Reliability of a visual test for the rapid detection of mucopolysaccharidoses: GAG-test(®)

Mucopolysaccharidoses (MPS) are a group of lysosomal storage disorders, characterized by the deficiency/absence of one of the enzymes involved in the intralysosomal degradation of glycosaminoglycans (GAGs). The quantitative determination of urinary GAGs using dimethylmethylene blue (DMB) shows high reliability. However, the logistics and staff for this method are not always available in primary care centers. Sending urine samples to reference laboratories increases the cost and delays the diagnosis. Thus, the aim of this article is to develop and evaluate a simple and low‐cost visual test (GAG‐test^®) for the screening of urine samples from patients under suspicion of suffering from MPS. The purpose is to narrow down the number of samples to be assayed through the quantitative method. A measure of 50 µl urine was added to 2 ml DMB solution. A color change from dark blue to purple indicates an excess of GAGs. The quantitative analyses showed a significant difference between controls' and patients' concentrations (P<0.05). After optimization of the composition, positive and negative results obtained with the qualitative test were able to discriminate between normal urines and those from patients suffering from mucopolysaccharidosis. Therefore, GAG‐test^® has proved to be a useful tool for the prior diagnosis of patients suffering from mucopolysaccharidosis, reducing the number of individuals with whom investigations should be continued. J. Clin. Lab. Anal. 25:179–184, 2011. © 2011 Wiley‐Liss, Inc.     

Toxo Chek — a simple and rapid immunoassay for the detection of antibodies to Toxoplasma gondii

Toxo Chek (E-Y Labs Inc.) is a recently developed immunoassay using filtration blotting for the detection of IgG class antibodies specific for Toxoplasma gondii in human serum. The disposable filtration device and all reagents are supplied in kit form, no further equipment is needed. The assay was evaluated in comparison to the dye test, the ‘gold standard’, on 499 routinely submitted sera. A sensitivity of 97.5%, specificity of 88.5% with a positive predictive value (PPV) of 85.4% and a negative predictive value (NPV) of 98.1% were obtained on initial testing. After retesting sensitivity improved to 99%, specificity to 93%, with a PPV of 90% and NPV of 99.3%. The false positive sera gave a similar profile to the negative control on immunoblotting, no unique bands were identified. The assay is of potential value for rapid initial screening in less specialized centres, particularly in screening antenatal sera.     

microRNA and the pathogenesis of ovarian cancer--a new horizon for molecular diagnostics and treatment?

Ovarian cancer is the leading cause of death among gynecologic malignancies and despite advances in treatment, more than 50% of all patients will experience recurrence, resulting in worse overall prognosis. Therefore, identification of novel biomarkers for ovarian cancer is of significant interest. microRNA (miRNA) constitute a class of small gene regulatory RNA molecules (18-24 nt) and by sequence complementarity, they negatively regulate messenger RNA (mRNA) translation of target genes. Rising data are available that miRNA are functionally involved in the pathogenesis of ovarian cancer. In this regard, recent advances in profiling studies revealed a variety of miRNA candidates, differently expressed in ovarian carcinomas and in disease-specific conditions like hypoxia or chemo-resistance. This review abstracts recent efforts on establishing miRNA as novel molecular biomarkers for ovarian cancer and depicts the existing preliminary framework for defining peripheral-blood derived miRNA as novel circulating biomarkers. Beside these clinical implications, we highlight the current functional understanding of miRNA alteration and discuss major challenges in miRNA profiling approaches. Finally, we briefly outline methodologies to therapeutically modulate miRNA expression in cancer and try to assess how miRNA can improve our conceptual understanding and the clinical management of ovarian cancer.     

Recent advances in the understanding of infectious mononucleosis: are prospects improved for treatment or control?

Symptomatic primary Epstein–Barr virus infection is known more commonly as infectious mononucleosis, an illness known for afflicting adolescents and younger adults as a febrile illness accompanied by pharyngitis and lymphadenopathy. Historically believed to be generally benign, infectious mononucleosis has been linked more recently to increased risks of developing Hodgkin’s lymphoma and multiple sclerosis. Advances in the understanding of host immune responses to Epstein–Barr virus have begun to elucidate the reasons why younger children typically experience subclinical infection whereas older individuals develop infectious mononucleosis. This review will highlight recent advances in the understanding of primary Epstein–Barr virus infection, and whether prospective treatments or vaccine strategies may affect native infection and its sequelae.     

Recent advances in the understanding of infectious mononucleosis: are prospects improved for treatment or control?

Symptomatic primary Epstein-Barr virus infection is known more commonly as infectious mononucleosis, an illness known for afflicting adolescents and younger adults as a febrile illness accompanied by pharyngitis and lymphadenopathy. Historically believed to be generally benign, infectious mononucleosis has been linked more recently to increased risks of developing Hodgkin's lymphoma and multiple sclerosis. Advances in the understanding of host immune responses to Epstein-Barr virus have begun to elucidate the reasons why younger children typically experience subclinical infection whereas older individuals develop infectious mononucleosis. This review will highlight recent advances in the understanding of primary Epstein-Barr virus infection, and whether prospective treatments or vaccine strategies may affect native infection and its sequelae.     

β-blockers in the management of hypertension: focus on nebivolol

Hypertension is a major cardiovascular risk factor but most patients remain asymptomatic for many years. Successful therapy not only needs to be effective, it also needs to be well tolerated. β-blockers are well established as effective antihypertensive agents. However, one major drawback to the currently available β-blockers, particularly the noncardioselective β-blockers, is their side-effect profile, including sexual dysfunction, fatigue, depression and metabolic abnormalities such as impaired glucose tolerance and lipid abnormalities. Nebivolol (Bystolic^®), a novel, highly cardioselective, third-generation β-blocker that recently received approval by the US FDA for the treatment of hypertension in the USA, is effective in treating blood pressure and has a favorable side-effect profile. Studies conducted in Europe, where nebivolol has been available for some time for the treatment of hypertension, have shown that nebivolol achieves blood pressure reductions comparable to other β-blockers but with fewer side effects. Additionally, nebivolol has demonstrated similar efficacy in blood pressure reduction when compared with calcium channel blockers and inhibitors of the renin–angiotensin system. When combined with hydrochlorothiazide there was an additive antihypertensive effect. Lastly, nebivolol exhibits a vasodilatory property that is related to its effect on nitric oxide, an intrinsic vasodilator produced in the vascular endothelium. Nebivolol enhances nitric oxide bioavailablility. Studies have also demonstrated nebivolol’s ability to function as an antioxidant and decrease markers of oxidative stress. These effects are believed to ultimately produce a modulation of the endothelial dysfunction typically seen in hypertension.     

Colonization of Extended-Spectrum-β-Lactamase- and NDM-1-Producing Enterobacteriaceae among Pregnant Women in the Community in a Low-Income Country: a Potential Reservoir for Transmission of Multiresistant Enterobacteriaceae to Neonates

The spread of extended-spectrum-β-lactamase-producing Enterobacteriaceae (ESBL-PE) in low-income countries, where the burden of neonatal sepsis is high, may have a serious impact on neonatal mortality rates. Given the potential for mother-to-child transmission of multiresistant bacteria, this study investigated the ESBL-PE rectal colonization among pregnant women at delivery in the community in Madagascar and estimated a prevalence of 18.5% (95% confidence interval, 14.5% to 22.6%). One strain of Klebsiella pneumoniae isolated was also a New Delhi metallo-β-lactamase-1 (NDM-1) producer.     

Urine molecular profiling distinguishes health and disease: new methods in diagnostics? Focus on UPLC-MS

Urine may be a waste product, but it contains an enormous amount of information. Its role in diagnostics cannot be underestimated. The combination of high-end analytical technology with multivariate statistics allows differential analysis of replicate samples with applications for both high- and low-molecular-weight analytes, namely proteins and metabolites. Global urine profiles measured with NMR or mass spectrometry (MS)-based methods distinguish, for example, individuals, health status and hormonal changes. They do not necessarily discriminate between substance classes but investigate method-determined subgroups, such as all compounds separated by reversed-phase liquid chromatography. For diagnostics, the identification of those compounds is of secondary interest because the overall features of the profile itself are used for sample comparison. The potential of this simple approach for clinical diagnostics is huge, since only minimal urine preparation (e.g., centrifugation and filtration for liquid chromatography-MS) is necessary and the experimental execution using, for instance, technologies such as ultra-performance liquid chromatography coupled with high-end MS can be standardized. However, concerted collaborative efforts are required to generate comparable datasets and to create the profile database necessary for diagnostic applications.     

Rapid molecular detection of methicillin-resistant Staphylococcus aureus: a cost-effective tool for infection control in critical care?

Control strategies for methicillin-resistant Staphylococcus aureus (MRSA) in critical care remain debated. Timely detection of MRSA carriers is crucial to an effective isolation policy. In this issue, Harbarth and colleagues report rapid MRSA screening among intensive care unit-admitted patients using a PCR assay. Preemptive isolation for all admissions until screened negative for MRSA was associated with a reduction of intensive care unit-acquired MRSA infections in one of two study units. The data provide preliminary evidence to the effectiveness of a MRSA control strategy combining rapid screening by a molecular method and preventive isolation. Further controlled studies are needed to evaluate the cost-effectiveness of this intervention.     

Are maternal and sonographic factors associated with the detection of a fetal echogenic cardiac focus?

OBJECTIVE: To evaluate the influence of maternal weight and the orientation of the fetal 4-chamber heart view on the detection of a fetal echogenic cardiac focus. METHODS: In this nested case-control study, 103 women undergoing anatomic surveys at 15 to 22 weeks between January 1, 1997, and June 15, 1999, were identified as having an echogenic cardiac focus via our computerized database. A control group was selected from among the same group of patients. Data were collected from the sonography reports, prenatal records, and sonographic images of 4-chamber heart views; maternal characteristics and sonographic details were recorded, including the orientation of the 4-chamber view (apical, basilar, and right and left lateral). RESULTS: Gravidas in the echogenic cardiac focus group were more likely to be of lower weight (68.0 +/- 14.4 versus 72.9 +/- 18.3 kg; P = .04), of lower body mass index (25.5 +/- 5.3 versus 27.3 +/- 6.2 kg/m2; P = .03), of younger age (24.4 +/- 6.5 versus 26.9 +/- 6.9 years; P = .01), and African American or Asian (37.9% versus 27.2% and 9.7% versus 2%; P = .01). Cases were scanned at earlier gestational ages (18.9 +/- 1.6 versus 19.5 +/- 1.7 weeks; P = .01). The focus group was more likely to have had an apical view of the fetal heart taken (80.8% versus 51.4%; P = .0001). Controls were more likely to have had a right lateral view taken (44.6% versus 20.8%; P = .002). No significant difference was found between groups in terms of any other maternal or sonographic variable studied. CONCLUSIONS: The echogenic cardiac focus group was more likely to have a lower body mass index and to be scanned with the apical fetal heart view. The orientation of the fetal 4-chamber heart view exerted the most statistically significant influence on detection rates for the echogenic cardiac focus, implying that the more technically facile the sonographic study, the more likely an echogenic cardiac focus will be found.     

The cobas® 6800/8800 System: a new era of automation in molecular diagnostics

Introduction: Molecular diagnostics is a key component of laboratory medicine. Here, the authors review key triggers of ever-increasing automation in nucleic acid amplification testing (NAAT) with a focus on specific automated Polymerase Chain Reaction (PCR) testing and platforms such as the recently launched cobas® 6800 and cobas® 8800 Systems. The benefits of such automation for different stakeholders including patients, clinicians, laboratory personnel, hospital administrators, payers, and manufacturers are described.

Areas Covered: The authors describe how molecular diagnostics has achieved total laboratory automation over time, rivaling clinical chemistry to significantly improve testing efficiency. Finally, the authors discuss how advances in automation decrease the development time for new tests enabling clinicians to more readily provide test results.

Expert Commentary: The advancements described enable complete diagnostic solutions whereby specific test results can be combined with relevant patient data sets to allow healthcare providers to deliver comprehensive clinical recommendations in multiple fields ranging from infectious disease to outbreak management and blood safety solutions.     

Clopidogrel’s role in the management of atherosclerotic disease: a focus on acute coronary syndromes

Recent advances in our understanding of the role of the platelet in the atherosclerotic process beyond the acute formation of arterial blood clots, such as inflammation, have highlighted the role of antiplatelet agents as being much more than just ‘blood thinners.’ Some of the most important cardiovascular trials performed in the last 20 years have studied antiplatelet therapies. However, despite their long history, current global health implications and proven benefit, there remain substantial gaps in our understanding as to how to best utilize the limited number of antiplatelet agents available. This article will discuss the mechanism of action of the antiplatelet class known as thienopyridines, the pharmacodynamics and pharmacokinetics of the thienopyridine agent clopidogrel (Plavix^®, Bristol-Myers Squibb and Sanofi Pharmaceuticals) as well as the literature supporting its clinical benefits and areas of ongoing research that will help clarify the optimal utilization of clopidogrel for the treatment of cardiovascular disease.     

Clinical and molecular analyses of bloodstream infections caused by IMP metallo-β-lactamase-producing Enterobacteriaceae in a tertiary hospital in Japan

Carbapenemase-producing Enterobacteriaceae infection has been reported worldwide and is a major threat to public health. However, reports of bloodstream infection (BSI) caused by metallo-β-lactamase (MBL), especially the IMP-type, are limited. Therefore, we aimed to investigate the clinical and microbial characteristics of patients with BSI caused by IMP-type MBL-producing Enterobacteriaceae (MBL-E) in a tertiary care hospital in Japan. The clinical data were collected from medical charts for all the patients. A next-generation sequencing approach and multilocus sequence typing were used to identify antimicrobial resistance genes. Six patients were enrolled and had severe conditions on admission. The sources of MBL-E BSI were as follows: catheter-related BSI, pyelonephritis, cholangitis, and bacterial peritonitis. No isolate was resistant to levofloxacin or aminoglycoside. Microbiological response rates were 100%. The all-cause 30-day mortality rate was 50%. Of the six isolates, three were Enterobacter hormaechei sequence type 78, one was Enterobacter cloacae Hoffman cluster IV ST997, and two were Klebsiella pneumoniae (ST134 and ST252). All isolates produced IMP-1 and carried bla_IMP-1 gene and various antimicrobial resistance genes. The results of this study showed that MBL-E BSI was fatal, although rare, in patients with severe diseases and long-term hospitalization. Further research is necessary to determine the appropriate treatment strategies for MBL-producing BSI.