Multiplexed femtomolar quantitation of human cytokines in a fluoropolymer microcapillary film

Sensitive quantitation of multiple cytokines can provide important diagnostic information during infection, inflammation and immunopathology. In this study sensitive immunoassay detection of human cytokines IL-1β, IL-6, IL-12p70 and TNFα is shown for singleplex and multiplex formats using a novel miniaturized ELISA platform. The platform uses a disposable plastic multi-syringe aspirator (MSA) integrating 8 disposable fluoropolymer microfluidic test strips, each containing an array of ten 200 mean i.d. microcapillaries coated with a set of monoclonal antibodies. Each MSA device thus performs 10 tests on 8 samples, delivering 80 measurements. Unprecedented levels of sensitivity were obtained with the novel fluoropolymer microfluidic material and simple colorimetric detection in a flatbed scanner. The limit of detection for singleplex detection ranged from 2.0 to 15.0 pg/ml, i.e. 35 and 713 femtomolar for singleplex cytokine detection, and the intra- and inter-assay coefficient of variation (CV) remained within 10%. In addition, a triplex immunoassay was developed for measuring IL-1β, IL-12p70 and TNFα simultaneously from a given sample in the pg/ml range. These assays permit high sensitivity measurement with rapid <15 min assay or detection from undiluted blood serum. The portability, speed and low-cost of this system are highly suited to point-of-care testing and field diagnostics applications

Assessment of serum vitamin D level in patients with systemic lupus erythematosus

Objective To evaluate the serum level of vitamin D in patients with systemic lupus erythematosus (SLE) and its relationship with disease activity. Patients and methods Forty patients suffering from SLE were enrolled in this study (group I). They were further divided into two subgroups according to the SLE disease activity index (SLEDAI) score: group Ia with respect to disease activity and group Ib with respect to disease remission. Another 20 age-matched and sex-matched healthy individuals were chosen as control group II. All patients underwent complete medical history taking and thorough clinical examination; the disease activity was assessed by the use of SLEDAI score. Serum vitamin D level in all patients and controls was measured. Results Vitamin D level was significantly higher in controls than in patients. The vitamin D deficiency was highly prevalent among patients with disease activity than in the remission group. There was highly significant inverse correlation between vitamin D level and SLEDAI score in the patient group. Vitamin D level correlated inversely with C reactive protein (CRP) and anti-dsDNA in the disease activity group, whereas it correlated positively with C3. Conclusion Vitamin D deficiency is prevalent in SLE patients more than in healthy controls; vitamin D deficiency is highly prevalent among patients with disease activity than in the remission group, and vitamin D level correlated inversely with disease activity, which suggest that inadequate vitamin D level, among other factors, probably contributed to the development of active disease in patients with SLE

Supplementary Material for: Identifying a Potential Summary Measure for Overall Caries Level in Children Examined with the International Caries Detection and Assessment System

<p><b><i>Objective:</i></b> To identify a potential summative summary measure that reflects a patient's overall caries level in children examined with the International Caries Detection and Assessment System (ICDAS). <b><i>Subjects and Methods:</i></b> Participants were 1- to 15-year-old children from Kuwait, Brazil, and Spain. Children's teeth were examined using the ICDAS. Multiple measures of central tendency and dentition-specific indices were considered as potential summative measures. The relationship between the summative measures and number of caries lesions was evaluated considering degrees of caries severity using Spearman's correlation analysis. The results were generated using the Kuwaiti sample and were cross-validated using the Brazilian and Spanish samples. <b><i>Results:</i></b> A total of 2,808 children participated in the present study. Total ICDAS score and mean ICDAS score showed a strong correlation with the number of caries lesions at different caries severity levels in primary, mixed, and permanent dentitions. The total ICDAS score of 51 buccal (B), 61B, 54 occlusal (O), 55O, 64O, 65O, 74O, 75O, 84O, and 85O surfaces in primary dentition and the total ICDAS score of 14O, 16 lingual (L), 16O, 24O, 26L, 26O, 36B, 36O, 37O, 46O, 46B, and 47O surfaces in permanent dentition or mixed dentition (if present) showed strong correlations with the number of caries lesions at different caries severity levels. <b><i>Conclusions:</i></b> Total ICDAS and mean ICDAS scores were the best summary measures of overall caries level at different dental stages. Total ICDAS scores of selectively examined 10 surfaces in primary dentition and 12 surfaces in permanent dentition can give an excellent summary measure for a patient's overall caries level with high diagnostic accuracy.</p