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Sterol analysis of complex matrices can be very laborious. To minimize the existing drawbacks, a new micro-method of sterols and squalene determination in cyanobacteria was developed and applied to monitor their production of Phormidium autumnale cultured heterotrophically. Sample extraction/saponification and GC analysis of the target compounds were optimized separately using Plackett-Burman design (PB) followed by a central composite rotational design (CCRD). The most influential variables were identified to maximize compound recovery. Chloroform presented the highest capability to extract all target compounds with a horizontal shaker table (HST) for homogenization in the saponification step. For the pretreatment, a small amount of chloroform was used for 90 min at 50 °C and 6 min for the saponification time. The sample introduction in the GC injector was studied by evaluating pressure and injector temperature. High response for sterols and squalene were obtained between 19 and 23 psi and at 310 °C of injection temperature. The new method was able to determine different sterol concentrations: 0.2-0.6 mg kg -1 of squalene, 5-18 mg kg -1 of stigmasterol, 6 mg kg -1 of cholesterol, and 3 mg kg -1 of β-sitosterol, showing high analytical performance and fulfilling all steps, thus proving to be a promising technique., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Anterior cruciate ligament intra-articular reconstruction may require extra-articular reinforcement in certain situations. As knee lateral region anatomical and biomechanical knowledge has increased with new research, it has been reported that the iliotibial band is important in the anterolateral stabilization of the knee. Possible indications for a "more anatomical" extra-articular tenodesis focusing on capsulo-osseous layer tensioning and distal Kaplan fibers reconstruction are reported, surgical approach details are described, and scientific data that gives support for this procedure are discussed., (© 2020 Published by Elsevier on behalf of the Arthroscopy Association of North America.)

The fluorescence quenching of TGA-CdSe quantum dots (QDs) was used for Cr(3+) quantification in vitamin supplements. The QD was electrochemically synthesized, demonstrating high reproducibility with control of particle size, thus making it a clean method, without the presence of reducing agents. Under ideal conditions, with the fluorescence band at 551 nm (excitation 365 nm), the maximum fluorescence quenching was observed at pH 4.0, with a time of 200 s for each data acquisition. Under optimum experimental conditions, linear quenching was observed for Cr(3+) in the range of 25.0-325.0 ng L(-1) (R=0.9996, n=6), a limit of detection of 5.67 ng L(-1), and relative standard deviation of 4.43% (n=10). The recovery test for Cr(3+) quantification in vitamin supplements presented results from 82% to 98%. These Cr(3+) determination results were compared to the same vitamin supplement sample using flame atomic absortion spectrometry (FAAS) method, and no significant differences were observed at 95% confidence level., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Cellular systems implanted into an injured nerve may produce growth factors or extracellular matrix molecules, modulate the inflammatory process and eventually improve nerve regeneration. In the present study, we evaluated the therapeutic value of human umbilical cord matrix MSCs (HMSCs) on rat sciatic nerve after axonotmesis injury associated to Vivosorb® membrane. During HMSCs expansion and differentiation in neuroglial-like cells, the culture medium was collected at 48, 72 and 96 h for nuclear magnetic resonance (NMR) analysis in order to evaluate the metabolic profile. To correlate the HMSCs ability to differentiate and survival capacity in the presence of the Vivosorb® membrane, the [Ca(2+)]i of undifferentiated HMSCs or neuroglial-differentiated HMSCs was determined by the epifluorescence technique using the Fura-2AM probe. The Vivosorb® membrane proved to be adequate and used as scaffold associated with undifferentiated HMSCs or neuroglial-differentiated HMSCs. In vivo testing was carried out in adult rats where a sciatic nerve axonotmesis injury was treated with undifferentiated HMSCs or neuroglial differentiated HMSCs with or without the Vivosorb® membrane. Motor and sensory functional recovery was evaluated throughout a healing period of 12 weeks using sciatic functional index (SFI), extensor postural thrust (EPT), and withdrawal reflex latency (WRL). Stereological analysis was carried out on regenerated nerve fibers. In vitro investigation showed the formation of typical neuroglial cells after differentiation, which were positively stained for the typical specific neuroglial markers such as the GFAP, the GAP-43 and NeuN. NMR showed clear evidence that HMSCs expansion is glycolysis-dependent but their differentiation requires the switch of the metabolic profile to oxidative metabolism. In vivo studies showed enhanced recovery of motor and sensory function in animals treated with transplanted undifferentiated and differentiated HMSCs that was accompanied by an increase in myelin sheath. Taken together, HMSC from the umbilical cord Wharton jelly might be useful for improving the clinical outcome after peripheral nerve lesion., (Copyright © 2012 International Society of Differentiation. Published by Elsevier B.V. All rights reserved.)

Rapid, accurate and inexpensive methods are essential to detect drug-resistant Mycobacterium tuberculosis and allow timely application of effective treatment and precautions to prevent transmission. The proportion method, the MTT and Alamar Blue redox methods, and the D29 mycobacteriophage assay, were compared for their ability to detect resistance to isoniazid and rifampicin. When tested against a panel of known M. tuberculosis strains, the redox methods and the D29 assay showed good sensitivity and specificity compared to the proportion method, suggesting that they could be useful alternatives for identifying multidrug resistance in M. tuberculosis.