Xenon inhalation increases norepinephrine release from the anterior and posterior hypothalamus in rats.

PURPOSE: To investigate the effect of xenon (Xe) and nitrous oxide (N(2)O) on norepinephrinergic neuronal activity in the rat medial preoptic area (mPOA) and posterior hypothalamus (PH) using microdialysis. METHODS: Sixty male Wistar rats were equally allocated to two groups: mPOA and PH. A microdialysis probe was implanted into the mPOA or the PH. In both groups, each animal was exposed to one of the following inhalations: 25% oxygen (control, n=6), 30% Xe (n=6), 60% Xe (n=6), 30% N(2)O (n=6) or 60% N(2)O (n=6). Norepinephrine concentration in the perfused artificial cerebrospinal fluid was measured by high pressure liquid chromatography at ten-minute intervals. After plotting the time-norepinephrine concentration curve, the area under the curve (AUC) in each group was calculated. RESULTS: In the mPOA, 30 and 60% Xe, but only 60% N(2)O significantly increased norepinephrine release. The AUC in the 30% Xe, 60% Xe or 60% N(2)O group was 160 +/- 9 (P <0.05), 288 +/- 42 (P <0.01) or 237 +/- 46 pg x min/sample (P <0.01), respectively, compared to that in the control group: 77 +/- 14 pg x min/sample. In the PH, only 60% Xe significantly increased norepinephrine release compared to control (AUC: 191 +/- 38 vs. 71 +/- 1 pg x min/sample, P <0.01). CONCLUSION: The present data suggest that Xe stimulates norepinephrinergic neurons more potently than N(2)O; 1.2 times more in the mPOA and 2.5 times more in the PH. This stimulant effect may contribute to the hypnotic and sympathotonic effects of Xe in rats.

[Combination of acute normovolemic hemodilution technique with preoperative autologous blood donation prevented allogeneic blood transfusion against 4000 g surgical blood loss in a patient undergoing left partial nephrectomy].

A 41-year-old male patient with well-controlled hypertension underwent a partial nephrectomy under total intravenous anesthesia with propofol, fentanyl and ketamine. To avoid allogeneic blood transfusion, preoperative autologous blood donation (400 g) a week before the surgery and acute normovolemic hemodilution (800 g) after induction of anesthesia were performed. As surgical blood loss was more than 4000 g, blood hemoglobin (Hb) level decreased to 6.4 g.dl-1. However, as intraoperative hemodynamics was relatively stable with no ischemic changes in ECG and arterial blood gas analysis did not show metabolic acidosis, autologous blood transfusion was withheld till hemostasis had been done. After returning the autologous blood, Hb increased to 9.4 g.dl-1. On the 2nd postoperative day, Hb decreased to 7.6 g.dl-1. As the patient's vital signs did not show any severe complications, blood transfusion was not performed. Then, the Hb level increased gradually to 13.9 g.dl-1, 3 month later without allogenic blood transfusion. In addition, any postoperative complications by low Hb level were not recognized so far. This case suggests that combination of autologous transfusion techniques may be effective to avoid allogeneic blood transfusion even against massive hemorrhage. However, to avoid disadvantage of these technique, we should always evaluate preoperative patient conditions.

Myristoleic acid, a cytotoxic component in the extract from Serenoa repens, induces apoptosis and necrosis in human prostatic LNCaP cells.

BACKGROUND: Prostatic tumors are well known to progress to hormonal therapy-resistant terminal states. At this stage, there are no chemotherapeutic agents to affect clinical outcome. An effective cell death inducer for these prostate cells may be a candidate as an attractive antitumor agent. The extracts from S. repens have been used to improve the state of prostatic diseases and we have attempted to identify the effective component from the extract. METHODS: Cell viability was examined in LNCaP cells, an in vitro model for hormonal therapy-resistant prostatic tumor. RESULTS: We found that exposure of the extract from S. repens resulted in cell death of LNCaP cells. We also identified myristoleic acid as one of the cytotoxic components in the extract. The cell death exhibited both apoptotic and necrotic nuclear morphology as determined by Hoechst 33342 staining. Cell death was also partially associated with caspase activation. CONCLUSIONS: It was demonstrated that the extract from S. repens and myristoleic acid induces mixed cell death of apoptosis and necrosis in LNCaP cells. These results suggest that the extract and myristoleic acid may develop attractive new tools for the treatment of prostate cancer.

Reversal of P-glycoprotein and multidrug-resistance protein-mediated drug resistance in KB cells by 5-O-benzoylated taxinine K.

A newly synthesized taxoid originally from the Japanese yew Taxus cuspidata, 5-O-benzoylated taxinine K (BTK) was examined for its ability to reverse P-glycoprotein (P-gp) and multidrug resistance protein (MRP)-mediated multidrug resistance. BTK reversed the resistance to paclitaxel, doxorubicin (ADM), and vincristine (VCR) of KB-8-5 and KB-C2 cells that overexpress P-gp by directly interacting with P-gp. BTK also moderately reversed the resistance to ADM of KB/MRP cells that overexpress MRP. However, BTK neither inhibited the transporting activity of MRP nor reduced intracellular glutathione levels in KB/MRP cells. BTK shifted the distribution of ADM in KB/MRP cells from punctate cytoplasmic compartments to the nucleoplasm and cytoplasm by inhibiting acidification of cytoplasmic organelles. These two functions of BTK make it able to reverse both P-gp- and MRP-mediated MDR. BTK in combination with ADM should be useful for treating patients with tumors that overexpress both P-gp and MRP.

Synthesis and biological evaluation of CX-659S and its related compounds for their inhibitory effects on the delayed-type hypersensitivity reaction.

In order to find novel nonsteroidal compounds possessing an inhibitory activity against delayed-type hypersensitivity (DTH) reactions, we conducted random screening using a picryl chloride (PC)-induced contact hypersensitivity reaction (CHR) in mice, and found compound 1 as a lead compound. Then we synthesized and evaluated an extensive series of 5-carboxamidouracil derivatives focused on both the uracil and the antioxidative moieties. Among them, we found that the hindered phenol moiety was necessary to exhibit the activities; especially, compounds 28a-28c having the partial structure of vitamin E were found to exert potent activities against the DTH reaction by both oral and topical administration. And compound 28c showed antioxidative activity against lipid peroxidation with an IC50 of 5.9 microM. Compound 28c (CX-659S) was chosen as a candidate drug for the treatment of cutaneous disorders such as atopic dermatitis and allergic contact dermatitis.

Inhibited hypothalamic histamine metabolism during isoflurane and sevoflurane anesthesia in rats.

BACKGROUND: Histamine is most densely distributed in the hypothalamus and has an important effect on consciousness or wakefulness. It has been little considered whether general anesthetics could exert their effects on hypothalamic histamine metabolism. The present study was conducted to investigate the effects of isoflurane and sevoflurane anesthesia on hypothalamic histamine metabolism. METHODS: Sixty male Wistar rats were divided equally into isoflurane and sevoflurane anesthesia groups. Each group was divided into three equal sub-groups: the control, anesthesia and recovery groups. The rats of the anesthesia and recovery groups were exposed to either 2% isoflurane or 3% sevoflurane for 30 min. The recovery group was kept in air for 30 min after anesthesia. The rats were decapitated to dissect out hypothalamus which was divided into the fore and rear portion. The contents of histamine and 1-methylhistamine, which is a main histamine metabolite, were determined by high-performance liquid chromatography. The obtained data were analyzed by one-way analysis of variance followed by Bonferoni's test. RESULTS: Histamine contents of the anterior and posterior hypothalamus in both isoflurane and sevoflurane groups increased significantly during the anesthesia and 1-methylhistamine contents of the anterior and posterior hypothalamus in sevoflurane group increased remarkably after anesthesia. The increases of histamine contents supposedly reflected inhibited histamine metabolism and the increases of 1-methylhistamine would be caused by acceleration of histamine degradation. CONCLUSIONS: Histamine metabolism was inhibited during both isoflurane and sevoflurane anesthesia and accelerated only in the posterior hypothalamus during the emergence from these anesthetics.

[Influence of fluid replacement on serum magnesium concentration and proper magnesium supplementation during general anesthesia].

We have studied the influence of fluid replacement on serum magnesium (Mg2+) concentrations, and studied proper Mg2+ supplementation during general anesthesia. Thirty eight patients undergoing elective surgery randomly received: Mg(2+)-free acetated Ringer solution (Group I, n = 15), acetated Ringer solution containing 0.5 mmol.l-1 of Mg2+ (Group II, n = 6), 1.0 mmol.l-1 of Mg2+ (Group III, n = 7), 2.0 mmol.l-1 of Mg2+ (Group IV, n = 6), or 4.0 mmol.l-1 of Mg2+ (Group V, n = 4). Measurements were made on serum and urine Mg2+ concentrations during anesthesia. In Group I, the serum Mg2+ concentrations decreased in correspondence with the water balance. It is suggested that dilution due to the fluid replacement induced the reduction in serum Mg2+ concentrations since the observed urine Mg2+ concentrations were negligible. In Group II-V, the reduction in serum Mg2+ concentrations was inhibited by Mg2+ supplementation, and the serum Mg2+ concentrations remained unchanged in Group IV. We conclude the Mg2+ supplementation is required during anesthesia when a large amount of fluid is infused.

Molecular cloning and characterization of a putative neural calcium channel alpha1-subunit from squid optic lobe.

The complete amino acid sequence of a putative calcium channel alpha1-subunit, SQCC1, from the optic lobe of the squid Loligo bleekeri has been deduced by cloning and sequence analysis of the complementary DNA. The open reading frame encodes 2206 amino acids, which corresponds to a molecular weight of 251,451. The deduced amino acid sequence shares general structural features with the other voltage-dependent calcium channels; it consists of four repeated units of homology. Each motif has five hydrophobic segments and one positively charged segment. The transcriptional products were detected in all nervous systems examined; optic lobe, cerebral ganglia and giant stellate ganglia. However, it was not detected in the mantle muscle, heart and stomach, indicating SQCC1 is a calcium channel alpha1-subunit specific for squid nervous system. SQCC1 is more closely related in its amino acid sequence patterns to dihydropyridine-insensitive calcium channels rather than dihydropyridine-sensitive ones.

Intraventricular administration of estradiol modulates rat prolactin secretion and synthesis.

The effect of estradiol (E2) on rat tuberoinfundibular dopaminergic (TIDA) neurons was examined in vivo, employing chronic intraventricular (i.c.v.) infusion technique using an osmotic mini-pump. The activity of TIDA neurons was assessed by the release and synthesis of prolactin (PRL) in the rat pituitary gland and by the changes in the 3, 4-dihydroxyphenylacetic acid (DOPAC) and dopamine (DA) levels and in the DOPAC/DA ratio in the rat hypothalamus. We also examined the [3H] E2 binding in the rat hypothalamus. Ovariectomized female Wistar rats with E2 replacement were treated with daily i.c.v. infusion of 1 microM of E2 or saline vehicle for 1, 3, and 7 days using the Alzet osmotic mini-pump and brain infusion kit. At 1 day of i.c.v. infusion of E2, the serum PRL level was significantly decreased compared with that in the vehicle group. Northern blot analysis of the total RNA isolated from the pituitary glands demonstrated a decrease in the PRL gene transcript level in the E2 group. At 3 days of E2 treatment, however, the serum PRL level was significantly increased compared with that of the vehicle-injected group and Northern blot analysis also demonstrated that the PRL gene transcript level was increased in the E2 group. At 7 days of E2 administration, there were no significant differences between the E2 and vehicle groups in either serum PRL or PRL gene transcript levels. There was a significant increase in the DOPAC/DA ratio after 1 day in the E2 group. However, no significant effects of E2 on this ratio were observed at 3 and 7 days of treatment. The DOPAC concentration in the E2 group was significantly increased at day 1 and significantly decreased at day 3, compared with that of the respective time in vehicle group. At day 7 there was no significant change in DOPAC concentration in either groups. The DA concentrations in the hypothalamus was not changed on any day in either group. Specific [3H] E2 binding was observed in the rat hypothalamus. These data suggest that E2 may have a biphasic effect on the accumulation of PRL gene transcripts and on the PRL secretion in the rat pituitary by first stimulating and then inhibiting the TIDA neuronal activity.

Promotion of neuronal differentiation of PC12h cells by natural lignans and iridoids.

We studied the effect of (+)- and (-)-syringaresinol, (+)-syringaresinol glucosides, syringin, aucubin and catalpol on neurite outgrowth of a cultured cell line of paraneuron, PC12h cells. Of these compounds, (+)-syringaresinol diglucoside and partly glucosidase-hydrolyzed aucubin were found to be the most potent in promotion of the neurite outgrowth and stimulated responses to a high concentration of KCl and to carbachol in the cells, as observed by increase of the concentration of cytosolic free calcium. It is suggested that some of these herb-derived compounds can induce neuronal differentiation in PC12h cells.

Prolactin stimulates [3H]dopamine release from dispersed rat tubero-infundibular dopaminergic neurons and dopamine decreases gonadotropin-releasing hormone release induced by calcium ionophore.

In order to investigate the involvement of prolactin-dopamine and dopamine-gonadotropin interactions in the hypothalamo-pituitary axis of hyperprolactinemia, in vitro studies were performed using primary cultures of dispersed rat hypothalamic heterogeneous cells containing tubero-infundibular dopaminergic neurons or gonadotropin-releasing hormone (GnRH) neurons. We observed that prolactin caused dose-dependent stimulation of [3H]dopamine release after a 16-h incubation. Staurosporin (10 nmol/l), an inhibitor of protein kinase C, significantly reduced the [3H]dopamine release induced by prolactin (1 mg/l). Incubation of tubero-infundibular dopaminergic neurons with prolactin (1 mg/l) had no effect on intracellular cyclic adenosine monophosphate accumulation. Dopamine (1 mumol/l) significantly (p < 0.01) reduced the release of GnRH induced by 50 mumol/l calcium ionophore from dispersed hypothalamic cells from the preoptic area, while prolactin had no effect on GnRH release. These data support the hypothesis that the antigonadotropic effect of prolactin on the hypothalamus is mediated by an inhibitory effect of dopamine on GnRH release.

Intraventricular administration of histidyl-proline-diketopiperazine [Cyclo(His-Pro)] suppresses prolactin secretion and synthesis: a possible role of Cyclo(His-Pro) as dopamine uptake blocker in rat hypothalamus.

Histidyl-proline-diketopiperazine [Cyclo (His-Pro) (CHP)] was discovered to be one of the metabolites of TRH. To understand the specific role of CHP in rat hypothalamic dopamine neurons, we examined the in vivo effects of intraventricular (icv) infusion of CHP on the release and synthesis of PRL in the rat pituitary and the 3,4-dihydroxyphenylacetic acid (DOPAC)/dopamine ratio in the rat hypothalamus. We also examined the in vitro effects of CHP on the release of [3H]dopamine from dispersed tuberoinfundibular dopamine neurons, [3H]dopamine reuptake in hypothalamic membrane fractions, and PRL release from rat pituitary cultured cells. Female rats were treated by icv infusion of 1 microM CHP daily for 1, 3, and 7 days, using Alzet osmotic pumps. After 1 day of treatment, the serum PRL concentration was significantly decreased. Northern blot analysis of the total RNA isolated from the pituitary glands of control animals using 32P-labeled PRL cDNA as a probe indicated the presence of PRL gene transcript, 1.0 kilobase in size, and its amount was decreased by CHP treatment. CHP did not affect [3H]dopamine release from dispersed tuberoinfundibular dopaminergic neurons at any concentration up to 1 microM. CHP did not inhibit PRL release from cultured pituitary cells at low concentrations (1-100 nM), but it stimulated PRL release at high concentrations (1 and 10 microM). We also examined the concentrations of dopamine and DOPAC in the rat hypothalamus when CHP was administered icv for 1 or 7 days. There was a significant decrease in the DOPAC/dopamine ratio after CHP treatment for 1 day. Furthermore, CHP caused dose-dependent inhibition of [3H]dopamine uptake by the rat hypothalamus similar to other dopamine uptake blockers, such as benztropine and GBR12909. These data suggest that icv administration of CHP might decrease both PRL secretion and accumulation of PRL gene transcripts in the pituitary by decreasing the DOPAC/dopamine ratio and inhibiting dopamine reuptake in the rat hypothalamus.

[Clinical study of total intravenous anesthesia with droperidol, fentanyl and ketamine--effects of nicardipine, diltiazem and nifedipine on intraoperative hypertension].

Effects of Ca ion channel blockers, nicardipine, diltiazem and nifedipine on intraoperative hypertension were evaluated clinically in ninety surgical patients who received various surgical procedures under total intravenous anesthesia with droperidol, fentanyl and ketamine. When the systemic blood pressure exceeded 160 mmHg systolic at least for ten minutes, even though they received a total dose of 10-15 micrograms.kg-1 of fentanyl, one of the following three antihypertension drugs was administered:nicardipine 0.5-1.0 micrograms i.v., diltiazem 5-10 mg i.v. and nifedipine 5-10 mg intranasally. The effects were evaluated by measuring the systemic blood pressure, heart rate and rate pressure product, before the administration as well as 5, 15, 20 and 30 minutes after the administration. Following the injection of the drug a significant 10-20% reduction in the mean systemic blood pressure was observed in the nifedipine group, while less decrease was observed in the diltiazem and nicardipine groups. The mean heart rate in the nifedipine group increased slightly, while a slight decreases was observed in the other two groups. Therefore the rate pressure product was reduced significantly in three groups, but there was no significant difference among them. No adverse episodes such as ischemic changes on E.K.G. and deterioration of the cardiovascular system were encountered in any patients. We conclude that any of these drugs, particularly nifedipine would be appropriate to control hypertension during total intravenous anesthesia with droperidol, fentanyl and ketamine.

Atrial and brain natriuretic peptides enhance dopamine accumulation in cultured rat hypothalamic cells including dopaminergic neurons.

Dopamine accumulation in hypothalamic cells by atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) was analyzed using newborn rat hypothalamic cells in culture. Both ANP and BNP caused a dose-dependent increase in [3H]-dopamine accumulation in the cells. ANP increased [3H]-dopamine accumulation significantly within 20 min. The effects of ANP and BNP on dopamine accumulation paralleled an increase in intracellular cGMP concentration. (Bu)2-cGMP and sodium nitroprusside, a stimulator of the soluble form of guanylate cyclase, also enhanced [3H]-dopamine accumulation. ANP had no effect on efflux of [3H] radioactivity after [3H]-dopamine uptake. These results suggest that a change in cGMP is one of the intermediate steps in dopamine accumulation in hypothalamic cells by ANP and BNP.

Vasoactive intestinal peptide enhances dopamine accumulation in primary cell culture of rat hypothalamus.

The effect of vasoactive intestinal peptide (VIP) and PHI-27 on dopamine accumulation in cultured rat hypothalamic cells was investigated. VIP enhanced [3H]dopamine accumulation dose dependently. This effect was significant at 10(-8)-10(-5) M VIP with a concomitant increase in intracellular cyclic AMP (cAMP), and reached its plateau level at 10(-6) M VIP. VIP increased [3H]dopamine accumulation significantly within 15 min. PHI-27 and dibutyryl cAMP ((Bu)2-cAMP) also enhanced [3H]dopamine accumulation. These results suggest that VIP enhances dopamine accumulation in hypothalamic cells by increasing intracellular cAMP.

Intraventricular administration of thyrotrophin-releasing hormone (TRH) suppresses prolactin secretion and synthesis: a possible involvement of dopamine release by TRH from rat hypothalamus.

The administration of thyrotrophin-releasing hormone (TRH) causes a variety of dopamine-related biological events. To understand the specific role of TRH on rat hypothalamic dopamine neurones, we examined the in-vivo effects of intraventricular (i.c.v.) infusion of TRH on the release and synthesis of prolactin in the rat pituitary gland and on the changes in binding of [3H]MeTRH and dopamine turnover rates in rat hypothalamus. We have also examined the in-vitro effects of TRH on the release of [3H]dopamine from dispersed tuberoinfundibular dopamine neurones. Female rats were treated with i.c.v. infusions of 1 mumol TRH/1 daily for 1, 3 and 7 days using Alzet osmotic pumps. Following 7 days of treatment the serum prolactin concentrations were significantly decreased. A reduction in hypothalamic TRH-binding sites (Bmax) was also apparent but the dissociation constant (Kd) was unaffected. Northern blot analysis of total RNA isolated from the pituitary glands of control animals using 32P-labelled prolactin cDNA as a probe indicated the presence of three species of prolactin gene transcripts of approximately 3.7, 2.0 and 1.0 kb in size, and these were decreased by TRH treatment. We examined the turnover rate of dopamine in the rat hypothalamus when TRH was administered i.c.v. for 7 days. There was a significant increase in 3,4-dihydroxyphenylacetic acid/dopamine ratio with TRH treatment. Moreover, exposure to TRH stimulated [3H]dopamine release from rat tuberoinfundibular neurones in a time- and dose-dependent manner. Dopamine receptor antagonists such as SCH23390 and (-)sulpiride, and other neuropeptides such as vasoactive intestinal peptide and oxytocin did not affect TRH-stimulated [3H]dopamine release.(ABSTRACT TRUNCATED AT 250 WORDS)

Interleukin 1 beta and tumour necrosis factor alpha stimulate the release of gonadotropin-releasing hormone and interleukin 6 by primary cultured rat hypothalamic cells.

The abilities of recombinant human interleukin 1 beta and tumour necrosis factor alpha to induce release of GnRH and interleukin 6 from primary culture of rat hypothalamic cells were examined. The effect of estradiol on the release of interleukin 6 by these cells was also tested. Both interleukin 1 beta and tumour necrosis factor alpha caused significant stimulation of GnRH secretion from the hypothalamic cells within 5 min. The hypothalamic cells secreted interleukin 6 spontaneously, and their secretion over 24 h was stimulated dose-dependently by interleukin 1 beta, tumour necrosis factor alpha and estradiol. These results suggest that interleukin 1 beta and tumour necrosis factor alpha stimulate the secretions of GnRH and interleukin 6 in the hypothalamus, and that these cytokines may be involved in the mechanism of GnRH secretion in the hypothalamus.

Adenosine 3',5'-cyclic monophosphate enhances dopamine accumulation in rat hypothalamic cell culture containing dopaminergic neurons.

The regulation of dopamine accumulation in cultured rat hypothalamic cells by adenosine 3',5'-cyclic monophosphate (cAMP) was investigated in cultures of newborn rat hypothalamic cells. Both dibutyryl cAMP, (Bu)2-cAMP, and forskolin enhanced [3H]dopamine accumulation in a dose-dependent manner. cAMP also enhanced [3H]dopamine accumulation, but to a lesser degree. Neither n-butyrate nor adenosine alone enhanced [3H]dopamine accumulation. (Bu)2-cAMP had no effect on basal efflux of [3H]radioactivity. The effect of (Bu)2-cAMP appeared on day 5 of culture, reached a maximum on day 6, and then rapidly decreased. These results suggest that dopamine uptake by cultured rat hypothalamic cells is regulated by intracellular cAMP.

Involvement of extracellular calcium and arachidonate in [3H] dopamine release from rat tuberoinfundibular neurons.

The mechanism of [3H]dopamine [( 3H]DA) release was investigated using primary cultures of dispersed cells from the rat tuberoinfundibular region, which contains tyrosine hydroxylase (TH)-like immunoreactive neurons. The calcium ionophore A23187 at 10 nM and above caused a significant and dose-dependent increase in [3H]DA release. In the presence of 50 microM A23187, [3H]DA release was detectable within 30 s and reached a plateau in 15 min. The induction of [3H]DA release by 50 microM A23187 was abolished by lowering the extracellular calcium concentration with 2 mM EDTA. Maitotoxin, another calcium-channel activator, also increased [3H]DA release at a concentration of 50 ng/ml. Exogenous additions of 100 mIU/ml phospholipase A2 and 10 microM arachidonate caused significant release of [3H]DA. Furthermore, A23187 stimulated [3H]arachidonate release from tuberoinfundibular dopaminergic (TIDA) neurons in a dose- and time-dependent manner. These results suggest that extracellular calcium and arachidonate are involved in the process of [3H]DA release from rat TIDA neurons.