The effect of nicotinamide on gene expression in a traumatic brain injury model.

Microarray-based transcriptional profiling was used to determine the effect of nicotinamide on gene expression in an experimental traumatic brain injury (TBI) model. Ingenuity Pathway Analysis (IPA) was used to evaluate the effect on relevant functional categories and canonical pathways. At 24 h, 72 h, and 7 days, respectively, 70, 58, and 76%, of the differentially expressed genes were up-regulated in the vehicle treated compared to the sham animals. At 24 h post-TBI, there were 150 differentially expressed genes in the nicotinamide treated animals compared to vehicle; the majority (82%) down-regulated. IPA analysis identified a significant effect of nicotinamide on the functional categories of cellular movement, cell-to-cell-signaling, antigen presentation and cellular compromise, function, and maintenance and cell death. The canonical pathways identified were signaling pathways primarily involved with the inflammatory process. At 72 h post-cortical contusion injury, there were 119 differentially expressed genes in the nicotinamide treated animals compared to vehicle; the majority (90%) was up-regulated. IPA analysis identified a significant effect of nicotinamide on cell signaling pathways involving neurotransmitters, neuropeptides, growth factors, and ion channels with little to no effect on inflammatory pathways. At 7 days post-TBI, there were only five differentially expressed genes with nicotinamide treatment compared to vehicle. Overall, the effect of nicotinamide on counteracting the effect of TBI resulted in significantly decreased number of genes differentially expressed by TBI. In conclusion, the mechanism of the effect of nicotinamide on secondary injury pathways involves effects on inflammatory response, signaling pathways, and cell death.

Therapeutic role for bismuth compounds in TNBS-induced colitis in the rat.

The 2,4,6-trinitrobenzene sulfonic acid (TNBS) -induced model of chronic inflammation of the rat colon was used to determine the efficacy of bismuth subsalicylate (BSS), bismuth subcitrate (CBS), and 5-aminosalicylic acid (5-ASA) administered in enema form. A novel bismuth compound 1, 2-bis[2-(1,3-dithiobismolane)thio]ethane [Bi2(EDT)3] was also tested. On day 1 colitis was induced with 50 mg TNBS/50% ethanol in female Sprague-Dawley rats, while controls received a saline enema. On day 3, twice-daily treatment with enemas of either saline, BSS, CBS, Bi2(EDT)3, or 5-ASA were initiated in the colitis and control rats. All rats were killed on day 14, and the colons excised, weighed, rated macroscopically, and then fixed for hematoxylin and eosin staining. Blinded microscopic scoring was used to determine injury and healing in all groups. Colon mass and macroscopic scores were increased (P < 0.05) in the group of rats treated with TNBS (N = 16) compared to saline controls (N = 12). Colon mass and macroscopic scores in controls treated with BSS (N = 4), CBS (N = 4), Bi2(EDT)3 (N = 4), and 5-ASA (N = 4) alone did not differ from saline control animals. Macroscopic scoring showed a decrease (P < 0.05) in the degree of damage in the group of rats treated with TNBS plus BSS (N = 15), TNBS plus Bi2(EDT)3 (N = 10) and TNBS plus CBS (N = 4) compared to the group of rats treated with TNBS plus saline (N = 16). A decrease (P < 0.05) in injury and an increase (P < 0.05, Kruskal-Wallis) in healing was observed in the groups of rats treated with TNBS plus BSS, TNBS plus CBS, and TNBS plus 5-ASA compared to the group of rats treated with TNBS plus saline. It appeared that Bi2(EDT)3 was not protective against injury at the microscopic level but that the novel Bi2(EDT)3 has an effective healing capacity at the macroscopic level. We conclude that BSS and CBS decrease injury and/or promote healing as effectively as 5-ASA in this model.

The inhibitory effect of ursodeoxycholic acid and pentoxifylline on platelet derived growth factor-stimulated proliferation is distinct from an effect by cyclic AMP.

This study assessed the ability of ursodeoxycholic acid (UDCA) and one of its metabolites, tauroursodeoxycholic acid (TUDCA), to inhibit platelet derived growth factor (PDGF) stimulated fibroproliferation and compared these results to the effect of pentoxifylline and its metabolite-1 [1-(5-hydroxyhexyl)-3,7-dimethylxanthine] and assessed the potential role of cyclic AMP in this process. Fibroproliferative activity was measured by the tritiated thymidine uptake assay in human fibroblast cultures. All four compounds: pentoxifylline, metabolite-1, UDCA and TUDCA inhibited the fibroproliferative activity stimulated by PDGF (8 ng/ml). Incubation of fibroblasts with dibutyryl cyclic AMP reduced proliferation stimulated by PDGF suggesting that the PDGF stimulated proliferation was sensitive to inhibition by a membrane permeable analogue of cyclic AMP. Incubation of myofibroblasts with dibutyryl cyclic AMP significantly inhibited PDGF stimulated proliferation suggesting that cyclic AMP can regulate PDGF stimulated proliferation in the myofibroblast. To determine if the effect of pentoxifylline on fibroproliferation was mediated by cyclic AMP, we used dideoxyadenosine, a potent inhibitor of adenylyl cyclase. The effect of pentoxifylline on fibroproliferation was not prevented by dideoxyadenosine, which inhibits formation of cyclic AMP, thus suggesting that the inhibitory effect of pentoxifylline on PDGF-stimulated proliferation of fibroblasts was not mediated by cyclic AMP, arguing against a role for cyclic AMP in this process. Combinations of UDCA (250 microM) plus pentoxifylline (120 microM) or UDCA (250 microM) plus TUDCA (250 microM) inhibited fibroproliferative activity stimulated by PDGF to a greater extent than either drug alone. As UDCA has been reported to decrease cyclic AMP these results argue against a role for cyclic AMP in this process. Finally the results suggest that UDCA may inhibit PDGF-stimulated proliferation via an inhibition of C-kinase.

Platelet-derived growth factor and pentoxifylline modulation of collagen synthesis in myofibroblasts.

Fibroblast proliferation and extracellular matrix accumulation are two major events occurring in fibrosis. Hepatic stellate cells are the major collagen-producing cells of the liver and are transformed into proliferative myofibroblasts following activation. Whether proliferation and extracellular matrix production are regulated by the same cytokines is not known. Monocyte-conditioned medium obtained from pigs with yellow phosphorus-induced hepatic fibrosis increased the collagen production by cultured procine myofibroblasts. Liver biopsies from these same fibrotic animals had increased levels of collagen alpha 1(I) and alpha 1(III) mRNA compared to control animals. Preincubation with platelet-derived growth factor (PDGF) B/B antibody significantly reduced the collagen-stimulating ability of the monocyte-conditioned medium. Recombinant PDGF stimulated proliferation in nonconfluent myofibroblasts and stimulated collagen production in confluent cultures of myofibroblasts without increasing cell number, suggesting that these events can occur independent of each other. Pentoxifylline and one of its active metabolites (metabolite-1) inhibited PDGF-stimulated collagen production in cultured porcine myofibroblasts. These results demonstrate the importance of PDGF in the pathogenesis of liver fibrosis and provide evidence that pentoxifylline interferes with PDGF-mediated events during experimental liver fibrosis.

Drug-metabolizing enzymes in rat liver myofibroblasts.

The myofibroblast is considered to be a key component in the pathogenesis of hepatic fibrosis. There is a need for therapeutic intervention in hepatic fibrosis, and, to date, the number of efficacious anti-fibrotic drugs is negligible. At best, the current therapeutic modalities reduce liver enzymes, an indicator of liver damage, but cannot reduce or prevent fibrosis. We have described the anti-fibrotic effect of pentoxifylline in an experimental model of hepatic fibrosis. Evidence suggests that, in addition to pentoxifylline itself, at least two of the metabolites of pentoxifylline are of therapeutic interest. We have reported that one of these metabolites (M-1) has a biological activity similar to that of its parent drug. The second metabolite (M-1R) has been reported to be more potent than the parent drug. Recent evidence suggests that inhibition of cytochrome P450 1A2 (CYP1A2) results in higher levels of pentoxifylline and M-1 and may be responsible for the production of the novel, potent metabolite (M-1R). We therefore investigated whether the myofibroblast, the cell with a crucial role in fibrosis, contains drug-metabolizing enzymes and thus may play a critical role in the anti-fibrotic actions of pentoxifylline. Our results showed that myofibroblasts contain aryl hydrocarbon hydroxylase activity, ethoxyresorufin O-deethylase activity, and methoxyresorufin O-demethylase activity. The results presented here also indicate that aryl hydrocarbon hydroxylase and methoxyresorufin O-demethylase activities can be increased by treatment of cells with dibenzanthracene, an inducer of CYP1A activities.

Effect of acute pentoxifylline treatment in an experimental model of colitis.

BACKGROUND: The effect of acute pentoxifylline treatment in an experimental model of colitis was assessed using the trinitrobenzene sulphonic acid (TNBS)-induced rat model of colitis. METHODS: Animals were treated with intracolonic injection (250 microliters) of TNBS (50 mg in 50% ethanol) to induce inflammation and ulcers. Animals received pentoxifilline (100 mg/kg intracolonically) or saline 24 and 48 h following TNBS treatment. Five days following TNBS treatment, colons were dissected and scored according to the morphology damage score. The colons were then rolled longitudinally, fixed in formalin and embedded in paraffin. The collagen content of colonic sections were determined by a Sirius red-Fast green technique. RESULTS: Animals treated with TNBS alone had significantly higher gross morphology damage scores compared to animals treated with saline. Pentoxifylline significantly reduced the gross morphology damage score in animals receiving TNBS. Colonic collagen levels were significantly elevated in TNBS-treated animals compared to animals receiving saline. Pentoxifylline treatment did not alter the collagen content of colons from TNBS-treated animals. CONCLUSION: TNBS treatment significantly elevates morphology damage score compared to controls. The results also suggest that colonic collagen was significantly elevated in animals treated with TNBS compared to controls. Pentoxifylline treatment was not sufficient to reduce the elevation in colonic collagen, although pentoxifylline treatment was sufficient to reduce the pathological changes due to TNBS, thus bringing the morphology damage score down to control levels.

Inhibition of fibroproliferation by pentoxifylline. Activity of metabolite-1 and lack of role of adenosine receptors.

We have reported previously that pentoxifylline and adenosine decrease platelet-derived growth factor- (PDGF) stimulated fibroproliferation. To determine the role of adenosine receptors in the inhibition of fibroproliferation observed with pentoxifylline, we used a non-selective adenosine receptor antagonist, 8-phenyltheophylline, and specific A1 and A2 adenosine receptor antagonists. If the A2 receptor, which is present on fibroblasts, mediates the inhibition of fibroproliferation which occurs with pentoxifylline, then pretreatment of fibroblasts with receptor antagonists prior to the addition of pentoxifylline should prevent the action of pentoxifylline. The results indicated that pretreatment of fibroblasts with 8-phenyltheophylline (100 microM) did not alter the inhibitory effect of pentoxifylline on PDGF-stimulated fibroproliferation. These results argue against a mechanism involving inhibition of adenosine reuptake as the mechanism for pentoxifylline's effect in this system. 8-Phenyltheophylline also did not alter the effect of pentoxifylline on baseline proliferation, suggesting that these effects of pentoxifylline are not mediated by adenosine receptors. Pentoxifylline is metabolized to several metabolites including 1-(5-hydroxyhexyl)-3,7-dimethylxanthine (metabolite-1). Metabolite-1 significantly reduced PDGF-stimulated fibroproliferation and was as effective as pentoxifylline. The combination of pentoxifylline and metabolite-1 had an additive effect. Metabolite-1 and pentoxifylline also reduced baseline proliferation. Preincubation of fibroblasts with 8-phenyltheophylline did not prevent the inhibitory action of metabolite-1 on PDGF-stimulated proliferation or on basal proliferation of fibroblasts, suggesting that the action of metabolite-1 on fibroproliferation was not mediated by adenosine receptors. Results using A1 and A2 adenosine receptor antagonists further suggest that the effect of pentoxifylline was not mediated by adenosine receptors.

Drug treatment of functional dyspepsia: a systematic analysis of trial methodology with recommendations for design of future trials.

OBJECTIVES: To evaluate drug treatment of functional dyspepsia (including Helicobacter pylori) and provide guidelines for future trials based on a critical systematic overview of published studies. METHODS: Data sources were a Medline search for articles published in English going back to 1966 and a manual search of four GI journals going back to 1980. Original randomized, double-blind, placebo-controlled trials were selected that enrolled at least 20 patients. Using a standardized, pretested data extraction form, studies were evaluated independently by two observers for study design, outcome measures, and results. RESULTS: Fifty two eligible studies were evaluated. Many studies suffered from important weaknesses in study design and execution. Only five studies used previously validated outcome measures. CONCLUSIONS: Because of suboptimal design and/or unclear presentation of the data, none of the trials provided unequivocal evidence that there is efficacious therapy for the treatment of functional dyspepsia.

Effect of pentoxifylline in rat and swine models of hepatic fibrosis: role of fibroproliferation in its mechanism.

Fibroproliferation was studied in two animal models of liver disease. Oral feeding of yellow phosphorus to pigs reproducibly results in fibrosis after 8 weeks of feeding, extensive fibrosis after 12 weeks and cirrhosis after 16 weeks of yellow phosphorus. Bile duct ligation was used to induce cirrhosis in the rat. Fibroproliferation was assessed as uptake of tritiated thymidine into fibroblasts which had been incubated with monocyte-conditioned medium obtained from monocytes of pigs treated with yellow phosphorus or bile duct-ligated rats and compared to the corresponding controls. Fibrosis was assessed by collagen content of liver sections obtained from the two animal models. The collagen content was determined by quantitation of Sirius red/Fast green-stained liver sections. In both animal models collagen content was significantly elevated at the conclusion of the treatment. Collagen content of liver sections of yellow phosphorus-treated animals were elevated (40 +/- 2.7, n = 15) compared to mineral oil-treated controls (23 +/- 1.2, n = 12) and collagen levels in the bile duct-ligated rat model liver sections were elevated (31.2 +/- 1.6, n = 6) compared to sham-operated controls (21.6 +/- 0.7, n = 6). The results of the fibroproliferation assay indicate that monocytes obtained from pigs treated with yellow phosphorus produce fibroproliferative factors during the development of fibrosis. This is in contrast to the bile duct-ligated rat model where no differences were observed in the production of fibroproliferative factors in the bile duct-ligated rats compared to sham operated controls suggesting that this may not be a key event in this model of fibrosis. Pentoxifylline treatment of the yellow phosphorus induced swine model of hepatic fibrosis has been associated with a marked improvement in fibrosis. In this study treatment of fibrotic pigs with pentoxifylline was associated with an improvement in liver function tests, a reduction of collagen content of liver sections, and reduction in fibroproliferation in pigs receiving yellow phosphorus treatment. Fibroproliferative factors were produced during the development of fibrosis in the swine model of fibrosis and their effect was blocked by pentoxifylline administered in vivo. This is in contrast to the bile duct-ligated rat model where pentoxifylline treatment was not associated with improvement in liver function tests or reduction of collagen content of liver sections and did not alter the fibroproliferative activity of monocyte-conditioned media. Taken together these results suggest that fibroproliferation and increased synthesis of collagen are key events in the yellow phosphorus-induced pig model of hepatic fibrosis and that the action of pentoxifylline in this animal model is likely to be related to its effects on fibroproliferation with a subsequent effect on collagen production. This is in contrast to the bile duct-ligated rat model where pentoxifylline does not prevent hepatic fibrosis.

Therapeutic potential for cytokine antagonists: Thalidomide and pentoxifylline in Hansen's disease.

Cytokine antagonists are a group of drugs defined by their actions on specific cytokines. Cytokine antagonists can inhibit action of cytokines by acting directly on receptors, by affecting production of cytokines or by binding to cytokines and preventing their subsequent action. Recent evidence suggests that Hansen's disease, which is characterized by reactional states, is associated with elevated serum levels of tumour necrosis factor-α (tnf-α) and interleukin-1ß during these reactional states. Thalidomide, a drug used to treat reactional states in Hansen's disease, has been reported to enhance degradation of tnf-α mrna. Pentoxifylline has also been reported to alter tnf-α mrna levels by inhibiting tnf-a transcription. Combination of these two drugs as cytokine antagonists may prove to be beneficial as therapeutic agents in the treatment of reactional states in Hansen's disease. Pentoxifylline may prove to be beneficial in the treatment of reactional states in Hansen's disease patients who are female and of childbearing age. Cytokine antagonists alone or in combination will likely fill a niche in future therapeutics.

Cytokine production and aging: overproduction of IL-8 in elderly males in response to lipopolysaccharide.

Interleukin-8 (IL-8) is a chemoattractant cytokine for polymorphonuclear neutrophils, and is found at the site of inflammation and infection. The levels of IL-8 from an elderly (ages 65-79) and young (ages 20-27) population were compared. Secretion of IL-8 was measured in monocyte conditioned medium (MCM), under both a spontaneous condition and with stimulation with detoxified LPS (10 mg/ml). Spontaneous production of IL-8 in the elderly group (39.4 +/- 8.3 ng/ml, n = 16) was found to be significantly lower than the control group (66.4 +/- 5.0 ng/ml, n = 17), P < 0.01. A sex difference was observed within the elderly population, with the male elderly producing 8.8 +/- 2.1 ng/ml of IL-8 and the elderly females producing levels of 57.8 +/- 9.1 ng/ml. There was a good correlation between IL-8 and IL-1 production in the elderly but differences between the elderly and young production of IL-1 did not reach statistical significance. IL-8 and TNF production did not correlate. Upon stimulation with the LPS, the male elderly levels increased eightfold (70.1 +/- 11.8 ng/ml) and was significantly different from the young male level, P < 0.01, while the female elderly showed no change with stimulation. No sex difference was observed in the control population. These results indicate that the spontaneous secretion of IL-8 in elderly males is lower than that of both elderly females and the young control group. However, upon stimulation with LPS, the elderly males are capable of an overproduction of IL-8 when compared to the young group and the elderly females. This overproduction may be the result of an in vivo priming in this healthy elderly group. The female elderly followed a pattern similar to the young group, showing no change upon stimulation with the detoxified LPS. Sex differences related to the immune system have been noted in the past with females having a more active immune system, and these results may be related to this difference.

In vitro effect of platelet-derived growth factor on fibroproliferation and effect of cytokine antagonists.

Platelet-derived growth factor (PDGF) stimulates fibroblast proliferation and increases collagen synthesis. Fibroproliferation, as assessed by tritiated thymidine uptake, was significantly stimulated by platelet-derived growth factor (8 ng/ml). Several drugs including dexamethasone (1 nM-20 microM), cysteamine (1.3-65 mM), N-acetylcysteine (0.6-15 mM), glutathione (1 microM-0.1 mM), glutathione peroxidase (0.1-1 Unit/ml), pentoxifylline (60 microM-36 mM), colchicine (0.025-250 nM) and aurothioglucose (1.15-23 microM) were assessed in the fibroproliferation assay for their ability to block the fibroproliferative effect of platelet-derived growth factor. Dexamethasone and aurothioglucose did not affect PDGF-stimulated fibroproliferation, while pentoxifylline, colchicine, cysteamine and N-acetylcysteine effectively reduced fibroproliferation stimulated by PDGF. The effect of pentoxifylline on PDGF stimulated fibroproliferation was compared to trapidil, theophylline and adenosine to assess mechanism of action. All four methylxanthines effectively inhibited PDGF stimulated fibroproliferation. Pentoxifylline was as effective as trapidil (IC50 = 129 microM and 141 microM, respectively), but pentoxifylline was more potent than theophylline (IC50 = 688 microM) and pentoxifylline was not as potent as adenosine (IC50 = 19 microM) in reducing PDGF-stimulated fibroproliferation.

Pentoxifylline prevents fibrosis in an animal model and inhibits platelet-derived growth factor-driven proliferation of fibroblasts.

Liver fibrosis is a complex process characterized by two major events: fibroproliferation and increased collagen synthesis. The exact role of cytokines in the pathogenesis of hepatic fibrosis remains to be established, but platelet-derived growth factor clearly stimulates proliferation of fibroblasts and increases collagen synthesis. In in vitro studies, pentoxifylline, a methylxanthine, significantly reduced platelet-derived growth factor-driven proliferation of fibroblasts. Platelet-derived growth factor has also been identified as a fibroproliferative factor produced spontaneously by monocytes obtained from patients with liver disease. Long-term administration of pentoxifylline (16 mg/kg orally, 5 days/wk for 12 wk) in an animal model of liver fibrosis prevented elevations in gamma-glutamyl transpeptidase and alkaline phosphatase levels and prevented the reduction in serum albumin level normally observed in this animal model of liver disease. The animal model used was a long-term, low-dose yellow phosphorus--induced model in pigs that reproducibly results in extensive fibrosis after 10 to 12 wk of treatment. Long-term administration of pentoxifylline also prevented the histological changes characteristic of fibrosis in this animal model. Collagen concentration was significantly elevated in liver sections obtained from animals receiving yellow phosphorus, compared with controls. Long-term pentoxifylline treatment resulted in significantly lower collagen concentrations in liver sections from animals receiving yellow phosphorus than in sections from animals receiving yellow phosphorus alone; this was supported by histological observation. Therefore administration of pentoxifylline prevented the biochemical and histological changes associated with an animal model of liver disease. Pentoxifylline will likely have an important therapeutic role in liver fibrosis.

Interleukin-1, platelet derived growth factor, free radicals and monocyte aryl hydrocarbon hydroxylase activity in liver disease. Role of cell communication.

Monocytes were isolated from blood of human origin and cultured in supplemented Leibovitz (L-15) medium for 24 hr. The medium was then decanted and filtered, and all subsequent tests were done on monocyte conditioned medium (MCM). The monocytes of patients with liver disease spontaneously secrete temperature-sensitive arylhydrocarbon hydroxylase (AHH) inhibitory factors detectable in the MCM. Anti-interleukin-1 antibody (IL-1Ab) reduced the AHH inhibitory activity of the MCM, suggesting that part of the AHH inhibitory activity was due to interleukin-1 (IL-1). Platelet derived growth factor did not affect AHH activity. Interleukin-1 beta was detectable in MCM but did not differ significantly between patients and normal volunteers. A time course experiment indicated that interleukin-1 beta inhibited hepatocyte AHH activity after only 2 hr of incubation. Catalase partially blocked the AHH inhibitory activity of MCM suggesting that activated oxygen intermediates are partially involved in the AHH inhibitory activity of the MCM. Simultaneous incubation of interleukin-1 beta and catalase did not prevent or augment the inhibitory action of IL-1 on AHH activity. IL-1 stimulates collagen synthesis and elevates serum procollagen type 3 peptide (P-III-P). Results indicated that serum P-III-P was elevated in blood sources producing temperature-sensitive AHH inhibitory factor.

Fibroproliferation in liver disease: role of monocyte factors.

Fibroproliferation was measured as the uptake of [3H]thymidine into fibroblasts. Human fibroblasts were incubated with 200 microliters monocyte-conditioned medium, the 0.22 microns filtrate from cultured monocytes, in Dulbecco's modified Eagle medium supplemented with controlled process serum replacement 2, a fetal calf serum substitute with low mitogenic activity. Increasing the numbers of fibroblasts resulted in a parallel increase in thymidine uptake to a maximal level. Fibroblasts (2 x 10(3] were plated into microwell plates and incubated with monocyte-conditioned medium for 72 hr. At 16 hr before harvest, 1 muCi [3H]thymidine was added. Cells were harvested with phosphate-buffered saline and washed, and the filters were counted. Fibroblasts incubated with Dulbecco's modified Eagle medium and controlled process serum replacement 2 showed minimal thymidine uptake. Fibroblasts incubated with Dulbecco's modified Eagle medium plus monocyte-conditioned medium from monocytes stimulated with 10 micrograms/ml lipopolysaccharides showed a sixfold increase in thymidine uptake over fibroblasts in Dulbecco's modified Eagle medium and controlled process serum replacement 2 alone. Fibroblasts incubated with Dulbecco's modified Eagle medium plus monocyte-conditioned medium from monocytes of patients with liver disease (n = 20) showed a 10-fold elevation in thymidine uptake compared with Dulbecco's modified Eagle medium and controlled process serum replacement 2. Results indicated that preincubation of monocyte-conditioned medium with either anti-interleukin-1 beta (12.5 half-maximal units, 4 degrees C, 16 hr) or catalase (1,870 IU, 25 degrees C, 1 hr) did not alter the fibroproliferative activity of the monocyte-conditioned medium, suggesting that neither interleukin-1 beta nor activated oxygen intermediates were involved in fibroproliferation.(ABSTRACT TRUNCATED AT 250 WORDS)

Cysteamine in combination with N-acetylcysteine prevents acetaminophen-induced hepatotoxicity.

N-Acetylcysteine (NAC) is protective against acetaminophen-induced hepatotoxicity primarily by providing precursor for the glutathione synthetase pathway, while cysteamine has been demonstrated to alter the cytochrome P-450 dependent formation of toxic acetaminophen metabolite. Mice administered acetaminophen (500 mg/kg) had elevations of serum alanine aminotransferase (ALT) to 273.0 +/- 37.5 and 555.8 +/- 193.4 U/mL at 12 and 24 h, respectively, after injection. Administration of cysteamine (100 mg/kg) or NAC (500 mg/kg) significantly reduced serum ALT activity (p less than 0.001). Reducing the dose of NAC or cysteamine by 50% greatly reduced their hepatoprotective effect while the co-administration of the reduced doses of NAC (250 mg/kg) and cysteamine (50 mg/kg) following acetaminophen overdose prevented elevation of serum ALT activity (39.2 +/- 1.17 and 32.5 +/- 5.63 U/mL at 12 and 24 h post-injection, p less than 0.001) and preserved normal mouse hepatic histology. Neither NAC (500 mg/kg), cysteamine (100 mg/kg), or the lower doses in combination of both agents were found to alter the half-life or peak levels of acetaminophen. Liver microsomal aryl hydrocarbon hydroxylase activity measured 24 h after drug administration was not significantly different between treatment groups and controls receiving only saline. These results indicate a possible role for the concomitant use of NAC and cysteamine in the prevention of hepatic necrosis following toxic doses of acetaminophen. Neither decrease in plasma acetaminophen levels nor depression of cytochrome P-450 enzyme activity appears to be the mechanism of protection when these doses of NAC, cysteamine, or both drugs together are administered with a toxic dose of acetaminophen in mice.

Monocyte aryl hydrocarbon hydroxylase (AHH) activity mimics Kupffer cell and hepatocyte AHH activity in an animal model of liver disease.

We have previously reported that monocyte aryl hydrocarbon hydroxylase (AHH) activity is depressed in patients with liver disease and is decreased more in cirrhosis than in early stage liver disease. To determine if monocyte AHH activity reflects liver AHH activity, we studied an animal model of cirrhosis, i.e., yellow phosphorus induced cirrhosis in the pig. AHH activity was detectable in monocytes isolated from peripheral blood of normal pigs (0.32 +/- 0.13 nmol.mg-1 P.h-1, n = 11) and was comparable to the level of AHH activity in hepatic Kupffer cells isolated from wedge or needle biopsies of livers of normal pigs (0.38 +/- 0.21, n = 7). The AHH level in pig Kupffer cells was approximately 10% of the AHH level in hepatocytes and microsomes. To induce liver disease, pigs were administered yellow phosphorus (0.6 mg/kg) 5 days per week for 16 weeks. At 4 weeks of treatment, monocyte AHH activity was not different from control and liver histology was normal. Depression of monocyte AHH activity was evident at 8 weeks of treatment when liver fibrosis was seen histologically. At 12 weeks of treatment when histology revealed extensive liver fibrosis and collagen levels were elevated, the level of monocyte AHH activity was decreased 67% compared with controls. Similar changes were observed at 12 weeks in Kupffer cell AHH activity (86% decrease) and hepatocyte AHH activity (70% decrease) compared with controls. These results suggest that monocyte AHH activity reflects liver AHH activity and may be a good indicator of change in liver enzyme function in liver disease in the pig model of cirrhosis.

Effect of aging and other factors on monocyte aryl hydrocarbon hydroxylase activity.

A measure of the activity of macrophage drug metabolizing enzymes through assay of peripheral monocytes was used to assess the hepatic enzymatic status and thereby evaluate age related changes in drug metabolism. Blood was obtained from elderly subjects (aged 74.8 +/- 5.2, mean +/- S.E., n = 16) and a young control group (aged 23.5 +/- 2.0, n = 27). Monocyte AHH activity was used as an index of liver drug metabolism, ALT activity as an index of liver function, monocyte media IL-1 and as an index of macrophage activation and serum IL-1 levels as a measure of endogenous pyrogenic activity. The medium collected from the cultured monocytes was also assessed for the presence of AHH inhibitory activity. Subjects provided information relating to their age, sex, alcohol consumption, cigarette smoking, recent infection, recent surgery, disease status and medications which could alter drug metabolism. Elderly patients were drawn both from independent seniors living at home and seniors visiting a geriatric day hospital and compared to a control group of young healthy volunteers. Using the experimental design AHH activity did not differ within experimental error between aged (0.832 +/- 0.32 nmol/mg prot. per h, n = 16) and young control subjects (0.452 +/- 0.17, n = 27). ALT activity did not differ between aged (2.83 I.U. +/- 0.46) and young (4.24 +/- 0.82). Monocyte AHH activity did not differ between males (0.45 +/- 0.14, n = 33) compared to females (0.65 +/- 0.18, n = 29), but was significantly higher in smokers (2.5 +/- 1.0, n = 5) compared to non-smokers (0.35 +/- 0.05, n = 52). Mild to moderate alcohol use showed no significant effect on AHH activity. There was no significant difference between the mean level of MCM inhibition of murine hepatocyte AHH between elderly (44.3 +/- 8.32%, n = 8) and control (31.5 +/- 6.21%, n = 15) subjects, but a larger proportion of the elderly population demonstrated such an effect. Serum IL-1 levels (range 0-55.9 pg/ml) were compared to MCM IL-1 and AHH inhibitory activity in the elderly and young group.

Monocyte arylhydrocarbon hydroxylase activity and mortality.

Monocyte arylhydrocarbon hydroxylase (AHH) activity is decreased in patients with liver disease and correlates with severity of disease. Patients with chronic liver disease (n = 34) were studied to determine if decreased monocyte AHH activity is associated with mortality. Monocyte AHH activity in the nonsurvivor group was 0.07 +/- 0.025 nmol/mgP/h (n = 11) and was significantly lower than the survivor group 0.198 +/- 0.031 (n = 23). Both groups were significantly lower than controls 0.41 +/- 0.053 (n = 19). Of the liver function tests, only serum albumin was different between the survivor group and the nonsurvivor group. Patients in the nonsurvivor group had significantly higher Child-Turcotte scores than the survivor group. These results suggest that the monocyte AHH activity may be a good index of survival in patients with liver disease, but the high degree of overlap between survivors and nonsurvivors suggests otherwise.

Restructuring a family practice obstetrics curriculum.

Maintaining a high-quality curriculum for family practice residency training in obstetrics has become increasingly difficult. In 1984 the faculty of the University of Vermont Department of Family Practice needed to upgrade its obstetric curriculum in a community where family practice obstetrics was nonexistent. The key steps to a new curriculum included the recruitment of family practice faculty with experience in obstetrics, expanded communication with the Department of Obstetrics and Gynecology, the development of baseline attending privileges in family practice obstetrics, the formation of educational tracks for residents, and the promotion of chart audits. Also important were faculty role modeling, intradepartmental meetings, intensive elective rotations, and community education. This case report of program development in family practice obstetrics may serve as a model to help other residency programs.