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Abscisic acid stimulates a calcium-dependent protein kinase in grape berry.

It has been demonstrated that calcium plays a central role in mediating abscisic acid (ABA) signaling, but many of the Ca2+-binding sensory proteins as the components of the ABA-signaling pathway remain to be elucidated. Here we identified, characterized, and purified a 58-kD ABA-stimulated calcium-dependent protein kinase from the mesocarp of grape berries (Vitis vinifera x Vitis labrusca), designated ACPK1 (for ABA-stimulated calcium-dependent protein kinase1). ABA stimulates ACPK1 in a dose-dependent manner, and the ACPK1 expression and enzyme activities alter accordantly with the endogenous ABA concentrations during fruit development. The ABA-induced ACPK1 stimulation appears to be transient with a rapid effect in 15 min but also with a slow and steady state of induction after 60 min. ABA acts on ACPK1 indirectly and dependently on in vivo state of the tissues. Two inactive ABA isomers, (-)-2-cis, 4-trans-ABA and 2-trans, 4-trans-(+/-)-ABA, are ineffective for inducing ACPK1 stimulation, revealing that the ABA-induced effect is stereo specific to physiological active (+)-2-cis, 4-trans-ABA. The other phytohormones such as auxin indoleacetic acid, gibberellic acid, synthetic cytokinin N-benzyl-6-aminopurine, and brassinolide are also ineffective in this ACPK1 stimulation. Based on sequencing of the two-dimensional electrophoresis-purified ACPK1, we cloned the ACPK1 gene. The ACPK1 is expressed specifically in grape berry covering a fleshy portion and seeds, and in a developmental stage-dependent manner. We further showed that ACPK1 is localized in both plasma membranes and chloroplasts/plastids and positively regulates plasma membrane H+-ATPase in vitro, suggesting that ACPK1 may be involved in the ABA-signaling pathway.

Abscisic Acid↗

Tryptophan biosynthesis and production of other related compounds from indolepyruvic acid by mixed ruminal bacteria, protozoa, and their mixture in vitro.

Tryptophan (Trp) biosynthesis and the production of other related compounds by mixed ruminal bacteria (B), protozoa (P), and their mixture (BP) in an in vitro system were quantitatively investigated by using 1 mM of indole-3-pyruvic acid (IPA) as substrate. Ruminal microorganisms were anaerobically incubated at 39 degrees C for 12 h. Trp and other related compounds in both the supernatants and the microbial hydrolyzates of the incubation were analyzed by HPLC. As a whole, about 334, 440, and 436 &mgr;M of Trp were produced from IPA in 12 h by B, P, and BP suspensions, respectively. In the B suspension, a greater portion of synthesized Trp (242 &mgr;M) from IPA was accumulated as free form in the medium, whereas a large amount of Trp (92 &mgr;M) was incorporated into cell protein in a 12-h incubation. On the other hand, in the P suspension, a large amount of Trp (475 &mgr;M) from IPA was also found as free form in the supernatant in a 12-h incubation. Protozoa did not incorporate Trp into cell protein, but they liberated endogenous Trp (34 &mgr;M) into the medium. The net productions of Trp from IPA were 344.3 and 447.7 &mgr;mol/g of microbial nitrogen in 12 h by B and P suspensions, respectively. The ability of P to synthesize Trp from IPA was about 30% higher than that of B in 12 h. Trp produced from IPA by B, P, and BP suspensions were simultaneously degraded into its related compounds, and among them, indoleacetic acid (IAA) was a major product found in all microbial suspensions. Productions of IAA were 124, 25, and 99 &mgr;M from IPA in 12 h by B, P, and BP suspensions, respectively. The formation of indolelactic acid (ILA) from IPA was observed for the first time in all microbial suspensions, and it was about 84, 24, and 54 &mgr;M in 12 h by B, P, and BP, respectively. Higher IAA and ILA productions were observed in B when compared with P. A small amount of skatole (SKT) (26 &mgr;M) was produced from IPA in B, whereas a sizable amount of SKT (38 &mgr;M) was found in BP after a 12-h incubation. p-Cresol (CRL) was also produced from IPA by both B (43 &mgr;M) and BP (65 &mgr;M) suspensions in 12 h, and this is also the first discovery to show the formation of CRL from IPA by B and BP suspensions. BP suspension was more active to produce both SKT and CRL from IPA, though P suspension has no ability to produce either SKT or CRL from IPA during a 12-h incubation.

Journal Article↗

Imipramine and desipramine in plasma and spinal fluid: relationship to clinical response and serotonin metabolism.

In a double-blind study of depressed patients treated with imipramine hydrochloride, levels of imipramine and desipramine were measured in plasma and in CSF. Levels of both drugs in CSF were approximately 10% of plasma levels, but the levels in the two body fluids were highly correlated. The levels of both drugs were approximately equal in plasma, but desipramine predominated in CSF (imipramine/desipramine ratio of 0.8). The imipramine-induced alteration in CSF levels of the serotonin metabolite (5-hydroxy-indoleacetic acid [5HIAA]) correlated with imipramine levels but not with desipramine. For the group of patients showing a clear antidepressant response, the mean drug levels were nearly double those of the nonresponder group, a difference that did not quite reach statistical significance in this relatively small sample. The desipramine levels showed no responder-nonresponder difference, while the ratio of imipramine/desipramine was significantly higher among the responders. On the average this particular patient group had relatively low pretreatment levels of 5HIAA in CSF, an observation that may partially account for the relatively low overall response rate to imipramine.

Adult↗

Cortisol in the CSF of depressed and suicidal patients.

Cortisol concentrations in CSF were measured by radioimmunoassay in healthy controls, depressed patients, patients who had attempted suicide but were not depressed, and obsessive-compulsive patients. The factors that contributed most to the variance in CSF cortisol levels were a diagnosis of depression, height, and important life changes during the six months preceding the investigation. Depression was by far the most important factor. The depressed patients had significantly higher CSF cortisol levels than the controls. In obsessive-compulsive and depressed patients treated with clomipramine hydrochloride, the levels were significantly correlated with mean urinary cortisol excretion. Of the three monoamine metabolites measured, only 5-hydroxy-indoleacetic acid level was weakly correlated with CSF cortisol level. This correlation was confined to the depressed patients and could be accounted for by the common correlation with height.

Adjustment Disorders↗

Pathophysiology of hypoalbuminemia associated with carcinoid tumor.

Decreased serum albumin levels are commonly observed in patients with carcinoid tumor, who also show several characteristic clinical and biochemical abnormalities. A large comparative study on a group of 96 carcinoid patients was performed with the purpose of identifying some of the mechanisms leading to hypoalbuminemia in patients with this form of cancer, and thereby to shed light on the cause of hypoalbuminemia of cancer in general. Serum albumin values were compared with a number of clinical parameters (including extent of liver metastases, severity of diarrhea, degree of right heart failure, and extent of gastrointestinal surgery) and of laboratory data (prothrombin time, BSP retention, serum transferrin concentration, hematocrit value, and daily urine excretion of 5-hydroxy-indoleacetic acid). In several patients the gastrointestinal protein loss was assessed by the 51Cr-albumin technique, whereas albumin renewal and distribution were evaluated by the use of 125I-albumin. The data obtained showed that the main factors in determining decreased serum albumin levels in patients with carcinoids are both reduced synthesis and increased loss of the protein. The hepatic synthetic defect appears to be related to a progressive decrease in the number of functioning liver cells; the origin of the gastrointestinal protein loss may be related to the obvious tumor involvement of the gut wall, as well as to the pharmacologically-induced diarrhea. Right heart failure occurring as a result of the carcinoid heart disease may be an additional cause for gastrointestinal protein loss in patients with carcinoid tumor.

Adult↗

Stereoselective disposition of etodolac enantiomers in synovial fluid.

The synovial fluid (SF) uptake of the chiral nonsteroidal anti-inflammatory drug, etodolac, was studied in six arthritic patients, 2 hours (n = 1) or 12 hours (n = 5) after a single 200 mg dose of racemate. Marked stereoselectivity was seen in both SF and plasma; concentrations of pharmacologically inactive R-etodolac were up to 10-fold greater than active S-etodolac. Concentrations of S-etodolac were greater in SF than in plasma (SF:plasma ratio = 1.98 +/- 0.8): No such difference was seen for R-etodolac (SF:plasma = 0.91 +/- 0.3). Considerable concentrations of conjugated enantiomers were present in SF. In vitro equilibrium dialysis studies in drug-spiked samples showed that the unbound fraction of both enantiomers in SF was greater than in plasma; both fluids bound R more extensively than S etodolac. Therapeutically active S-etodolac has greater concentrations in synovial fluid than plasma during the post-distributive phase, which may be of possible clinical relevance.

Aged↗

Plasma and urinary serotonin and 5-HIAA in children treated with lamotrigine for intractable epilepsy.

Alteration of monoamine levels by some antiepileptic drugs (AEDs) was elucidated in this study. Lamotrigine (LTG) is a new AED, acting the sodium-channels. LTG was given as add-on therapy to 16 patients aged 4.5-18 yrs with intractable epilepsy and comedicated with carbamazepine or valproate. An equal group of epileptics with comparable clinical characteristics and treatment served as control. Plasma and urinary (24 h-samples) serotonin and 5-HIAA were determined before onset of LTG therapy and after 2-3 months. HPLC and electrochemical detection was used for the determination of serotonin (5-HT) and 5-hydroxy indoleacetic acid (5-HIAA). No significant effect of LTG on both urinary 5-HT and 5-HIAA levels was found, whereas plasma 5-HT concentrations significantly decreased in comparison with levels before LTG starting and relevant values in controls. This findings was noted in 7/16 children with favourable response to LTG. Increased serotonin catabolism may be result of LTG action.

Adolescent↗

Tryptophan availability and the control of 5-hydroxytryptamine and tryptamine synthesis in human CNS.

The data presented here suggest that control of human brain 5HT synthesis by precursor availability is similar to that in the rat. Plasma tryptophan controls the brain level, although the plasma-brain relationship is modified by other large neutral amino acids in plasma. In normal circumstances brain tryptophan is an important factor controlling the synthesis of 5HT and tryptamine in human brain. However, the elevated brain tryptophan in patients with chronic liver disease does not lead to an increase in the rate of 5HT metabolism. In human brain the rate of tryptamine synthesis is normally aobut 10-20% of the rate of 5HT synthesis. Tryptamine metabolism is more sensitive than 5HT metabolism to changes in brain tryptophan. This is especially apparent after a tryptophan load. Our results suggest that tryptophan administration increases indoleamine function, as well as indoleamine synthesis, in depressed patients. Whether physiological variations in brain tryptophan in normal people are responsible for variations in indoleamine function is an open question.

Animals↗

Single dose pharmacokinetics of sumatriptan in healthy volunteers.

Sumatriptan is classified as a vascular 5HT1 receptor agonist and is effective in the acute treatment of migraine and cluster headache. Sumatriptan is available as an injection for subcutaneous administration and as a tablet for oral administration. The pharmacokinetics of sumatriptan differ depending on the route of administration. The mean subcutaneous bioavailability is 96% compared to 14% for the oral tablet. The lower bioavailability following oral administration is due mainly to presystemic metabolism. The inter-subject variability in plasma sumatriptan concentrations is greater following oral administration and a faster rate of absorption of drug into the systemic circulation is achieved following subcutaneous dosing. The pharmacokinetics of sumatriptan are linear up to a subcutaneous dose of 16 mg. Following oral dosing up to 400 mg, the pharmacokinetics are also linear, with the exception of rate of absorption, as indicated by a dose dependent increase in time to peak concentration. Sumatriptan is a highly cleared compound that is eliminated from the body primarily by metabolism to the pharmacologically inactive indoleacetic acid analogue. Both sumatriptan and its metabolite are excreted in the urine. Although the renal clearance of sumatriptan is only 20% of the total clearance, it exceeds the glomerular filtration rate, indicating that sumatriptan undergoes active renal tubular secretion. Sumatriptan has a large apparent volume of distribution (170 l) and an elimination half-life of 2 h. Oral doses of sumatriptan were administered as a solution of dispersible tablets and subcutaneous dosing was by injection into the arm. In clinical practice, sumatriptan is administered as a film coated tablet or by subcutaneous injection into the thigh.

Adolescent↗

Temporal changes in medial basal hypothalamic catecholamines in male Syrian hamsters exposed to short photoperiod.

The gonadal and accessory organ atrophy following transfer of male hamsters from long (LP) to short photoperiod (SP) is preceded by reduced prolactin secretion and involves reductions in hypothalamic LHRH release and catecholamine turnover. These experiments examined the temporal aspects of changes in medial basal hypothalamic/median eminence (MBH/ME) catecholamine turnover rates in male hamsters undergoing SP-induced gonadal atrophy. Hamsters were sacrificed at three, six, nine and twelve weeks of SP exposure. MBH/ME catecholamines and indoleamines were determined by high performance liquid chromatography coupled with electrochemical detection. Reductions in serum prolactin (PRL) levels and increased MBH/ME dopamine (DA) turnover rates were observed at three and six weeks of SP exposure. Both steady state concentrations and turnover rates of norepinephrine (NE) and DA were depressed after nine and twelve weeks of SP exposure, at which time testicular and accessory organ atrophy had occurred. Serotonin (5-HT) and 5-hydroxy-3-indoleacetic acid (5-HIAA) concentrations were insignificantly changed during the period of SP treatment but the 5-HIAA/5-HT ratio was significantly increased after six weeks of SP exposure. It was concluded that increased MBH/ME DA turnover represents an initial, SP-induced neuroendocrine event. This increase in DA turnover probably contributes to the reduced PRL secretion which precedes, and may play a role in the ensuing gonadal and accessory organ atrophy.

Animals↗

Metabolism of 5-hydroxytryptamine and levels of tricyclic antidepressant drugs in rat brain after acute and chronic treatment.

Because tricyclic antidepressants (TAD) are usually given chronically to patients, both their acute and their chronic effects on 5-hydroxytryptamine (5-HT) metabolism were studied. The probenecid method was used and, in addition to 5-hydroxy-indoleacetic acid (5-HIAA), some other indole compounds in brain were measured. Simultaneously, TAD levels in brain and plasma were determined. Dimethylated as well as monomethylated TADs were administered, both at 10 and 25 mg/kg i.p. Treatment with either 10 mg/kg during 14 days or 25 mg/kg given acutely resulted in a similar brain level of TAD, so any differences found could be attributed to differences in administration schedule. Drug levels in brain and plasma differed considerably after chronic and acute treatments but no major differences in the effect on 5-HIAA level in the brain were found, although accumulation of 5-HIAA following probenecid treatment was mostly lowered after treatment with dimethylated TAD. The TAD level in rat brain was not decisive for the effect on central 5-HT turnover. The monomethylated TAD affected the 5-HT turnover very little, not only acutely but also chronically.

Animals↗

Effect of repeated amiflamine administration on serotonergic and noradrenergic neurotransmission: electrophysiological studies in the rat CNS.

Amiflamine is a selective and reversible inhibitor of monoamine oxidase (MAO) type A which exerts a preferential effect on serotonin (5-HT) catabolism. The present studies were undertaken to compare the effects of repeated administration of amiflamine (2 mg/kg, twice daily) on several aspects of the functioning of the 5-HT and norepinephrine (NE) systems in the rat CNS. The activity of MAO-A and B was assessed in forebrain slices and the whole brain contents of the neurotransmitters and their metabolites were determined by HPLC after 2-, 7- and 21-day treatments. MAO-A was inhibited by about 50% 2 h after the last dose and its activity was back to normal in rats sacrificed 12 h after the last dose. The activity of MAO-B was unaffected two or 12 h after the last dose. Whole brain concentration of 5-HT was increased to a greater degree than that of NE following repeated administration of amiflamine. These increases in 5-HT and NE were accompanied by decreased levels of their respective metabolites 5-hydroxy-indoleacetic acid and 3-methoxy-4-hydroxyphenylethyleneglycol. The firing activity of dorsal raphe 5-HT neurons, but not that of NE neurons, was markedly decreased 2-6 h after the last dose of a 2-day treatment. However, 2-6 h after a 21-day treatment, the firing activity of 5-HT neurons was back to normal, whereas that of NE neurons was decreased by 30%.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗