Lamivudine, adefovir and tenofovir exhibit long-lasting anti-hepatitis B virus activity in cell culture

In this work, we investigated the anti-hepatitis B virus (HBV) activity of lamivudine, adefovir, tenofovir, penciclovir and lobucavir after short-term (i.e. 24 or 48 h) or continuous (9 days) exposure of the HBV-containing cell line, HepG2 2.2.15, to these drugs. Lamivudine maintained significant anti-HBV activity when added for only 24 or 48 h to the cell cultures compared to when the drug was present for the whole period (9 days) on the cells, i.e. 50% effective concentration (EC₅₀) values for the inhibition of HBV DNA synthesis were 0.07 ± 0.02 μg ml⁻¹ after 24 h of incubation, 0.02 ± 0.01 μg ml⁻¹ after 48 h of incubation and 0.0016 ± 0.001 μg ml⁻¹ after 9 days of incubation. Similarly, the nucleoside phosphonate analogues, adefovir and tenofovir, retained significant anti-HBV activity when added for only a short period of time to the cells. The EC₅₀ values were 12 ± 1 μg ml⁻¹ (24 h) and 1.0 ± 0.2 μg ml⁻¹ (48 h) vs 0.003 ± 0.001 μg ml⁻¹ (9 days) for adefovir, and 6.5 ± 1.1 μg ml⁻¹ (24 h) and 0.8 ± 0.1 μg ml⁻¹ (48 h) vs 0.03 ± 0.02 μg ml⁻¹ (9 days) for tenofovir. In contrast, penciclovir and lobucavir lost most of their anti-viral activity when present on the cells for 48 h or less.     

Application of the intravenous glucose tolerance test and the minimal model to patients with insulinoma: insulin sensitivity (Si) and glucose effectiveness (Sg) before and after surgical excision

Background  The unmodified frequently sampled intravenous glucose tolerance test (FSIGT) has not previously been used to assess insulin/glucose kinetics in patients with insulinoma.
Objective  To measure insulin sensitivity (Si) and glucose effectiveness (Sg) by means of the FSIGT in patients with insulinoma, before and after surgical removal of the tumour.
Subjects and methods  FSIGTs were performed in five patients, before and approximately 3 months post-surgery, and in 11 controls. Si and Sg were estimated using Minimal Model computer analysis of dynamic glucose and insulin data.
Results  Si was lower in insulinoma patients before, compared with after surgery (3·37 ± 0·62 vs. 6·24 ± 1·09 SE [×10⁻⁴] min⁻¹µU⁻¹ ml, P  < 0·05). Sg was similar in patients pre- and post-surgery (3·0 ± 0·67 vs. 2·4 ± 0·6 [×10⁻²] min⁻¹, NS).
Conclusions  Insulin sensitivity improves after excision of an insulinoma. Glucose effectiveness is not influenced by chronic hyperinsulinaemia and hypoglycaemia.     

Intrahepatic transfusion-transmitted virus detected by in situ hybridization in patients with liver diseases

Transfusion-transmitted virus (TTV) has been identified from patients with post-transfusion hepatitis of unknown aetiology, but the clinical relevance remains unclear. The aim of this study was to evaluate TTV in liver. We studied 15 patients with hepatitis non-A–E and 44 with hepatitis B virus (HBV). The nested polymerase chain reaction (PCR) with primers corresponding to the conserved region of the published TTV genome was employed to amplify TTV fragments in serum, and in situ hybridization was used to detect TTV in biopsied liver specimens. TTV DNA was detected in serum from six (40%) of 15 patients with hepatitis of unknown aetiology and from 16 (36.4%) of 44 patients with chronic hepatitis B, respectively. The intrahepatic viral fragment was detected in 17 (77.3%) of 22 patients with TTV in serum. There was no statistical difference in the prevalence of TTV infection between the two groups (hepatitis non-A–E 40% vs HBV 25%, P  > 0.75). When patients in both groups, with and without TTV, were compared, no differences were found in serum alanine aminotransferase (ALT) levels (hepatitis non-A–E: 131.5 ± 66.6 vs 244.2 ± 257.4, P =0.955; HBV: 240.1 ± 418.9 vs 214.6 ± 276.7 U l⁻¹, P =0.761) or histological activity index (grade) score (hepatitis non-A–E: 6.4 ± 5.5 vs 5.6 ± 5.9, P =0.689; HBV: 5.6 ± 3.7 vs 5.5 ± 3.7, P =0.345). HBV DNA levels in patients with and without TTV co-infection did not differ significantly (300 ± 776.4 μg ml⁻¹ vs 97.1 ± 160.5 μg ml⁻¹, P =0.980). Hence, TTV does exist in liver, but plays no role in hepatitis or aggravation of liver damage when co-infected with HBV.     

Pathogen reduction technology (Mirasol®) treated single-donor platelets resuspended in a mixture of autologous plasma and PAS

Background and Objectives  Mirasol^® pathogen reduction technology (PRT) for platelet concentrates uses riboflavin and ultraviolet light. Previously, we described increased metabolism and activation for PRT platelets stored in 100% plasma. To improve platelet quality, we resuspended platelets in a mixture of plasma and platelet additive solution (PAS).
Materials and Methods  Single-donor platelets were resuspended in plasma and split into an untreated control and a PRT-treated single product. One hundred and fifty millilitre PAS (SSP+) was added to both. Over 7 days, we assayed pH, glucose consumption-, lactate production rate and CD62p with and without TRAP.
Results  On day 5, PRT units showed a significantly lower pH (7·087 ± 0·105 vs. 7·288 ± 0·200) accompanied by a higher lactate production (0·104 ± 0014 vs. 0·063 ± 0·017 mmol/10¹²/h) and glucose consumption rate (0·039 ± 0005 vs. 0·028 ± 0·009 mmol/10¹² platelets/h). CD62p expression was higher in treated units (44·5 ± 13·0 vs. 16·5 ± 7·6%).
Conclusion  In comparison to PRT platelets resuspended in 100% plasma, a mixture of plasma and PAS improves pH and platelet metabolism but not platelet activation. Prolonged shelf-life for up to 7 days may be possible.     

Improved leucoreduction of red blood cell units prepared after a 24-h hold with the platelet-rich plasma method using newly developed filters

Background and Objectives  A previous study indicated that the extension of whole blood (WB) storage from 8 to 24 h at 20–24 °C before the processing of platelet-rich plasma (PRP)-depleted red blood cell (RBC) units had a negative effect on the efficacy of leucoreduction filters. In this study, we further characterized the phenomenon and tested the leucoreduction capacity of two newly developed filters.
Materials and Methods  Whole blood was stored at 20–24 °C and processed at 4-h intervals between 8 and 24 h postcollection. Components were leucoreduced before storage. Efficacy of novel filters to leucoreduce 24-h-hold PRP-depleted RBC units was also evaluated.
Results  Using a conventional filter, the mean residual white blood cell (WBC) counts in leucoreduced PRP-depleted RBCs were comparable in units prepared within 12 h from collection but gradually increased upon extended preprocessing storage from 0·36 ± 0·03 at 12 h to 0·46 ± 0·21, 0·76 ± 0·54 and 1·72 ± 1·76 × 10⁶ per unit at 16, 20 and 24 h, respectively. However, the mean residual WBC content in 24-h-hold RBCs was reduced to 0·60 ± 0·39 × 10⁶ and 0·46 ± 0·13 × 10⁶ per units using RC2D and the prototypes B-1582 rev B filters, respectively.
Conclusion  For PRP-depleted RBC units, the extension of the WB room temperature storage from 8 to 24 h before processing is likely to require the introduction of newly developed filters having an increased leucoreduction capacity in order to meet the maximal residual WBC guideline in the RBCs.     

Chronic consumers of boiled coffee have elevated serum levels of lipoprotein(a)

Objectives. Lipoprotein(a) consists of an LDL-particle attached to apolipoprotein(a), which is made by the liver. Diterpenes present in boiled coffee raise serum levels of LDL cholesterol and of the liver enzyme alanine aminotransferase in man. We investigated the association between intake of boiled coffee and serum levels of lipoprotein(a).
Design, setting and subjects. Healthy Norwegians 40–42 years of age, who habitually consumed five or more cups of boiled coffee per day ( n =150) were compared with matched filter coffee consumers ( n =159) in a cross-sectional study, as part of the Norwegian National Health Screening in 1992.
Results. The median lipoprotein(a) level was 13.0 mg  dL⁻¹ (10th and 90th percentile: 2.5 and 75.0 mg  dL⁻¹, respectively) on boiled and 7.9 mg dL^-1 (10th and 90th percentile: 1.9 and 62.5 mg dL⁻¹, respectively) on filter coffee ( P =0.048). Means±SE were 25.8±2.4 mg dL⁻¹ and 19.6±2.0 mg dL⁻¹, respectively ( P =0.04). Although not statistically significant, subjects consuming nine or more cups of coffee per day had higher lipoprotein(a) levels than those drinking five to eight cups per day in both coffee groups.
Conclusion. Chronic consumers of unfiltered, boiled coffee have higher serum levels of lipoprotein(a) than filter coffee drinkers.     

Effect of metformin, orlistat and pioglitazone treatment on mean insulin resistance and its biological variability in polycystic ovary syndrome

Context  Mean insulin resistance (IR) is greater and it is also more variable in overweight women with polycystic ovarian syndrome (PCOS) compared to weight matched controls. Whilst treatment will reduce the mean IR, it is not known if the IR variability is also reduced.
Objective  To compare the change in IR and its variability before and after treatment with insulin sensitization through metformin and pioglitazone, compared to that induced by weight loss with orlistat.
Design  Randomized, open labelled parallel study.
Setting  Endocrinology outpatient clinic at a referral centre.
Patients  Thirty obese PCOS patients [BMI 36·0 ± 1·2 kg/m² (mean ± SEM)] participated in the study.
Intervention  The change in biological variability (BV) was assessed by measuring IR (homeostasis model assessment method) at 4-day intervals on 10 consecutive occasions before and 12 weeks after randomization to metformin, pioglitazone or orlistat.
Outcome measured  The primary end point of the study was a change in BV of IR.
Results  Treatment with pioglitazone, orlistat and metformin reduced the overall IR by 41·0 ± 4·1%, 19·7 ± 6·4% and 16·1 ± 6·8% ( P =  0·005, P  = 0·013, P  = 0·17, respectively) and IR variability by 28·5 ± 18·0%, 41·8 ± 11·6% and 23·7 ± 17·0 ( P =  0·20, P  = 0·015 and P  = 0·28, respectively). Free androgen index reduced significantly with all treatments.
Conclusion  Only orlistat reduced both IR and its variability significantly, though all three drugs were effective in reducing hyperandrogenism within the 12-week period of the study.     

Impact of GH substitution on exercise capacity and muscle strength in GH-deficient adults: a meta-analysis of blinded, placebo-controlled trials

Background  Several but not all trials suggest that GH replacement in GH-deficient adults improves aerobic exercise capacity, whereas its effect on muscle strength is more dubious. However, a denominator of these studies is a low sample size.
Objective  We systematically reviewed and analysed all randomized, double-blind, placebo-controlled trials on the effects of GH administration on aerobic exercise capacity and muscle strength in GH-deficient adults.
Study selection  Fifteen trials were identified from four databases. We conducted an analysis of effects on aerobic exercise capacity, performed on either a treadmill or a bicycle ergometer, muscle strength assessed by a dynamometer, and muscle mass assessed by computerized tomography.
Results  The total number of patients included was 306 and the duration of treatment ranged from 3 to 12 months. GH replacement significantly increased aerobic exercise capacity [8·9 ± 0·8%, ( P  < 0·001)] including VO₂ max [0·17 ± 0·02 l/min ( P  < 0·001)], as well as muscle volume [7·1 ± 1·6%, ( P  < 0·001)]. In contrast, muscle strength measured in 113 patients was not significantly increased [3·2 ± 2·2% ( P  = 0·15)].
Conclusion  GH replacement in GH-deficient adults is associated with a significant positive effect on aerobic exercise capacity and muscle mass.     

Augmentation index in resistance to thyroid hormone (RTH)

Objective  Resistance to thyroid hormone (RTH) is associated with a varied clinical presentation. The cardiac effects of RTH have been described but vascular function has yet to be fully evaluated in this condition. We have measured the arterial function of those with RTH to assess any vascular changes.
Design  An observational study.
Patients  Twelve RTH patients were recruited from the thyroid clinic (mean value ± SD), age 40·8 ± 18·7 years; BMI 27·2 ± 4·2 kg/m² and compared with 12 healthy, euthyroid, age-matched controls (age 41·4 ± 19·3; BMI 24·8 ± 4·4 kg/m²) with no history of cardiovascular disease. No interventional measures were instituted.
Measurements  Arterial stiffness was measured using pulse wave analysis at the radial artery. Thyroid function, fasting lipids and glucose were also measured on the same occasion in both patients and controls.
Results  The corrected augmentation index, a surrogate marker of arterial stiffness was significantly higher in patients compared with controls (21·0% ± 14·1% vs. 5·4% ± 18·2%, P  < 0·03). Low density lipoprotein cholesterol (LDL-cholesterol) levels were also significantly elevated in patients compared with controls (3·0 ± 0·6 vs. 2·1 ± 0·5 mmol/l; P  < 0·002).
Conclusion  RTH patients show evidence in this study of increased augmentation index consistent with an increase in arterial stiffness compared with euthyroid controls. They also demonstrate elevated LDL-cholesterol levels. Both these measures may lead to increased cardiovascular risk.     

PPARγ Pro12Ala Pro/Pro and resistin SNP-420 G/G genotypes are synergistically associated with plasma resistin in the Japanese general population

Objective  The Ala allele of the Pro12Ala polymorphism (rs1801282) of peroxisome proliferator-activated receptor γ (PPARγ) is protective against type 2 diabetes (T2DM). Resistin, secreted from adipocytes, causes insulin resistance in rodents. Resistin gene expression is reduced by the PPARγ ligand. We previously reported that subjects with the G/G genotype of a resistin gene single nucleotide polymorphism (SNP) at –420 (rs1862513) had the highest circulating resistin levels, followed by C/G and C/C. The aim of this study was to determine the relationship among PPARγ Pro12Ala polymorphism, resistin SNP-420, and plasma resistin.
Design, patients and measurements  We cross-sectionally analysed 2077 community-dwelling subjects attending an annual medical check-up. Genotypes were determined by TaqMan analysis. Fasting plasma resistin was measured using ELISA.
Results  Plasma resistin appeared to be higher in subjects with the Pro/Pro genotype of PPARγ than those with Pro/Ala and Ala/Ala genotypes (mean ± SE, 11·6 ± 0·2 vs. 10·4 ± 0·5 μg/l). Multiple regression analysis, adjusted for age, gender, BMI, and resistin SNP-420, revealed that the Pro/Pro genotype was a positive predictor of plasma resistin (PPARγ ,  Pro/Pro vs. Pro/Ala + Ala/Ala, unstandardized regression coefficient (β) = 1·03, P  = 0·0384). The effects of the Pro/Pro genotype of PPARγ (Pro/Pro vs. Pro/Ala + Ala/Ala) and the G/G genotype of resistin SNP-420 (G/G vs. C/C) on plasma resistin were synergistic (β = 4·76, P  = 0·011).
Conclusions  The PPARγ Pro12Ala Pro/Pro and resistin SNP-420 G/G genotypes were synergistically associated with plasma resistin, when adjusted for age, gender, and BMI, in the Japanese general population.     

Interleukin-6 (IL-6) –572C→G promoter polymorphism is associated with type 2 diabetes risk in Koreans

Objective  Increased levels of inflammatory markers, such as interleukin-6 ( IL -6), are associated with type 2 diabetes (T2DM). We investigated the association of IL-6 gene polymorphisms with T2DM and circulating levels of IL -6 in Koreans.
Subjects  A total of 1477 subjects with normal glucose tolerance and 476 T2DM patients were included.
Measurements  We examined IL-6 – 174G→C, –572C→G, –597G→A and –1363G→T promoter region polymorphisms. The main outcome measures were the odds ratio (OR) on T2DM risk and serum concentrations of IL -6 and high-sensitivity C-reactive protein (hs-CRP).
Results  Homozygosity for the rare G allele IL-6 – 572C→G was associated with a higher risk of T2DM [OR 1·69 (95%CI 1·11–2·58), P  = 0·015]. Serum IL -6 concentrations were associated with the IL-6 – 572C→G genotype in control subjects (G/G: 2·33 ± 0·41: C/G: 1·53 ± 0·09: C/C: 1·72 ± 0·08 ng/l, P  = 0·023). Also in the control group, subjects homozygous for the rare G allele showed significantly higher concentrations of hs-CRP than C/C and C/G carriers (G/G: 13·6 ± 2·9: C/G: 9·2 ± 0·6: C/C: 7·8 ± 0·4 mg/l, P  = 0·003). The C-allele at the IL-6 – 174 SNP was very rare (< 0·01) and –597G→A and –1363G→T were monomorphic in this population.
Conclusions  Our data demonstrate that the IL-6 – 572G/G genotype is associated with higher serum IL -6 and hs-CRP concentrations and with increased risk for T2DM.     

Hyperhyaluronanemia in Alcoholic Hepatitis Is Associated with Increased Levels of Circulating Soluble Intercellular Adhesion Molecule-1

The purpose of this study was to evaluate the role of the sinusoidal endothelial cell (SEC) during the clinical course of alcoholic hepatitis. Twenty consenting patients (mean age: 49.4 ± 11.0 years) with moderate or severe hepatitis were studied. The patients were selected and characterized according to their history of drinking and laboratory profile, including serum aminotransferases, bilirubin, total white blood cell and neutrophil count, and prothrombin times. C-reactive protein and interleukin-6 were also measured as markers of the hepatic acute phase response. A marker of the SEC functional state, the circulating level of hyaluronan, was measured in parallel with the circulating levels of soluble intercellular adhesion molecule (sICAM)-1 over a 6-month observation period. All patients were hospitalized for the first month and encouraged to abstain from drinking for the duration of the study. The initial increased levels of both hyaluronan (542 ± 32 ng ml⁻¹ serum) and sICAM-1 (488 ± 70 ng ml⁻¹ serum), gradually fell during the 6-month observation period, eventually reaching values close to those seen in healthy subjects. A positive correlation was obtained between changes in these two markers of SEC function/activation on the one hand, and between these two tests and bilirubin, on the other hand. These data indicate that abnormalities of SEC function/activation, as reflected by serum hyaluronan and sICAM-1, are prominent in alcoholic hepatitis, and these alterations improve within relatively short periods of time after cessation of alcohol consumption.     

Hepatic 11beta-HSD1 mRNA expression in fatty liver and nonalcoholic steatohepatitis

Context  Nonalcoholic fatty liver disease represents the hepatic manifestation of the metabolic syndrome. Nonalcoholic steatohepatitis (NASH) is the progressive form of liver injury. The pathophysiology that leads to NASH is not well understood.
Objective  We hypothesize that an altered cortisol metabolism in the liver may be a pathogenetic factor.
Design and patients  75 patients (28 men, 47 women) underwent liver biopsy for elevation in liver enzymes. Histological diagnosis identified normal liver in eight, fatty liver in 20, NASH grade 1 in 22, grade 2 in nine, grade 3 in three patients, and other forms of hepatitis or cirrhosis in 13 patients. We quantified hepatic 11β-hydroxysteroid dehydrogenase type1 (11β-HSD1) and hexose-6-phosphate-dehydrogenase (H6PDH) mRNA expression by real-time PCR. In addition, analysis of 24 h urinary excretion of cortisol metabolites using GCMS was performed and compared with healthy controls.
Results  11β-HSD1 mRNA expression correlated significantly ( R ²= 0·809; P  < 0·001) with H6PDH mRNA expression, negatively with waist-to-hip ratio in women ( R ²= 0·394; P = 0·005), but not with urinary (THF + 5α-THF)/THE ratio, total cortisol metabolite excretion, age, BMI, degree of fatty liver or NASH stages. Total cortisol metabolite excretion was increased in patients with fatty liver or NASH compared with healthy controls.
Conclusions  Our data suggest that expression of hepatic 11β-HSD1 and H6PDH are closely interlinked. 11β-HSD1 gene expression does not seem to be involved in the pathogenesis of fatty liver or NASH. However, those patients showed an increased 5α- and 5β-reduction of cortisol leading to an increased cortisol turnover rate and an activation of the HPA axis.     

High density lipoprotein-cholesterol levels increase with age in American women but not in Hong Kong Chinese women

Objectives  High-density lipoprotein (HDL) cholesterol is a powerful cardiovascular risk factor. Important gender and ethnic differences in plasma HDL levels exist and warrant investigation.
Design  Cross-sectional survey in two different general populations.
Patients  7700 participants of the National Health and Nutrition Examination Survey (NHANES) 1999–2002 and 1944 participants of the Hong Kong Cardiovascular Risk Factor Prevalence Study-2 (CRISPS2) 2000–2004.
Measurements  Plasma HDL levels.
Results  Plasma HDL levels were higher in women than in men in both populations. In the United States women, it increased with age, whereas in Chinese women, it declined with age and converged with male HDL levels. In the United States, 37·1 ± 1·2% men and 38·9 ± 1·1% women had low HDL levels. In Hong Kong, 34·3 ± 1·6% men and 34·5 ± 1·5% women had low HDL levels. In Americans, the independent predictors of low HDL levels were lower age, being non-Mexican Hispanic, waist circumference, triglycerides and not drinking alcohol in men, and lower age, being Hispanic, waist circumference, triglycerides, current smoking and not drinking alcohol in women. In Hong Kong Chinese, the independent predictors of low HDL levels were body mass index, triglycerides, current smoking and not drinking alcohol in men, and lower age, waist circumference, triglycerides, diabetes and former smoking in women.
Conclusions  The decline in plasma HDL with age in Chinese women is opposite to that seen in American women. The increased cardiovascular risk in elderly Chinese women requires further study.     

Usefulness of troglitazone administration to obese hyperglycaemic patients with near-normoglycaemia

Aims:   We assessed the effectiveness of 400 mg/day of troglitazone administered to hyperglycaemic patients with near-normoglycaemia who were obese and who had hyperinsulinaemia.
Results:   The area under the plasma glucose curve in oral glucose tolerance tests (OGTT) significantly decreased from 39.8 ± 19.4–20.5 ± 10.2 mg/dL · h and the area under the insulin-response curve from 31.8 ± 22.5–12.2 ± 5.7 μU/ml · h 4 months after the start of treatment. The level of HbA1c significantly improved from 6.6 ± 0.2 to 6.3 ± 0.2% (p < 0.05) by 1 month after administration, and that of serum 1,5-anhydroglucitol (1,5-AG) from 12.6 ± 1.1–18.3 ± 2.5 μ/ml (p < 0.05). In some cases, recovery of the first-phase insulin secretion was observed.
Conclusions:   These findings suggest that the administration of this insulin sensitizer is useful in the treatment of obese Japanese subjects with borderline or mild diabetics accompanied by hyperinsulinaemia.     

Association between plasma renin activity and metabolic cardiovascular risk factors in essential hypertension

Objectives. To study the relationships between plasma renin activity and metabolic cardiovascular risk factors in patients with essential hypertension.
Subjects and design. Patients with uncomplicated essential hypertension ( n =36) with a diastolic blood pressure of 95–115 mmHg were studied. Assessment of plasma renin activity (PRA) related to urinary sodium excretion was used to define subgroups with high ( n =12), medium ( n =16) and low renin profiles ( n =8).
Main outcome measures. Fasting plasma lipid levels were determined. Glucose, insulin and C-peptide responses to standard oral glucose tolerance test (OGTT) were measured.
Results. Patients with high PRA had higher levels of plasma cholesterol (6.13±0.81 versus 4.67±0.7 mmol L^-1, P <0.05) and triglycerides (2.14±0.18 versus 0.98±0.13 mmol L^-1, P <0.05), than the low PRA group. HDL-cholesterol levels were lower in the high renin group than in the low renin group (1.05±0.04 versus 1.26±0.09 mmol L^-1, P <0.05). Insulin and C-peptide sums were higher in high PRA group (33.8±1.2 versus 25.1±0.9 and 2.6±0.3 versus 1.9±0.4 ng L^-1, P <0.05), than in the low PRA group.
Conclusions. Essential hypertensive patients with a high renin profile display more pronounced dyslipidaemia and higher levels of plasma insulin than patients with a low renin profile. This may be one explanation for higher incidence of cardiovascular disease previously reported in high PRA group.     

Drug Delivery at the Aortic Valve Tissues of Healthy Domestic Pigs with a Paclitaxel-Eluting Valvuloplasty Balloon

Background: Restenosis occurs invariably within 1 year following balloon valvulopasty in aortic valve stenosis. The mechanism of restenosis seems to involve a dynamic cellular component that could be a target for drug inhibition. We investigated the feasibility of local drug delivery at the aortic valve tissues of healthy pigs with a paclitaxel-eluting balloon.
Methods: Aortic valvuloplasty was performed in eight anesthetized domestic pigs using paclitaxel-eluting balloons (3 μg/mm² balloon surface area). They were assigned to two or four times 15-second balloon inflations and were sacrificed 30 minutes after final balloon inflation.
Results: The aortic annulus to balloon diameter ratio was 1.15 ± 0.07. The mean paclitaxel concentration in the aortic valve leaflets was 0.91 ± 1.36 μg/mL (0.34 ± 0.05 μg/mL in the two-inflation group, 1.48 ± 1.86 μg/mL in the four-inflation group, P = 0.23). The percentage of the total paclitaxel dose recovered in the aortic valve leaflets was 18 ± 11⁻⁶% (13 ± 6⁻⁶% and 25 ± 14⁻⁶% in the two- and four-inflation group, P = 0.16).
Conclusion: Local drug delivery at the aortic valve leaflets of healthy pigs with a paclitaxel-eluting balloon is feasible and concentrations within the therapeutic window are detected 30 minutes after the procedure. The antirestenotic potential of this treatment should be studied.     

Mutations in the ABCC8 (SUR1 subunit of the KATP channel) gene are associated with a variable clinical phenotype

Objective  Previous studies of the association between autoimmune Addison's disease (AAD) and a nonsynonymous single nucleotide polymorphism (SNP) in the PTPN22 gene (C1858T, pR620W; SNP ID no. rs2476601) have shown conflicting results. We aimed to examine this association using additional cohorts of AAD subjects from the UK and Poland.
Design  DNA samples were obtained from UK and Polish AAD subjects ( n  = 251 and 87, respectively) and ethnically matched healthy controls ( n  = 429 and 236, respectively). Genotyping for the C1858T PTPN22 marker was performed by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) assay. Meta-analysis of the results, together with those from three other populations, was performed using RevMan v5·0 software.
Results  In 251 UK AAD subjects the frequency of the PTPN22 1858T allele was 12·2% compared to 7·8% in healthy UK controls; P  = 0·008. Similarly, in 87 Polish AAD subjects the PTPN22 1858T allele was found in 19·5% of alleles compared to 11·7% in healthy Polish subjects; P  = 0·010. A meta-analysis, combining these result with published data for three other populations, involving 797 AAD subjects and 2032 controls in total, showed that the 1858T allele was associated with AAD susceptibility with a pooled odds ratio (OR) of 1·44 [95% confidence interval (CI) 1·21–1·72; P  = 5·6 × 10⁻⁵], under a fixed-effects model.
Conclusion  This study confirms the association between the PTPN22 1858T allele and AAD in an expanded UK cohort and in the previously unstudied Polish population. This meta-analysis allows for the first time a reliable estimate of the strength of effect of this autoimmune disease susceptibility allele across different European Caucasian populations.     

Characterization of the anaemia associated with Graves' disease

Background  Graves' disease (GD) is associated with hyperthyroidism. Thyrotoxicosis adversely affects multiple organ systems including haematopoiesis. Anaemia occurring specifically in GD has not been systematically studied previously.
Objective  To define the prevalence and characteristics of the anaemia associated with GD.
Design  Eighty-seven newly diagnosed patients with GD were recruited. Haematological indices, thyroid function and inflammatory parameters were examined at presentation and following successful treatment of hyperthyroidism.
Setting  Tertiary care academic referral centre.
Results  Thirty-three per cent of subjects presented with anaemia. The prevalence of anaemia not attributable to other causes (GD anaemia) was 22%. GD anaemia affected 41·6% (10/24) of men compared to 17·5% of women (11/63). Mean erythropoietin (EPO) levels (15·5 ± 5·3 mIU/ml) were within normal reference limits but significantly higher ( P =  0·004) than those of the non-anaemic controls. Hgb correlated inversely with EPO ( P =  0·05) and CRP ( P =  0·04) levels, a relationship that persisted after multivariate adjustment for TT3 or TT4. With antithyroid therapy for 16 ± 6·3 weeks, Hgb levels normalized in 8 out of 9 subjects with GD anaemia (10·7 ± 0·8 to 13·5 ± 1·3 g/dl, P  = 0·0001). After normalization of Hgb, mean MCV and TIBC were significantly increased, and median ferritin and mean EPO were significantly decreased.
Conclusions  GD anaemia is common, resembles the anaemia of chronic disease, and is associated with markers of inflammation. It corrects promptly with return to the euthyroid state following treatment.     

A population pharmacokinetic model for pegylated-asparaginase in children

We analysed 1221 serum activity measurements in 168 children from the Berlin-Frankfürt-Münster acute lymphoblastic leukaemia studies, ALL-BFM (Berlin-Frankfürt-Münster) 95 and ALL-BFM REZ, in order to develop a pharmacokinetic model describing the activity-time course of pegylated (PEG)-asparaginase for all dose levels. Patients received 500, 750, 1000 or 2500 U/m² PEG-asparaginase on up to nine occasions. Serum samples were analysed for asparaginase activity and data analysis was done using nonlinear mixed effects modelling (NONMEM Vers. VI, Globomax, Hanouet, MD, USA). Different linear and nonlinear models were tested. The best model applicable to all dosing groups was a one-compartmental model with clearance (Cl) increasing with time according to the formula: Cl=Cl_i * e ^{(0·0793 * t )} where Cl_i = initial clearance and t  = time after dose. The parameters found were: volume of distribution ( V ) 1·02 ± 26% l/m², Cl_i 59·9 ± 59% ml/d per m² (mean ± interindividual variability). Interoccasion variability was substantial with 0·183 l/m² for V and 44·7 ml/d per m² for Cl, respectively. A subgroup of the patients showed a high clearance, probably due to the development of inactivating antibodies. This is the first model able to predict the activity-time course of PEG-asparaginase at different dosing levels and can therefore be used for developing new dosing regimens.