Direct replacement of antibodies with molecularly imprinted polymer (MIP) nanoparticles in ELISA - development of a novel assay for vancomycin

A simple and straightforward technique for coating microplate wells with molecularly imprinted polymer nanoparticles (nanoMIPs) to develop ELISA type assays is presented here for the first time. NanoMIPs were synthesized by a solid phase approach with immobilized vancomycin (template) and characterized using Biacore 3000, dynamic light scattering and electron microscopy. Immobilization, blocking and washing conditions were optimized in microplate format. The detection of vancomycin was achieved in competitive binding experiments with a HRP-vancomycin conjugate. The assay was capable of measuring vancomycin in buffer and in blood plasma within the range 0.001-70 nM with a detection limit of 0.0025 nM (2.5 pM). The sensitivity of the assay was three orders of magnitude better than a previously described ELISA based on antibodies. In these experiments nanoMIPs have shown high affinity and minimal interference from blood plasma components. Immobilized nanoMIPs were stored for 1 month at room temperature without any detrimental effects to their binding properties. The high affinity of nanoMIPs and the lack of a requirement for cold chain logistics make them an attractive alternative to traditional antibodies used in ELIS

Statystyka w badaniach medycznych

S\u142owa kluczowe: statystyka w badaniach medycznych; metody analizy statystycznej \u2013 statystyka opisowa, testowanie hipotez i testy statystyczne; korelacja \u2013 badanie zale\u17cno\u15bci; biometria; pomiary biologiczn

Recommendation of RILEM TC 212-ACD: acousticemission and related NDE techniques for crack detectionand damage evaluation in concrete. Measurement method for acoustic emission signals in concrete

The text presented hereafter is a draft for general consideration

The mitochondrial receptor complex

The receptor complex in the mitochondrial outer membrane, which consists of at least seven different proteins, is responsible for the recognition and translocation of cytosolically synthesized preproteins. Two of its subunits, MOM19 and MOM72, function as surface receptors for preproteins. Four other subunits (MOM38, MOM30, MOM8, and MOM7) have been suggested to constitute the general insertion pore (GIP). Here we report on the structure and function of MOM22. MOM22 is anchored in the outer membrane by a single transmembrane segment. The highly negatively charged N-terminal domain is exposed to the cytosol and the C-terminal domain to the intermembrane space. MOM22 appears to be a central component of the receptor complex, required for the transfer of preproteins from the receptors to the GIP. We speculate that the negatively charged domain of MOM22 is involved in the transfer of positively charged signal sequences of preproteins.

Recommendation of RILEM TC 212-ACD: acousticemission and related NDE techniques for crack detectionand damage evaluation in concrete.Test method for damage qualification of reinforced concrete beams by acousticemission

The text presented hereafter is a draft for general consideration

Aim and shoot: molecule-imprinting polymer coated MoO3 for selective SERS detection and photocatalytic destruction of low-level organic contaminants

A sensitive and selective SERS sensor with easy and excellent recyclability is highly demanded because of its great potential application in complex detection environments. Here, using methylene blue (MB) as a model target, a semiconductor-based SERS substrate composed of a MoO3 nanorod core and a uniform molecule-imprinting polymethacrylic acid shell (MIP) with a thickness of 4 nm was designed and fabricated (MoO3@MIP) to achieve selective detection. The key to the successful coating of the ultrathin uniform MIP shell lies in the pretreatment of a MoO3 core with nitric acid, providing sufficient surficial hydroxyls for the anchoring of a polymer precursor. The molecule-imprinted voids for MB were formed simply via light irradiation as a result of photocatalytic degradation by a MoO3 semiconductor. This core–shell MIP composite shows a high SERS selectivity towards low-level MB in a mixed MB/CV solution. The enhanced factor (EF) is high, at 1.6 × 104. More importantly, the selective detection allows the further photocatalytic recycling of MoO3@MIP in an “aim-and-shoot” way, which well preserves the detection selectivity and sensitivity towards MB at least for 4 cycles. Based on decreased sensitivity with the increasing shell thickness (10–24 nm), a MIP-gating charge transfer mechanism is proposed to demonstrate the high EF instead of the molecule-enrichment effect. This “aim-and-shoot” strategy is expected to push forward the prosperous application of selective SERS for trace detection in versatile environments

Zmiany w składzie tkankowym obserwowane w trakcie terapii ciężkich zaburzeń funkcji tarczycy

  Introduction: Hyper- and hypothyroidism are accompanied by altered metabolic rate, thermogenesis, and body weight. The aim of this study was to estimate the relation between treatment-induced changes in thyroid function, and the accompanying body composition in patients with either severe hypo- or hyperthyroidism. Material and methods: Body composition analysis and hormonal assessment were measured at the initial diagnosis of thyroid disorder, after three-month treatment, and finally after complete recovery from hyperthyroidism (n = 18) or hypothyroidism (n = 27). Nonparametric Spearman correlation was used to analyse the relation between thyroid hormones and body composition as well as their respective changes. Results: In hypothyroid patients applied treatment significantly reduced (p < 0.05) total body weight, mainly due to a decrease in fat mass, whereas in hyperthyroid patients it caused a weight gain, with a simultaneous increase in muscle, water and fat mass. Total body weight and fat mass were significantly correlated with thyroid hormones’ concentrations in all patients. Changes of fat, water, or muscle mass were strongly correlated with the changes in the patients’ hormonal status. Conclusions: Body composition is related to the concentration of thyroid hormones in thyroid dysfunction. Treatment-induced changes in thyroid hormones concentrations are correlated with the magnitude of the change of body weight, including muscle, water, and fat amount. (Endokrynol Pol 2016; 67 (4): 359–366)    Wstęp: Zarówno nadczynność, jak i niedoczynność tarczycy charakteryzują się zaburzeniami podstawowej przemiany materii, termogenezy oraz masy ciała. Celem pracy była ocena związku między zmianami funkcji tarczycy oraz zmianami składu tkankowego ciała u pacjentów w trakcie terapii ciężkich zaburzeń funkcji tarczycy. Materiał i metody: Badanie składu ciała oraz badanie biochemicznych wykładników funkcji tarczycy przeprowadzono u 18 chorych z pełnoobjawową nadczynnością tarczycy oraz u 27 chorych z niedoczynnością tarczycy, w okresie rozpoznania choroby, po około trzech miesiącach leczenia oraz po całkowitym wyrównaniu funkcji tarczycy, w okresie eutyreozy. Ponadto przeprowadzono analizę związku między zmianami funkcji tarczycy oraz zmianami w składzie ciała przeprowadzono nieparametryczną analizę Spearmana. Wyniki: W grupie chorych leczonych z powodu niedoczynności tarczycy zaobserwowano statystycznie istotny spadek masy, głównie kosztem masy tkanki tłuszczowej (p< 0,05), podczas gdy w grupie chorych z pierwotnie rozpoznaną nadczynnością tarczycy stwierdzono istotny wzrost masy tkanki tłuszczowej, mięśniowej oraz wody całkowitej (p < 0,05). W obu grupach zaobserwowano ponadto istotne korelacje między stężeniem hormonów tarczycy a masą tkanki tłuszczowej na wszystkich etapach leczenia (p < 0,05). Jednocześnie zmiany wszystkich parametrów składu tkankowego ciała (Δ) silnie korelowały ze zmianami biochemicznych wykładników funkcji tarczycy(Δ) (p < 0,05). Wnioski: W zaburzeniach funkcji tarczycy obserwuje się silne zależności między składem tkankowym ciała a stężeniem hormonów tarczycy. Zmiany w składzie tkankowym ciała są jednocześnie silnie skorelowane ze zmianami stężenia hormonów tarczycy obserwowanymi w trakcie terapii. (Endokrynol Pol 2016; 67 (4): 359–366)

The cytosolic DnaJ-like protein Djp1p is involved specifically in peroxisomal protein import

The Saccharomyces cerevisiae DJP1 gene encodes a cytosolic protein homologous to Escherichia coli DnaJ. DnaJ homologues act in conjunction with molecular chaperones of the Hsp70 protein family in a variety of cellular processes. Cells with a DJP1 gene deletion are viable and exhibit a novel phenotype among cytosolic J-protein mutants in that they have a specific impairment of only one organelle, the peroxisome. The phenotype was also unique among peroxisome assembly mutants: peroxisomal matrix proteins were mislocalized to the cytoplasm to a varying extent, and peroxisomal structures failed to grow to full size and exhibited a broad range of buoyant densities. Import of marker proteins for the endoplasmic reticulum, nucleus, and mitochondria was normal. Furthermore, the metabolic adaptation to a change in carbon source, a complex multistep process, was unaffected in a DJP1 gene deletion mutant. We conclude that Djp1p is specifically required for peroxisomal protein import

Long-term renal outcome in children with OCRL mutations: retrospective analysis of a large international cohort

BACKGROUND: Lowe syndrome (LS) and Dent-2 disease (DD2) are disorders associated with mutations in the OCRL gene and characterized by progressive chronic kidney disease (CKD). Here, we aimed to investigate the long-term renal outcome and identify potential determinants of CKD and its progression in children with these tubulopathies. METHODS: Retrospective analyses were conducted of clinical and genetic data in a cohort of 106 boys (LS: 88 and DD2: 18). For genotype-phenotype analysis, we grouped mutations according to their type and localization. To investigate progression of CKD we used survival analysis by Kaplan-Meier method using stage 3 CKD as the end-point. RESULTS: Median estimated glomerular filtration rate (eGFR) was lower in the LS group compared with DD2 (58.8 versus 87.4 mL/min/1.73 m(2), P < 0.01). CKD stage II-V was found in 82% of patients, of these 58% and 28% had moderate-to-severe CKD in LS and DD2, respectively. Three patients (3%), all with LS, developed stage 5 of CKD. Survival analysis showed that LS was also associated with a faster CKD progression than DD2 (P < 0.01). On multivariate analysis, eGFR was dependent only on age (b = -0.46, P < 0.001). Localization, but not type of mutations, tended to correlate with eGFR. There was also no significant association between presence of nephrocalcinosis, hypercalciuria, proteinuria and number of adverse clinical events and CKD. CONCLUSIONS: CKD is commonly found in children with OCRL mutations. CKD progression was strongly related to the underlying diagnosis but did not associate with clinical parameters, such as nephrocalcinosis or proteinuria