Kinetics of drug action in disease states. VI. Effect of experimental diabetes on phenobarbital concentrations in rats at onset of loss of righting reflex

To investigate the effect of diabetes on the sensitivity of the central nervous system to the hypnotic action of a barbiturate, studies were conducted on adult female Lewis rats made diabetic by injection of either streptozotocin or alloxan. The animals then received a slow i.v. infusion of phenobarbital (PB) until the onset of a defined pharmacologic effect [loss of righting reflex (LRR)] and the PB concentrations at that time in serum (total and unbound drug), brain and cerebrospinal fluid (CSF) were determined. In the experiments on rats with streptozotocin-induced diabetes, animals not treated with insulin had significantly lower serum concentrations of total PB at onset of LRR than did animals treated with insulin and nondiabetic control rats. Otherwise, there were no significant differences in PB concentrations between untreated diabetic and control animals. Additional experiments on untreated diabetic rats showed that, as in normal rats, the PB concentrations in CSF (but not in serum and brain) at onset of LRR were independent of PB infusion rate over a 10-fold range, indicating that PB equilibrates very rapidly between CSF and receptor sites. Experiments in rats with alloxan-induced diabetes showed no significant differences between untreated diabetic, insulin-treated diabetic, alloxan-nonresponding and nondiabetic control rats with respect to PB concentrations at onset of LRR in serum (total and unbound drug), brain and CSF and in serum protein binding. These results show that the central nervous system response to the hypnotic effect of PB is not significantly affected in two different experimental models of diabetes.     

Aortic root replacement with a freestyle stentless valve for aortitis syndrome with ascending aortic aneurysm and aortic regurgitation.

A 47-year-old woman who had been diagnosed as having aortitis syndrome underwent aortic root replacement for an ascending aortic aneurysm and aortic regurgitation. Because the patient has been treated with steroids for more than 20 years, a Freestyle stentless valve was used to avoid the risk of valve detachment. There were no complications observed during the postoperative course. Although long-term follow-up will be necessary to observe the valve durability, the Freestyle stentless valve seems to be useful for aortic root replacement in patients at high risk of valve detachment due to aortitis syndrome.     

Antidepressants increase glial cell line-derived neurotrophic factor production through monoamine-independent activation of protein tyrosine kinase and extracellular signal-regulated kinase in glial cells

Recent studies show that neuronal and glial plasticity are important for therapeutic action of antidepressants. We previously reported that antidepressants increase glial cell line-derived neurotrophic factor (GDNF) production in rat C6 glioma cells (C6 cells). Here, we found that amitriptyline, a tricyclic antidepressant, increased both GDNF mRNA expression and release, which were selectively and completely inhibited by mitogen-activated protein kinase kinase inhibitors. Indeed, treatment of amitriptyline rapidly increased extracellular signal-regulated kinase (ERK) activity, as well as p38 mitogen-activated protein kinase and c-Jun NH2-terminal kinase activities. Furthermore, different classes of antidepressants also rapidly increased ERK activity. The extent of acute ERK activation and GDNF release were significantly correlated to each other in individual antidepressants, suggesting an important role of acute ERK activation in GDNF production. Furthermore, antidepressants increased the acute ERK activation and GDNF mRNA expression in normal human astrocytes as well as C6 cells. Although 5-hydroxytryptamine (serotonin) (5-HT), but not noradrenaline or dopamine, increased ERK activation and GDNF release via 5-HT2A receptors, ketanserin, a 5-HT2A receptor antagonist, did not have any effect on the amitriptyline-induced ERK activation. Thus, GDNF production by amitriptyline was independent of monoamine. Both of the amitriptyline-induced ERK activation and GDNF mRNA expression were blocked by genistein, a general protein tyrosine kinase (PTK) inhibitor. Actually, we found that amitriptyline acutely increased phosphorylation levels of several phosphotyrosine-containing proteins. Taken together, these findings indicate that ERK activation through PTK regulates antidepressant-induced GDNF production and that the GDNF production in glial cells may be a novel action of the antidepressant, which is independent of monoamine.     

Enhancement of Rho/Rho-kinase system in regulation of vascular smooth muscle contraction in tachycardia-induced heart failure

OBJECTIVE: The Rho/Rho-kinase system regulates Ca(2+) sensitivity in vascular smooth muscle. A new drug, Y-27632, specifically inhibits Rho-kinase and hence decreases the phosphorylation of myosin light chain, thus reducing contraction. Here, we compare the effects of Y-27632 and nifedipine on the vasoconstrictor response of the femoral artery in heart failure. METHODS: Heart failure (HF) was produced by chronic rapid RV pacing (250 bpm, 28 days, six dogs). Indo1-AM was loaded into endothelium-denuded femoral artery segments for measuring intracellular [Ca(2+)]. Tension and changes in intracellular [Ca(2+)] [the change in the ratio (418 nm/468 nm) of Indo1 fluorescence (F(ratio))] were simultaneously measured in Krebs-Ringer solution. RESULTS: In HF: (i) norepinephrine (10 microM) produced greater tension (784+/-52 g/cm(2)) than in control (502+/-64 g/cm(2)) despite a similar increase in F(ratio), indicating increased Ca(2+) sensitivity in vascular smooth muscle; (ii) nifedipine attenuated this enhanced response by only a maximum of 27% at 1 micromol/l with a 56% reduction in F(ratio); (iii) Y-27632 attenuated it by a maximum of 80% at 100 micromol/l without a significant change in F(ratio); (iv) RhoA protein and mRNA expression levels in the femoral artery were up-regulated by +110% and +56%, respectively, while those of Rho-kinase were unchanged. CONCLUSIONS: The Ca(2+)-sensitizing mechanism involving the Rho/Rho-kinase system may be deeply involved in the enhanced arterial vasoconstriction seen in HF. Since Y-27632 attenuated this response in small arteries, it shows potential as a novel, potent vasodilator for the treatment of HF.     

Change in serum G-CSF levels in patients with Graves' disease by treatment with methimazole]

We evaluated the determination of serum G-CSF in the diagnosis of granulocytopenia due to methimazole (MMI) in 54 patients with Graves' disease, while they were being treated with MMI, by way of measuring WBC counts and serum levels of G-CSF, thyroid hormones, IgE, and interleukin-2. Serum TSH was measured by immunoradiometric assay, serum G-CSF was done by enzyme immunoassay, thyroid hormones and IgE were done by radioimmunoassay, and serum Interleukin-2 was done by enzyme-linked immunosorbent assay. The population whose G-CSF levels were higher than the minimum detectable level (30pg/ml) was 6 (30%) in normal subjects, 4 (22%) in patients with untreated Graves' disease, 2 (12%) in patients with treated euthyroid Graves' disease, 3 (23%) in patients with Graves' disease who had gone through agranulocytosis, and 2 (33%) in patients with Graves' disease complicated with granulocytopenia. There was no significant change in WBC counts for 4 weeks, but there was a significant difference between WBC counts before treatment and those at 8 weeks after treatment. We observed no significant change of serum G-CSF levels in patients with Graves' disease under treatment. However, there were significantly high levels of serum G-CSF and significantly low counts of WBC in patients with Graves' disease complicated with granulocytopenia induced by MMI, compared with those in normal subjects, patients with untreated Graves' disease, patients with treated euthyroid Graves' disease, and patients with euthyroid Graves' disease who had gone through agranulocytosis.(ABSTRACT TRUNCATED AT 250 WORDS)     

Androgen regulation of a specific gene in hamster flank organs

Summary Flank organs of Golden Syrian hamster containing large sebaceous glands are a useful model for studying androgen-dependent gene expression. Recently, we have isolated an androgen-dependent gene from a cDNA library constructed from male mRNAs of flank organs. In the present study, using this gene as a probe, we examined the kinetics of loss and re-expression of the mRNA of this gene. Dot blot hybridization, the current method, can approximately quantify mRNA levels. Castration caused a marked decrease in the mRNA within a few days to undetectable levels. Topical application of testosterone re-activated the mRNA levels, with the earliest activation within 24 h. The results of various topical steroid applications showed a full re-activation of the mRNA by testosterone and dihydrotestosterone, partial re-activation by androstanedione and androstenedione, and no activation by androstanediol or dehydroepiandrosterone. Simultaneous application of testosterone and progesterone inhibited the expression of mRNA but application of dihydrotestosterone and progesterone did not. The current dot blot hybridization technique appears to be a more direct approach for studying androgen action on DNAs than other previously used methods such as morphometry or enzyme assay.