Substance p plays a critical role in photic resetting of the circadian pacemaker in the rat hypothalamus.

Glutamate is considered to be the primary neurotransmitter in the retinohypothalamic tract (RHT), which delivers photic information from the retina to the suprachiasmatic nucleus (SCN), the locus of the mammalian circadian pacemaker. However, substance P (SP) also has been suggested to play a role in retinohypothalamic transmission. In this study, we sought evidence that SP from the RHT contributes to photic resetting of the circadian pacemaker and further explored the possible interaction of SP with glutamate in this process. In rat hypothalamic slices cut parasagittally, electrical stimulation of the optic nerve in early and late subjective night produced a phase delay (2.4 +/- 0.5 hr; mean +/- SEM) and advance (2.6 +/- 0.3 hr) of the circadian rhythm of SCN neuronal firing activity, respectively. The SP antagonist L-703,606 (10 microm) applied to the slices during the nerve stimulation completely blocked the phase shifts. Likewise, a cocktail of NMDA (2-amino-5-phosphonopentanoic acid, 50 microm) and non-NMDA (6,7-dinitroquinoxaline-2,3-dione, 10 microm) antagonists completely blocked the shifts. Exogenous application of SP (1 microm) or glutamate (100 microm) to the slices in early subjective night produced a phase delay ( approximately 3 hr) of the circadian firing activity rhythm of SCN neurons. Coapplication of the NMDA and non-NMDA antagonist cocktail (as well as L-703,606) resulted in a complete blockade of the SP-induced phase delay, whereas L-703,606 (10 microm) had no effect on the glutamate-induced delay. These results suggest that SP, as well as glutamate, has a critical role in photic resetting. Furthermore, the results suggest that the two agonists act in series, SP working upstream of glutamate.

Effect of sulfation substrates/inhibitors on N-(3,5-dichlorophenyl)succinimide nephrotoxocity in Fischer 344 rats.

The agricultural fungicide N-(3,5-dichlorophenyl)succinimide (NDPS) is an acute nephrotoxicant in rats. Our previous studies suggested that sulfate conjugation of NDPS metabolites might be a bioactivation step mediating NDPS nephrotoxicity. In this study, effects of substrates and/or inhibitors of sulfation on NDPS nephrotoxicity were examined to explore further the role of sulfation in NDPS nephrotoxicity. Male Fischer rats (4-8/group) were administered one of the following intraperitoneal (ip) pretreatment (dose, pretreatment time) prior to NDPS (0.6 mmol/kg) or NDPS vehicle (sesame oil, 2.5 ml/kg): (1) no pretreatment, (2) dehydroepiandrosterone (DHEA) (0.5 mmol/kg, 1 h), or (3) 2,6-dichloro-4-nitrophenol (DCNP) (0.04 mmol/kg, 1 h). Following NDPS or NDPS vehicle administration, renal function was monitored at 24 and 48 h. Pretreatment with DHEA, a typical substrate for and an inhibitor of hydroxysteroid (alcohol) sulfotransferase, resulted in marked protection against NDPS nephrotoxicity. A selective inhibitor of phenol sulfotransferase, DCNP, afforded little attenuation in NDPS nephrotoxicity. These results suggest that alcohol sulfate conjugates of NDPS metabolites, rather than phenolic sulfate conjugates, may be a penultimate or ultimate nephrotoxicant species mediating NDPS nephrotoxicity. The marked, but not complete, protection by DHEA also suggests that there are other metabolites or mechanisms responsible for NDPS nephrotoxicity.

Effects of sodium sulfate on acute N-(3,5-dichlorophenyl)succinimide (NDPS) nephrotoxicity in the Fischer 344 rat.

The agricultural fungicide N-(3,5-dichlorophenyl)succinimide (NDPS) induces acute polyuric renal failure in rats. Results of previous studies have suggested that NDPS may induce nephrotoxicity via conjugates of NDPS metabolites. Thus, the purpose of this study was to examine if administered sodium sulfate could alter NDPS nephrotoxicity. Male Fischer 344 rats (four rats per group) were administered a single intraperitoneal (i.p.) injection of sodium sulfate (0.035, 0.07, 0.35 or 3.5 mmol/kg) or sodium chloride (7.0 mmol/kg) 20 min before NDPS (0.2, 0.4 or 0.8 mmol/kg) or NDPS vehicle (sesame oil, 2.5 ml/kg) and renal function monitored at 24 and 48 h. High dose sodium sulfate (3.5 mmol/kg) markedly attenuated NDPS nephrotoxicity, while sodium chloride had no effect on NDPS-induced renal effects. NDPS nephrotoxicity was also attenuated by a pretreatment dose of 0.35 mmol/kg sodium sulfate, while 0.07 mmol/kg sodium sulfate pretreatment potentiated NDPS 0.2 mmol/kg to produce nephrotoxicity without markedly attenuating NDPS 0.4 mmol/kg to induce renal effects. A dose of 0.035 mmol/kg sodium sulfate did not potentiate NDPS 0.2 mmol/kg to induce nephrotoxicity. These results suggest that sulfate conjugates of NDPS metabolites might contribute to NDPS nephrotoxicity.

Nephrotoxicity of N-(3-bromophenyl)-2-hydroxysuccinimide: role of halogen groups in the nephrotoxic potential of N-(halophenyl) succinimides.

Among N-(halophenyl)succinimides. N-(3,5-dichlorophenyl)succinimide (NDPS) is a potent nephrotoxicant as well as an agricultural fungicide. Although two chloride groups on the phenyl ring are essential to induce optimal nephrotoxicity, the role of halogen groups in NDPS nephrotoxicity is not clear. In this study, N-(3-bromophenyl)-2-hydroxysuccinimide (NBPHS) was prepared as a monohalophenyl derivative of N-(3,5-dichlorophenyl)-2-hydroxysuccinimide (NDHS), an oxidative and nephrotoxicant metabolite of NDPS. The nephrotoxic potential of NBPHS was evaluated in vivo and in vitro to determine the role of halogen groups in N-(halophenyl)succinimide nephrotoxicity. Male Fischer 344 rats (four/group) were administered a single intraperitoneal (i.p.) injection of NBPHS (0.1, 0.4 or 0.8 mmol/kg) or vehicle (25% dimethyl sulfoxide in sesame oil) and renal function monitored for 48 h. Administration of NBPHS (0.8 mmol/kg) induced nephrotoxicity, while very mild changes or no changes in renal function were observed following administration of 0.4 mmol/kg or 0.1 mmol/kg of NBPHS, respectively. Nephrotoxicity induced by NBPHS (0.8 mmol/kg) was characterized by diuresis, transiently increased proteinuria, glucosuria and hematuria elevated kidney weight and reduced tetraethylammonium (TEA) uptake by renal cortical slices, and was not as marked as nephrotoxicity induced by NDHS (0.1 mmol/kg) or NDPS (0.4 mmol/kg). In the in vitro studies the effects of NBPHS on organic ion accumulation, pyruvate-stimulated gluconeogenesis, and lactate dehydrogenase (LDH) release were measured using renal cortical slices. NBPHS decreased p-aminohippurate (PAH) and TEA accumulation at NBPHS bath concentrations of 0.05 mM and 0.5 mM and 0.5 mM or greater, respectively. Renal gluconeogenesis was inhibited by NBPHS at 1 mM bath concentration, while LDH leakage was not increased at NBPHS bath concentrations up to 1 mM. The results demonstrate that NBPHS is a mild nephrotoxicant in vivo and in vitro, but does not have cytotoxic effects to renal tissues at the concentrations tested. From these results, it appears that halogen groups are essential to the nephrotoxic potential of N-(halophenyl)-2-hydroxysuccinimides or N-(halophenyl)succinimides and play an important role in the mechanism of NDPS nephrotoxicity following NDHS formation.

Hyperbaric oxygen effect on active Na+ transport across isolated toad skin.

The effect of hyperbaric oxygen (HBO) on Na+ transport across the isolated toad (Bufo marinus) skin was studied by measuring the transepithelial short-circuit current (ISC) and resistance (R) at 5, 8, and 10 ATA PO2 and 15 ATA normoxia during steady state conditions. The imposition of 5, 8, and 10 ATA PO2 for 2 h resulted in 45, 52, and 85% decrease in ISC, respectively. This decrease in ISC was always accompanied by an increase in R. When amiloride (10(-4) M) was added to the bathing medium, ISC decreased to zero within 15 min regardless of the PO2 level, indicating that the HBO-induced decrease in ISC is caused by an inhibition of amiloride-sensitive Na+ transport. Addition of both superoxide dismutase (SOD) and catalase to the medium bathing both sides of the skin markedly attenuated the HBO effect on ISC and R. Applying HBO to the serosal or mucosal surface independently produced similar effects on ISC. However, the presence of antioxidant enzymes (SOD and catalase) with 10 ATA PO2 prevented the toxic HBO effect only from the serosal side; no protection by these antioxidant enzymes was observed from the mucosal side. These findings are consistent with a view that free radicals are involved in the HBO-induced inhibition of ISC. However, further studies involving the site(s) of radical generation as well as site(s) of toxic action are needed to understand the cellular and molecular mechanism of HBO toxicity.

Suppressed renin release during hyperoxia in the conscious dog.

Recent studies have documented a marked inhibition of renal prostaglandin E2 excretion during hyperbaric oxygenation (HBO) in the conscious dog. Numerous other studies have shown that pharmacological inhibition of prostaglandin synthesis is associated with diminished renin release. The current experiments were designed to test whether HBO is also accompanied by reduced renin secretory rate (RSR) in conscious dogs. Dogs were implanted with electromagnetic flow probes on the left renal artery as well as with chronic arterial and renal venous catheters. Renal blood flow, peripheral plasma renin activity, and renal venous renin activity were measured, which allowed calculation of RSR for the left kidney. The dogs were studied at normal atmospheric pressure while breathing room air, followed by exposure to 100% oxygen at 2 or 3 ATA. The dogs were then returned to room air. The RSR decreased from 153.6 +/- 51.6 (ng . h-1) . min-1 to 72 +/- 21 (ng . h-1) . min-1 during HBO. Upon return to normoxia, RSR rose to 345 +/- 140.4 (ng . h-1) . min-1. This renin response to HBO seems best related to the previously documented inhibition of renal prostaglandin synthesis. These experiments provide nonpharmacological evidence that there may be a functional relationship between the renin-angiotensin system and renal prostaglandin release in awake animals.

Antidiuresis and inhibition of PGE2 excretion by hyperoxia in the conscious dog.

Experiments were performed to determine the influence of varying inspired oxygen tension on renal prostaglandin E2 (PGE2) excretion, renal hemodynamics, and water and electrolyte excretion in the conscious dog. Hypoxic exposure (PIO2 = 56 torr) resulted in a 13% increase in renal blood flow (RBF), while hyperoxic breathing with PIO2 of 700, 1426, or 2139 torr, all resulted in significant 5--7% decline in RBF, a response that was significantly attenuated compared to the striking renal vasoconstriction caused by hyperoxia in anesthetized dogs. Hyperbaric oxygenation (HBO) (1426 torr O2, 2139 torr O2) was associated with unexpected decreases in urine flow (V) of 61% and 70%, respectively. The antidiuresis and mild hemodynamic adjustments were correlated with a 67% decline in urinary PGE2 excretion (UPGE2 x V) when the dogs breathed 700 torr O2, while exposure to 1426 torr O2 and 2139 torr O2 diminished UPGE2 x V by 92% and 99%, respectively. Plasma antidiuretic hormone (ADH) concentration, measured during exposure to 1426 torr O2, was unchanged. In addition, this nonpharmacologic hyperbaric decline in PGE2 excretion was not associated with any changes in sodium excretion of renin secretion, in contrast to the usual depression of these variables with pharmacologic PG inhibition (indomethacin). The HBO antidiuresis may be a consequence of an increased medullary osmotic gradient secondary to reduced vasa recta blood flow. Alternatively, this antiduresis could occur as a consequence of a lowering of the normal functional antagonism existing between PGE2 and ADH, such that the influence of endogenous ADH is potentiated.

Physiological responses to head-out immersion in water at 11 ATA.

Cardiorespiratory, thermal, and renal responses to a 30-min head-out immersion in 15 degree C water were studied at 1-ATA air and 11-ATA helium-oxygne environments in four male subjects wearing dry suits. Cardiorespiratory responses to immersion (reductions in heart rate, expiratory reserve volume, vital capacity, and thoracic impedance; and increases in stroke volume, cardiac output, and inspiratory capacity) were comparable at both pressures. However, thermal responses to immersion (a reduction in mean skin temperature and increases in skin heat flux and suit conductance) were significantly greater at 11 ATA compared to those at 1 ATA. The rate of urinary excretion of norepinephrine increased significantly during and after immersion at 11 ATA but not at 1 ATA. In contrast, the urinary excretion of epinephrine was not altered by pressure or immersion. The immersion diuresis was greater and lasted longer at 11 ATA than at 1 ATA although there was no difference in the endogenous creatinine excretion . This diuresis was accompanied by a significant natriuresis which was more marked at 1 ATA than at 11 ATA. At 1 ATA, the urinary excretion of both aldosterone and antidiuretic hormone (ADH) decreased during immersion. At 11 ATA, the rate of excretion of these hormones before immersion was lower compared to that at 1 ATA and did not change significantly during immersion. These results indicate that immersion in a hyperbaric helium-oxygen environment presents a greater cold stress than at 1-ATA air, and also that immersion diuresis and natriuresis at high pressure may be induced by a factor other than inhibition of aldosterone and ADH.

Hana Kai II: a 17-day dry saturation dive at 18.6 ATA. I. Objectives, design, and scope.

The dive (Hana Kai II) described in these papers was designed to determine the effects on man of a prolonged exposure to a dry helium-oxygen hyperbaric environment. Comprehensive studies on energy balance, body fluid balance, cardiorespiratory functions, maximal oxygen uptake, psychological performance, and physiological responses to cold were performed at a simulated depth of 580 ft (18.6 ATA) over a 30-day period in March-April 1975. Following a 3-day predive control period at 1 ATA air (period 1), 5 male divers spent 17 days at 18.6 ATA in a helium-oxygen environment (periods 2-6), and returned to 1 ATA air after 7 days of decompression (periods 7-8). They stayed an additional 3 days inside the chamber for postdive control measurements (period 9). The chamber temperature was maintained at 25-27 degrees C during periods 1 and 9, 30-31 degrees C during periods 2-5, and 27-28 degrees C during period 6. At 18.6 ATA, the PO2 and PCO2 of the chamber gas were maintained at approximately 225 and 2 mmHg, respectively. In this introductory paper, physical and physiological characteristics of individual subjects, the major daily activity schedule, and the scope of investigation are presented.

Hana kai ii: a 17-day dry saturation dive at 18.6 ATA. II. Energy balance.

Since previous saturation dives have caused loss of body weight despite apparently adequate-to-high food intake, a complete study of energy balance was undertaken during the saturation dive Hana Kai II. Over a 30-day period in the hyperbaric chamber (3 days of predive control, 1 day of compression, 16 days at 18.6 ATA, 7 days of decompression, and 3 days of postdive control), all food, urine, and feces for five men were analyzed by bomb calorimetry; 24-h energy expenditure (M) was measured from continuous VO2, VCO2, and urine N. Body weight was taken daily; body composition was assessed from density, total body water, and skinfold thickness. Food intake was high throughout the 30 days (about 3500 kcal/day) while fecal and urinary losses were a normal 6-8% of intake. Energy expenditure was increased a little by the hyperbaric condition, but averaged only 2431 kcal/day for the 30 days and yet there was an average loss of adipose tissue of 0.8 kg for each man for the entire period. Nitrogen balance was positive. There was no evidence of heat gain or loss. The energy balance, total fuel compared with energy expenditure, required an additional 919 kcal/man-day for 30 days, an unidentified term which is not measured by conventional techniques.

Hana kai ii: a 17-day dry saturation dive at 18.6 ATA. V. Maximal oxygen uptake.

Cardiorespiratory responses of four men to submaximal and maximal cycling exercise were observed during 17 days at 18.6 ATA. Inspired gas at pressure consisted of hyperoxic (PO2 = 232 mmHg) and normoxic (PO2 = 159 mmHg) helium mixtures with relative gas densities of 3.8 and 2.8, respectively. The average of pre- and postdive VO2max (1 ATA air), which were not significantly different, was 3.10 liters - min-1. During 5 min of submaximal exercise at 50% of VO2max, no significant difference in work rate, VO2, VCO2, VE, respiratory rate, heart rate (HR), stroke volume, blood pressures, or rectal temperature was noted at 18.6 ATA compared to 1 ATA with either gas mixture. Submaximal HR tended to decrease by 5 to 10 beats - min-1 at pressure, and in hyperoxia the VO2/HR ratio was significantly higher. Maximal exercise was performed to exhaustion at work rates requiring about 120% of VO2max. Significant increased in VO2max of 0.10 liter - min-1 (3%) and in endurance time of 2 min (48%) were found during hyperoxic gas breathing, whereas normoxic values at 18.6 ATA were similar to those at 1 ATA. Significant reductions in maximal HR of 8 beats - min-1 (4%) were observed with both gas mixtures at pressure, and VE was significantly decreased by 36 liters - min-1 (26%) in hyperoxia and 29 liters - min-1 (21%) in normoxia. No change was found in the calculated cardiac output. Maximal voluntary ventilation, which was measured only for the hyperoxic gas, fell significantly by 80 liters - min-1 (40%). Results indicate that aerobic power and endurance performance were affected by oxygen pressure. Normoxic work capacity, however, was not decreased at 18.6 ATA, despite marked reductions in HR and VE.

Heart rate response to breath holding at 18.6 ATA.

The heart rate (HR) responses to breath-holding (BH) with the without face immersion (FI) in 31 or 27 degrees C water was studied in 1 ATA air and hyperbaric He-O2 environments in 4 male subjects during a dry saturation dive to simulated depth of 580 ft (18.6 ATA). When a 60 sec BH or FI was performed while leaning forward, there was a significant linear correlation between the maximal bradycardial response (delta HRmax) and ambient pressure for simple BH (r-0.08, P less than 0.05) and 31 degrees C FI (r = 0.91, P less than 0.01), but not for 27 degrees C FI. A similar trend was seen during 30 sec BH's while seated erect. The facial cold-dependent component of the FI bradycardia was not significantly altered by pressure. In general, there were significant correlations between the initial HR and the initial thoracic conductive volume (TCV; measured by the four-electrode Minnesota impedance cardiograph), and between the initial TCV and delta HRmax observed during seated erect BH's. Since the TCV was generally higher at depth, it is suggested that a mechanical effect due to increased TCV at depth, possibly related to increased gas density, is at least partly responsible for the pressure dependence of BH bradycardia.