intensity and compared with the astringent intensity of a standardized brew of black tea. The differences in temporal profiles of potential reference standards across taste attributes were evident and intensity ratings were found to be dependent upon the stimulus and its concentration. Both concentrations of tannic acid were evaluated as the highest in perceived bitterness. For the juices, a strong sour taste was perceived in addition to astringency. It was concluded that the best reference standard for the astringency of black tea is a solution of 0.7 g/L alum as it is low in perceived bitterness and sourness.

Notes: We developed an alternative method for the determination of 6-n propylthiouracil (PROP) taster status that is based on the comparison of intensity ratings of PROP solutions with intensity ratings of visual, tactile, and kinesthetic (physical) stimuli. We then compared the classification obtained with this method to those derived from traditional approaches. One hundred and forty-two subjects rated the perceived intensity of 0.000032, 0.00018, 0.00032, 0.0018, and 0.0032 M PROP, and 0.01, 0.032, 0.1, 0.32, and 1.0 M NaCl, as well as five levels each of perceived roughness of sandpaper, grayness of shades, and thickness of honey and water mixtures on the Labeled Magnitude Scale (LMS). Ratings for 0.00032 M PROP and 0.1 M NaCl were also compared in a one solution test. Subjects were classified as PROP nontasters, tasters or supertasters by all three procedures. Seventy-six percent of panelists were classified in a similar way by the physical stimuli and NaCl procedures, while 70% percent of panelists were classified similarly by the physical stimuli procedure and the one solution test, as well as by the NaCl procedure and the one solution test. Only 65% of panelists were classified similarly by all three procedures. Further, in replicate ratings of PROP solutions, supertasters’ evaluations of PROP at all concentration levels decreased significantly (P 〈 0.01) from the first to the second rating. It is evident from these findings that the method used to determine PROP taster status can greatly influence the outcome of any investigation of PROP taster status on other phenotypes.

Notes: The objective of this study was to verify key aroma-active compounds responsible for reconstituted fresh skim milk powder (SMP) aroma using threshold analysis, odor activity values, and model systems. Twelve odor-active compounds of SMP and one odor-active compound from fluid milk were selected based on flavor dilution factors from gas chromatography-olfactometry. Thresholds for the 13 odor-active compounds were identified using five-set ascending forced choice threshold analysis in odor-free water and fluid skim milk. Model systems were prepared using rehydrated milk retentate (RMR). The aroma of each model was evaluated by descriptive sensory analysis and by difference-from-control testing using a trained panel. The aroma of reconstituted fresh SMP and liquid skim milk were used as controls. Models containing a mixture of twelve of the thirteen chemicals had the most similar odor characteristics to rehydrated SMP aroma (9.0/10) indicating that these compounds constitute the character impact odorants of rehydrated fresh SMP.

Notes: The sensory characteristic “beany” is a limiting factor in introducing soy products to many consumers in western cultures. This research described sensory properties of beany flavor and described sensory odor characteristics of 19 chemicals that have been associated with beany aroma and flavor in previous literature. Seven concentrations varying from 1 to 100,000 ppm of each chemical (if soluble in propylene glycol) were examined by a five member trained descriptive panel. The panel described beany as having multiple attributes. Musty/earthy, musty/dusty, sour aromatics, and characterizing attributes such as green/pea pod, nutty, or brown collectively imparted beany aroma and flavor of most beans. Starchy flavor and powdery texture also were intrinsically associated with beany in bean product samples. Some chemicals tested were not beany at any level tested. Three alcohols, two ketones, one aldehyde, and one pyrazine exhibited beany characteristics (green/peapod or brown) only at low concentrations, generally 1-10 ppm.

Notes: This paper deals with the analysis of drivers for self-defined craveability assessed in an Internet-based, conjoint analysis task. The stimuli comprised 36 descriptions of restaurant hamburgers, including product features, benefits, restaurant names, and emotional reactions that might ensue after eating the hamburger. Elements were combined into concepts by experimental design, and the resulting concepts evaluated by 145 respondents, on the attribute of craveability. Models relating the presence/absence of concept elements to ratings revealed that statements about the hamburger itself were the most powerful, but that no single element was highest across all the respondents. Segmentation of the 145 individuals by the pattern of their individual utilities revealed four key segments. These are Elaborates who may be sensory-oriented and respond strongly to product descriptions; Classics who like the notion of a grilled hamburger; Imaginers who respond to restaurant name and advertising copy; and Emotionals who respond to statements about food to descriptions how the eater feels after consumption.

Notes: Judges were required to rate the total intensity of NaCl solutions and the sweetness of orange flavored beverages using a variety of unstructured category and line scales. Discrimination errors were noted for each type of scale. No strong differences were observed in discrimination between category and line scales although a 20-point category scale incurred fewer discrimination errors than 9-point category and line scales. Allowing judges to retaste stimuli and review and modify their scores reduced discrimination errors.

Notes: The objectives of this study were to compare oral and nonoral sensory evaluation for discrimination of texture of acid skim milk gels and to establish whether nonoral attributes could be correlated to the oral perception of texture. Trained panelists (n = 13) identified 11 nonoral (visual and in-hand) and 4 oral attributes during a preliminary profiling session that could be used to discriminate textures (P 〈 0.001) in a range of acid gels prepared with different solid contents and heat treatment of the milks. Both methods of sensory appraisal were found to discriminate between gels. Correlation analysis showed high interrelationship between individual oral and nonoral attributes (P 〈 0.01). Principal component analysis revealed that all 4 oral attributes could be combined into one single attribute (PC1), with equal relative importance of the individual attributes in explaining the variance in the oral sensory data set. Canonical correlation analysis revealed good correlation between the oral and nonoral set of attributes (R2 〉 87.5%).

Notes: The flavor of eight samples of commercial strawberry yogurt was studied by Free-Choice Profile analysis (FCP). Generalized Procrustes Analysis (GPA) applied to FCP allowed differentiation between samples and highlighted flavor attributes responsible for the observed differences. The relation between sensory and physicochemical datasets was studied by means of GPA. Those samples with higher carbohydrate content were perceived as sweeter, having stronger strawberry flavor, and with more dairy and yogurt flavors. Samples with higher titratable acidity, ash and protein content were perceived as more acidic and higher in intensity of “faulty” or “defective” flavors. Higher moisture content was associated with lower intensity of “dairy” flavors (creamy, dairy, and yogurt) and greater intensity of rancid flavor. It is concluded that, though not often used to this end, GPA is a suitable method to study the relationship of sensory and instrumental measurements.

Notes: In a recent paper, we described the development and application of a labeled affective magnitude (LAM) scale for assessing liking/disliking (Schutz and Cardello 2001). Here we present the exact numerical scale-point locations corresponding to the verbal labels of the scale, so that investigators can easily construct the LAM scale for use with either paper or computer-based ballots.

Notes: This article describes an exploration of the concept of ‘oral freshness’ for as far as it is important in oral care. It intends to consider the most important mouth sensations and cognitive connotations, including (but not restricted to) the well-known effect of menthol in toothpaste.Two aspects were given attention:(1) The attributes that together form the concept of oral freshness were investigated using a ‘personal construct approach’(Kelly 1955). This method consists of unbiased, structured interviews with subjects, and ultimately yields attribute dimensions that the subjects have in common.(2) The intensity of the freshness sensation over time was investigated using the Time-Intensity method. Subjects gave repeated freshness judgments every few seconds after the intake of a freshness-related stimulus, and continued to do so after the stimulus had left their mouth. In addition we asked for ‘overall’ freshness judgments directly after the TI-measurements and after 2 months.The results confirm that ‘oral freshness’ is a complex concept. We identified 6 attributes, some of which were perceptual/physiological and some cognitive in nature: ‘water’-ness, cool/cold-ness, taste (menthol-ness), clean-ness, smell, energy (texture and touch, e.g., bubbles). Different persons will generally give different importance weights to each of these 6 attributes.Time-intensity measurements show that temperature, menthol-content and, to some extent, the presence of bubbles (energy) influence the momentous sensation of freshness. It appears, however, that in the way freshness is remembered after a few months, mainly cleanness and taste (menthol-ness) are important. Apparently, people tend to value (temporary) sensory input during the actual freshness experience, but tend to remember the freshness result in the longer run.

Notes: Time-intensity (TI) methodology and a trained panel were used to characterize the perceived bitterness of Yerba mate (YM) Ilex paraguariensis infusions. Two sampling procedures (sip and spit; sip and swallow) and two conditions for residence time in mouth (free or fixed duration of 5 s until spit or swallow) were evaluated.At a fixed duration the maximum bitterness as well as the time to reach it showed a significant YM concentration dependence. No change on bitterness was observed by swallowing or spitting YM infusions except a larger rate of decay of the response (vr) at spit condition. Dynamic bitterness at free sampling time showed that the decision period to spit or swallow the YM infusion approximately duplicated the fixed one of 5 s.

Notes: The effect of fat and fat replacers (FR) on the flavor release from cherry ice cream was investigated. Protein based FR, Simplesse (“S”) and carbohydrate based FR, Litesse (“L”) either partially or completely substituted milk fat in ice cream. Overall and temporal flavor profiles of ice cream during eating were obtained using chemical and sensory analyses. The performance of FR depended on the level of fat it was substituting and the amount of fat present in the ice cream. In the reduced fat ice cream group, both FRs failed to mimic the characteristics of 6% fat ice cream, mainly because of the low retention of cherry flavor. In the full fat ice cream group, the sensory properties of ice cream containing “L” were closer to the full-fat (12%) ice cream than those with “S”. The critical flavor volatile compounds that strongly impact the sensory flavor attributes were identified.

Notes: Response surface methodology was used to optimize formulations of chocolate peanut spread. Thirty-six formulations with varying levels of peanut (25-90%), chocolate (5-70%) and sugar (5-55%) were processed using a three-component constrained simplex lattice design. The processing variable, roast (light, medium, dark) was also included in the design. Response variables, measured with consumers (n = 60) participating in the test, were spreadability, overall acceptability, appearance, color, flavor, sweetness and texture/mouthfeel, using a 9-point hedonic scale. Regression analysis was performed and models were built for each significant (p 〈 0.01) response variable. Contour plots for each attribute, at each level of roast, were generated and superimposed to determine areas of overlap. Optimum formulations (consumer acceptance rating of ≥ 6.0 for all attributes) for chocolate peanut spread were all combinations of 29-65% peanut, 9-41% chocolate, and 17-36% sugar, adding up to 100%, at a medium roast. Verification of two formulations indicated no difference between predicted and observed values.

Notes: This paper presents the results from a study of consumer reactions to different attributes of red and white wine, with stimuli presented in the form of small, experimentally designed, test concepts (conjoint measurement). The results show that possibly four consumer mind-sets exist, with each mind-set comprising a different set of communication drivers for concept acceptance. Most subjects fit into one of three key categories; “Classics” (who want their wines to be traditional), “Imaginers” (who want to get into the drinking experience through knowing details about their wines) or “Elaborates” (who want their wines to produce many sensations and flavors). A fourth group (No Frills), were interested in simple and safe wines without any detail of flavor or origin. This segment was found in the red wine study, but not in the white wine study.

Notes: This paper describes a method that can be used to investigate consumers’ reported reasons for using a particular product. This method is applied to ratings of degree of agreement with statements about reasons for product use. The method is illustrated using data on self-reported reasons for smoking among adolescents. The approach used is based on a probabilistic model of similarity (Ennis et al. 1988) and provides a display of the density of respondents’ individual motivations concurrent with their perceptions of the statements. Factor analysis and its derived factor scores provide complementary information which is used to understand the interdependence of smoking motivations with the age of respondents and their degree of smoking. The probabilistic similarity model has many applications in studying consumer motivations such as those involved in the consumption of particular food and personal care products.

Notes: The addition of hazelnut paste to dark chocolate formulations altered the sensory profile. The flavors chocolate, burnt, rancid, hazelnut, green and metallic, and the texture attributes hardness, graininess, consistency of melt, viscosity and onset of melt determined by descriptive analysis were significantly (P 〈 0.05) influenced by the addition of up to 10% hazelnut paste. The shelf-life of dark chocolates containing hazelnut exceeded 10 months. The textural attributes of hardness, consistency of melt and onset of melt were highly correlated to instrumental measurements.

Notes: Sensory analysis was applied to the study of thermal perception, in order to determine and characterize end-users’ preferences concerning five indoor thermal environments generated by different heating systems. In this review, we developed a sensory profile method to describe and quantify the sensory characteristics of the thermal environments. A consumer test was also performed in order to investigate the hedonic judgment of 119 end users. The results show that trained assessors are able to describe and quantify sensory differences between the environments with high reliability. Consumers’ preferences are influenced by both their thermal sensations in the environments, as expressed by the sensory panel, and their personal opinions of the corresponding heating systems.

Notes: Survival analysis methodology was used to estimate the shelf life of alfajor (a chocolate-coated individually wrapped cake) at 20 and 35C by using results obtained from consumers when asked if they would accept or reject samples with different storage times. Sensory acceptability (measured by consumers), off-flavor (measured by a trained panel) and moisture content were linearly related to time. These correlations were used to estimate values at the shelf-life times calculated for 25 and 50% rejection probability. Survival analysis provided the following shelf-life estimation: 74 days at 20C and 33 days at 35C for a 25% of rejection, 87 days at 20C and 39 days at 35C for a 50% of rejection. An alfajor stored at 20C having an acceptability value below 4.9 (1–9 hedonic scale) and off-flavor intensity above 5.3 (0–10 scale) would be rejected by 25% of the consumers. Chemical data were not good shelf-life predictors.

Notes: Consumers rated a set of toothbrushes and a set of potato chips on 9-point and 21-point hedonic scales under two experimental protocols: a traditional approach and rank-rating. The hedonic data were analyzed in the usual way by using anova and multiple comparisons and also by ranking the data and using an R-index analysis. The hypothesis that the latter analysis would elicit fewer significant differences was not confirmed.

Notes: The profile of nine samples of Chardonnay wines was used to compare the performance of an expert panel and a trained descriptive panel using a common sensory terminology. Each panel (expert and trained) consisted of eight members, and the descriptors used by both groups were generated by the trained assessors. Thirteen out of 17 attributes evaluated turned out significant for wine discrimination among experts, and seven out of 17 for trained subjects. The trained panel reached a higher level of consensus, while the experts were more discriminative among attributes. The principal components analysis (PCA) corresponding to the expert group explained 88.68% of the total variance among the samples of the first two components. A third component was needed for the group of trained assessors (87.66% variance). The PCA showed the same wine grouping for both panels, indicating that differences were perceived in a similar way, despite that the sensory dimensions of these differences was not the same in all the cases.

Notes: Hispanics and non-Hispanics Americans participated in four sets of studies dealing with health-oriented foods, with the goal to identify how they respond to food categories relevant in any cuisine. Each study comprised a conjoint analysis using 36 elements followed by a self-profiling questionnaire to learn more about Hispanic and non-Hispanic food responses to concepts for four different product categories (morning bread, bottled water, healthful salad dressing and popcorn as a snack). The study identified patterns of preference of marketing statements in the areas of health information, sensory characteristics, emotional benefits and endorsements. Conjoint utility values generated by both Hispanics and non-Hispanic respondents correlated highly, suggesting that their food attitudes are quite similar for“non-signature”products despite the differences in their original cuisines.

Notes: In a descriptive sensory analysis, a principal component analysis (PCA) is usually applied to analyze products with attributes. To visualize the variability of a mean point (i.e., the center of gravity of the product), we introduced confidence ellipses. In this particular application, resampling methods are performed. Twenty products composed of five varieties of cider were studied. We would like to know if these five varieties are different. Confidence ellipses reveal differences between varieties foreshadowed in PCA.

Notes: The amount of training necessary to adequately “train” a descriptive panel is a matter of contention. The objectives of this research were to compare the performance of descriptive panelists after short-term (4 h), moderate (60 h) and extensive training (120 h). Seven screened panelists were chosen to evaluate three commercial tomato pasta sauces after each training period. Panelist performance improved with increased training. Sample differences were observed in all texture attributes and some flavor attributes, even after the shortest training time (4 h). However, more differences were found with at least 60 h of product training. After 120 h of training, product differences for all texture attributes and most of the flavor attributes studied could be ascertained by the trained panelists. These results suggested that only limited training may be necessary to find differences among products for many texture attributes and some flavor attributes. However, extensive training may be required to reduce variation among panelists and increase the discriminant abilities of panelists.

Notes: This study tests the possibility that sensory attributes are affected by the method by which a food is transferred to the mouth. For example, hot liquids appear to be considerably hotter when taken through a straw than when taken using a cup. Pre-weighed samples of two vanilla flavoured dairy custards were presented, in random order, to 16 trained panellists using a spoon, an 11-mm diameter straw or a modified straw that had a 4-mm diameter constriction within it. Panellists rated products using a subset of attributes generated by a quantitative descriptive analysis panel. The amount ingested was measured by re-weighing each sample after assessment. Each experiment was repeated three times. There were significant differences in the amount ingested for the two products. The delivery method had no effect on the sensory attributes of the products other than for thickness and melting, where products taken with the spoon were rated as less viscous than when taken through a straw. The constriction in the straw had no effect on either the amount ingested or on any of the sensory attributes. This study demonstrates that resistance to sucking does not affect perception of thickness.

Notes: Roasted peanuts were stored at 20 treatment combinations of water activities (0.33, 0.44, 0.54, 0.67, 0.75) and temperatures (23, 30, 35, 40C), and evaluated after storing for 0, 20, 40, 60, 80, 100, and 110% of estimated shelf life, ranging from 0 to 91 days. Regression models indicated that increasing storage time and storage water activity resulted in decreasing crispness, crunchiness, hardness, roasted peanutty, sweet aromatic, salty, bitter and sweet attributes and increasing fracturability, chewiness, tooth packing and cardboard flavor. Storage temperature did not contribute to regression models of textural properties of roasted peanuts. Increasing storage temperature resulted in a faster rate of decrease for roasted peanutty and faster rate of increase of cardboard flavor. Roasted peanuts stored between 0.33 and 0.41 a w at 23C are predicted to have the least change in sensory properties after 68 and 91 days respectively.

Notes: We investigated whether the perception of the crispness and staleness of potato chips can be affected by modifying the sounds produced during the biting action. Participants in our study bit into potato chips with their front teeth while rating either their crispness or freshness using a computer-based visual analog scale. The results demonstrate that the perception of both the crispness and staleness was systematically altered by varying the loudness and/or frequency composition of the auditory feedback elicited during the biting action. The potato chips were perceived as being both crisper and fresher when either the overall sound level was increased, or when just the high frequency sounds (in the range of 2 kHz−20 kHz) were selectively amplified. These results highlight the significant role that auditory cues can play in modulating the perception and evaluation of foodstuffs (despite the fact that consumers are often unaware of the influence of such auditory cues). The paradigm reported here also provides a novel empiric methodology for assessing such multisensory contributions to food perception.

Notes: The effect of three pleasant fine women's fragrances on the mood of women was assessed. The three fragrances had similar hedonic values but different sensory characteristics. The fragrances elicited nonspecific effects on mood. All three fragrances’ impact on mood was influenced by initial mood and lasted up to three hours, i.e., as long as the scent was still noticeable. Trends revealed that all three fragrances decreased negative affect and increased vigor. Also, independent of their hedonic value, the three fragrances elicited specific mood patterns. With the “woody citrus coniferous” scent, respondents experienced higher hostility and tension. The “floral chypre citrus” fragrance decreased anger and confusion. The “floral woody” scent had the lowest Total Mood Disturbance score and lowered depression, tension and confusion.

Notes: Amarone is a red wine produced with a technique already known to the ancient Romans, based on withered grapes and a slow fermentation. Amarone is now considered to be one of the great Italian wines and it is exported all over the world. Sensory and compositional analyses were carried out to describe the profile of the Amarone over four vintages (1998–2001) at different stages of aging. All wines were analyzed for titratable and volatile acidity, pH, alcohol content, residual sugars, t-resveratrol, antioxidant power, color, tonality, total polyphenols and anthocyans. Twelve trained people evaluated aroma and flavor attributes of all the wines in triplicate in a sensory session. Principal component analysis (PCA) of chemical, physical and sensory data for Amarone wine allowed us to highlight the differences between the wines. In general, the 1998 and 1999 vintages had higher aroma and flavor of Wood and Alcohol than the 2000 vintage, which, on the other hand, had higher aroma and flavor of Liquor and Cherry in alcohol. In contrast, the younger wine (2001 vintage) was the more Astringent and had many polyphenols and anthocyans, distinctive features for a red wine that has been in the cask for a short time.

Notes: The structures of the Ca2+-ATPase (SERCA1a) have been determined for five different states by X-ray crystallography. Detailed comparison of the structures in the Ca2+ bound form and unbound (but thapsigargin bound) form reveals that very large rearrangements of the transmembrane helices take place accompanying Ca2+ dissociation and binding and that they are mechanically linked with equally large movements of the cytoplasmic domains. The meanings of the rearrangements of the transmembrane helices and those of the cytoplasmic domains as well as the mechanistic roles of phosphorylation are now becoming clear. Furthermore, the roles of critical amino acid residues identified by extensive mutagenesis studies are becoming evident in terms of atomic structure.

Notes: The bacterial TolC protein plays a common role in the expulsion of diverse molecules, which include protein toxins and antibacterial drugs, from the cell. TolC is a trimeric 12-stranded alpha/beta barrel, comprising an alpha-helical trans-periplasmic tunnel embedded in the outer membrane by a contiguous beta-barrel channel. This structure establishes a 140 A long single pore fundamentally different to other membrane proteins and presents an exit duct to substrates, large and small, engaged at specific inner membrane translocases. TolC is open to the outside medium but is closed at its periplasmic entrance. When TolC is recruited by a substrate-laden translocase, the entrance is opened to allow substrate passage through a contiguous machinery spanning the entire cell envelope, from the cytosol to the external environment. Transition to the transient open state is achieved by an iris-like mechanism in which entrance alpha-helices undergo an untwisting realignment, thought to be stabilized by interaction with periplasmic helices of the translocase. TolC family proteins are ubiquitous among gram-negative bacteria, and the conserved entrance aperture presents a possible cheomotherapeutic target in multidrug-resistant pathogens.

Notes: Telomeres are essential for genome stability in all eukaryotes. Changes in telomere functions and the associated chromosomal abnormalities have been implicated in human aging and cancer. Telomeres are composed of repetitive sequences that can be maintained by telomerase, a complex containing a reverse transcriptase (hTERT in humans and Est2 in budding yeast), a template RNA (hTERC in humans and Tlc1 in yeast), and accessory factors (the Est1 proteins and dyskerin in humans and Est1, Est3, and Sm proteins in budding yeast). Telomerase is regulated in cis by proteins that bind to telomeric DNA. This regulation can take place at the telomere terminus, involving single-stranded DNA-binding proteins (POT1 in humans and Cdc13 in budding yeast), which have been proposed to contribute to the recruitment of telomerase and may also regulate the extent or frequency of elongation. In addition, proteins that bind along the length of the telomere (TRF1/TIN2/tankyrase in humans and Rap1/Rif1/Rif2 in budding yeast) are part of a negative feedback loop that regulates telomere length. Here we discuss the details of telomerase and its regulation by the telomere.

Notes: Protein S-palmitoylation is the thioester linkage of long-chain fatty acids to cysteine residues in proteins. Addition of palmitate to proteins facilitates their membrane interactions and trafficking, and it modulates protein-protein interactions and enzyme activity. The reversibility of palmitoylation makes it an attractive mechanism for regulating protein activity, and this feature has generated intensive investigation of this modification. The regulation of palmitoylation occurs through the actions of protein acyltransferases and protein acylthioesterases. Identification of the protein acyltransferases Erf2/Erf4 and Akr1 in yeast has provided new insight into the palmitoylation reaction. These molecules work in concert with thioesterases, such as acyl-protein thioesterase 1, to regulate the palmitoylation status of numerous signaling molecules, ultimately influencing their function. This review discusses the function and regulation of protein palmitoylation, focusing on intracellular proteins that participate in cell signaling or protein trafficking.

Notes: Cytochrome P450 enzymes are heme-containing monooxygenases that are named after an absorption band at 450 nm when complexed with carbon monoxide. They catalyze a wide variety of reactions and are unique in their ability to hydroxylate nonactivated hydrocarbons. P450 enzymes are involved in numerous biological processes, which include the biosynthesis of lipids, steroids, antibiotics, and the degradation of xenobiotics. In line with the variety of reactions catalyzed, the size of their substrates varies significantly. Some P450s have open active sites (e.g., BM3), and some have shielded active sites that open only transiently (e.g., P450cam), whereas others bind the substrate only when attached to carrier proteins (e.g., Oxy proteins). Structural aspects of both organic and gaseous ligand binding and electron transfer are described.

Notes: ATP-binding cassette (ABC) transporters couple ATP hydrolysis to the uptake and efflux of solutes across the cell membrane in bacteria and eukaryotic cells. In bacteria, these transporters are important virulence factors because they play roles in nutrient uptake and in secretion of toxins and antimicrobial agents. In humans, many diseases, such as cystic fibrosis, hyperinsulinemia, and macular dystrophy, are traced to defects in ABC transporters. Recent advances in structural determination and functional analysis of bacterial ABC transporters, reviewed herein, have greatly increased our understanding of the molecular mechanism of transport in this transport superfamily.

Notes: Just as Darwinian evolution in nature has led to the development of many sophisticated enzymes, Darwinian evolution in vitro has proven to be a powerful approach for obtaining similar results in the laboratory. This review focuses on the development of nucleic acid enzymes starting from a population of random-sequence RNA or DNA molecules. In order to illustrate the principles and practice of in vitro evolution, two especially well-studied categories of catalytic nucleic acid are considered: RNA enzymes that catalyze the template-directed ligation of RNA and DNA enzymes that catalyze the cleavage of RNA. The former reaction, which involves attack of a 2'- or 3'-hydroxyl on the alpha-phosphate of a 5'-triphosphate, is more difficult. It requires a comparatively larger catalytic motif, containing more nucleotides than can be sampled exhaustively within a starting population of random-sequence RNAs. The latter reaction involves deprotonation of the 2'-hydroxyl adjacent to the cleavage site, resulting in cleaved products that bear a 2',3'-cyclic phosphate and 5'-hydroxyl. The difficulty of this reaction, and therefore the complexity of the corresponding DNA enzyme, depends on whether a catalytic cofactor, such as a divalent metal cation or small molecule, is present in the reaction mixture.

Notes: The genetic code is established by the aminoacylation of transfer RNA, reactions in which each amino acid is linked to its cognate tRNA that, in turn, harbors the nucleotide triplet (anticodon) specific to the amino acid. The accuracy of aminoacylation is essential for building and maintaining the universal tree of life. The ability to manipulate and expand the code holds promise for the development of new methods to create novel proteins and to understand the origins of life. Recent efforts to manipulate the genetic code have fulfilled much of this potential. These efforts have led to incorporation of nonnatural amino acids into proteins for a variety of applications and have demonstrated the plausibility of specific proposals for early evolution of the code.

Notes: DNA damage is a relatively common event in the life of a cell and may lead to mutation, cancer, and cellular or organismic death. Damage to DNA induces several cellular responses that enable the cell either to eliminate or cope with the damage or to activate a programmed cell death process, presumably to eliminate cells with potentially catastrophic mutations. These DNA damage response reactions include: (a) removal of DNA damage and restoration of the continuity of the DNA duplex; (b) activation of a DNA damage checkpoint, which arrests cell cycle progression so as to allow for repair and prevention of the transmission of damaged or incompletely replicated chromosomes; (c) transcriptional response, which causes changes in the transcription profile that may be beneficial to the cell; and (d) apoptosis, which eliminates heavily damaged or seriously deregulated cells. DNA repair mechanisms include direct repair, base excision repair, nucleotide excision repair, double-strand break repair, and cross-link repair. The DNA damage checkpoints employ damage sensor proteins, such as ATM, ATR, the Rad17-RFC complex, and the 9-1-1 complex, to detect DNA damage and to initiate signal transduction cascades that employ Chk1 and Chk2 Ser/Thr kinases and Cdc25 phosphatases. The signal transducers activate p53 and inactivate cyclin-dependent kinases to inhibit cell cycle progression from G1 to S (the G1/S checkpoint), DNA replication (the intra-S checkpoint), or G2 to mitosis (the G2/M checkpoint). In this review the molecular mechanisms of DNA repair and the DNA damage checkpoints in mammalian cells are analyzed.

Notes: Opioid receptors belong to the large superfamily of seven transmembrane-spanning (7TM) G protein-coupled receptors (GPCRs). As a class, GPCRs are of fundamental physiological importance mediating the actions of the majority of known neurotransmitters and hormones. Opioid receptors are particularly intriguing members of this receptor family. They are activated both by endogenously produced opioid peptides and by exogenously administered opiate compounds, some of which are not only among the most effective analgesics known but also highly addictive drugs of abuse. A fundamental question in addiction biology is why exogenous opioid drugs, such as morphine and heroin, have a high liability for inducing tolerance, dependence, and addiction. This review focuses on many aspects of opioid receptors with the aim of gaining a greater insight into mechanisms of opioid tolerance and dependence.

Notes: Mechanical processes are involved in nearly every facet of the cell cycle. Mechanical forces are generated in the cell during processes as diverse as chromosomal segregation, replication, transcription, translation, translocation of proteins across membranes, cell locomotion, and catalyzed protein and nucleic acid folding and unfolding, among others. Because force is a product of all these reactions, biochemists are beginning to directly apply external forces to these processes to alter the extent or even the fate of these reactions hoping to reveal their underlying molecular mechanisms. This review provides the conceptual framework to understand the role of mechanical force in biochemistry.

Notes: The fastest simple, kinetically two-state protein folds a million times more rapidly than the slowest. Here we review many recent theories of protein folding kinetics in terms of their ability to qualitatively rationalize, if not quantitatively predict, this fundamental experimental observation.

Notes: I had the good luck to start research at the dawn of molecular biology when it was possible to ask fundamental questions about the nature of the nucleic acids and how information is transferred in living systems. The search for answers led me into many different areas, often with the question of how molecular structure leads to biological function. Early work in this period provided some of the roots supporting the current explosive developments in life sciences. Here I give a brief account of my development, describe some contributions, and provide a hint of the exhilaration in discovering new things. Most of all, I had the good fortune to have inspiring teachers, stimulating colleagues, and excellent students.

Notes: Eukaryotic mRNAs are primarily degraded by removal of the 3' poly(A) tail, followed either by cleavage of the 5' cap structure (decapping) and 5'-〉3' exonucleolytic digestion, or by 3' to 5' degradation. mRNA decapping represents a critical step in turnover because this permits the degradation of the mRNA and is a site of numerous control inputs. Recent analyses suggest decapping of an mRNA consists of four central and related events. These include removal, or inactivation, of the poly(A) tail as an inhibitor of decapping, exit from active translation, assembly of a decapping complex on the mRNA, and sequestration of the mRNA into discrete cytoplasmic foci where decapping can occur. Each of these steps is a demonstrated, or potential, site for the regulation of mRNA decay. We discuss the decapping process in the light of these central properties, which also suggest fundamental aspects of cytoplasmic mRNA physiology that connect decapping, translation, and storage of mRNA.

Notes: The prion hypothesis proposes that proteins can act as infectious agents. Originally formulated to explain transmissible spongiform encephalopathies (TSEs), the prion hypothesis has been extended with the finding that several non-Mendelian traits in fungi are due to heritable changes in protein conformation, which may in some cases be beneficial. Although much remains to be learned about the specific role of cellular cofactors, mechanistic parallels between the mammalian and yeast prion phenomena point to universal features of conformation-based infection and inheritance involving propagation of ordered beta-sheet-rich protein aggregates commonly referred to as amyloid. Here we focus on two such features and discuss recent efforts to explain them in terms of the physical properties of amyloid-like aggregates. The first is prion strains, wherein chemically identical infectious particles cause distinct phenotypes. The second is barriers that often prohibit prion transmission between different species. There is increasing evidence suggesting that both of these can be manifestations of the same phenomenon: the ability of a protein to misfold into multiple self-propagating conformations. Even single mutations can change the spectrum of favored misfolded conformations. In turn, changes in amyloid conformation can shift the specificity of propagation and alter strain phenotypes. This model helps explain many common and otherwise puzzling features of prion inheritance as well as aspects of noninfectious diseases involving toxic misfolded proteins.

Notes: Pyridoxal phosphate (PLP)-dependent enzymes are unrivaled in the diversity of reactions that they catalyze. New structural data have paved the way for targeted mutagenesis and mechanistic studies and have provided a framework for interpretation of those results. Together, these complementary approaches yield new insight into function, particularly in understanding the origins of substrate and reaction type specificity. The combination of new sequences and structures enables better reconstruction of their evolutionary heritage and illuminates unrecognized similarities within this diverse group of enzymes. The important metabolic roles of many PLP-dependent enzymes drive efforts to design specific inhibitors, which are now guided by the availability of comprehensive structural and functional databases. Better understanding of the function of this important group of enzymes is crucial not only for inhibitor design, but also for the design of improved protein-based catalysts.

Notes: The superfamily of intermediate filament (IF) proteins contains at least 65 distinct proteins in man, which all assemble into ~10 nm wide filaments and are principal structural elements both in the nucleus and the cytoplasm with essential scaffolding functions in metazoan cells. At present, we have only circumstantial evidence of how the highly divergent primary sequences of IF proteins lead to the formation of seemingly similar polymers and how this correlates with their function in individual cells and tissues. Point mutations in IF proteins, particularly in lamins, have been demonstrated to lead to severe, inheritable multi-systemic diseases, thus underlining their importance at several functional levels. Recent structural work has now begun to shed some light onto the complex fine tuning of structure and function in these fibrous, coiled coil forming multidomain proteins and their contribution to cellular physiology and gene regulation.

Notes: Protein S-palmitoylation is the thioester linkage of long-chain fatty acids to cysteine residues in proteins. Addition of palmitate to proteins facilitates their membrane interactions and trafficking, and it modulates protein-protein interactions and enzyme activity. The reversibility of palmitoylation makes it an attractive mechanism for regulating protein activity, and this feature has generated intensive investigation of this modification. The regulation of palmitoylation occurs through the actions of protein acyltransferases and protein acylthioesterases. Identification of the protein acyltransferases Erf2/Erf4 and Akr1 in yeast has provided new insight into the palmitoylation reaction. These molecules work in concert with thioesterases, such as acyl-protein thioesterase 1, to regulate the palmitoylation status of numerous signaling molecules, ultimately influencing their function. This review discusses the function and regulation of protein palmitoylation, focusing on intracellular proteins that participate in cell signaling or protein trafficking.

Notes: Great advances have been made in the past three decades in understanding the molecular mechanics underlying protein synthesis in bacteria, but our understanding of the corresponding events in eukaryotic organisms is only beginning to catch up. In this review we describe the current state of our knowledge and ignorance of the molecular mechanics underlying eukaryotic translation. We discuss the mechanisms conserved across the three kingdoms of life as well as the important divergences that have taken place in the pathway.

Notes: The signal recognition particle (SRP) directs integral membrane and secretory proteins to the cellular protein translocation machinery during translation. The SRP is an evolutionarily conserved RNA-protein complex whose activities are regulated by GTP hydrolysis. Recent structural investigations of SRP functional domains and interactions provide new insights into the mechanisms of SRP activity in all cells, leading toward a comprehensive understanding of protein trafficking by this elegant pathway.

Notes: From a process involved in cell wall synthesis in archaea and some bacteria, N-linked glycosylation has evolved into the most common covalent protein modification in eukaryotic cells. The sugars are added to nascent proteins as a core oligosaccharide unit, which is then extensively modified by removal and addition of sugar residues in the endoplasmic reticulum (ER) and the Golgi complex. It has become evident that the modifications that take place in the ER reflect a spectrum of functions related to glycoprotein folding, quality control, sorting, degradation, and secretion. The glycans not only promote folding directly by stabilizing polypeptide structures but also indirectly by serving as recognition "tags" that allow glycoproteins to interact with a variety of lectins, glycosidases, and glycosyltranferases. Some of these (such as glucosidases I and II, calnexin, and calreticulin) have a central role in folding and retention, while others (such as alpha-mannosidases and EDEM) target unsalvageable glycoproteins for ER-associated degradation. Each residue in the core oligosaccharide and each step in the modification program have significance for the fate of newly synthesized glycoproteins.

Notes: Great advances have been made in the past three decades in understanding the molecular mechanics underlying protein synthesis in bacteria, but our understanding of the corresponding events in eukaryotic organisms is only beginning to catch up. In this review we describe the current state of our knowledge and ignorance of the molecular mechanics underlying eukaryotic translation. We discuss the mechanisms conserved across the three kingdoms of life as well as the important divergences that have taken place in the pathway.

Notes: Just as Darwinian evolution in nature has led to the development of many sophisticated enzymes, Darwinian evolution in vitro has proven to be a powerful approach for obtaining similar results in the laboratory. This review focuses on the development of nucleic acid enzymes starting from a population of random-sequence RNA or DNA molecules. In order to illustrate the principles and practice of in vitro evolution, two especially well-studied categories of catalytic nucleic acid are considered: RNA enzymes that catalyze the template-directed ligation of RNA and DNA enzymes that catalyze the cleavage of RNA. The former reaction, which involves attack of a 2'- or 3'-hydroxyl on the alpha-phosphate of a 5'-triphosphate, is more difficult. It requires a comparatively larger catalytic motif, containing more nucleotides than can be sampled exhaustively within a starting population of random-sequence RNAs. The latter reaction involves deprotonation of the 2'-hydroxyl adjacent to the cleavage site, resulting in cleaved products that bear a 2',3'-cyclic phosphate and 5'-hydroxyl. The difficulty of this reaction, and therefore the complexity of the corresponding DNA enzyme, depends on whether a catalytic cofactor, such as a divalent metal cation or small molecule, is present in the reaction mixture.

Notes: Secreted signaling proteins function in a diverse array of essential patterning events during metazoan development, ranging from embryonic segmentation in insects to neural tube differentiation in vertebrates. These proteins generally are expressed in a localized manner, and they may elicit distinct concentration-dependent responses in the cells of surrounding tissues and structures, thus functioning as morphogens that specify the pattern of cellular responses by their tissue distribution. Given the importance of signal distribution, it is notable that the Hedgehog (Hh) and Wnt proteins, two of the most important families of such signals, are known to be covalently modified by lipid moieties, the membrane-anchoring properties of which are not consistent with passive models of protein mobilization within tissues. This review focuses on the mechanisms underlying biogenesis of the mature Hh proteins, which are dually modified by cholesteryl and palmitoyl adducts, as well as on the relationship between Hh proteins and the self-splicing proteins (i.e., proteins containing inteins) and the Hh-like proteins of nematodes. We further discuss the cellular mechanisms that have evolved to handle lipidated Hh proteins in the spatial deployment of the signal in developing tissues and the more recent findings that implicate palmitate modification as an important feature of Wnt signaling proteins.

Notes: Researchers have long predicted that complex carbohydrates on cell surfaces would play important roles in developmental processes because of the observation that specific carbohydrate structures appear in specific spatial and temporal patterns throughout development. The astounding number and complexity of carbohydrate structures on cell surfaces added support to the concept that glycoconjugates would function in cellular communication during development. Although the structural complexity inherent in glycoconjugates has slowed advances in our understanding of their functions, the complete sequencing of the genomes of organisms classically used in developmental studies (e.g., mice, Drosophila melanogaster, and Caenorhabditis elegans) has led to demonstration of essential functions for a number of glycoconjugates in developmental processes. Here we present a review of recent studies analyzing function of a variety of glycoconjugates (O-fucose, O-mannose, N-glycans, mucin-type O-glycans, proteoglycans, glycosphingolipids), focusing on lessons learned from human disease and genetic studies in mice, D. melanogaster, and C. elegans.

Notes: The prion hypothesis proposes that proteins can act as infectious agents. Originally formulated to explain transmissible spongiform encephalopathies (TSEs), the prion hypothesis has been extended with the finding that several non-Mendelian traits in fungi are due to heritable changes in protein conformation, which may in some cases be beneficial. Although much remains to be learned about the specific role of cellular cofactors, mechanistic parallels between the mammalian and yeast prion phenomena point to universal features of conformation-based infection and inheritance involving propagation of ordered beta-sheet-rich protein aggregates commonly referred to as amyloid. Here we focus on two such features and discuss recent efforts to explain them in terms of the physical properties of amyloid-like aggregates. The first is prion strains, wherein chemically identical infectious particles cause distinct phenotypes. The second is barriers that often prohibit prion transmission between different species. There is increasing evidence suggesting that both of these can be manifestations of the same phenomenon: the ability of a protein to misfold into multiple self-propagating conformations. Even single mutations can change the spectrum of favored misfolded conformations. In turn, changes in amyloid conformation can shift the specificity of propagation and alter strain phenotypes. This model helps explain many common and otherwise puzzling features of prion inheritance as well as aspects of noninfectious diseases involving toxic misfolded proteins.

Notes: Eukaryotic mRNAs are primarily degraded by removal of the 3' poly(A) tail, followed either by cleavage of the 5' cap structure (decapping) and 5'-〉3' exonucleolytic digestion, or by 3' to 5' degradation. mRNA decapping represents a critical step in turnover because this permits the degradation of the mRNA and is a site of numerous control inputs. Recent analyses suggest decapping of an mRNA consists of four central and related events. These include removal, or inactivation, of the poly(A) tail as an inhibitor of decapping, exit from active translation, assembly of a decapping complex on the mRNA, and sequestration of the mRNA into discrete cytoplasmic foci where decapping can occur. Each of these steps is a demonstrated, or potential, site for the regulation of mRNA decay. We discuss the decapping process in the light of these central properties, which also suggest fundamental aspects of cytoplasmic mRNA physiology that connect decapping, translation, and storage of mRNA.

Notes: Cytochrome P450 enzymes are heme-containing monooxygenases that are named after an absorption band at 450 nm when complexed with carbon monoxide. They catalyze a wide variety of reactions and are unique in their ability to hydroxylate nonactivated hydrocarbons. P450 enzymes are involved in numerous biological processes, which include the biosynthesis of lipids, steroids, antibiotics, and the degradation of xenobiotics. In line with the variety of reactions catalyzed, the size of their substrates varies significantly. Some P450s have open active sites (e.g., BM3), and some have shielded active sites that open only transiently (e.g., P450cam), whereas others bind the substrate only when attached to carrier proteins (e.g., Oxy proteins). Structural aspects of both organic and gaseous ligand binding and electron transfer are described.

Notes: Recent applications of neutron reflectometry to the study of wet interfaces are described. An outline is given of the basic principles that allow the techniques to determine composition and structure in a variety of situations. These are the adsorption of surfactant molecules at air/liquid and solid/liquid interfaces, the shape of the segment-density profiles of different types of polymer, including block copolymers and polyelectrolytes, adsorption in mixed surfactant and polymer/surfactant systems, and interfacial systems of biophysical interest.

Notes: Charge transport at conjugated polymer interfaces with metals and inorganic semiconductors is reviewed. Experiments on the equilibrium properties and DC current-voltage behavior of four specific classes of interfaces-metal-doped conjugated polymer, inorganic semiconductor-doped conjugated polymer, metal-intrinsic conjugated polymer, and metal-intrinsic conjugated polymer/electrolyte-are discussed. To facilitate this discussion, classic models of equilibration at ideal interfaces between electronic conductors and free-electron transport are introduced and their limitations discussed. Particular emphasis is placed on the charge distributions and interfacial potential profiles expected at various types of electroactive interfaces.

Notes: Recent progress in the development of semiclassical methods to describe quantum effects in molecular dynamics is reviewed. Focusing on rigorous semiclassical methods that are based on the initial-value representation of the semiclassical propagator, we discuss several promising schemes that have been developed in the past few years to extend the applicability of semiclassical approaches to complex molecular systems. In particular, integral-filtering techniques and forward-backward methods are surveyed. Furthermore, recently proposed approaches that allow the semiclassical description of nonadiabatic molecular dynamics are discussed. The potential and efficiency of these methods is illustrated by selected applications.

Notes: Time-dependent density functional theory (TDDFT) can be viewed as an exact reformulation of time-dependent quantum mechanics, where the fundamental variable is no longer the many-body wave function but the density. This time-dependent density is determined by solving an auxiliary set of noninteracting Schrodinger equations, the Kohn-Sham equations. The nontrivial part of the many-body interaction is contained in the so-called exchange-correlation potential, for which reasonably good approximations exist. Within TDDFT two regimes can be distinguished: (a) If the external time-dependent potential is "small," the complete numerical solution of the time-dependent Kohn-Sham equations can be avoided by the use of linear response theory. This is the case, e.g., for the calculation of photoabsorption spectra. (b) For a "strong" external potential, a full solution of the time-dependent Kohn-Sham equations is in order. This situation is encountered, for instance, when matter interacts with intense laser fields. In this review we give an overview of TDDFT from its theoretical foundations to several applications both in the linear and in the nonlinear regime.

Notes: The review describes the studies of the magneto-optical properties of II-VI and III-V semiconductor nanocrystals (NCs) capped with organic or inorganic epitaxial shells. The investigations focused on the chemical identification of localization sites (core, shell, or interface) of photogenerated carriers in spherical NCs and elucidated the influence of the surface/interface quality on the optical properties of the materials. Optically detected magnetic resonance (ODMR) spectroscopy was used for the study of the proposed physical properties. The ODMR method provides the means to identify the surface/interface sites and correlate them with specific optical transition. In addition, this method reveals information about the spin multiplicity of band edge and trapped states and the electron-hole exchange interaction, determines the spectroscopic g-factors, distinguishes between the radiative and nonradiative characteristic of a trapping site, and evaluates the spin-lattice relaxation times.

Notes: This review describes some of the biochemical and toxicological properties of CYP2E1, especially as it relates to alcohol metabolism and toxicity and the establishment of human hepatoma HepG2 cell lines that overexpress human CYP2E1. Ethanol, polyunsaturated fatty acids, and iron were found to be cytotoxic in HepG2 cells that overexpress CYP2E1. GSH appears to be essential in protecting HepG2 cells against the CYP2E1-dependent cytotoxicity, and GSH levels were elevated owing to a twofold increase in activity and expression of glutamate cysteine ligase. We suggest that this up-regulation of GSH synthesis was an adaptive response to attenuate CYP2E1-dependent oxidative stress and toxicity. Induction of a state of oxidative stress appears to play a central role in the CYP2E1-dependent cytotoxicity. Mitochondrial membrane potential decreased in the CYP2E1-expressing HepG2 cells, and this decrease shared similar characteristics with the developing toxicity. Alcohol-dependent liver injury is likely to be a multifactorial process involving several mechanisms. We believe that the linkage between CYP2E1-dependent oxidative stress, mitochondrial injury, and GSH homeostasis contribute to the toxic actions of ethanol on the liver.

Notes: Pharmacokinetic (PK) and pharmacodynamic (PD) studies have proven to be powerful and instructive tools, particularly in elucidating important aspects of human pharmacology. Nevertheless, they remain imperfect tools in that they only allow researchers to indirectly extrapolate, through computational modeling, the dynamic processes of drug action. Furthermore, neither tool alone provides a complete nor necessarily relevant picture of drug action. This review explores the utility and applications of PK and PD in the study of drugs, provides examples of lessons learned from their application to studies of human pharmacology, points out some of their limitations, and advances the thesis that these tools ideally should be employed together in an integrated approach. As we continue to apply these tools across the continuum of age and disease, they provide a powerful means to enhance our understanding of drug action, drug interactions, and intrinsic host factors that influence pharmacologic response.

Notes: Chemical carcinogenesis follows a multistep process involving both mutation and increased cell proliferation. Oxidative stress can occur through overproduction of reactive oxygen and nitrogen species through either endogenous or exogenous insults. Important to carcinogenesis, the unregulated or prolonged production of cellular oxidants has been linked to mutation (induced by oxidant-induced DNA damage), as well as modification of gene expression. In particular, signal transduction pathways, including AP-1 and NFkappaB, are known to be activated by reactive oxygen species, and they lead to the transcription of genes involved in cell growth regulatory pathways. This review examines the evidence of cellular oxidants' involvement in the carcinogenesis process, and focuses on the mechanisms for production, cellular damage produced, and the role of signaling cascades by reactive oxygen species.

Notes: Numerous lines of evidence demonstrate that calpains, a family of 14 Ca2+-activated neutral cysteine proteases, are involved in oncotic cell death in a variety of models. At this time, the biochemistry of most calpains and the specific roles of different calpains in physiology and pathology remain to be determined. A number of calpain substrates have been identified in cellular systems, including cytoskeletal proteins, and recent studies suggest that calpains mediate the increase in plasma membrane permeability to ions and the progressive breakdown of the plasma membrane observed in oncosis through the proteolysis of cystokeletal and plasma membrane proteins. Further, a number of reports provide evidence that the mitochondrial dysfunction observed in oncosis may be mediated by a mitochondrial calpain of unknown identity. Finally, a number of diverse calpain inhibitors have been developed that show cytoprotective properties in cellular systems and in vivo following diverse insults. It is suggested that future research be directed toward elucidation of the role(s) of specific calpain isozymes in physiological and pathological conditions; identifying and linking specific calpain substrates with altered cellular functions; and developing cell-permeable, potent, isozyme-selective calpain inhibitors.

Notes: GABAA receptors are molecular substrates for the regulation of vigilance, anxiety, muscle tension, epileptogenic activity, and memory functions, and the enhancement of GABAA receptor-mediated fast synaptic inhibition is the basis for the pharmacotherapy of various neurological and psychiatric disorders. Two kinds of GABAA receptor-targeted mutant mice have been generated: (a) knockout mice that lack individual GABAA receptor subunits (alpha1, alpha5, alpha6, beta2, beta3, gamma2, delta, and rho1) and (b) knockin mice that carry point mutations affecting the action of modulatory drugs [alpha1(H101R), alpha2(H101R), alpha3(H126R), alpha5(H105R), and beta3(N265M)]. Whereas the knockout mice have provided information primarily with respect to the regulation of subunit gene transcription, receptor assembly, and some physiological functions of individual receptor subtypes, the point-mutated knockin mice in which specific GABAA receptor subtypes are insensitive to diazepam or some general anesthetics have revealed the specific contribution of individual receptor subtypes to the pharmacological spectrum of diazepam and general anesthetics.

Notes: G protein-coupled receptors (GPCRs) modulate diverse physiological and behavioral signaling pathways by virtue of changes in receptor activation and inactivation states. Functional changes in receptor properties include dynamic interactions with regulatory molecules and trafficking to various cellular compartments at various stages of the life cycle of a GPCR. This review focuses on trafficking of GPCRs to the cell surface, stabilization there, and agonist-regulated turnover. GPCR interactions with a variety of newly revealed partners also are reviewed with the intention of provoking further analysis of the relevance of these interactions in GPCR trafficking, signaling, or both. The disease consequences of mislocalization of GPCRs also are described.

Notes: Cytochrome P450 (CYP) and UDP-glucuronosyltransferase (UGT), which both exist as enzyme "superfamilies," are together responsible for the metabolism of most hepatically cleared drugs. There is currently intense interest in the development of techniques that permit identification of the CYP and UGT isoform(s) involved in the metabolism of a newly discovered drug, and hence prediction of factors likely to alter elimination in vivo. In addition, the quantitative scaling of kinetic parameters for a metabolic pathway assumes importance for identifying newly discovered drugs with undesirable in vivo pharmacokinetic properties. Although qualitative and quantitative in vitro-in vivo correlation based on data generated using human liver tissue or recombinant enzymes have been applied successfully to many drugs eliminated by CYP, these strategies have proved less definitive for glucuronidated compounds. Computational (in silico) modeling techniques that potentially provide a facile and economic alternative to the in vitro methods are now emerging. This review assesses the utility of in vitro and in silico approaches for the qualitative and quantitative prediction of drug glucuronidation parameters and the challenges facing the development of generalizable models.

Notes: The developing central nervous system (CNS) is more vulnerable to injury than the adult one. Although a great deal of research has been devoted to subtle effects of developmental exposure, such as neurobehavioral changes, this review instead focuses on a number of chemicals that have been shown, in several experimental models as well as humans, to cause morphological changes in the developing nervous system. Chemicals that are discussed include methylmercury (MeHg), lead (Pb), antiepileptic drugs, and ethanol. Additionally, the issue of silent neurotoxicity, i.e., persistent morphological and/or biochemical injury that remains clinically unapparent until later in life, is discussed.

Notes: The epidermal growth factor (EGF)-related peptides bind the ErbB receptors, inducing the formation of different homo- and heterodimers. Receptor dimerization promotes activation of the intrinsic kinase, leading to phosphorylation of specific tyrosines located in the ErbB's cytoplasmic region. These phosphorylated residues serve as docking sites for a variety of signaling molecules whose recruitment stimulates intracellular signaling cascades, which ultimately control diverse genetic programs. Particular ligand-receptor complexes have essential roles in embryonic development as well as in the adult. Finally, ErbB receptors are being pursued as therapeutic targets because aberrant ErbB activity has been observed in many human cancers. In this review, we discuss these data in more detail, illustrating the importance of tightly regulated ErbB signaling throughout life.

Notes: Reactive (low pKa) cysteine residues in proteins are critical components in redox signaling. A particularly reactive and versatile reversibly oxidized form of cysteine, the sulfenic acid (Cys-SOH), has important roles as a catalytic center in enzymes and as a sensor of oxidative and nitrosative stress in enzymes and transcriptional regulators. Depending on environment, sometimes the sulfenic acid provides a metastable oxidized form, and other times it is a fleeting intermediate giving rise to more stable disulfide, sulfinic acid, or sulfenyl-amide forms.

Notes: New cells are continuously generated from immature proliferating cells throughout adulthood in many organs, thereby contributing to the integrity of the tissue under physiological conditions and to repair following injury. In contrast, repair mechanisms in the adult central nervous system (CNS) have long been thought to be very limited. However, recent findings have clearly demonstrated that in restricted areas of the mammalian brain, new functional neurons are constantly generated from neural stem cells throughout life. Moreover, stem cells with the potential to give rise to new neurons reside in many different regions of the adult CNS. These findings raise the possibility that endogenous neural stem cells can be mobilized to replace dying neurons in neurodegenerative diseases. Indeed, recent reports have provided evidence that, in some injury models, limited neuronal replacement occurs in the CNS. Here, we summarize our current understanding of the mechanisms controlling adult neurogenesis and discuss their implications for the development of new strategies for the treatment of neurodegenerative diseases.

Notes: The importance of reviewing and studying sex-based differences in pharmacologic parameters is demonstrated by the increasing data on gender variation in drug efficacy and toxicity profiles. Sex-based differences in the four major factors that contribute to interindividual pharmacokinetic variability-bioavailability, distribution, metabolism, and elimination-are theorized to stem from variations between men and women in factors such as body weight, plasma volume, gastric emptying time, plasma protein levels, cytochrome P450 activity, drug transporter function, and excretion activity. Sex-determined variations in pharmacodynamics have traditionally been more difficult to study, but a number of recent studies have explored these differences. This review examines the biologic basis of differences in pharmacokinetics and pharmacodynamics between the sexes and summarizes studies that have addressed these differences. As an example, sex-based variation in the efficacy and toxicity of antiretroviral therapy in human immunodeficiency virus (HIV)-infected patients is explored more thoroughly to illustrate some of the factors underlying sex-based differences in drug therapy.