Protective effect of L-carnitine on platelet apoptosis during storage of platelet concentrate

BackgroundPlatelet apoptosis is considered as one of the important factors involved in platelet storage lesion (PSL) and affect the quality of platelets during storage. The beneficial effect of L-carnitine (LC) on platelet apoptosis during platelet concentrates (PCs) storage has not been fully investigated. The aim of this study was to evaluate the effects of LC on platelets of PC regarding their apoptosis markers during storage.MethodsTen PCs from healthy donors were investigated in this study. PCs were prepared by platelet rich plasma (PRP) method and stored at 22 ± 2° C with gentle agitation during storage. The effects of LC (15 mM) on the platelet apoptosis were assessed by analyzing different indicative presence or absence of LC. Sampling was performed to evaluate apoptosis markers during platelet storage.ResultsThe results indicated significantly higher mitochondrial membrane potential for LC-treated platelets than the untreated on the days 2 and 5 of storage (P_day2 = 0.001, P_day5 = 0.001). Phosphatidylserine (PS) exposure significantly increased on the untreated compared with LC-treated platelets on the second and third days of storage (P_day2 = 0.014, P_day3 = 0.012). Also, active caspase 3 was lower in the LC- treated platelets than the control group on the day 5 of storage (P_day5 = 0.004). Cytosolic cytochrome C was so significantly lower in LC-treated compared to the untreated platelets during storage time (P_day2 = 0.002, P_day3 = 0.001, Pday5 = 0.001).ConclusionThe results of this study indicate that the use of LC as an additive solution in platelets may be useful to reduce PSL by decreasing platelet apoptosis via mitochondrial pathway and increase platelet quality during storage.     

Platelet transfusion practice and related transfusion reactions in a large teaching hospital

BackgroundPlatelet transfusion practice varies widely since many aspects of platelet concentrate (PC) use have not been definitively determined. The objectives of this retrospective study were to present platelet transfusion practice and evaluate PC and patient characteristics, as well as their association with transfusion reaction (TR) rate.Material and methodsPlatelet transfusions over a 5-year period were analysed regarding PC characteristics (the ABO and RhD compatibility, product type, and storage duration), patient characteristics (most responsible diagnosis, age, and gender), and TR type.ResultsA total of 46,351 PCs were transfused: 76.4% whole blood-derived (WBD) and 23.6% single donor apheresis (SDA). Three thousand seven hundred seventy-six patients received platelet transfusions: 24.7% paediatric and 75.3% adult patients, 79.6% outpatients and 20.4% inpatients. As much as 63.1% of all transfused PCs were fresh (stored for ≤ 3 days), 98.0% ABO-identical, and 87.3% of all PCs given to RhD? patients were RhD?. PCs were mainly transfused to haemato-oncology (76.8%) and cardiovascular surgery patients (6.5%). Overall, 84 (0.18%) TRs were reported, with allergic TRs (ATRs) being the most common. Although PC ABO compatibility and storage duration, as well as patient age and gender, showed differences in TR rate, only the use of PCs in platelet additive solution (PAS) showed a statistically significant reduction of TRs (P < 0.001).ConclusionTransfusion practice at the University Hospital Centre Zagreb resulted in almost all patients receiving ABO and RhD identical PCs, and most of them were fresh PCs. The most important factor affecting the incidence of TRs was platelet storage solution. The use of PAS effectively reduced the rate of TRs, particularly allergic TRs.     

Microbial translocation is associated with residual viral replication in HAART-treated HIV+ subjects with <50 copies/ml HIV-1 RNA

BackgroundRecent data have shown that plasma levels of lipopolysaccharide (LPS) are a quantitative indicator of microbial translocation in HIV infected individuals.ObjectivesTo assess the impact of residual viral replication on plasma LPS in HAART-treated HIV+ subjects with <50 copies/ml HIV-1 RNA and to evaluate LPS changes during repeated HAART interruptions not exceeding 2-month duration.Study designLPS was measured in 44 HIV+ subjects at T0 (during HAART) and at day 15 of the first and fourth HAART interruption. Ten uninfected, healthy donors were studied as well. Residual plasma HIV-1 RNA was measured at T0 by an ultra-ultrasensitive method with limit of detection of 2.5 copies HIV-1 RNA/ml. Subjects with less than 2.5 copies/ml (fully suppressed – FS) were compared to those with 2.5–50 copies/ml (partially suppressed – PS).ResultsAt T0, plasma LPS levels were comparable in FS and uninfected subjects, whereas in PS they were higher than in uninfected subjects (p = 0.049). After 4 HAART interruptions, they did not change significantly. However, LPS values were lower in FS than in PS (p = 0.020). An inverse correlation was found between CD4 and LPS levels (p = 0.044) in PS group only.ConclusionsA reduced degree of microbial translocation was seen in subjects with a more complete suppression of viral replication. Repeated HAART interruptions had no significant impact on plasma LPS levels.     

HHV-8 DNA replication correlates with the clinical status in AIDS-related Kaposi’s sarcoma

BackgroundThe value of plasma levels of human herpesvirus 8 (HHV-8) DNA as a marker of clinical status in acquired immunodeficiency syndrome-related Kaposi’s sarcoma (AIDS-KS) remains to be elucidated.ObjectivesTo investigate the relationship between the plasma HHV-8 DNA viral load and the clinical status of AIDS-KS.Study DesignA total of 378 blood samples were obtained from 62 patients with AIDS-KS followed longitudinally. All patients received antiretroviral therapy (ART) or anti-neoplastic therapy. The patients were divided into four groups according to their clinical status: onset disease (OD), progressive disease (PD), stable or partial remission (S/PR) and complete remission (CR).ResultsPlasma HHV-8 DNAaemia was detected in all samples obtained from patients with OD or PD (100%); in contrast, HHV-8 DNAaemia was found only in a minority of patients with CR (8%) and was invariably undetectable in patients with stable CR. HHV-8 DNA detection in plasma was strongly associated with an unfavourable outcome (odds ratio = 231.9; p < 0.0001). Conversely, neither the HIV-1 viral load nor peripheral CD4⁺ T-cell counts were associated with the KS clinical status, though both parameters did affect HHV-8 DNAaemia levels (p < 0.0001). Multivariate analysis confirmed that HHV-8 DNAaemia was strongly and independently correlated with both clinical status (p < 0.05) and HIV-1 plasma viraemia (p = 0.027).ConclusionsThe strong association of plasma HHV-8 DNAaemia with onset or progressive disease is compatible with an active role of replicating virus in clinically active AIDS-KS. An accurate evaluation of the plasma HHV-8 load might be useful for monitoring AIDS-KS under antiretroviral or antineoplastic therapy.     

High concentrations of anti-caspase-8 antibodies in Chilean patients with type 1 diabetes

IntroductionDeregulation of apoptosis across the Fas–FasL pathway is an increasingly relevant phenomenon in the pathogenic mechanisms associated with autoimmune diseases. Caspase-8 initiates the activation of the apoptotic process and interacts directly with Fas in the membrane of the T lymphocyte.ObjectivesTo standardize an Elisa essay to measure the concentration of anti-caspase-8 antibodies in plasma of Type 1 Diabetes (T1D) patients and analyze their possible distribution and association with characteristics of the disease.Methods and subjects124 patients newly diagnosed with T1D and 132 controls: children and youngsters. ELISA test was standardized to detect anti-caspase-8 antibodies in plasma. It correlated the concentration of this antibody with classical markers of autoimmunity as anti-IA-2 and anti-GAD65, and the clinical characteristics at onset of diabetes mellitus. The statistical analysis was performed using logistic regression.ResultsPatients with T1D showed a higher concentration of anti-caspase-8 antibodies regarding the controls (87.5 ng/ml versus 24.3 ng/ml, p < 0.0001, values expressed as median). The proportion of patients with T1D and high concentrations of anti-caspase-8 (percentile 50–75) was significantly different from the control group (p < 0.0001). Anti-caspase-8 showed a strong association with positive anti-GAD65 (OR = 3.48, p < 0.035) and ketoacidosis (OR = 10.74, p < 0.0001) events, with glycemia and age at diagnosis as contributing variables.ConclusionThis is the first report in the literature of levels of anti-caspase-8 antibodies in T1D through ELISA. The high concentration in patients with T1D, and its strong correlation with anti-GAD65 auto-antibodies, suggests a potential role of anti-caspase-8 auto-antibodies as surrogate marker autoimmunity in T1D patients.     

Microbial translocation is associated with residual viral replication in HAART-treated HIV+ subjects with <50 copies/ml HIV-1 RNA

BackgroundRecent data have shown that plasma levels of lipopolysaccharide (LPS) are a quantitative indicator of microbial translocation in HIV infected individuals.ObjectivesTo assess the impact of residual viral replication on plasma LPS in HAART-treated HIV+ subjects with <50 copies/ml HIV-1 RNA and to evaluate LPS changes during repeated HAART interruptions not exceeding 2-month duration.Study designLPS was measured in 44 HIV+ subjects at T0 (during HAART) and at day 15 of the first and fourth HAART interruption. Ten uninfected, healthy donors were studied as well. Residual plasma HIV-1 RNA was measured at T0 by an ultra-ultrasensitive method with limit of detection of 2.5 copies HIV-1 RNA/ml. Subjects with less than 2.5 copies/ml (fully suppressed – FS) were compared to those with 2.5–50 copies/ml (partially suppressed – PS).ResultsAt T0, plasma LPS levels were comparable in FS and uninfected subjects, whereas in PS they were higher than in uninfected subjects (p = 0.049). After 4 HAART interruptions, they did not change significantly. However, LPS values were lower in FS than in PS (p = 0.020). An inverse correlation was found between CD4 and LPS levels (p = 0.044) in PS group only.ConclusionsA reduced degree of microbial translocation was seen in subjects with a more complete suppression of viral replication. Repeated HAART interruptions had no significant impact on plasma LPS levels.     

Correlation between HIV-1 viral load quantification in plasma,dried blood spots,and dried plasma spots using the Roche COBAS Taqman assay

BackgroundThe use of simplified methods for viral load determination could greatly increase access to treatment monitoring of HIV patients in resource-limited countries.ObjectiveThe aim of the present study was to optimize and evaluate the performance of the Roche COBAS Taqman assay in HIV-RNA quantification from dried blood spots (DBS) and dried plasma spots (DPS).Study designEDTA blood samples from 108 HIV-infected women were used to prepare 129 DBS and 76 DPS on Whatman 903 card. DBS and DPS were stored at ?20 °C. HIV-1 RNA was extracted from DBS/DPS using the MiniMAG system (bioMerieux). Amplification and detection were performed using the Roche COBAS TaqMan assay. Plasma viral load results were used as standard.ResultsThere was a high correlation between measures of viral load in plasma and in DBS/DPS (r = 0.96 and 0.85 respectively, P < 0.001). Overall, viral load values in DBS and DPS tended to be lower than in plasma with mean (SD) differences of 0.32 log (0.22) for DBS and of 0.35 (0.33) for DPS. Detection rates were 96.4% for DBS and 96.1% for DPS in samples with corresponding plasma values >3.0 log copies/ml. Samples with HIV-RNA below 50 copies/ml were correctly identified in 18/19 DBS and in 7/7 DPS.ConclusionsBoth DBS and DPS provided results highly correlated to the plasma values. High detection rate was obtained with both DBS and DPS when HIV-RNA was >3.0 log copies/ml. Our results support the use of DBS/DPS to detect virologic failure in resource-limited settings.     

Association of plasma viremia with HIV-1 RNA levels in cervicovaginal lavage secretions in HIV-1 seropositive ART naïve women from Pune,India

BackgroundCoherent drug/microbicide/vaccine development research would benefit through a precise knowledge of HIV dissemination and its persistence in the female genital tract. Understanding relationship between plasma viremia and cervicovaginal HIV shedding may help to unveil mechanisms underlying transmission, compartmentalization and pathogenesis.ObjectivesTo study the association between HIV-1 RNA levels in the plasma and CVL specimens.Study designWhole blood, plasma and CVL specimens were collected from 36 ART naïve HIV-1 seropositive women qualifying the study inclusion criteria. Absolute CD4 counts, plasma and CVL HIV-1 RNA levels were estimated using commercially available kits (BD MultiSET™ Kit, Becton Dickinson, US and Abbott RT, Abbott Molecular, Germany). Correlation between plasma and CVL viral load was estimated by the Spearman's rank correlation coefficient. Additionally, the correlation between CVL viral load and absolute CD4 counts was studied.ResultsHIV-1 viral load in the CVL specimens was successfully quantified using the Abbott RT. Twenty-seven of 36 women (75%) had detectable HIV-1 RNA levels in plasma and CVL specimens. The CVL viral load did not show any correlation with plasma viral load (ρ = 0.281, p = 0.096) and showed a ‘moderate correlation’ (ρ = −0.563, p = 0.0004) with absolute CD4 counts.ConclusionsAlbeit, the Abbott RT is designed for estimating plasma HIV-1 RNA levels, the study reports its use for estimating HIV-1 RNA levels in the CVL specimens as well. In accordance with the previous studies, our results suggest that plasma and CVL viral load are not correlated and plasma viremia might not solely predict cervico vaginal HIV shedding.     

Improving chemotherapy processes with a protocol-based information system: A pre and post-implementation study

BackgroundThe medical application domain has been a great challenge for information technology solutions for decades, especially when the target process has been complex and multidisciplinary such as chemotherapy processes.ObjectiveTo evaluate the impact of a homegrown protocol based information system on the efficiency of chemotherapy workflow processes in an outpatient setting.MethodsA day care unit of the Hematology/Oncology outpatient clinic of Erasmus Medical Center was the setting for this study. The study consisted of comparison of pre- and post-implementation of four workflow efficiency related external indicators: turn-around times of a commonly administered chemotherapy course (Paclitaxel–Carboplatin), chemotherapy course administration postponing rate, the rate of recording course administration time, and patient admission rate of the outpatient clinic. The data was gathered retrospectively from patient charts and information systems’ log files. For the purpose of turn-around-time 109 Paclitaxel–Carboplatin chemotherapy courses of pre-implementation were compared to 118 those of post-implementation. For the other indicators: 247 chemotherapy courses pre-implementation were compared to 324 courses post-implementation. The process maps of pre- and post-implementation were also compared to each other.ResultsThe implementation of the system improved the process by removing repetition and sequencing of the tasks. Following the implementation, chemotherapy postponing decreased by 17.2% (Z = ?4.723, P = .000) and there were 5.7% less records with missing administration time (Z = ?3.047, P = .002). The admission rate increased 1.9 patient per working day (t(94) = ?5.974, P = .000). The overall turn-around-time reduced 18.9 min following the implementation (t(169) = 3.48, P = .001). In a multivariate multiple regression model the reduction in turn-around time was related to the implementation of the system (Pillai's Trace = 0.159, F(4,161) = 7.613, P = .000).ConclusionInformation systems based on treatment protocols can reduce communication and synchronization needs between the stakeholders in a complex workflow process. These systems can help reengineering the process and improve workflow efficiency by removing unnecessary sequencing and repetitions of tasks.     

Comparative effectiveness of dried plasma HIV-1 viral load testing in Brazil using ViveST for sample collection

BackgroundUtilization of dried plasma for HIV-1 viral load testing would significantly decrease sample shipping costs.ObjectivesTo describe the precision and reproducibility of ViveST^® (ST) as a transportation method for shipping specimens for HIV-1 viral load (VL) testing.Study designThirty clinical plasma samples were used to generate replicate samples with HIV VL values of 4 log₁₀, 3 log₁₀ and 2 log₁₀ copies/mL for reproducibility testing and an additional 299 samples with HIV VL <50 copies/mL (99); 1.7 log₁₀ to 3.99 log₁₀ (100); and 4 log₁₀ to 5.99 log₁₀/mL (100) were used to compare ViveST to frozen plasma samples using the VERSANT^® HIV-1 RNA 3.0 Assay. Results were compared using Student t-test, Pearson correlation and Bland–Altman analyses.ResultsMean intra-assay variance among frozen and dried plasma triplicates was 0.15 log₁₀ and 0.09 log₁₀ copies/mL respectively (n = 10, P = NS). Compared to frozen plasma, there was a mean reduction of 0.3 log₁₀, 0.27 log₁₀, and 0.35 log₁₀ copies/mL at the 4 log₁₀, 3 log₁₀, and 2 log₁₀ copy/mL samples respectively (n = 30, all comparisons, P < 0.01). Overall correlation between 299 frozen and ViveST samples was r = 0.97, where 12 of 99 undetectable frozen VL were positive with ST, and 12 of 200 frozen detectable VL were undetectable with ViveST (mean VL 2.1, 1.9 log₁₀ copies/mL respectively).ConclusionsHIV-1 viral load results using ViveST were reproducible, correlated well with frozen plasma, though yielding minimally lower values. Our data suggest that dried plasma for HIV-1 VL testing using ViveST has promise for use in HIV clinical practice.     

Clinical implications of hepatitis B viral infection in Epstein–Barr virus-associated nasopharyngeal carcinoma

BackgroundLittle is known about the clinical implication of hepatitis B virus (HBV) infection in Epstein–Barr virus (EBV)-associated nasopharyngeal carcinoma (NPC).ObjectiveThis study aimed to investigate the clinical characteristics and prognostic factors in patients with newly-diagnosed NPC with HBV infection.Study designA total of 722 patients with pathologically-diagnosed NPC who received comprehensive treatment at First People's Hospital of Foshan between June 2006 and December 2011 were enrolled in this retrospective study; 79 and 643 patients were HBsAg(+) and HBsAg(−), respectively. The correlations between HBV (HBsAg status and HBV DNA load) and EBV DNA were analyzed, further long-term survival and prognostic factors also were explored.ResultsWe reported NPC patients with HBsAg(+) represented worse outcome, and distant-failure especially liver metastasis was more common in these patients. HBV infection was more frequent in younger patients and male patients. No correlation was observed between the pre-treatment plasma EBV DNA load (cutoff, 1500 copies/ml) and HBsAg status (positive or negative; r = −0.036, P = 0.392), or the pre-treatment plasma EBV DNA load and HBV DNA load (r = 0.042, P = 0823).ConclusionsBoth HBV and EBV infection is an independent negative prognostic factor for long-term survival, distant metastasis, especially liver metastasis, was more common in NPC patients with HBsAg(+), and it seemed no link between EBV DNA load and HBsAg status in NPC.     

CMV plasma DNAemia and risk of cancer among HIV-infected patients: A case–control study nested in the ANRS CO3 Aquitaine Cohort,France, 2002–2007

BackgroundCMV reactivation, which enhances immune senescence, could be associated with a higher risk of cancer.ObjectivesWe compared the prevalence of positive CMV DNAemia in HIV-infected patients with and without cancer.Study designThis case–control study, nested in the ANRS-CO3 Aquitaine Cohort, included patients with a first diagnosis of cancer (2002–2007) as cases. Two controls were matched per case.Cancer risk was estimated using conditional logistic regression models, an Odds Ratio (OR) of 2 could be detected with 80% power. The variables considered were: ≥1 positive CMV DNAemia, CD4+ and CD8+ counts, HIV plasma load. Plasma CMV DNA was retrospectively quantified within the 3-year period preceding the endpoint.ResultsThe 143 cases (93 non-AIDS-related and 50 AIDS-related cancers) and 284 controls had a median age of 47 years (IQR: 41–56). At the time of diagnosis or censorship, for cases and controls, median values were respectively, for CD4+ count: 327 cells/mm³ (IQR: 164–514) and 416 (IQR: 275–582), and for HIV plasma load: 2.6 log₁₀ copies/mL (IQR: 1.7–4.7) and 1.7 log₁₀ copies/mL (IQR: 1.7–3.3). We performed 2056 CMV PCR; 14 cases (9.8% [95% CI: 4.9–14.7]) and 19 controls (6.7% [CI: 3.8–9.6]) presented ≥1 positive PCR. CMV DNAemia was not associated with the risk of cancer (unadjusted and adjusted p-values = 0.19 and 0.54, respectively). HIV load >500 copies/mL was independently associated with a higher risk of cancer (OR = 2.02; p = 0.002; 95% CI: 1.29–3.17).ConclusionThis large case–control study did not show any differential exposure to positive CMV plasma DNAemia between cancer cases and controls.     

The role of ficolins and MASPs in hereditary angioedema due to C1-inhibitor deficiency

Background and ObjectiveHereditary angioedema due to C1-inhibitor deficiency (HAE-C1-INH) causes disturbances in the complement system. However, the influence of HAE-C1-INH on the lectin pathway of complement is unresolved. Thus, we studied the main initiator molecules, enzymes and regulators in the lectin pathway in patients with HAE-C1-INH.MethodsThe serum concentrations of ficolin-2, ficolin-3, MBL, MASP-2, MASP-3, and MAP-1 were measured during symptom-free periods in 91 patients with HAE-C1-INH, and in 100 healthy controls using sandwich ELISAs.ResultsCompared with controls, the levels of ficolin-2 (p < 0.0001) and MASP-2 (p = 0.0238) were reduced, while the levels of MBL and MASP-3 were elevated (p = 0.0028 and p < 0.0001, respectively) in HAE-C1-INH patients. Ficolin-3 and MAP-1 levels did not differ significantly between the two groups. Ficolin-2 correlated with MASP-3 in patients (r = 0.3443, p = 0.0008), while these parameters showed an opposite relationship in controls (r = −0.4625, p < 0.0001). In the patients, ficolin-3 correlated with MASP-2 (r = 0.3698, p = 0.001). Ficolin-2, -3, and MAP-1 correlated negatively with the annual requirement of plasma derived C1-INH concentrate (r = −0.2863, p = 0.0059; r = −0.2654, p = 0.0110 and r = −0.2501, p = 0.0168, respectively). Ficolin-3 showed a negative correlation with the annual number of attacks (r = −0.2478, p = 0.0179).ConclusionsWe found significant differences between patients and controls in the levels of some of the molecules belonging to the lectin complement pathway. Low concentrations of particularly ficolin-2 and -3 were inversely correlated with the severity of HAE-C1-INH, while this was not observed for MBL. This suggests a previously unrecognized involvement of the ficolin-dependent lectin complement pathway in the pathophysiology of HAE-C1-INH.     

High levels of plasma interferon gamma and +874T/A gene polymorphism is associated with HIV-TB co-infection

ObjectiveTuberculosis (TB) is one of the most frequent opportunistic infections in HIV patients leading to increased morbidity and death rate. This study was carried out to investigate the role of the cytokines IFN-γ and TNF-α level and their single nucleotide polymorphisms (SNPs) in HIV-TB co-infection.Methods247 HIV-TB (124 HIV-pulmonary TB, 123 HIV-extra pulmonary TB), 126 HIV positive individuals without tuberculosis and 129 healthy subjects (HS) were included to measure plasma levels of IFN-γ and TNF-α by sandwich ELISA and One way ANOVA statistical analysis was carried out among the groups. The SNPs of TNF-α-308 G/A, -238 G/A and IFN-γ + 874 T/A were also investigated using amplification refractory mutation system polymerase chain reaction (ARMS-PCR). The frequencies between the groups were compared by Pearson’s chi square statistical analysis.ResultsPlasma IFN-γ and TNF-α were significantly elevated in HIV-TB and TB (p < 0.05) as compared to those in HS group. There was significant association between IFN-γ + 874 ‘A’ allele and AA genotype in HIV-TB groups compared to HS and HIV (p < 0.05) and no such association was found for TNF-α-308 and -238. The plasma cytokine levels of TNF-α and IFN-γ reveals no significant association with levels of IFN-γ + 874 T/A, TNF-α -308 G/Aand-238 G/A genotypes in any of the study groups.ConclusionIn conclusion, the present study revealed elevated plasma IFN-γ and its +874 ‘A’ allele are associated with HIV-TB co-infection indicating 1.6 times increased risk for TB susceptibility. Elevated TNF-α levels in TB and HIV-TB suggest its involvement in TB pathogenesis.     

The MHC2TA 1614 C&gt;G gene polymorphism is associated with risk of developing acute coronary syndrome

BackgroundInflammation plays an essential role in the development and progression of atherosclerotic lesions. The major histocompatibility complex class II trans-activator (MHC2TA) is considered an important molecule in the inflammatory process regulation. The aim of the present study was to evaluate the role of MHC2TA gene polymorphisms as susceptibility markers for acute coronary syndrome (ACS).MethodsThree polymorphisms (−168 A>G, 1614 C>G, and 2536 G>A) of the MHC2TA gene were analyzed by 5′ exonuclease TaqMan genotyping assays in a group of 297 patients with ACS and 283 healthy controls. Haplotypes were constructed after linkage disequilibrium analysis.ResultsThe 1614 C allele and CC genotype were associated with risk of developing ACS (PC = 0.014, OR = 1.37 and PC = 0.006, OR = 1.90, respectively). Based on Hosmer–Lemeshow Goodness of Fit test, the recessive model was selected to estimate risk between ACS patients and controls adjusted by cardiovascular risk factors using a multiple logistic analysis. In this case, the OR adjusted was 1.78 for the 1614 CC genotype (P = 0.023). The analysis of linkage disequilibrium showed one risk haplotype (ACG) and one protective haplotype (AGG) for developing ACS (P = 0.02, OR = 1.5 and P = 0.04, OR = 0.72, respectively).ConclusionThe results suggest that MHC2TA 1614 gene polymorphism could be involved in the risk of developing ACS.     

ESI-MS/MS measurement of free carnitine and its precursor γ-butyrobetaine in plasma and dried blood spots from patients with organic acidurias and fatty acid oxidation disorders

BackgroundIn patients with fatty acid oxidation disorders (FAODs) and organic acidurias (OAs) “secondary carnitine deficiency” occurs. In OAs carnitine supplementation is widely performed and dose is often adjusted to blood-free carnitine levels. Dried blood spots (DBS) are mostly used to measure carnitine status, however measurements in plasma are discussed to be more accurate. The concentration and the predictive value of the carnitine precursor γ-butyrobetaine in blood during carnitine deficiency are unknown.MethodsFree carnitine and γ-butyrobetaine were quantified by tandem mass spectrometry in plasma and DBS from supplemented patients with OAs (n = 18) and unsupplemented patients with FAODs (n = 66) and were compared with healthy controls (n = 50).ResultsCarnitine concentrations in plasma were significantly higher than in DBS. In contrast, γ-butyrobetaine concentrations in plasma were significantly lower than in DBS. Supplemented patients had high free carnitine concentrations in combination with high γ-butyrobetaine concentrations. Unsupplemented carnitine palmitoyltransferase I-deficient patients had exceptionally high free carnitine concentrations without elevated γ-butyrobetaine, however, carnitine in plasma was much lower than in DBS. In patients with low carnitine, γ-butyrobetaine in plasma is no evidence of induced carnitine biosynthesis.ConclusionsParallel measurements in plasma and DBS demonstrated that numerous patients with low values in DBS had normal values when measured in plasma, suggesting plasma to be the more appropriate medium to use for carnitine status monitoring. In contrast, diagnosis of CPT-I deficiency may be missed when analysis is performed in plasma. Carnitine supplementation presumably inhibits γ-butyrobetaine dioxygenase and results in high γ-butyrobetaine.     

Comparison of graft healing in anterior cruciate ligament reconstruction with and without a preserved remnant in rabbits

BackgroundThe remnant of the native anterior cruciate ligament (ACL) might contribute to the biological integration of the graft in ACL reconstruction. The aim of this study was to explore whether the preserved remnant enhanced graft healing in ACL reconstruction.MethodsForty New Zealand rabbits underwent bilateral anterior cruciate ligament reconstructions. One knee was treated with a 2-mm remnant preserved on the tibial side (remnant-preservation, RP group) while the contralateral knee underwent a complete removal of the remnants by cauterization (remnant-resection, RR group) in each animal. Gross observations combined with microangiography, histological evaluation, and uniaxial load testing were performed after 4, 8, and 12 weeks.ResultsThe vascular density on the graft surface was statistically higher in the RP group as compared to that of the RR group at 4 (P = 0.002) and 8 weeks (P = 0.020). Additionally, the accelerated intra-articular and intra-tunnel graft integration were histologically observed in the RP group. Histological scores in the RP group were statistically higher than the RR group at 4 weeks (P = 0.028 for the intra-articular healing and P = 0.046 for the intra-tunnel healing) and 8 weeks (P = 0.031 for the intra-articular healing and P = 0.014 for the intra-tunnel healing). The ultimate failure load (P = 0.017), yield load (P = 0.025), and stiffness (P = 0.004) were statistically higher in the RP group as compared to those of the RR group, with corresponding significant differences in the failure mode (P = 0.020) between the two groups at 8 weeks.ConclusionsThe preserved remnant enhanced ACL graft healing with improved biomechanical properties in the rabbit model.Level of evidenceLevel II.     

Determinants of vitamin D status in healthy men and women aged 40–80 years

ObjectivesTo determine the contribution of life style and health related factors on vitamin D status in middle-aged and older men and women.Study designA cross-sectional single-center study in 400 male subjects (40–80 years) and 402 postmenopausal female subjects (56–73 years), conducted in a University Medical Center in the central part of the Netherlands (52 degrees northern latitude).Main outcome measuresMedical history, vitamin D, calcium and alcohol intake, physical activity, Body Mass Index, Blood pressure, smoking, total fat body mass and total lean body mass were measured using DEXA. Laboratory analysis included 25-hydroxyvitamin D (25OHD) and sex hormones.ResultsThirty-six percent of men and 51% of women had 25OHD less than 50 nmol/L. In summertime men had significant higher 25OHD as compared to women (81.5 vs 53.3 nmol/L, P = .000) but this difference disappeared come winter. In a saturated model, male gender (B = .16, P = .008), and season (summer vs winter B = .30, P = .000) remained statistically significant. In men, physical activity and season explained 21% of the variance. In women, household physical activity (B = .13, P = .03), sport physical activity (B = .02, P = .02) and estradiol (B = ?.003, P = .048) remained in the model,.ConclusionIn healthy middle-aged and older men and postmenopausal women, male gender and season were important predictors of vitamin D status. In men, physically activity and season, explained 21% of the variance in vitamin D status. In women, physical activity and estradiol explained 9.3% of the variance in vitamin D.