Diazoxide Attenuates Postresuscitation Brain Injury in a Rat Model of Asphyxial Cardiac Arrest by Opening Mitochondrial ATP-Sensitive Potassium Channels

Objective. We investigated whether and how diazoxide can attenuate brain injury after cardiopulmonary resuscitation (CPR) by selective opening of mitochondrial ATP-sensitive potassium (mitoKATP) channels. Methods. Adult male Sprague-Dawley rats with induced cerebral ischemia ( per group) received an intraperitoneal injection of 0.1% dimethyl sulfoxide (1 mL; vehicle group), diazoxide (10 mg/kg; DZ group), or diazoxide (10 mg/kg) plus 5-hydroxydecanoate (5 mg/kg; DZ + 5-HD group) 30 min after CPR. The control group (sham group, ) underwent sham operation, without cardiac arrest. Mitochondrial respiratory control rate (RCR) was determined. Brain cell apoptosis was assessed using TUNEL staining. Expression of Bcl-2, Bax, and protein kinase C epsilon (PKCε) in the cerebral cortex was determined by Western blotting and immunohistochemistry. Results. The neurological deficit scores (NDS) in the vehicle group decreased significantly at 24 h and 48 h after CPR. Diazoxide significantly improved NDS and mitochondrial RCR after CPR at both time points; 5-HD cotreatment abolished these effects. Diazoxide decreased TUNEL-positive cells following CPR, upregulated Bcl-2 and PKCε, downregulated Bax, and increased the Bcl-2/Bax ratio; 5-HD cotreatment reversed these effects. Conclusions. Diazoxide attenuates postresuscitation brain injury, protects mitochondrial function, inhibits brain cell apoptosis, and activates the PKC pathway by opening mitoKATP channels.

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ProFold: Protein Fold Classification with Additional Structural Features and a Novel Ensemble Classifier

Protein fold classification plays an important role in both protein functional analysis and drug design. The number of proteins in PDB is very large, but only a very small part is categorized and stored in the SCOPe database. Therefore, it is necessary to develop an efficient method for protein fold classification. In recent years, a variety of classification methods have been used in many protein fold classification studies. In this study, we propose a novel classification method called proFold. We import protein tertiary structure in the period of feature extraction and employ a novel ensemble strategy in the period of classifier training. Compared with existing similar ensemble classifiers using the same widely used dataset (DD-dataset), proFold achieves 76.2% overall accuracy. Another two commonly used datasets, EDD-dataset and TG-dataset, are also tested, of which the accuracies are 93.2% and 94.3%, higher than the existing methods. ProFold is available to the public as a web-server.

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Simultaneous bioimaging recognition of Al3+ and Cu2+ in living-cell, and further detection of F− and S2− by a simple fluorogenic benzimidazole-based chemosensor

Publication date: 1 December 2016
Source:Talanta, Volume 161
Author(s): Haiyang Liu, Bibo Zhang, Chunyan Tan, Feng Liu, Jiakun Cao, Ying Tan, Yuyang Jiang
A simple Schiff base (BMSA) prepared from salicylaldehyde and 2-(1H-benzo[d]imidazol-2-yl)aniline was evaluated as an efficient fluorescent chemosensor for the selective recognition of Al³⁺and Cu²⁺ over other common metal ions. This sensor could detect Al3⁺ in CH3OH/PBS with distinct emission red-shift (the detection limit 0.31μM)and Cu²⁺in CH3OH/Tris-HCL (the detection limit 0.54μM) with obvious fluorescence quenching. The obtained BMSA-Al³⁺ and BMSA-Cu²⁺ complexes could act as cascade sensors for detecting F⁻ and S²⁻, respectively. The recognizing behavior of BMSA toward Al³⁺and Cu²⁺ has been investigated in detail through Job's Plot, FT-IR NMR, and HRMS analysis. Moreover, this chemosensor was verified to be of low cytotoxicity and good imaging characteristics for the detection of Al³⁺ and Cu²⁺, and further for the recognition of F⁻ and S²⁻ in living cells, suggesting that BMSA was proved to be a useful tool for tracking Al3⁺/Cu²⁺and F⁻/S²⁻ ions in vivo.

Graphical abstract

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Neural networks applied to characterize blends containing refined and extra virgin olive oils

Publication date: 1 December 2016
Source:Talanta, Volume 161
Author(s): Regina Aroca-Santos, John C. Cancilla, Enrique S. Pariente, José S. Torrecilla
The identification and quantification of binary blends of refined olive oil with four different extra virgin olive oil (EVOO) varietals (Picual, Cornicabra, Hojiblanca and Arbequina) was carried out with a simple method based on combining visible spectroscopy and non-linear artificial neural networks (ANNs).The data obtained from the spectroscopic analysis was treated and prepared to be used as independent variables for a multilayer perceptron (MLP) model. The model was able to perfectly classify the EVOO varietal (100% identification rate), whereas the error for the quantification of EVOO in the mixtures containing between 0% and 20% of refined olive oil, in terms of the mean prediction error (MPE), was 2.14%. These results turn visible spectroscopy and MLP models into a trustworthy, user-friendly, low-cost technique which can be implemented on-line to characterize olive oil mixtures containing refined olive oil and EVOOs.

Graphical abstract

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Discrimination and quantification of two isomeric antineoplastic drugs by rapid and non-invasive analytical control using a handheld Raman spectrometer

Publication date: 1 December 2016
Source:Talanta, Volume 161
Author(s): L.M.M. Lê, A. Tfayli, J. Zhou, P. Prognon, A. Baillet-Guffroy, E. Caudron
Raman spectroscopy is a rapid, non-destructive and non-invasive method that is a promising tool for real-time analytical control of drug concentrations. This study evaluated a handheld Raman device to discriminate and quantify two isomeric drugs used to treat cancer. Doxorubicin (DOXO) and epirubicin (EPIR) samples were analyzed at therapeutic concentrations from 0.1 to 2mg/mL (n=90) and 0.08–2mg/mL (n=90) by non-invasive measurements using a portable Raman spectrometer.The discrimination of these two molecules was demonstrated for all concentrations (n=180) by qualitative analysis using partial least square discriminant analysis (PLS-DA) with 100% classification accuracy, sensitivity and specificity and 0% error rate. For each molecule, quantitative analyses were performed using PLS regression. The validity of the model was evaluated using root mean square error of cross validation (RMSECV) and prediction (RMSEP) that furnished 0.05 and 0.02mg/mL for DOXO and 0.17 and 0.16mg/mL for EPIR after pretreatment optimization. Based on the accuracy profile, the linearity range was from 1.256 to 2.000mg/mL for DOXO (R²=0.9988) and from 0.553 to 2.000mg/Ml for EPIR (R²=0.9240) and repeatability (CV% max of 1.8% for DOXO and 3.2% for EPIR) and intermediate precision (CV% max of 2.8% for DOXO and 4.5% for EPIR) were both acceptable.Despite the narrow validated concentration range for quantitative analysis, this study shows the potential of a handheld Raman spectrometer coupled to chemometric approaches for real-time quantification of cytotoxic drugs, as well for discriminating between two drugs with similar UV absorption profiles. Finally, the use of a handheld spectrometer with the possibility of a direct measurement of substances in containers is a potentially valuable tool for combining patient safety with security of healthcare workers.

Graphical abstract

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The association between arterial stiffness and tongue manifestations of blood stasis in patients with type 2 diabetes

Diabetes mellitus (DM) is a hypercoagulable state and is associated with highly increased risk of vascular complications. In the theory of traditional Chinese medicine (TCM), these vascular complications are c...

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Fluorescence Activated Cell Sorting (FACS) and Gene Expression Analysis of Fos-expressing Neurons from Fresh and Frozen Rat Brain Tissue

Here we present a Fluorescence Activated Cell Sorting (FACS) protocol to study molecular alterations in Fos-expressing neuronal ensembles from both fresh and frozen brain tissue. The use of frozen tissue allows FACS isolation of many brain areas over multiple sessions to maximize the use of valuable animal subjects.

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