Differential effects of nicotine treatment and ethanol self-administration on CYP2A6, CYP2B6 and nicotine pharmacokinetics in African green monkeys.

In primates, nicotine is metabolically inactivated in the liver by CYP2A6 and possibly CYP2B6. Changes in the levels of these two enzymes may affect nicotine pharmacokinetics and influence smoking behaviors. This study investigated the independent and combined effects of ethanol self-administration and nicotine treatment (0.5 mg/kg b.i.d. s.c.) on hepatic CYP2A6 and CYP2B6 levels (mRNA, protein, and enzymatic activity), in vitro nicotine metabolism, and in vivo nicotine pharmacokinetics in monkeys. CYP2A6 mRNA and protein levels and in vitro coumarin (selective CYP2A6 substrate) and nicotine metabolism were decreased by nicotine treatment but unaffected by ethanol. CYP2B6 protein levels and in vitro bupropion (selective CYP2B6 substrate) metabolism were increased by ethanol but unaffected by nicotine treatment; CYP2B6 mRNA levels were unaltered by either treatment. Combined ethanol and nicotine exposure decreased CYP2A6 mRNA and protein levels, as well as in vitro coumarin and nicotine metabolism, and increased CYP2B6 protein levels and in vitro bupropion metabolism, with no change in CYP2B6 mRNA levels. Chronic nicotine resulted in higher nicotine plasma levels achieved after nicotine administration, consistent with decreased CYP2A6. Ethanol alone, or combined with nicotine, resulted in lower nicotine plasma levels by a mechanism independent of the change in these enzymes. Thus, nicotine can decrease hepatic CYP2A6, reducing the metabolism of its substrates, including nicotine, whereas ethanol can increase hepatic CYP2B6, increasing the metabolism of CYP2B6 substrates. In vivo nicotine pharmacokinetics are differentially affected by ethanol and nicotine, but when both drugs are used in combination the effect more closely resembles ethanol alone.

Independent and combined effects of ethanol self-administration and nicotine treatment on hepatic CYP2E1 in African green monkeys.

Cytochrome P450 2E1 metabolizes ethanol and also bioactivates many toxins and procarcinogens. Elevated levels of hepatic CYP2E1 are associated with an increased susceptibility to chemical toxicity and carcinogenesis. This study investigated the induction of hepatic CYP2E1 by ethanol and nicotine, alone and in combination, in a nonhuman primate model. Monkeys that self-administered ethanol and that received subcutaneous injections of nicotine (0.5 mg/kg b.i.d.), alone and in combination, were compared with control animals (four groups, n = 10/group). Chlorzoxazone (CZN) was used as a probe drug to phenotype in vivo CYP2E1 activity before and after chronic ethanol and/or nicotine exposure. CYP2E1 protein levels and in vitro chlorzoxazone metabolism were assessed in liver microsomes. Average daily ethanol consumption was ≈3.0 g/kg (blood ethanol levels ≈24 mM) and was unaffected by nicotine treatment. Ethanol self-administration and nicotine treatment, alone and in combination, significantly increased in vivo CZN disposition compared with that in control animals. The effect of ethanol was only observed at higher levels of intake. Ethanol and nicotine increased CYP2E1 protein levels and in vitro CZN metabolism, with combined exposure to both drugs resulting in the greatest increase. The effect of ethanol was also dependent on level of intake. Chronic exposure to ethanol and nicotine induced hepatic CYP2E1 activity and protein levels, particularly when both drugs were used in combination and when ethanol intake was high. These results have important implications for public health, given the association between elevated CYP2E1 and disease, and the large proportion of individuals who are exposed to ethanol and nicotine, often in combination.

Respiratory muscle blood flows during physiological and chemical hyperpnea in the rat.

Whether the diaphragm retains a vasodilator reserve at maximal exercise is controversial. To address this issue, we measured respiratory and hindlimb muscle blood flows and vascular conductances using radiolabeled microspheres in rats running at their maximal attainable treadmill speed (96 +/- 5 m/min; range 71-116 m/min) and at rest while breathing either room air or 10% O(2)-8% CO(2) (balance N(2)). All hindlimb and respiratory muscle blood flows measured increased during exercise (P < 0.001), whereas increases in blood flow while breathing 10% O(2)-8% CO(2) were restricted to the diaphragm only. During exercise, muscle blood flow increased up to 18-fold above rest values, with the greatest mass specific flows (in ml. min(-1). 100 g(-1)) found in the vastus intermedius (680 +/- 44), red vastus lateralis (536 +/- 18), red gastrocnemius (565 +/- 47), and red tibialis anterior (602 +/- 44). During exercise, blood flow was higher (P < 0.05) in the costal diaphragm (395 +/- 31 ml. min(-1). 100 g(-1)) than in the crural diaphragm (286 +/- 17 ml. min(-1). 100 g(-1)). During hypoxia+hypercapnia, blood flows in both the costal and crural diaphragms (550 +/- 70 and 423 +/- 53 ml. min(-1). 100 g(-1), respectively) were elevated (P < 0.05) above those found during maximal exercise. These data demonstrate that there is a substantial functional vasodilator reserve in the rat diaphragm at maximal exercise and that hypoxia + hypercapnia-induced hyperpnea is necessary to elevate diaphragm blood flow to a level commensurate with its high oxidative capacity.

A physiological description of critical velocity.

Although critical velocity (CV) provides a valid index of aerobic function, the physiological significance of CV is not known. Twelve individuals performed exhaustive runs at 95% to 110% of the velocity at which VO2max was attained in an incremental test. VO2max was elicited in each run. Using the time to exhaustion at each velocity, CV was calculated for each participant. Using the time to achieve VO2max at each velocity, which was shorter at higher velocities, a parameter we have designated as CV' was calculated for each participant. During exercise at or below CV', VO2max cannot be elicited. CV (238+/-24 m x min(-1)) and CV' (239+/-25 m x min(-1)) were equal (t = 0.60, p = 0.56) and correlated (r = 0.97, p < 0.01). These results demonstrate that CV is the threshold intensity above which exercise of sufficient duration will lead to attainment of VO2max.

An alternative method to determine maximal accumulated O2 deficit in runners.

An accepted measure of anaerobic capacity is the maximal O2 deficit. But it is not feasible to use O2 deficit if > or =10 submaximal runs are needed to extrapolate the O2 demand of high velocity running (Medbø et al. 1988). Recently, an alternative method to determine O2 deficit was proposed (Hill 1996) using only results of supramaximal cycle ergometer tests. The purpose of this study was to evaluate this alternative method with data from treadmill tests. Twenty-six runners ran at 95%, 100%, 105%, and 110% of their velocity at VO2max. Times to exhaustion, velocity, and accumulated oxygen uptake (VO2) from each individual's four tests were fit to the following equation using iterative nonlinear regression: accumulated VO2 = (O2 demand x velocity x time)-O2 deficit. The mean value s derived for O2 demand and O2 deficit were 0.198+/-0.031 ml x kg(-1) x m(-1) and 42+/-22 ml x kg(-1). SEE for the parameters were 0.007+/-0.007 ml x kg(-1) x m(-1) and 8+/-10 ml x kg(-1), respectively. Mean R2 was 0.998+/-0.003. It was concluded that O2 deficit can be determined from all-out treadmill tests without the need to perform submaximal tests.

Greenhouse and field evaluations of entomopathogenic nematodes (Nematoda:Heterorhabditidae and Steinernematidae) for control of cabbage maggot (Diptera:Anthomyiidae) on cabbage.

Entomopathogenic nematodes--Heterorhabditis bacteriophora Poinar (Oswego strain), Steinernema carpocapsae (Weiser) (NY001 strain), Steinernema carpocapsae (25 strain), Steinernema feltiae Filipjev (= Neoaplectana carpocapsae Weiser) (369 strain), Steinernema feltiae (27 strain), and Steinernema riobravus Cabanillas and Poinar (355 strain)--were examined for pathogenicity against cabbage maggot, Delia radicum (L.), larvae in the greenhouse and field. Applications (per plant) of 3,000 and 4,000 infective juveniles of S. feltiae (369 strain), 30,000 infective juveniles of H. bacteriophora (Oswego strain), and 300 and 30,000 infective juveniles of S. feltiae (27 strain) reduced the number of D. radicum that developed to pupae on potted cabbage plants. H. bacteriophora (Oswego) at applications of 3,000 and 30,000 infective juveniles per plant and S. feltiae (27 strain) at applications of 30,000 (but not 3,000) infective juveniles per plant significantly reduced root damage caused by larvae of D. radicum. Logarithmically increased dosages between 100 and 100,000 infective juveniles per plant of S. feltiae (27 strain) linearly reduced the number of D. radicum pupae that developed on potted cabbage plants and the damage caused to the roots by D. radicum larvae. Root and stem dry weights of cabbage plants infested with D. radicum were significantly greater for plants inoculated with 100,000 infective juveniles of S. feltiae (27 strain) than for plants not inoculated with nematodes. Nematode inoculation did not prevent significant losses in root or stem dry weights at dosages less than 100,000 infective juveniles per plant. Soil surface applications of 100,000 and 200,000 infective juveniles per plant of S. feltiae (27 strain) were more effective than subsurface applications in preventing damage by natural or augmented populations of D. radicum larvae on cabbage in the field. However, mortality rates of wax moth larvae exposed to soil samples treated with S. feltiae (27 strain) suggested that this nematode showed greater persistence when applied beneath rather than on the soil surface.

Field evaluation of a DNA hybridization assay for nuclear polyhedrosis virus in gypsy moth (Lepidoptera: Lymantriidae) larvae.

DNA hybridization assays were used to detect the presence of viral DNA in gypsy moth (Lymantria dispar L.) larvae collected weekly from high density populations or reared from field-collected egg masses. DNA was extracted from larvae, bound to nitrocellulose filters, and hybridized with digoxigenin-labeled L. dispar NPV (LdNPV) DNA probes. The virus incidence determined from DNA hybridization assays was compared with that determined with conventional microscopic examination of larvae for polyhedral inclusion bodies. Among neonates reared from field-collected egg masses, average mortality from LdNPV (15.4%) within 10 d after hatch was not significantly different from the percentage of extracts containing LdNPV DNA (14.8%) found among larvae frozen 5 d after hatch before any mortality occurred. Field-collected larvae were split into two groups: half were frozen immediately and probed for LdNPV DNA and the other half were reared on artificial diet. The proportion containing LdNPV DNA closely approximated the proportion that died within 6 d of collection, but the proportion that died within 13 d of collection was underestimated.