Residual Blood Samples Research Articles

Analytical performance evaluation of the Elecsys® Cyclosporine and Elecsys® Tacrolimus assays on the cobas e411 analyzer

BackgroundCyclosporine (CsA) and tacrolimus (TAC) are immunosuppressant drugs that are often used to treat autoimmune diseases and as transplantation therapy; therefore, their concentrations need to be monitored carefully. We herein evaluated the analytical performance of the Elecsys® Cyclosporine and Elecsys® Tacrolimus assay kits, which have been newly developed to measure CsA and TAC concentrations in the whole blood. MethodsWe used residual whole blood samples from autoimmune disease and transplantation patients who were being treated with CsA or TAC. CsA concentrations were measured using an affinity chrome-mediated immunoassay (ACMIA) and an electrochemiluminescence immunoassay (ECLIA). TAC concentrations were measured using a chemiluminescence immunoassay (CLIA) and ECLIA. We investigated assay precision, linearity, lower limit of quantitation (LOQ), stability of calibration, influence of interference substances and the hematocrit, correlation of ACMIA with ECLIA, and correlation of CLIA with ECLIA. ResultsWithin-assay coefficients of variation were 1.8−3.6% (CsA: 94−1238 ng/mL) and 2.9−3.9% (TAC: 2.1−17.8 ng/mL), whereas day-to-day coefficients of variation ranged between 3.0−4.1% (CsA) and 2.8−3.9% (TAC). The limits of quantitation were defined as the concentration at which the CV was approximately 10%. Each lower LOQ obtained was 16 ng/mL (CsA), and 0.95 ng/mL (TAC). CsA and TAC calibrations were stable for at least 21 days. Neither the presence of conjugated bilirubin, unconjugated bilirubin, chyle, and rheumatoid factor nor the hematocrit affected these assays. A method comparison using a standardized major axis regression analysis of ACMIA and ECLIA was r=0.995, y=0.924x −1.175, n=200 (CsA), while that of CLIA and ECLIA was r=0.994, y=1.080x −0.197, n=200 (TAC). ConclusionsThe analytical performances of the Elecsys® Cyclosporine and Elecsys®Tacrolimus assays were acceptable. Furthermore, CyA and TAC concentrations may be simultaneously measured using a single pretreatment which is of benefit if patients have to undertake conversion between these two drugs. Additionally, it benefits the workflow in the clinical laboratory. Thus, the Elecsys® Cyclosporine and Elecsys® Tacrolimus assays may be suitable for routine therapeutic drug monitoring.

Could daratumumab be used to treat severe allergy?

In the management of allergic diseases, targeting IgE using biologicals is a current research focus. This proof of concept shows that plasma cell depletion using daratumumab results in decreased total and allergen-specific IgE levels.

Vzhe-039, a Novel Structurally-Enhanced Allosteric Hemoglobin Effector Inhibits Sickling of SS Erythrocytes In Vitro, and Exhibits Improved Pharmacologic Properties In Vivo

Allosteric effectors of hemoglobin (AEH) remain promising as a viable therapeutic approach for the management of sickle cell disease (SCD). AEH bind to hemoglobin (Hb) in a transiently covalently manner, increasing Hb affinity for oxygen (O2), with concomitant inhibition of polymerization of deoxygenated sickle Hb (HbS) and erythrocyte sickling. As part of our ongoing efforts to surmount well-known AEH druggability challenges, we have designed and synthesized VZHE-039, a novel substituted benzaldehyde with potent in-vitro anti-polymerization activity, as well as more favorable in-vivopharmacokinetic and pharmacodynamic (PK/PD) properties. Here, we report the results of our early findings.The anti-sickling properties of VZHE-039 were tested in-vitro by incubating 0.5, 1, and 2 mM concentrations with blood suspensions from a subject with homozygous SCD (hematocrit: 20%) under hypoxic conditions (4% O2/96% N2) at 37°C for 2 h. At conclusion, aliquots were drawn into a fixative (2% glutaraldehyde solution), and sickling was assessed by microscopy. Aliquot samples were also subjected to cation-exchange HPLC analyses to measure the degree of Hb modification (Hb adduct formation), as well as standard O2 equilibrium curves (OEC) to assess p50 shifts. Subsequently, we conducted in-vivo PK/PD studies in wild-type mice that received single doses of VZHE-039 via the intravenous (I.V.: 25 and 50 mg/kg), intraperitoneal (I.P.: 100-150 mg/kg) and oral (P.O.: 100-200 mg/kg) routes. Serial blood samples were collected up to 24 h after I.V. and P.O., and up to 6 h after I.P. administration, respectively; aliquots were hemolyzed, de-proteinized and subjected to reverse-phase HPLC-UV assay to quantify VZH-039 blood concentrations. Residual blood samples were also hemolyzed, and clarified lysates were assayed for in-vivoHb adduct formation, and the corresponding change in Hb oxygen affinity (Δp50, %).The results of our in-vitrostudies demonstrated a concentration-dependent inhibition of SS cell sickling: 39±1.4%, 68.6±4.9%, and 89.9±5.2%, at 0.5, 1 and 2 mM of VZHE-039, respectively. HbS was modified correspondingly (46±7.4%, 83.6±6.4%, and 96.4±6.2%), and this adduct formation was correlated linearly with the left shift in OEC (Δp50 values of 34.3±8.7%, 63.6±2.4%, and 76.6±2.5%).Noncompartmental PK analysis showed that, after I.V. administration, systemic PK was dose-independent with a CLtot of 0.5 mL/min/kg, resulting in a terminal half-life of 9 h. After I.P. administration, AUCtrap increased supra-proportionally with dose; the limited sampling schedule suggested an I.P. bioavailability of >20%. After P.O. administration, AUCtrap increased supra-proportionally with dose as well due to an increase in oral bioavailability to ~ 10%, with peak blood concentrations up to ~ 0.25 mM; the terminal half-lives were prolonged (relative to I.V.) to 14 h. The PD effect-time course of change in %Hb adduct paralleled (and was reasonably linearly related to) blood concentrations for all routes. Baseline-corrected peak PD effects after I.V. administration were 15.7% and 31.9%, after 25 and 50 mg/kg respectively; after P.O. administration of 200 mg/kg, the peak change was 17.6%. Overall, VZHE-039 exhibited a low (metabolic) clearance, presumably reflecting high-affinity and sustained Hb (and possibly albumin) binding, leading to the observed extended terminal half-life and duration of action. While its oral bioavailability is low (likely due to limited GI solubility) and there is evidence of saturable first-pass effects and possibly enterohepatic recycling, oral in-vivo exposures were associated with PD effects that were consistent with the results from the in-vitrostudies.In conclusion, the results of our current studies establish VZHE-039 as a novel, potent anti-sickling agent, confirm its proposed mechanism of action in-vitro and in-vivo, and suggest that therapeutically-relevant blood concentrations may be achieved after daily oral doses. Therefore, VZHE-039 is our new lead drug candidate that may require further modifications and studies to (a) improve its GI solubility and oral bioavailability; (b) elucidate atomic level structural interactions and kinetics of its Hb binding; and (c) formally investigate its biological activity after repeat-doses in a SCD mouse model. DisclosuresNo relevant conflicts of interest to declare.

Retinol binding protein and vitamin D associations with serum antibody isotypes, serum influenza virus-specific neutralizing activities and airway cytokine profiles.

Vitamin A supports the induction of immunoglobulin (Ig)A responses at mucosal surfaces in mice, but much less is known about the influence of vitamins on antibody isotype expression in humans. To address this knowledge gap, we examined 46 residual blood samples from adults and children, some of whom were experiencing influenza virus infections of the respiratory tract. Assays were performed for retinol binding protein (RBP, a surrogate for vitamin A), vitamin D (a related vitamin) and antibody isotypes. Results showed that all but two tested samples exhibited RBP and/or vitamin D insufficiencies or deficiencies. Vitamin D correlated with blood IgM and IgG3, while RBP correlated with IgG4 and IgA. RBP also correlated positively with age and with influenza virus-specific antibody neutralization titres. Individuals with low blood RBP levels exhibited the highest frequencies of over-expressed cytokines and growth factors in nasal wash samples, an indication of inflamed mucosal tissues. While cause-effect relationships were not discerned, results support a hypothesis that vitamins directly influence B cell isotype expression in humans, and by so doing may help protect mucosal surfaces from respiratory viral disease.

Prevalence and knowledge of hepatitis C in a middle-aged population, Dunedin, New Zealand.

To determine the prevalence of infection with hepatitis C virus (HCV) in those most at risk of advanced liver disease and to identify gaps in knowledge of HCV. Questionnaires were mailed to randomly selected residents aged 40-59 to assess the extent of their general knowledge about HCV. The questionnaire assessed demographics, the extent of general knowledge about viral hepatitis, potential risks for infection and the prevalence of risk factors associated with increased progression of liver fibrosis. Anonymised residual laboratory blood samples from 40-59 years old people from Dunedin taken in hospital or in the community, were tested for HCV antibodies and alanine transaminase (ALT), aspartate transaminase (AST), gamma-glutamyl transpeptidase (GGT). Linear regression was performed to examine whether the demographics sex, age, socio-economic status, qualification level and occupation sector (categorical variables) were predictors of level of general knowledge about hepatitis. For the demographics that were found to be significant predictors of score outcome, multiple regression analysis was used to determine independent effects. χ (2) tests were used to compare our selected sample and our responder population demographics, to the demographics of the entire 40-59 years old population in Dunedin using the 2006 NZ census data. Exact confidence intervals for the proportion positive for HCV and HBV were calculated using the binomial distribution. The response rate to the mailed questionnaire was 431/1400 (30.8%). On average 59.4% questions were answered correctly. Predictors for higher scores, indicating greater knowledge about symptoms and transmission included sex (female, P < 0.01), higher level of qualification (P < 0.000) and occupation sector (P < 0.000). Sharing intravenous drug utensils was a known risk factor for disease transmission (94.4%), but the sharing of common household items such as a toothbrush was not. 93% of the population were unaware that HCV infection can be asymptomatic. 25% did not know that treatment was available in New Zealand and of those who did know, only 40% assumed it was funded. Six hundred and eighty-two residual anonymised blood samples were tested for HCV antibodies, ALT, AST and GGT. The prevalence for HCV was 4.01%, 95%CI: 2.6%-5.8%. Liver function tests were not useful for identifying likelyhood of HCV infection. Prevalence of HCV in our population is high, and the majority have limited knowledge of HCV and its treatment.

In utero Exposure to Cigarette Smoking, Environmental Tobacco Smoke and Reproductive Hormones in US Girls Approaching Puberty

Background/Aims: Evidence is unclear whether prenatal smoking affects age at menarche and pubertal development, and its impact upon hormones has not been well studied. We aim to identify potential pathways through which prenatal smoking and environmental tobacco smoke (ETS) affect reproductive hormones in girls approaching puberty. Methods: We examined the association between prenatal smoking, current ETS and luteinizing hormone (LH) and inhibin B (InB) in 6- to 11-year-old girls in the 3rd National Health and Nutrition Examination Survey, 1988-1994. Parents/guardians completed interviewer-assisted questionnaires on health and demographics at the time of physical examination. Residual blood samples were analyzed for reproductive hormones in 2008. Results: Of 660 girls, 19 and 39% were exposed to prenatal smoke and current ETS, respectively. Accounting for multiple pathways in structural equation models, prenatally exposed girls had significantly lower LH (β = -0.205 log-mIU/ml, p < 0.0001) and InB (β = -0.162, log-pg/ml, p < 0.0001). Prenatal smoking also influenced LH positively and InB negatively indirectly through BMI-for-age. ETS was positively associated with LH, but not with InB. Conclusion: Exposure to maternal smoking may disrupt reproductive development manifesting in altered hormone levels near puberty.

Measles vaccine coverage estimates in an outbreak three years after the nation-wide campaign in China: implications for measles elimination, 2013.

BackgroundChina is approaching measles elimination, but indigenous measles still circulates. County L in China has reported measles-containing vaccine (MCV) coverage rates >95% since 2000. Despite high reported coverage, a large measles outbreak occurred among young children in L County. We measured MCV coverage using 5 different methods during an investigation on this outbreak and compared our estimates with reported rates.MethodsReported coverage rates are determined by aggregating clinic-based data across the county: doses administered in each clinic divided by the number of children registered in each clinic. Our methods estimated coverage for the 2010–2012 birth cohort, and were (1) administrative method: doses administered in clinics divided by the birth cohort recorded in the Statistical Year Book, (2) house-to-house convenience-sample survey of children living near cases, (3) vaccination clinic records review, (4) determination of a convenience sample of measles outbreak cases’ vaccination statuses and using the field vaccine efficacy outbreak equation to estimate population coverage, and (5) a seroprevalence survey using a convenience sample of residual blood samples from hospitals.ResultsThe measles outbreak totaled 215 cases, representing an incidence of 195.8 per million population. Our estimated MCV coverage rates were: (1) administrative method: 84.1%-87.0% for MCV1 and 80.3%-90.0% for MCV2, (2) in-house survey: 83.3% of 9–17 month children received MCV1, and 74.5% of 24–47 month children received MCV2, (3) clinic record review: 85.5% of 9–17 month children received MCV1, and 73.2% of 24–59 month children received MCV2, (4) field VE method: 83.6% of 9–47 month children received one or more MCV doses, and (5) serology: seropositive rates were <80% in the 12–17 and 18–23 month age cohorts.ConclusionsCompared with reported coverage >95%, our 5 coverage assessments all showed substantially lower coverage. China should evaluate guidelines for reporting vaccination coverage and identify feasible improvements to the assessment methods.

Validation of a New (1 → 3)-β-D-Glucan Test Kit, Fungitec G Test MKⅡ " Nissui "

Using 415 residual blood samples subjected to (1→3) -β-D-glucan assay, we studied the correlation of measurement results between Fungitec G Test MKII "Nissui" (Nissui Pharmaceutical Co., Ltd., Tokyo) and its predecessor, Fungitec G Test MK (Seikagaku Corporation, Tokyo), which is now out of production. Their major difference is that MK II uses reagents derived from blood cells of Limulus polyphemus, the American horseshoe crab, whereas MK used those of Tachypleus tridentatus, an Asian horseshoe crab. Passing-Bablok analysis showed a linear regression with nearly one-to-one correspondence (slope=1.065, intercept=-0.287) between the two test kits over the regular range of measurements (4.0 pg/ml -500 pg/ml ). This was also true when data were subdivided and analyzed in the low to medium (≦150 pg/ml ) and in the low range (≦50 pg/ml ). There were several cases, however, that showed a wide discrepancy between the pair of measurements. This discrepancy is believed to be due in part to the difference between Limulus and Tachypleus in their reactivity to β-glucans with diverse side chains. Despite of this, the even distribution on either side of the regression line of those samples that are associated with deep fungal infection and the abrupt disappearance of such samples below 20 pg/ml attest that MK II "Nissui" is an acceptable successor of MK.

Toward Optimal Selection of Cellular Products for Hematology/Oncology Patients Eliminating Adverse Effects

Context: To provide critical cellular support to hem/onc patients undergoing chemotherapy, US hospitals consumed w675,000 single donor platelets (34.4% of the 2010 total transfused) and w2,100,000 red cells (15.1% of the 2010 total transfused). In the emerging paradigm of genomics and personalized care, the selection of cellular products for the treatment of oncology patients in accordance with their immunomolecular signature has the potential to reduce antibody-mediated destruction of cells and improve treatment outcomes. Turning this strategy into a logistical reality calls for three ingredients: the identification of patients at high risk of mounting an adverse immune reaction; the clinical decision to provide optimal cellular products; and real-time access to an adequate supply of such products. Objective: To clinically validate a novel process of classifying hematology patients and candidate donors on the basis of an immunomolecular attribute profile comprising platelet (HLA class I and HPA) as well as red cell antigen profiles for a total of w100markers (and the potential of scaling this process so as to permit the patient profiling and classification of 1,000 candidate donors per 24hr period). Design: Residual blood samples from MDACC blood bank were collected and analyzed by our novel process combining: the parallel sequencing for pools of samples at informative sites; haplotype recovery; antigen and epitope profile prediction; and classification by “machine learning”. Results: HLA class I and HPA profiles of samples from 296 hematologic and non-hematologic oncology patients, and from 160 candidate platelet donors were determined, and classified by HLA class I and HPA profiles; the process achieved 100% concordance with high-resolution HLA types available for 42 patients; in addition, we demonstrated the direct recovery of haplotype pairs and extended the method to red cell antigens including RH variants. Conclusions: Our new “virtual cell sorting” process offers an effective approach to the large-scale classification of patients and candidate donors, and optimal selection of cells, on the basis of immunomolecular signatures.

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Introduction: The use of biomarkers to distinguish bacterial infection from non-infectious processes has great potential as a means for an earlier and more appropriate treatment strategy, although currently used markers have shown inconsistent results. In addition to procalcitonin (PCT), interleukin 27 (IL-27), a heterodimeric cytokine produced by antigen presenting cells, has recently exhibited early promise in a preliminary study, demonstrating high specificity and positive predictive values in bacterial infection. Methods: The acquisition of a blood culture at any point during admission by the primary PICU team was used to define "clinical suspicion of infection" and trigger the procurement of residual blood samples. These samples (N=63), acquired with a waiver of informed consent following IRB approval, were then used for both IL-27 and procalcitonin (PCT) biomarker assay testing. Further analysis of biomarker levels was then performed with respect to a definition of bacterial infection reached by a blinded intensivist chart review and based upon all gathered microbiologic and clinical data surrounding the time of enrollment. Formal performance comparisons included calculations of ROC curves for IL-27 and PCT individually in addition to a combination strategy utilizing an optimal decision tree generated by CART analysis. Further test characteristics (sensitivity, specificity, etc.) were then generated based upon optimal cut-off points. Results: Ages of included patients ranged from 3 months to 34 years with mean = 9.4 yrs, median = 9.0 yrs, and IQR = 3 to 14 yrs. Of the 63 patients, 19/63 were diagnosed with bacterial infection and were 44/63 labeled uninfected. ROC curve analysis for IL-27 and PCT yielded AUCs of 0.59 (0.42 to 0.75) and 0.71 (0.53 to 0.90) respectively. The CART-derived decision tree incorporating both IL-27 and PCT yielded an AUC of 0.79 (0.66 to 0.92), specificity of 86% (71%, 94%), sensitivity of 68% (43%, 86%), PPV of 68% (43%, 86%) and NPV of 86% (71%, 94%) . Conclusions: With a planned enrollment of 700 patients, these preliminary data and analyses are part of a larger study to investigate the value of both IL-27 and a combination strategy in this clinical context. Existing data suggest however that the ability to predict bacterial infection is enhanced when using both PCT and IL-27 in combination rather than either biomarker alone.

Effect of Phenylbutazone, Flunixin Meglumine and Firocoxib on ex vivo Cyclo‐Oxygenase Activity in Horses Undergoing Elective Surgery

AimsNonsteroidal anti‐inflammatory drugs (NSAIDs) inhibit the production of prostaglandins and other inflammatory mediators by inhibiting the activity of the cyclo‐oxygenase enzymes (COX). Two major isoforms of COX enzymes exist: COX‐2, which is expressed during the inflammatory response, and COX‐1, which is responsible for the physiological production of prostaglandin that regulates tissue homeostasis. The study aims to evaluate the effect of firocoxib ex vivo in the horse as, to the authors' knowledge, published studies assess its effect only in vitro.MethodsHorses (n = 18) undergoing elective surgery were recruited and allocated to treatment groups depending on clinician preference (1) phenylbutazone (4.4 mg/kg bwt i.v. b.i.d.), (2) flunixin meglumine (FM, 1.1 mg/kg bwt i.v. b.i.d.) and (3) firocoxib (FIR, 0.1 mg/kg bwt i.v. s.i.d.). Residual blood samples were collected prior to NSAIDs (T0), 2 h after NSAIDs (T2), and 24 h following surgery (T24). The COX activity was measured using validated immune‐enzymatic assays. A Kruskall–Wallis test was used to determine the effect of time and treatment on COX‐1 and COX‐2 activity. Bonferroni corrections were used to identify the level of significance accounting for multiple comparisons (P&lt;0.017).ResultsAt T2 and T24, the relative COX‐1 activity was significantly greater in horses receiving firocoxib compared with horses receiving either phenylbutazone (P&lt;0.008) or flunixin meglumine (P&lt;0.005). At T2 and T24, COX‐1 activity was reduced (compared with baseline) in horses receiving phenylbutazone or flunixin meglumine. The effect on COX‐2 activity was not significantly different between drugs (P = 0.471).Conclusions and practical significanceCyclo‐oxygenase selectivity of firocoxib is demonstrated ex vivo. Firocoxib is as effective as phenylbutazone or flunixin meglumine in modulating the production of prostaglandins by COX‐2 isoenzyme, whilst the physiological action of COX‐1 isoenzyme is preserved with firocoxib, but not with phenylbutazone and flunixin ex vivo.Ethical animal researchStudy approved by Ethics and Welfare Committee ‐ School of Veterinary Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow. Sources of funding: Funding provided by John Crawford Endowment Fund, and Mrs I.J. Gates Charity Fund, University of Glasgow. Competing interests: None.

Waiver of consent in noninterventional, observational emergency research: the PROMMTT experience.

In the PRospective Observational Multicenter Major Trauma Transfusion (PROMMTT) study, waiver of consent was used because previous literature reported low response rates and subsequent bias. The goal of this article was to examine the rationale and tradeoffs of using waiver of consent in PROMMTT. PROMMTT enrolled trauma patients receiving at least 1 U of red blood cells within 6 hours after admission at 10 US Level 1 trauma centers. Local institutional review boards (IRBs) from all sites approved the study. Site 8 was required by their IRB to attempt consent but was allowed to retain data on patients unable to be consented. Of 121 subjects enrolled at Site 8, 55 consents were obtained (46%), and no patient or legally authorized representative refused to give consent. Of the patients, 36 (30%) died, and 6 (5%) were discharged before consent could be attempted. Consent was attempted but not possible among 24 patients (20%). Of the 10 clinical sites, 6 of the local IRBs approved collection of residual blood samples, 1 had previous approval to collect timed blood samples under a separate protocol, and 3 reported that their local IRBs would not approve collection of residual blood under a waiver of consent. Waiver of consent was used in PROMMTT because of the potential adverse impact of consent refusals; however, there were no refusals. If the IRB for Site 8 had required withdrawal of patients unable to consent and destruction of their data, a serious bias would likely have been introduced. Other tradeoffs included a reduction in sites participating in residual blood collection and a smaller than expected amount of residual blood collected among sites operating under a waiver of consent. Noninterventional emergency research studies should consider these potential tradeoffs carefully before deciding whether waiver of consent would best achieve the goals of a study.

‘Once the stuff's left my body, it's not me’: service users' views on unlinked anonymous testing of blood for HIV

This paper reports on the qualitative component of a mixed-methods study on unlinked anonymous testing for HIV in genitourinary medicine (GUM) clinics in two English cities. Unlinked anonymous testing is a system of monitoring population prevalence by testing residual blood samples taken for diagnostic purposes after they have been unlinked and anonymised from their source. Little is known about how individuals feel about their blood being tested in this way without their explicit consent, nor is it clear whether the process of unlinking blood affects how people feel about the use of their bodily material for public health surveillance purposes. We report participants' views on these issues, drawing on in-depth interviews with 20 GUM clinic users. The majority thought it preferable for blood samples to be used for population surveillance rather than being discarded. For most, blood and bodily tissue were not seen to represent personal identity even though participants understood that information about them could be gleaned from their analysis. The provision of information, rather than a strict consent process, was advocated as many felt that transactions between health professionals and patients should be as transparent as possible.

Policies for Handling Residual Newborn Blood Samples for Human Health Research

Newborn screening prevents thousands of premature illnesses and deaths every year and is one of the most successful public health programs in the US. Lately, there has been considerable controversy regarding the retention and use of residual newborn blood samples after their initial use in screening without obtaining parental informed consent. The concerns have focused on the confidentiality, privacy, and autonomy rights of parents to decide whether to permit the secondary use of these samples for research. This ongoing controversy regarding the preservation and use of dried blood samples in research has led to the destruction of several million archived samples and potentially more in the near future if no further actions are taken. Left out of the discussion, however, have been the benefits of making the residual blood …

Sero-epidemiology of hepatitis B markers in the population of Tuscany, Central Italy, 20 years after the implementation of universal vaccination

Italy was one of the first industrialized countries to introduce a program of universal vaccination against hepatitis B in 1991. Twenty years later we verified the impact of universal immunisation on the epidemiology of hepatitis B infection by analyzing the prevalence of specific viral markers (anti-HBs, anti-HBc and HBsAg). The ELISA tests were performed on residual blood samples collected by 0.05% of the resident population aged 1-50 years in Tuscany (Italy). About 63% of subjects aged < 30 years were anti-HBs positive compared to about 25% in older subjects, without differences between genders. About 22% of subjects over 40 years were anti-HBc-positive compared to 5% in the younger age groups. The number of HBsAg-positive subjects was almost 10 fold higher in the unvaccinated age groups than in the cohorts involved in the universal vaccination program. The results of our study show the persisting high anti-HBs reactivity in vaccinated cohorts, while HBV markers related to natural exposure or persistent infection remain remarkably higher in older age groups. This sero-epidemiological study supports with prevalence data the downward incidence trend of acute hepatitis B highlighted by epidemiological surveillance systems, and corroborates the forecast for elimination of hepatitis B in Italy in a few decades.

Seroepidemiology of Varicella-Zoster Virus in Korea

This study was designed to assess the immune status of the Korean population against varicella-zoster virus (VZV) through a seroepidemiologic study. Residual blood samples were collected from diagnostic laboratories throughout Korea. Samples were collected in October 2009 to March 2010 from persons 0-79 yr of age and were tested by ELISA (Enzygnost®; Dade Behring, Schwalbach, Germany). Total seroprevalence in subjects 1-79 yr of age was 89.6%. Seroprevalence increased as age increased from 67.3% in subjects 1-4 yr of age to 94.2% in subjects 10-14 yr of age and in subjects over 20 yr of age seroprevalence ranged from 98.0% to 100%. In children under 1 yr of age, passive immunity waned after birth with none of the subjects having antibodies from 7 months of age and over. Among subjects 1-79 yr of age, susceptible subjects to VZV were mainly under 20 yr of age. These results provide information in understanding the dynamics of varicella disease in Korea, which is important in building up strategies for disease control.

Polymorphic mononuclear neutrophils CD64 index for diagnosis of sepsis in postoperative surgical patients and critically ill patients

Surface neutrophil CD64 expression is upregulated in patients with bacterial infection. As it was suggested that the CD64 index could be used to detect sepsis in hospitalized patients, we questioned whether the CD64 index could discriminate between septic patients and postoperative surgical patients, defined as systemic inflammatory response syndrome (SIRS), both admitted at the intensive care unit (ICU). Furthermore, we wondered whether the CD64 index was an improved diagnostic compared to standard assays used at the laboratory. For this, outclinic (OC) patients were included as controls. The Leuko64™ assay was used to determine the CD64 index in residual EDTA blood samples from selected septic patients (n=25), SIRS patients (n=19), and OC patients (n=24). Additionally, WBC count, neutrophilic and eosinophilic granulocyte count, C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) were measured simultaneously. The CD64 index was higher in septic patients compared to both the SIRS and OC group (p<0.0001). In addition, the WBC count, neutrophil count, ESR and CRP were also higher in septic patients than the OC group (p<0.0001). However, only the WBC count, eosinopenia, and ESR were comparable between the SIRS and the sepsis group and proved to be discriminative to the OC group (p<0.05). The CD64 index demonstrated higher sensitivity and specificity than CRP, WBC count, neutrophilic and eosinophilic granulocyte count, and ESR. A high CD64 index was found in septic intensive care patients, while a low CD64 index was observed in OC and SIRS patients, demonstrating that the CD64 index can be used for routine diagnostics in the ICU setting.

Prevalence of canine haemotropic mycoplasma infections in Sydney, Australia

Haemotropic mycoplasmas (haemoplasmas) are epierythrocytic, wall-less bacteria that cannot be cultivated in vitro (Messick 2003). Two species have been identified in dogs: Mycoplasma haemocanis (Mhc) and ‘ Candidatus Mycoplasma haematoparvum’...

Lymphoma Vaccine Therapy: Next Steps After a Positive, Controlled Phase III Clinical Trial

Many of the efforts toward developing vaccines against human malignancies have been frustrated by the lack of identification of a tumor-specific antigen that would allow tumor cells to be distinguished from normal cells. Idiotypic determinants of the surface immunoglobulin (Ig) associated with a given patient's B-cell lymphoma are unique to that tumor, and can thus serve as a tumor-specific marker. When conjugated to the immune carrier keyhole limpet hemocyanin (KLH), vaccination with an idiotype protein vaccine has been able to improve length of freedom from disease relapse in patients with follicular lymphoma (FL) in a minimal residual disease (MRD) state after induction therapy, as demonstrated in a recent randomized, controlled phase III trial. In addition to predictive biomarker discovery, using residual autologous tumor and blood samples from patients vaccinated on the phase III trial, we have now developed a next generation idiotype DNA vaccine with the goal of reducing vaccine production time while maintaining efficacy. A first-in-human study is planned to evaluate its use in patients with asymptomatic phase lymphoplasmacytic lymphoma.

Assessing public attitudes on the retention and use of residual newborn screening blood samples: A focus group study

This paper discusses attitudes and opinions of a diverse group of participants toward the retention and use of residual newborn blood samples for research. Data were drawn from focus groups based in six states in the USA, and results provide support for the retention and use of residual newborn blood samples for research when parental permission is asked beforehand. However, there were a number of concerns that also warrant attention for the development of policy and maintaining trust with the public, such as timing of permission, use of samples already stored, level of personal control of sample use and education. The results demonstrate the complexity of the topic and the ethical ambiguities associated with the retention and use of residual newborn blood samples.