Metabolic disorders associated with chronic hepatitis C: impact of genotype and ethnicity.

BACKGROUND & AIM: Patients with hepatitis C virus (HCV) infection, especially those with genotypes 1 and 4, have an increased risk of developing metabolic disorders. The aim of this study was to evaluate the associations among metabolic disorders, ethnicity and genotype in a large cohort of patients with chronic hepatitis C (CHC). PATIENTS AND METHODS: All consecutive patients with CHC who were seen in our hepato-gastroenterology unit between January 2002 and September 2008 were included. Demographical data and variables related to the metabolic syndrome were collected. Insulin resistance was assessed using the homeostasis model for the assessment of insulin resistance test (HOMA-IR) test. RESULTS: Among the 454 CHC patients, the prevalence of the metabolic syndrome was 12.4%. The HOMA-IR test was performed in 140 patients, and 35.0% had insulin resistance. There were more Black Africans among the patients with genotypes 1/4 than among those with genotypes 2/3 (32.0 vs 1.2%, P<0.0001). Insulin resistance was more common in patients with genotypes 1/4 than in those with genotypes 2/3 (17 vs 1.7%, P=0.0001 and 43.3 vs 16.3%, P=0.001, respectively). Genotypes 1/4 were more frequently present in patients with insulin resistance than in those without insulin resistance (85.7 vs 60.5%, P=0.001). By logistic regression, genotypes 1/4 [odds ratio (OR)=2.79; 95% confidence interval (CI): 1.09-7.12, P=0.032] and older age (OR=1.03; 95% CI: 1.004-1.06, P=0.024) were independently associated with the presence of insulin resistance. CONCLUSIONS: In CHC, insulin resistance is independently associated with the presence of genotypes 1/4. Ethnicity is not independently associated with metabolic disorders in patients with CHC.

Evaluation of an automated capillary electrophoresis system in the screening for hemoglobinopathies.

BACKGROUND: The biochemical diagnosis of hemoglobinopathies (thalassemias and hemoglobin (Hb) variants) is based on the separation and the quantification of Hb fractions. HPLC is the most commonly used method but capillary electrophoresis (CE) methods have also been developed successfully. The Capillarys II system is the first fully automated CE system that allows the quantification of Hb A2 and Hb F and the separation of Hb variants. We evaluated the ability of this system to separate and identify Hb variants and to quantify Hb A2 and Hb F. MATERIAL AND METHODS: The separation of 18 different Hb variants was studied and the imprecision on migration times was calculated for the three most frequent ones. The total imprecision on Hb A2 and Hb F quantification was determined. The results obtained for 44 patients were compared with those given by HPLC. The interference on Hb A2 measurement due to the presence of Hb S was studied. RESULTS: Fourteen out of the 18 variants tested, including all variants of clinical importance, were separated from Hb A. Imprecision on migration times was less than 1%. For Hb A2 quantification, imprecision was less than 3.5% and for Hb F, less than 7.0%. The comparison with HPLC showed an acceptable agreement between both methods but a systematic negative bias for Hb A2 and both proportional and systematic biases for Hb F. No interference from the presence of Hb S on the quantification of Hb A2 was observed. CONCLUSIONS: The fully automated Capillarys Hemoglobin method allows the detection and the separation of the most common Hb variants. It provides also a precise, quick, and very easy quantification of Hb F and Hb A2, even in the presence of Hb S. It is very suitable for routine investigation of hemoglopinopathies.

An estimation of the incidence and demographic picture of the major hemoglobinopathies in Belgium (from a confidential inquiry).

An estimation of the incidence and demographic picture of the major hemoglobinopathies in Belgium has been approached through a confidential inquiry sent to 228 pediatric and adult hematological departments. Forty-two percent of responses showed that 417 patients are known in Belgium: 83% with sickle cell disease, 13% with beta-thalassemia (beta-thal) major, 2% with beta-thal intermedia, and 1% with Hb H disease. Twenty-five percent of the sickle cell disease patients and 54% of those suffering from a beta-thal major were older than 20 years. Three hospitals ensure the follow-up of 70% of the patients and are situated in Brussels, Belgium; a follow-up of less than 20 patients was reported at 21 centers. These results confirm that sickle cell disease is the major hemoglobinopathy in Belgium; it concerns mostly pediatricians but adult hematologists are also confronted with these pathologies. Therefore, it is necessary to implement integrated programs of prevention and treatment.

Association between CD14 polymorphisms and serum soluble CD14 levels: effect of atopy and endotoxin inhalation.

BACKGROUND: A prerequisite for activation of the innate immune response by endotoxin is its binding to CD14. OBJECTIVE: The aim of this study was to evaluate the role of CD14 polymorphisms, atopy, and inhaled endotoxin in modulating serum CD14 levels. METHODS: Healthy volunteers (n = 88) were genotyped for CD14 polymorphisms at the -1619, -1359, and -159 loci, relative to the transcription start site. Subjects inhaled 20 mug of endotoxin, and white blood cell, C-reactive protein, LPS-binding protein, and soluble CD14 (sCD14) levels were measured before and after exposure. RESULTS: Homozygotes for the -1619G, -1359G, and -159T alleles had higher baseline levels of sCD14 than carriers of the CD14/-1619AA (P = .015), -1359GT/TT (P = .015), or -159CC (P = 0.017) genotypes. sCD14 levels increased within 24 hours of endotoxin inhalation (P < .0001 for all biomarkers); however, the association between CD14 polymorphisms and sCD14 levels was no longer present after exposure. The atopic status of an individual did not alter these associations. CD14 polymorphisms were not associated with levels of white blood cells, C-reactive protein, and LPS-binding protein before or after endotoxin challenge. CONCLUSION: These data suggest that CD14 promoter polymorphisms and inhaled endotoxin modulate sCD14 levels.

The determination of the levels of circulating galectin-1 and -3 in HNSCC patients could be used to monitor tumor progression and/or responses to therapy.

To evaluate galectin-1, -3 and -7 serum levels as diagnostic and/or prognostic markers for head and neck squamous cell carcinomas (HNSCCs). ELISA was employed to test sera from 102 patients with HNSCCs and from 38 healthy control volunteers for galectin-1, -3 and -7 serum levels. Serum galectin levels were assayed by ELISA and the levels of galectin expression in HNSCCs were determined by means of immunohistochemistry. HNSCCs display significant immunohistochemical amounts of galectin-7, but this galectin cannot be detected in the blood of HNSCC patients. Galectin-3 levels differ significantly (p=0.03) in healthy volunteers and HNSCC patients. Using a threshold value of 4.3 ng/ml, galectin-3 serum level enabled a significant level of discrimination (p=0.03) to be established between the cancer patients and the healthy volunteers, with 90% level of specificity and 36% level of sensitivity. The discrimination was even better when using a threshold value of 13.5 ng/ml for galectin-1 (p=0.001), with 100% level of specificity and 22% level of sensitivity. A subgroup of stage IV HNSCC patients displayed significantly reduced levels of circulating galectin-1 (p=0.003) and galectin-3 (p=0.001) after treatment as opposed to before. Galectin-3 concentrations in sera from the patients with a metastatic disease were significantly (p=0.01) higher than in sera from the patients with localized tumors. The determination of circulating levels of galectin-1 and -3 could be used to monitor the progression of their disease or their response to therapy.

Evaluation of oral corticosteroids and phosphodiesterase-4 inhibitor on the acute inflammation induced by inhaled lipopolysaccharide in human.

BACKGROUND: Endotoxins are pro-inflammatory substances present in the environment. In man, inhalation of its purified derivative lipopolysaccharide (LPS) induces inflammation related to macrophages and neutrophils. Corticosteroids and phosphodiesterase (PDE)-4 inhibitors have inhibiting effects on macrophages and neutrophils, respectively. This study investigated the effect of prednisolone and of the PDE-4 inhibitor cilomilast on the LPS-induced acute inflammation. METHODS: The study was a placebo-controlled, double-blind crossover design. On three occasions, at 2 weeks interval, 16 healthy subjects inhaled 50 microg LPS after a 6-day treatment with cilomilast (15 mg bd), prednisolone (10 mg bd) or placebo. For the assessment of the inflammatory response, induced sputum was obtained before inclusion and 6h post-LPS while blood samples were collected before, 6 and 24 h post-LPS. RESULTS: Inhaled LPS induced an increase in sputum neutrophils (p<0.0001), logMMP-9 (p<0.05), logMMP-9/TIMP-1 (p<0.01) and logTNF-alpha (p<0.02). At the blood level there were significant rise in neutrophilia (p<0.001), E-selectin (p<0.02), C-reactive protein (CRP) (p<0.001) and LPS-binding protein (p<0.001). There was both a slight, but not significant, increase in body temperature and decrease in forced expiratory volume in 1 s (FEV(1)). Neither prednisolone nor cilomilast had protective effect on the LPS-induced airways' inflammation. The LPS-induced CRP acute-phase protein of inflammation (0.58+/-0.13 and 3.52+/-0.41 mg/dL, before and after LPS, respectively) was significantly inhibited by a pre-treatment with prednisolone (1.39+/-0.32 mg/dL, p<0.01) and attenuated (2.65+/-0.30 mg/dL, p=0.09) with cilomilast. CONCLUSION: In healthy subjects, while the LPS-induced airways' inflammation was not modified either by oral prednisolone or by PDE-4 inhibitor cilomilast (at actual dosage), the LPS-induced acute phase of blood inflammation was reduced by prednisolone.

The rare HLA-DQA1*03-DQB1*02 haplotype confers susceptibility to type 1 diabetes in whites and is preferentially associated with early clinical disease onset in male subjects.

The heterozygous combination of DQA1*03-DQB1*0302 (DQ8) and DQA1*05-DQB1*0201 (DQ2) confers the highest known HLA-DQ-linked risk for type 1 diabetes, suggesting a role for transcomplementation. The trans-heterodimer encoded by DQA1*03 and DQB1*02 is also rarely observed in cis in whites. Islet antibody-positive diabetic patients (P; n = 2,238) and control subjects (C; n = 2,223) of white descent were genotyped by a HLA-DQA1-DQB1 dot-blot method. The presence of the DQA1*03-DQB1*02 haplotype was observed in 22 patients (1%) versus 6 controls (0.3%) (odds ratio [OR] = 3.7, p = 0.005). It was more prevalent in whites of Northern African descent, but both in European (n = 3,813) and in Northern African whites (n = 648), the DQA1*03-DQB1*02 haplotype tended to be associated with diabetes (respectively, P 0.3% vs. C 0.03%, OR = 12.2, p = 0.005; and P 2.1% vs. C 0.6%, OR = 3.8, p = 0.03). DRB1 typing revealed that DQA1*03-DQB1*02 is usually associated with the DRB1*0405 risk allele in European patients and with DRB1*0405, DRB1*07 and DRB1*09 in Northern African whites. Like in DQ2/DQ8-positive patients, the presence of DQA1*03-DQB1*02 is preferentially associated with younger age at clinical onset than in other genotypes, but unlike in subjects carrying DQ2/DQ8, earlier clinical manifestation was mostly restricted to male subjects, often carrying DR3 and/or DQB1*02 on the other chromosome. These results are compatible with an effect of cis-encoded heterodimers or with previously suggested interactions of X-linked genetic factors with (DR3-)DQB1*02 haplotypes.

The place of capillary electrophoresis techniques in screening for haemoglobinopathies.

BACKGROUND: Capillary zone electrophoresis (CZE) methods are new laboratory diagnostic tools. The screening of haemoglobin (Hb) variants by a battery of two automated CZE methods at alkaline and acid pH, followed by micellar electrokinetic capillary chromatography (MECC), has been evaluated. METHODS: Three hundred and ninety-two patients' samples with an abnormal haemoglobin fraction, detected either by isoeletric focusing (IEF) or by automated cation-exchange exchange high-performance liquid chromatography (automated HPLC), were tested by both CZE methods. Their performances were compared with IEF and automated HPLC techniques. The place of MECC has been evaluated. RESULTS: Using both CZE methods, the clinically relevant variants (HbS, HbC, etc.) as well as 15 of 20 clinically silent variants tested were separated from HbA. Using the alkaline CZE method, a presumptive identification of Hb Bart's, HbH and Hb Constant Spring was obtained. Complementary testing by MECC has demonstrated it to be an aid in distinguishing the globin chain mutations before confirmation by DNA or protein structure analysis. CONCLUSION: Using only one device, alkaline CZE, acid CZE and MECC methods offer high resolution, automated sampling and rapid analysis. They are good tools for screening of haemoglobinopathies and can replace conventional techniques.

The nutritional iodine supply of Belgian neonates is still insufficient.

UNLABELLED: Belgium used to be affected by mild iodine deficiency. Improvement in iodine nutrition has been recently documented in schoolchildren in Belgium in spite of the absence of any systematic programme of iodine supplementation. The question arises as to whether this 'silent iodine prophylaxis' affected also the neonates. A total of 185 random urine samples were collected from 90 full term and 65 preterm neonates in Brussels on day 5 and repeated on day 30 in 30 preterms who were bottle-fed with iodine-enriched formula-milk. The iodine content was also determined in 58 samples of breast-milk on day 5. The median urinary iodine on day 5 in full term neonates was 86 micro g/l, which is markedly higher than the figure of 48 micro g/L reported 15 years previously in neonates in the same area but still much lower than normal for this age group (150-200 micro g/l). The mean iodine content of breast-milk was 78 micro g/l, which is unchanged as compared to 15 years ago and is about 66% of normal. Finally, the median urinary iodine increased from 60 micro g/l on day 5 to 150 micro g/l on day 30 in preterms bottle-fed with iodine-enriched formula-milk. CONCLUSION: the status of iodine nutrition has also improved spontaneously in Belgian neonates but has not yet normalised. Lactating and probably pregnant women remain clearly iodine deficient. The iodine-enriched formula-milk for preterms is efficient in correcting their iodine deficiency. National measures are urgently required for correction of iodine deficiency in Belgium.

Immediate effects of an 8-h advance shift of the rest-activity cycle on 24-h profiles of cortisol.

To investigate the adaptation of plasma cortisol profiles to an abrupt phase advance of the rest-activity cycle, eight normal young subjects were submitted in a sleep laboratory to an 8-h advance shift of their sleep-wake and dark-light cycles. The shift was achieved by advancing bedtimes from 2300-0700 to 1500-2300. Blood samples were obtained at 20-min intervals for 68 consecutive hours. The shift resulted within 6-9 h in a 3- to 4-h advance of timings of the nadir of the cortisol profile and of the end of the quiescent period but had no immediate effect on the timing of cortisol acrophase. The quiescent period of cortisol secretion was shortened and fragmented. Thus a major advance shift achieved without enforcing sleep deprivation results in a rapid partial adaptation of the temporal profiles of cortisol but also in a marked disruption of the cortisol quiescent period. Sleep onset was consistently followed by a decrease in cortisol concentrations. Conversely, both sleep-wake and dark-light transitions were consistently associated with cortisol secretory pulses.