Immunohistochemical demonstration of vascular endothelial growth factor in vestibular schwannomas correlates to tumor growth rate

OBJECTIVE: Vascular endothelial growth factor (VEGF) is one of the most potent mediators of angiogenesis, which is a mandatory process during tumor growth. The present objectives were to determine expression of VEGF in vestibular schwannomas by immunohistochemistry and to examine a possible correlation with symptom duration, tumor size, or growth rate. STUDY DESIGN: Retrospective patient file review; immunohistochemistry and light microscopy of vestibular schwannomas removed by surgery. METHODS: Vestibular schwannomas from 18 patients were immunolabelled using a polyclonal antibody against VEGF, followed by light microscopy and blinded semiquantitation of VEGF expression. Fifteen patients had a well-defined tumor growth rate defined by repeated preoperative magnetic resonance imaging scans. RESULTS: All tumors showed expression of VEGF in the Schwann cell cytoplasm, with a more intense staining of the perinuclear region of some cells. The staining intensity varied from tumor to tumor, and semiquantitation revealed a significant correlation between VEGF expression and tumor growth rate, but not symptom duration or tumor size. CONCLUSION: VEGF is expressed in vestibular schwannomas and the level of expression correlates positively with tumor growth rate, but not with tumor size and symptom duration. We conclude that VEGF seems to be a factor involved in the growth of vestibular schwannomas.     

Contrasting effects of interleukin-2 secretion by rat glioma cells contingent upon anatomical location: accelerated tumorigenesis in the central nervous system and complete rejection in the periphery

Rat T9.F glioma cells were transduced with the interleukin (IL)-2 gene. Clone T9.F/IL2/#12 secreted a high level of IL-2 (15 ng/10⁶ cells/48 h). Enhanced tumor progression and reduced survival was observed when T9.F/IL2/#12 cells were implanted intracranially. Subcutaneous injection of T9.F/IL2/#12 cells induced a palpable nodule, which regressed in approximately 15 days, resulting in tumor-specific protection. Lymphocytes from T9.F/IL2/#12 primed rats specifically respond to T9.F antigens but lacked cytotoxicity towards T9.F cells. Intracranial T9.F/IL2/#12 tumors were markedly infiltrated by CD4⁺ and CD8⁺ T cells, natural killer (NK)-T cells and myeloid progenitor cells, whereas subcutaneous T9.F/IL2/#12 tumors contained an elevated level of NK cells.     

Impact of two common polymorphisms in the PPARgamma gene on glucose tolerance and plasma insulin profiles in monozygotic and dizygotic twins: thrifty genotype,thrifty phenotype,or both?

The Pro12Ala polymorphism in the PPARgamma2 gene has been associated with reduced risk of type 2 diabetes and insulin resistance. Recently, an association between dizygotic twinning and PPARgamma gene polymorphisms has been proposed. We investigated the phenotypic appearance of the two polymorphisms (Pro12Ala and exon 6 C-->T) in PPARgamma among elderly twins (207 monozygotic [MZ] and 342 dizygotic [DZ]) and evaluated whether they could explain previously reported differences in plasma glucose and insulin profiles among MZ and DZ twins. We demonstrated a significant impact of the Pro12Ala polymorphism on glucose tolerance, diabetic status, homeostasis model assessment for insulin resistance, and plasma insulin profiles in twins. No impact of the silent exon 6 polymorphism on glucose homeostasis or plasma insulin profiles was found. Independent of the polymorphisms, we observed a significant impact of zygosity status per se on the plasma insulin profile after oral glucose ingestion, with the MZ twins being more hyperinsulinemic, indicating insulin resistance, than the DZ twins. Nonsignificantly higher glucose concentrations were observed among MZ compared with DZ twins. We demonstrated an association between the Ala allele and reduced risk of diabetes and insulin resistance in twins. However, the differences in metabolic profiles among MZ and DZ twins were not explained by differences in frequencies of the genetic variants and may be due to intrauterine environmental factors operating in twins independent of genotype. Accordingly, our study simultaneously supports a role for both the intrauterine environment (thrifty phenotype) and for genetics (thrifty genotype) in the etiology of insulin resistance and perhaps glucose intolerance in twins.     

Effect of smoking on erythorbic acid pharmacokinetics

Considerable evidence exists that smoking significantly lowers the concentration of plasma antioxidants. While for most antioxidants this effect appears to result mainly from altered dietary habits, ascorbic acid has recently been shown to be depleted by smoking per se. However, the direct cause of ascorbate depletion remains unclear. Erythorbic acid is a stereoisomer of ascorbic acid commonly used as antioxidant in foodstuffs and has the same redox properties as ascorbic acid. We therefore investigated if erythorbic acid could be used as a non-isotopic marker of smoking-induced oxidative stress. In a sample of smokers (n 10) and non-smokers (n 10), the pharmacokinetics of erythorbic acid were followed after a single oral dose (1 g) and subsequently, the effect of a 2-week ascorbic acid supplementation (0.5 g/d) on erythorbic acid kinetics was studied in a double-blind, placebo-controlled fashion. There were no significant effects of smoking or supplementation on relative bioavailability (difference in area under curve, AUC 0-infinity) of erythorbic acid (smokers 357 (sd 119), non-smokers 414 (sd 142) micromol.h/l; P=0.34). Time to reach maximum plasma concentration (Tmax) was significantly less in smokers (P=0.03). If the relative pharmacokinetics of erythorbic acid between smokers and non-smokers compares with those of AA, our present results do not suggest that altered pharmacokinetics is likely to play a major role in the ascorbic acid depletion consistently observed in smokers.     

Feeding minipigs fish oil for four weeks lowers postprandial triacylglycerolemia

We wanted to establish a minipig model for the study of postprandial lipemia and plasma lipid clearance after fish oil consumption. Seven minipigs were fed a fish oil-enriched nonpurified diet and a control diet for 4 wk in a randomized cross-over study. After each intervention period, each pig was challenged with a gastric fat load (2 g fat/kg body) and an intravenous fat bolus (0.1 g/kg body) on separate days. Frequent blood samples were collected for 6 h after the gastric fat load and for 40 min after the intravenous bolus. The fish oil-enriched diet was associated with lower triacylglycerol, glycerol and nonesterified fatty acid concentrations in the hours after the gastric fat load than the control diet (P < 0.05). In contrast, the triacylglycerol disappearance rate after the intravenous fat bolus was not affected by fish oil (P = 0.19). In conclusion, dietary fish oil supplementation attenuates postprandial lipemia in minipigs similarly to what occurs in humans. Minipigs could serve as a useful model for future studies of this phenomenon. We observed no significant effect of fish oil supplementation on plasma triacylglycerol clearance and thus were unable to identify the mechanism explaining the attenuated lipemia in minipigs.     

The impact of genes and pre- and postnatal environment on the metabolic syndrome. Evidence from twin studies

Our population-based Danish twin study demonstrated a genetic influence on several of the components included in the metabolic syndrome, i.e. glucose intolerance, overall obesity, systolic and diastolic blood pressure and low levels of HDL-cholesterol. Abdominal obesity, insulin resistance and hypertriglyceridaemia had, on the other hand, a relatively higher environmental aetiological component. Furthermore we demonstrated a difference in aetiology among male and female twins indicating an influence of sex on several of the components in the metabolic syndrome. Studies have demonstrated an impact of the intrauterine environment (i.e. low birth weight) for the development of the components in the metabolic syndrome. The validity of conclusions drawn from classical twin studies has therefore been questioned due to the different prenatal circumstances characterising monozygotic (MZ) and dizygotic (DZ) pregnancies. Due to a potentially more adverse intrauterine environment among MZ compared to DZ twins, MZ twins may be more prone to develop various metabolic abnormalities. Our findings of a higher glucose and insulin profiles after oral glucose ingestion, and recently lower insulin-stimulated glucose uptake--indicating glucose intolerance and insulin resistance--together with higher levels of total-cholesterol and triglycerides among MZ compared to DZ twins demonstrate an effect of zygosity (i.e. intrauterine environment) on these metabolic variables and therefore question the assumption of equal pre- and postnatal environment in MZ and DZ twins. Our studies provide further evidence for a prenatal component in the aetiology of the components included in the syndrome and question the validity of classical twin studies on phenotypes with a known prenatal aetiological component. However, our present knowledge is currently far too insufficient to discard the results from classical twin studies concerning the relative role of genes versus environment for the development of the metabolic and haemodynamic components included in the metabolic syndrome.     

Comparison of chlorzoxazone one-sample methods to estimate CYP2E1 activity in humans

Objective Comparison of a one-sample with a multi-sample method (the metabolic fractional clearance) to estimate CYP2E1 activity in humans.Methods Healthy, male Caucasians (n=19) were included. The multi-sample fractional clearance (Cl_fe) of chlorzoxazone was compared with one-time-point clearance estimation (Cl_est) at 3, 4, 5 and 6 h. Furthermore, the metabolite/drug ratios (MRs) estimated from one-time-point samples at 1, 2, 3, 4, 5 and 6 h were compared with Cl_fe.Results The concordance between Cl_est and Cl_fe was highest at 6 h. The minimal mean prediction error (MPE) of Cl_est as a percentage of actual mean Cl_fe was –4.2% at 6 h. Furthermore, regarding Cl_fe, there was a negligible difference (P=0.56) of bias between Cl_est at 3 h (MPE=–8.9%) and 6 h (MPE=–4.2%). The best concordance between MR and Cl_fe was found at 3 h (r=0.74; P<0.001).Conclusion All three single-dose-sample estimates, Cl_est at 3 h or 6 h, and MR at 3 h, can serve as reliable markers of CYP2E1 activity. The one-sample clearance method is an accurate, renal function-independent measure of the intrinsic activity; it is simple to use and easily applicable to humans.     

The analgesic effect of tramadol after intravenous injection in healthy volunteers in relation to CYP2D6

Tramadol analgesia results from a monoaminergic effect by tramadol itself and an opioid effect of its metabolite (+)-M1 formed by O-demethylation of tramadol by CYP2D6. In this study we sought to determine the impact of (+)-M1 on the analgesic effect of tramadol evaluated by experimental pain models. The effect of an IV injection of 100 mg tramadol on experimental pain was studied 15-90 min after dosing in volunteers, 10 extensive metabolizers with CYP2D6 and 10 poor metabolizers without CYP2D6 in 2 placebo-controlled trials. The pain tests included detection and tolerance threshold to single electrical sural nerve stimulation, pain summation threshold to repetitive electrical sural nerve stimulation (temporal summation), and the cold pressor test. In extensive metabolizers, tramadol reduced discomfort experienced during the cold pressor test (P = 0.002). In poor metabolizers, the pain tolerance thresholds to sural nerve stimulation were increased (P = 0.04). (+)-M1 could be detected in the serum samples from all extensive metabolizers except one, but (+)-M1 was below the limit of determination in all poor metabolizers. The opioid effect of (+)-M1 appears to contribute to the analgesic effect of tramadol, but the monoaminergic effect of tramadol itself seems to create an analgesic effect.