Geometric Flow Control Lateral Flow Immunoassay Devices (GFC-LFIDs): A New Dimension to Enhance Analytical Performance.

The nitrocellulose (NC) membrane based lateral flow immunoassay device (LFID) is one of the most important and widely used biosensor platforms for point-of-care (PoC) diagnostics. However, the analytical performance of LFID has limitations and its optimization is restricted to the bioassay chemistry, the membrane porosity, and the choice of biolabel system. These bottom neck technical issues resulted from the fact that the conventional LFID design principle has not evolved for many years, which limited the LFID for advanced biosensor applications. Here we introduce a new dimension for LFID design and optimization based on geometric flow control (GFC) of NC membranes, leading to highly sensitive GFC-LFID. This novel approach enables comprehensive flow control via different membrane geometric features such as the width (w) and the length (l) of a constriction, as well as its input angle (θ 1) and output angle (θ 2). The GFC-LFID (w=0.5 mm, l=7 mm, θ 1 = 60°, θ 2 = 45°) attained a 10-fold increase in sensitivity for detection of interleukin-6 (IL-6), compared with conventional LFID, whereas reducing by 10-fold the antibody consumption. The GFC-LFID detects IL-6 over a linear range of 0.1-10 ng/mL with a limit of detection (LoD) of 29 pg/mL, which even outperforms some commercial IL-6 LFIDs. Such significant improvement is attained by pure geometric control of the NC membrane, without additives, that only relaying on a simple high throughput laser ablation procedure suitable for integration on regular large-scale manufacturing of GFC-LFIDs. Our new development on GFC-LFID with the combination of facile scalable fabrication process, tailored flow control, improved analytical performance, and reduced antibodies consumption is likely to have a significant impact on new design concept for the LFID industry.

Lack of association between filaggrin gene mutations and onset of psoriasis in childhood.

BACKGROUND: Atopic dermatitis (AD; OMIM#603165) and psoriasis (OMIM#177900) are two common inflammatory skin disorders. Both are genetically complex, multifactorial and do not follow a Mendelian pattern of inheritance. Both diseases share several genetic susceptibility loci such as the epidermal differentiation complex (EDC) on chromosome 1q21. Within the EDC, mutations in the filaggrin (FLG) gene are strongly associated with AD whereas no association has been replicated with psoriasis. However, reduced levels of filaggrin have been reported in psoriatic skin. Further, filaggrin deficiency was shown to be a modifying factor for the phenotype in another epidermal skin disorder, X-linked recessive ichthyosis. Altogether, this raises the question if FLG mutations may modify the disease course in other epidermal skin diseases such as psoriasis. Psoriasis is a highly heterogeneous disease and so far genetic studies have not taken the distinct sub-phenotype childhood onset into account. OBJECTIVE: To determine if FLG mutations modify the onset of psoriasis. MATERIALS AND METHODS: A total of 241 children with onset of psoriasis below 15 years of age and 314 healthy controls were identified at the Dermatology clinic, Karolinska University Hospital and diagnosed by the same dermatologist (JL). Blood samples were taken and medical history was recorded. FLG was genotyped in all patients and controls using allelic discrimination (n = 555) and sequencing (n = 20). RESULTS AND CONCLUSIONS: No association between FLG mutations and early onset of psoriasis was demonstrated (P = 0.57) and no novel mutations were detected, indicating that FLG loss-of-function variants do not have a strong effect on the onset of psoriasis in childhood.

Immunohistochemical determination of thioredoxin and glutaredoxin distribution in the human cervix, and possible relation to cervical ripening.

Thioredoxin (Trx) and glutaredoxin (Grx) are dithiol redox enzymes, catalyzing general thiol-disulfide oxidoreductions apart from being hydrogen donors for ribonucleotide reductase, an enzyme essential for DNA synthesis. In mammals, isoenzymes of Trx and Grx are found in the cytoplasm (Trx1 and Grx1) or in mitochondria (Trx2 and Grx2). Trx and Grx play a role in cellular defence against oxidative stress and in redox regulation of cellular function. The localization and levels of human Trx1 and human Grx1 have been determined in the human cervix by immunohistochemistry and image analysis. Cervical biopsies were obtained from five non-pregnant, five term pregnant and five postpartum women. The levels of both Trx1 and Grx1 were increased in the nuclei (after translocation from the cytoplasm) of stromal cells in cervices from the term pregnant group as compared to the non-pregnant group, but the levels in the postpartum group did not differ significantly from those of the other two groups. These results are in agreement with our previous data on the mRNA expression of these two redox enzymes. The increased levels of the redox enzymes in term pregnancy suggest that they can be regulating factors involved in the process of cervical ripening, e.g. transcription factors and enzymes. Secreted Trx may participate in removing inhibitors of collagen-degrading metalloproteinases.