Common genetic variants of the vitamin D binding protein (DBP) predict differences in response of serum 25-hydroxyvitamin D [25(OH)D] to vitamin D supplementation

BackgroundTo determine the effect of vitamin D binding protein (DBP) genotypes on 25-hydroxyvitamin D [25(OH)D] changes with vitamin D supplements, we studied 98 adults receiving 600 or 4000 IU/d vitamin D₃ for one year.MethodsThe DBP functional variant, T436K, was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).ResultsMean 25(OH)D increases were 97% for TT (n = 48), 151% for TK (n = 31) and 307% (n = 6) for KK genotypes (p = .004).ConclusionsAs with baseline 25(OH)D, T436K genotype predicts 25(OH)D changes after long-term vitamin D supplementation.     

Plasma S100β is not a useful biomarker for tumor burden in neurofibromatosis

ObjectivesNeurofibromatosis 1 (NF1), NF2, and schwannomatosis are characterized by a predisposition to develop multiple neurofibromas and schwannomas. Currently, there is no blood test to estimate tumor burden in patients with these disorders. We explored whether S100β would act as a biomarker of tumor burden in NF since S100β is a classic immunohistochemical marker of astrocytes, oligodendrocytes and Schwann cells and a small study showed S100β concentrations correlate with the volume of vestibular schwannomas.Design and methodsWe calculated whole-body tumor burden in subjects with NF1, NF2, and schwannomatosis using whole-body MRI (WBMRI) and measured the concentration of S100β in plasma using ELISA. We used chi-square tests and Spearman rank correlations to test the relationship between S100β levels and whole-body tumor burden.Results127 consecutive patients were enrolled in the study (69 NF1 patients, 28 NF2 patients, and 30 schwannomatosis patients). The median age was 40 years, 43% were male, and median whole-body tumor volume was 26.9 mL. There was no relationship between the presence of internal tumors and the presence of detectable S100β in blood for the overall group or for individual diagnoses (p > 0.05 by chi-square for all comparisons). Similarly, there was no correlation between whole-body tumor volume and S100β concentration for the overall group or for individual diagnoses (p > 0.05 by Spearman for all comparisons).ConclusionsPlasma S100β is not a useful biomarker for tumor burden in the neurofibromatoses. Further work is needed to identify a reliable biomarker of tumor burden in NF patients.     

Seasonal vitamin D changes and the impact on health risk assessment

ObjectiveTo investigate seasonal variation of vitamin D levels in 148,821 serum samples during a 2 year time period in a northern-latitude city in the United States.MethodsTotal vitamin D assay testing by chemiluminescence was performed on the DiaSorin Liaison. Vitamin D results were extracted from the laboratory information system without patient identification during 2011 and 2012 and separated by season and vitamin D results: less than 10 ng/mL (deficient), 10–20.0 ng/mL (insufficient), 20.1–30 ng/mL (borderline), 30.1–40 ng/mL (sufficient), 40.1–100 ng/mL, and greater than 100 ng/mL.ResultsThe seasonal winter period constituted the months of January through March; spring, April through June; summer, July through September; and fall, October through December. The data set analyzed included 36,643 samples during the winter, 38,299 in spring, 36,141 in summer, and 37,738 in fall and demonstrated an expected rise and fall in vitamin D levels.ConclusionThis retrospective epidemiological study demonstrates seasonal variation of vitamin D levels at clinical decision points. Although not unexpected, this variation has an impact on studies relating low vitamin D levels to higher rates of cancer, cardiovascular disease, multiple sclerosis, diabetes, autoimmune disease, and a host of other health risk assessments.     

Simultaneous quantification of four vitamin D metabolites in human serum using high performance liquid chromatography tandem mass spectrometry for vitamin D profiling

ObjectivesFor quantification of 25-hydroxyvitamin D₃ (25OH-D₃), 25-hydroxyvitamin D₂ (25OH-D₂), 3-epi-25-hydroxyvitamin D₃ (3-epi-25OH-D₃) and 24R,25-dihydroxyvitamin D₃ (24R,25(OH)₂-D₃) in human serum a high performance liquid chromatography tandem mass spectrometry (HPLC–MS/MS) method was developed and validated.Design and methodsAfter protein precipitation further purification is achieved with on-line sample preparation using a reversed phase (RP) C-4 column. Chromatographic separation is realized by a RP-column with core shell material and pentafluorophenyl (PFP) selectivity. Atmospheric pressure chemical ionization in the positive ion mode with multi‐reaction monitoring is used for analyte detection.ResultsBaseline separation of the analytes is achieved below 10 min. The method is linear over the range 4.0–265.3 nmol/L for 25OH-D₃, 3.9–183.6 nmol/L for 25OH-D₂, 2.0–133.8 nmol/L for 3-epi-25OH-D₃ and 2.8–129.9 nmol/L for 24R,25(OH)₂-D₃ (r² > 0.998). The limit of quantification is 4.0 nmol/L for 25OH-D₃, 3.9 nmol/L for 25OH-D₂, 2.0 nmol/L for 3-epi-25OH-D₃ and 2.8 nmol/L for 24R,25(OH)₂-D₃. The CVs for the intra-day and inter-day precision are < 5% and < 4%, respectively. Metabolite levels for a set of 50 human serum samples have been determined and resulted in the detection of considerable amounts of 3-epi-25OH-D₃ and 24R,25(OH)₂-D₃.ConclusionsThis highly specific HPLC–MS/MS method is suitable for vitamin D profiling. There is a correlation between 25OH-D₃ and 24R,25(OH)₂-D₃. Serum concentration of 24R,25(OH)₂-D₃ increases disproportionally with increasing concentration of 25OH-D₃.     

Independent association between 1,25-dihydroxyvitamin D, 25-hydroxyvitamin D and the renin–angiotensin system: The Ludwigshafen Risk and Cardiovascular Health (LURIC) study

BackgroundAntihypertensive and tissue-protective properties of vitamin D metabolites are increasingly attributed to the inhibition of renin synthesis by 1,25-dihydroxyvitamin D [1,25(OH)2D] in the kidney.MethodWe aimed to document a potential association between 25-hydroxyvitamin D [25(OH)D], 1,25(OH)2D and the circulating renin–angiotensin system (RAS) in a large cohort of patients referred (n = 3316) to coronary angiography.ResultsOf the 3316 subjects, 3296 (median age: 63.5 (56.3–70.6) years; 30.2% women) had a baseline measurement of 25(OH)D [median: 15.6(10.1–23.0) µg/L)], 1,25(OH)2D [median: 33.2(25.2–42.9) pg/mL], plasma renin concentration [PRC; median: 11.4(6.0–24.6) pg/mL] and angiotensin 2 [median: 20.0(12.0–35.0) ng/L]. Multivariate adjusted ANCOVA showed a steady increase of PRC values across declining deciles of 25(OH)D and 1,25(OH)2D values (P = 0.013 and P = 0.045), respectively. Additionally, mean angiotensin 2 values increased significantly across decreasing 25(OH)D and 1,25(OH)2D values (P = 0.020 and P = 0.024, respectively). In contrast, multivariate adjusted ANCOVA revealed no significant associations between aldosterone, aldosterone-to-renin ratio and 25(OH)D/1,25(OH)2D values. In multivariate stepwise regression analyses both, 25(OH)D and 1,25(OH)2D emerged as independent predictors of plasma renin and angiotensin 2 concentrations.ConclusionsOur data showed for the first time in humans that both, lower 25(OH)D and 1,25(OH)2D values are independently related to an upregulated circulating RAS.     

Evaluation of the new restandardized 25-hydroxyvitamin D assay on the iSYS platform

Background25-hydroxyvitamin D [25(OH)D] is the most reliable biomarker of vitamin D status, but until now 25(OH)D assays have suffered from inter-laboratory and inter-assay discrepancies. In the setting of the international Vitamin D Standardization Program, Immunodiagnostic Systems (IDS) recently reformulated and restandardized the 25(OH)D immunoassay available on the automated iSYS platform. In the present study, we evaluated this new generation of the 25(OH)D immunoassay (IS-2500).MethodsRepeatability and within-laboratory imprecision were verified according to the Clinical and Laboratory Standards Institute EP15-A3. Results from the sera of 63 patients were compared with those obtained with the previous iSYS method (IS-2700S) using Passing-Bablok and Bland-Altman analysis. The prevalence and bias-adjusted kappa (PABAK) coefficient was calculated to assess the agreement of vitamin D status provided by the two iSYS immunoassays. Fourteen Vitamin D External Quality Assessment Scheme (DEQAS) samples were used to evaluate inaccuracy.ResultsUsing the EP15-A3 protocol, repeatability and within-laboratory imprecision obtained with the new iSYS method were lower than 6% and 8%, respectively. These results are consistent with the manufacturer's claims. In more adverse conditions (50 measurements over 15 days with multiple calibrations), the within-laboratory imprecision was 14.8% (39 nmol/L) and 7.7% (155 nmol/L). 25(OH)D concentrations measured with the new assay showed a strong correlation with those provided by the previous version (r = 0.969, p < 0.0001). The Passing-Bablok regression equation was as follows: new assay = 1.079 x (previous assay) - 3.6 nmol/L. The PABAK coefficient of 0.810 reflected almost perfect agreement between the two immunoassays to classify patients according to their vitamin D status (85.7% of agreement). Using DEQAS samples, the mean inaccuracy bias was lower than 5% when the new iSYS method was compared with LC-MS/MS methods and the NIST reference measurement procedure.ConclusionThe new generation of the iSYS immunoassay evaluated in this study meets requirements for routinely measuring 25(OH)D levels in clinical laboratories.     

Association of R230C ABCA1 gene variant with low HDL-C levels and abnormal HDL subclass distribution in Mexican school-aged children

BackgroundThe effect of ABCA1 genetic variation on HDL-C levels has been widely documented, although studies in children are scarce. We recently found a frequent non-synonymous ABCA1 variant (R230C) exclusive to populations with Native American ancestry, associated with low HDL-C levels and other metabolic traits in adults.MethodsWe genotyped R230C variant in 1253 healthy unrelated Mexican school-aged children aged 6–15 years (595 boys and 658 girls) to seek associations with HDL-C levels and other metabolic traits. HDL subclass distribution was analyzed in a subgroup of 81 age, gender and BMI-matched children.ResultsIndividuals carrying the C230 allele showed a significantly lower HDL-C levels (P = 2.9 × 10^? 8), and higher TC/HDL-C ratio, BMI, BMI z-score and percent fat mass (P = 0.001, 0.049, 0.032 and 0.039, respectively). HDL size was smaller in R230C heterozygotes as compared to R230R homozygotes (P < 0.05). Moreover, the proportion of HDL_2b was lower, while the proportion of HDL_3a and HDL_3b particles was higher in R230C heterozygous and/or C230C homozygous individuals as compared to R230R homozygotes (P < 0.05).ConclusionsOur data suggest that the R230C ABCA1 gene variant plays an important role in HDL-C level regulation and HDL subclass distribution in healthy Mexican school-aged children.     

The fatty acid composition of plasma cholesteryl esters and estimated desaturase activities in patients with nonalcoholic fatty liver disease and the effect of long-term ezetimibe therapy on these levels

BackgroundThe aim of this study was to investigate the relationship between fatty acid composition of plasma cholesteryl esters (CEs) and estimated desaturase activity and the development and progression of nonalcoholic fatty liver disease (NAFLD). The study also assessed the effect of ezetimibe on CE levels.MethodsPlasma CEs fatty acid composition was analyzed in 3 groups: patients with a NAFLD activity score (NAS) ≤ 4 (n = 31) or NAS ≥ 5 (n = 32) and normal controls (n = 25). The estimated desaturase activities were calculated using ratios of 16:1n?7/16:0 (D9-16D), 18:1n?9/18:0 (D9-18D), 18:3n?6/18:2n?6 (D6D) and 20:4n?6/20:3n?6 (D5D).ResultsCompared with controls, the levels of palmitate, palmitoleate, γ-linoleate, D9-16D and D6D were significantly increased, whereas levels of linoleate and D5D were significantly decreased. Patients with NAS ≥ 5 had significantly higher palmitate levels than patients with NAS ≤ 4. The levels of these fatty acids, especially palmitate and palmitoleate, correlated with NAFLD-related lipid, metabolic, and inflammatory parameters. Long-term therapy with ezetimibe caused significant improvements in the levels of these fatty acids, estimated desaturase activity index and NAFLD-related parameters.ConclusionsOur results suggest that fatty acids and desaturase activity associate with the development and progression of NAFLD, and that ezetimibe may be a novel treatment for this disorder.     

Exposure to parental verbal abuse is associated with increased gray matter volume in superior temporal gyrus

ObjectiveExposure to parental verbal aggression (PVA) during childhood increases risk for the development of psychopathology, particularly mood and anxiety disorders. Other forms of childhood abuse have been found to be associated with alterations in brain structure. The aim of this study was to ascertain whether exposure to PVA was associated with discernible effects on brain morphology.MethodsOptimized voxel-based morphometry was performed on 21 unmedicated, right-handed subjects (18–25 years) with histories of PVA and 19 psychiatrically healthy controls of comparable age and gender. Group differences in gray matter volume (GMV) – covaried by age, gender, parental education, financial stress, and total GMV – were assessed using high-resolution, T1-weighted, volumetric MRI data sets (Siemens 3T trio scanner).ResultsGMV was increased by 14.1% in the left superior temporal gyrus (STG, BA 22) (P = 0.004, corrected cluster level). GMV in this cluster was associated most strongly with levels of maternal (ß = 0.544, P < 0.0001) and paternal (ß = 0.300, P < 0.02) verbal aggression and inversely associated with parental education (ß = ? 0.577, P < 0.0001).ConclusionPrevious studies have demonstrated an increase in STG GMV in children with abuse histories, and found a reduction in fractional anisotropy in the arcuate fasciculus connecting Wernicke's and frontal areas in young adults exposed to PVA. These findings and the present results suggest that the development of auditory association cortex involved in language processing may be affected by exposure to early stress and/or emotionally abusive language.     

Bone turnover markers in Spanish adult men: The Camargo Cohort Study

BackgroundThis cross-sectional study was performed to determine the reference ranges for two bone turnover markers—aminoterminal propeptide of type I collagen (P1NP) and C-terminal telopeptide of type I collagen (β-CTX)—in normal adult Spanish men as measured in serum by automated methods.MethodsA community-based population of 660 healthy men ≥ 50 years was evaluated. Fasting serum levels of P1NP, β-CTX, 25-hydroxyvitamin D, and intact parathyroid hormone were measured on the Elecsys 2010 automated analyzer (Roche). BMD at lumbar spine, femoral neck and total hip was determined by DXA.ResultsMean age of participants was 65 ± 9 years. Logarithmic transformation of both markers was performed to allow for normal distribution. Mid-95% ranges for P1NP and β-CTX were 15–78 ng/ml and 0.069–0.760 ng/ml, respectively. Median and interquartile range of serum P1NP and β-CTX were 33.5 [25.5;44.4] ng/ml and 0.27 [0.19;0.38] ng/ml, respectively. Mean values of P1NP (37.1 ± 16.7 ng/ml) were similar to those previously described. β-CTX mean values (0.300 ± 0.171 ng/ml) were also similar to those quoted by the manufacturers in men younger than 70 years, but slightly lower than those reported in subjects older than 70 years. Both markers were higher among osteoporotic men. After excluding from the analysis those men who were found to have BMD below ?2.5 T-score, 25OHD serum level below 30 ng/ml or serum PTH above 65 pg/ml, P1NP and β-CTX ranges were 17–71 ng/ml and 0.070–0.681 ng/ml, again respectively.ConclusionsValues obtained from this well-characterized population study provide reference ranges for serum automated P1NP and β-CTX in normal Spanish adult men.     

BMI and hyperinsulinemia in children

ObjectivesOverweight is associated with hyperinsulinemia in adults and children. The aim of our study was to test if body mass index (BMI) alone or in combination with waist circumference (WC) predicts hyperinsulinemia in individual children.MethodsIn 466 healthy German schoolchildren aged 7 to 12 years enrolled in the population based PEP Family Heart Study, anthropometric data, standard laboratory data and fasting serum insulin concentrations were assessed.ResultsAmong children with hyperinsulinemia (fasting serum insulin concentrations  > 85th age specific percentile), 56% were not overweight (BMI-for-age   < 85th percentile). Among overweight children, 54% were normoinsulinemic. Increased waist circumference (WC for age and gender  > 85th percentile) was not a better predictor of hyperinsulinemia, neither alone nor in combination with BMI.ConclusionOverweight and abdominal obesity are poor indicators of hyperinsulinemia in individual German school children aged 7 to 12 years. Insulin measurement seems to be necessary to reliably detect hyperinsulinemia.     

Analytical measurement of serum 25-OH-vitamin D3, 25-OH-vitamin D2 and their C3-epimers by LC–MS/MS in infant and pediatric specimens

ObjectivesTo develop a simple and sensitive LC–MS/MS procedure for quantification of serum 25-OH-vitamin D₃ (25-OH-D₃), 25-OH-vitamin D₂ (25-OH-D₂), and their C3-epimers.MethodsSerum 25-OH-vitamin D metabolites were extracted with MTBE and quantified by LC–MS/MS. Commercially available calibrators and QC materials were employed. The ion-transition 401.2 → 365.2 was monitored for 25-OH-D₃ and C3-epi-25-OH-D₃, 407.2 → 371.3 for d₆-25-OH-D₃, 413.2 → 331.2 for 25-OH-D₂ and C3-epi-25-OH-D₂ and 419.2 → 337.1 for, d₆-25-OH-D₂. As a proof-of-principle, 25-OH-D₃ and C3-epi-25-OH-D₃ were quantified in 200 pediatric subjects (0–20 years of age). Cholecalciferol supplements were examined as a potential source of C3-epimer.ResultsThe assay provided an LLOQ of ≤ 2.8 nmol/L for all 25-OH-D metabolites, with a linear response up to 400 nmol/L. The CV was < 10% for 25-OH-D_2/3 and < 15% for C3-epi-25-OH-D₃. C3-epi-25-OH-D₃ was quantified in all subjects, with higher concentrations observed in infants ≤ 1 year of age (11.44 nmol/L vs. 4.4 nmol/L; p < 0.001). Within the first year of life, 25-OH-D₃ concentrations increased linearly, while C3-epi-25-OH-D₃ concentrations remained constant. At 12 months of age, C3-epi-25-OH-D₃ concentration dropped by almost 50% (11.4 nmol/L in infants ≤ 1 year of age vs. 5.4 nmol/L in infants 1–2 years of age; p < 0.001). Liquid vitamin D₃ supplements did not contain appreciable amounts of C3-epi-D₃.ConclusionsThe proposed LC–MS/MS procedure is suitable for quantifying 25-OH-D₃ metabolites. Although the C3-epimer is present in all pediatric subjects, it is significantly elevated in individuals ≤ 1 year of age and drops at 12 months of age. Oral vitamin D supplements are unlikely to be a significant source of C3-vitamin D epimer.     

Lactobacillus rhamnosus induced epithelial cell apoptosis,ameliorates inflammation and prevents colon cancer development in an animal model

Background/AimProbiotics have been suggested as prophylactic measure in colon carcinogenesis. This study aimed at determining the potential prophylactic activity of Lactobacillus rhamnosus GG CGMCC 1.2134 (LGG) strain on colorectal carcinogenesis via measuring its effect on Nuclear factor kappa B (NFκB) inflammatory pathway and apoptosis.Materials and methods64 Sprague Dawley rats were grouped into four as follows; Group 1 (Healthy control), Group 2 (LGG), Group 3 (cancer control Dimethyl hydrazine (DMH)) and Group 4 (LGG + DMH). LGG was administered orally to LGG and LGG + DMH groups. Colon carcinogenesis was chemically induced in LGG + DMH and DMH groups by weekly injection of 40 mg/kg DMH. Animals were sacrificed after 25 weeks of experiment and tumor characteristics assessed. The change in expression of NFκB-p65, COX-2, TNFα, Bcl-2, Bax, iNOS, VEGFα, β-catenin, Casp3 and p53 were evaluated by western blotting and qRT-PCR.ResultsLGG treatment significantly reduced tumor incidence, multiplicity and volume in LGG + DMH treatment group compared to DMH cancer control group. Also, LGG treatment reduced the expression of β-catenin and the inflammatory proteins NFκB-p65, COX-2 and TNFα; the anti-apoptotic protein Bcl-2, but increased the expression of the pro-apoptotic proteins Bax, casp3 and p53 compared with DMH group.ConclusionLGG have a potential protection effect against colon carcinogenesis; inducing apoptosis and ameliorating inflammation, and may hold a promise as bio-therapeutic dietary agent.     

High-throughput liquid–liquid extraction and LCMSMS assay for determination of circulating 25(OH) vitamin D3 and D2 in the routine clinical laboratory

BackgroundIncreasing focus on vitamin D as essential to health has underscored the need for accurate and precise high-throughput measurement of serum 25(OH)D.MethodsSerum was denatured in acetonitrile containing hexadeuterated 25(OH)D3 as internal standard (IS) and automatically applied to filter plates packed with inert diatomous earth material for subsequent heptane extraction. Extracts were chromatographed on a C12 HPLC column, and detected on a triple quadropole mass spectrometer.ResultsThe inter-assay precision was 9.4% and 8.8% respectively at 32.0 and 59.7 nmol/l for 25(OH)D3 and 8.6% and 8.0% at 23.4 and 64.4 nmol/l for 25(OH)D2. The detection limit was 10 nmol/l for both metabolites. Three percent of samples contained > 50 nmol/l 25(OH)D2. Total run time was 4 min. We have performed more than 200,000 routine samples and the method performs well in external control schemes.ConclusionWe describe a robust, high-throughput, LLE-LCMSMS method for accurate and precise quantitation of 25(OH)D3 and 25(OH)D2 in serum. The use of diatomaceous earth material for extraction of vitamin D in 96-well format enables fast, simple and efficient sample preparation. The method offers a cost-effective alternative to immunological methods for use in the routine clinical biochemical laboratory.     

Validity of participant-reported diagnoses in an online patient registry: A report from the NF1 Patient Registry Initiative

BackgroundWith increased internet accessibility worldwide, it is now possible to assemble individuals with rare diseases through web-based patient registries. However, the validity of participant-reported medical diagnoses is unknown. The objective of this study was to evaluate the accuracy of participant-reported Neurofibromatosis Type 1 (NF1) diagnoses among participants in the NF1 Patient Registry Initiative (NPRI).MethodsSubjects enrolled in the NPRI from 5/17/2011 to 7/7/2014 were included. Medical records (MRs) were obtained for participants who returned medical record release forms (MRRFs) during the study period. Participants were classified as having definite, probable, suspected, or no NF1 diagnosis based on MR information. To assess whether a returned MRRF served as a reliable marker of MR-documented NF1, we calculated the positive predictive value (PPV) as the proportion of individuals with MR-documented NF1 among those from whom MRs were obtained. We further examined whether a returned MRRF predicted the number of reported NF1 clinical signs in multivariable linear regression analyses.ResultsA total of 1456 individuals were included in the analyses. Of 416 individuals who returned MRRFs, 205 MRs were reviewed within the study period. The PPV ranged from 72.0 to 98.5% when including definite or definite/probable/suspected cases, respectively. The mean number of reported NF1 clinical signs was similar between those who returned (mean = 3.3 ± 1.2) and did not return (mean = 3.2 ± 1.3) their MRRFs. MRRF return was not a significant predictor of the number of NF1 clinical signs after adjusting for covariates.ConclusionThese data strongly suggest that individuals enrolling in the NPRI accurately report their NF1 diagnosis.     

Outcome and risk factors in children after traumatic cardiac arrest and successful resuscitation

IntroductionProspective collected data of the TraumaRegister DGU^® were analyzed to derive survival rates and predictors for non-survival in the children who had suffered traumatic cardiorespiratory arrest. Different time points of resuscitation efforts (only preclinical, in the emergency room (ER) or preclinical + ER) were analyzed in terms of mortality and neurological outcome.MethodsThe database of the TraumaRegister DGU^® comprising 122,742 patients from 1993 to 2013 was analyzed. The main focus of this survey was on the paediatric group defined by an age ≤14 years who could be compared to adults. Different statistical analysis (univariate and multivariate analysis, logistic regression) were performed with mortality as the target variable. Differences between the paedatric group and adults were analysed by Fisher's exact test.ResultsData after preclinical and/or ER resuscitation from 152 children and 1690 adults were analyzed. A good or moderate outcome (GOS 5 + 4) was found in 19.4% of the children's group compared to 12.4% of the adults (p = 0.02).Analysis of the GOS 5 + 4 subgroups after preclinical resuscitation only revealed that these outcomes were achieved by 19.4% of the paediatric group and 13.2% of the adults (p = 0.24), after ER-only resuscitation by 37.0% of the children and 19.6% of the adults (p = 0.046), and after preclinical and ER resuscitation by only 10.9% of the children compared to 2.5% of the adults (p = 0.006). Taking only survivors into account, 84.8% of the children and 62% of the adults had a GOS 4 + 5.The highest risk for mortality in the logistic regression model was associated with preclinical intubation, followed by GCS 3, blood transfusion and severe head injury with AIS ≥3 and ISS.ConclusionsCPR in children after severe trauma seems to yield a better outcome than in adults, and appears to be more justified than the current guidelines would imply. Resuscitation in the ER is associated with better neurological outcomes compared with resuscitation in a preclinical context or in both the preclinical phase and the ER. Our children's outcomes seem to be better than those in most of the earlier studies, and the data presented might support algorithms in the future especially for paediatric resuscitation.     

Prediction equation of hip external rotators maximum torque in healthy adults and older adults using the measure of hip extensors maximum torque

BackgroundThe use of predictive equation of muscular torque can reduce physical effort and time spent during evaluation.ObjectivesTo establish, validate, and test the accuracy of a prediction equation to estimate the hip external rotators (HER) torque in adults and older adults by means of hip extensors (HEX) torque measurement.MethodsEighty-three healthy adults (development set) were assessed to test the association of HEX and HER torques and to establish the prediction equation. A separate 36 adults and 15 older adults (validation sets) were assessed to test the ability of the equation to estimate HER torque. Hip isometric strength was assessed by a handheld dynamometer.ResultsSimple linear regression analysis revealed that HEX torque was associated with HER torque (r = 0.80; p < 0.0001), resulting in the following prediction equation: HER_torque= −0.02 + (0.58 * HEX_torque). Paired t-test revealed no difference between directly measured and predicted values of HER torque in adults (mean difference = 0.02; 95% CI = −0.115, 0.072) and older adults (mean difference = 0.05; 95% CI = −0.02, 0.12).ConclusionThe HEX and HER torques were strongly correlated. The prediction equation was valid, accurate, and can be used to estimate HER muscle strength in healthy adults and older adults, requiring only the direct measurement of HEX torque.     

Urine 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG), a specific marker of oxidative stress,using direct,isocratic LC–MS/MS: Method evaluation and application in study of biological variation in healthy adults

BackgroundUrine 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) is a specific biomarker of oxidative stress. We evaluated a modified LC–MS/MS assay for urine 8-oxodG and determined biological variation in healthy adults.MethodUntreated urine was injected into an isocratic LC–MS/MS system (positive-ion MRM mode). Urine 8-oxodG in 51 healthy volunteers was measured; within- and between-day variations in 23 healthy volunteers were investigated.ResultsDose–response was linear to 452 nmol/l; limit of detection = 2.3 nmol/l; within-run and between-run CVs were < 3.0% and < 4.7%, respectively; recovery = 97%–101%; accuracy = 97.7–103.5%. Urine 8-oxodG (median, mean [SD]): 1.70, 1.70[0.60]nmol/mmol creatinine (n = 51). Men had higher (p = 0.027) concentrations than women matched for age and body mass index: mean [SD]: 1.90[1.60]; n = 26 vs. 1.50[0.55]; n = 25. Within- and between-day variations were wide but random. No significant differences were seen overall across time-points within 1 day or at the same time-point across 5 consecutive days.ConclusionsThe method has advantages of speed and relative simplicity as it does not require sample pre-treatment for 8-oxodG extraction, the use of internal standard or gradient LC elution and has high linearity, specificity, precision and recovery. Biological variation in urine 8-oxodG is wide, but no within- or between-day differences at the group concentration were seen in healthy adults.     

High frequency ultrasound sacral images in the critically ill: Tissue characteristics versus visual evaluation

PurposeHigh frequency ultrasound (HFUS) systems may identify tissue injury. We compared HFUS tissue characteristics (dermal thickness and dermal density) with visual image examination.MethodsLongitudinal study in critically ill mechanically ventilated adults, from three ICUs (Surgical Trauma, Medical Respiratory, Neuroscience) enrolled within 24 hours of airway intubation. Sacral HFUS images were obtained daily for up to seven days. Expert evaluation of the best image per day was completed and compared to HFUS generated tissue characteristics (dermal thickness and dermal density).ResultsOf the113 subjects with 1614 comparisons analysed, 73.2% to 84% were normal, and 6.3% to 11.8% of the comparisons had injury present but no change was noted in the injury observed. There were no significant differences in one-day comparisons among type of injury and mean dermal thickness (p = 0.6645) or dermal median intensity (adjusted p = 0.06-0.17). All other day-to-day comparisons were similarly non-significant.ConclusionsWe found no association among dermal density, dermal thickness and visual examination of changes in sacral HFUS images for any day-to-day comparison. The use of sacral HFUS as a screening tool for the development of tissue injury is in its infancy. Additional comparative studies should be conducted to identify its future clinical usefulness.     

Serum levels of basic fibroblastic growth factor (bFGF) in children with vascular anomalies: Another insight into endothelial growth

ObjectivesDysregulation of angiogenesis has been proposed to play a central role in hemangioma pathogenesis. The aim of the study was to determine the peripheral and local serum levels of bFGF in patients with hemangiomas and vascular malformations (VM).Design and methodsThe study group consisted of 52 children with infantile hemangioma, 14 with VM and 36 healthy patients. bFGF serum levels were analyzed by an ELISA assay. Urinary bFGF was determined in 11 individuals with hemangioma.ResultsThe serum peripheral bFGF concentrations in children with proliferating hemangiomas were lower than in healthy controls (p = 0,03). There was no correlation between the measured cytokine level and hemangioma size, as well as patients’ age. The serum local bFGF levels in 29 children with hemangiomas were higher than in the peripheral blood (p = 0.022). Urinary bFGF in hemangioma patients did not differ statistically from healthy controls.Conclusions(1) Determination of bFGF serum levels is not helpful in differentiating the phases of hemangioma growth and distinguishing hemangiomas from VM; (2) serum levels of bFGF cannot distinguish between extrinsic and intrinsic theories of endothelial cell proliferation in hemangiomas.