Miniaturized Rapid Electrochemical Immunosensor Based on Screen Printed Carbon Electrodes for Mycobacterium tuberculosis Detection.

In 2019, over 21% of an estimated 10 million new tuberculosis (TB) patients were either not diagnosed at all or diagnosed without being reported to public health authorities. It is therefore critical to develop newer and more rapid and effective point-of-care diagnostic tools to combat the global TB epidemic. PCR-based diagnostic methods such as Xpert MTB/RIF are quicker than conventional techniques, but their applicability is restricted by the need for specialized laboratory equipment and the substantial cost of scaling-up in low- and middle-income countries where the burden of TB is high. Meanwhile, loop-mediated isothermal amplification (LAMP) amplifies nucleic acids under isothermal conditions with a high efficiency, helps in the early detection and identification of infectious diseases, and can be performed without the need for sophisticated thermocycling equipment. In the present study, the LAMP assay was integrated with screen-printed carbon electrodes and a commercial potentiostat for real time cyclic voltammetry analysis (named as the LAMP-Electrochemical (EC) assay). The LAMP-EC assay was found to be highly specific to TB-causing bacteria and capable of detecting even a single copy of the Mycobacterium tuberculosis (Mtb) IS6110 DNA sequence. Overall, the LAMP-EC test developed and evaluated in the present study shows promise to become a cost-effective tool for rapid and effective diagnosis of TB.

Conditional reduction of the loss value versus reinforcement learning for biassing a de-novo drug design generator.

Deep learning has demonstrated promising results in de novo drug design. Often, the general pipeline consists of training a generative model (G) to learn the building rules of valid molecules, then using a biassing technique such as reinforcement learning (RL) to focus G on the desired chemical space. However, this sequential training of the same model for different tasks is known to be prone to a catastrophic forgetting (CF) phenomenon. This work presents a novel yet simple approach to bias G with significantly less CF than RL. The proposed method relies on backpropagating a reduced value of the cross-entropy loss used to train G according to the proportion of desired molecules that the biased-G can generate. We named our approach CRLV, short for conditional reduction of the loss value. We compared the two biased models (RL-biased-G and CRLV-biased-G) for four different objectives related to de novo drug design.CRLV-biased-G outperformed RL-biased-G in all four objectives and manifested appreciably less CF. Besides, an intersection analysis between molecules generated by the RL-biased-G and the CRLV-biased-G revealed that they can be used jointly without losing diversity given the low percentage of overlap between the two to further increase the desirability. Finally, we show that the difficulty of an objective is proportional to (i) its frequency in the dataset used to train G and (ii) the associated structural variance (SV), which is a new parameter we introduced in this paper, calling for novel exploration techniques for such difficult objectives.

Pesticides in fruits and vegetables from the Souss Massa region, Morocco.

This survey reports the monitoring of multi-pesticide residues of some fruits and vegetables sold in the local markets, sampled in 2018-2019, in the Souss Massa region in Morocco. A QuEChERS-LC-MS/MS method for 202 pesticides, belonging to different classes (carbamates, organophosphorus and organonitrogen pesticides) was applied and 51 samples were randomly bought from the local market, belonging to different products (tomato, cucumber, coriander, apricot, parsley, potato, zucchini, green bean, lettuce, strawberry and orange) and analysed for pesticide residues, which were detected in 69% of the samples, below the maximum residue limits (MRLs) for some pesticides which represent 14% of the targeted compounds. The most frequently detected compounds were acetamiprid, acibenzolar-s-methyl, abamectin, azoxystrobin, bifenazate, bitertanol, bromuconazole, butoxycarboxim, cyromazine, difenoconazole, epoxiconazole, fenbuconazole, fluometuron, linuron, metaflumizone, metconazole, metribuzin, myclobutanil, pirimicarb, pyraclostrobin, propamocarb, rotenone, trichlorfon, tebuconazole, tetraconazole, thiamethoxam and thiophanate-methyl. The obtained results provide a value to the situation of pesticide residues in Morocco.

Analysis of the major chiral compounds of Artemisia herba-alba essential oils (EOs) using reconstructed vibrational circular dichroism (VCD) spectra: En route to a VCD chiral signature of EOs.

An unprecedented methodology was developed to simultaneously assign the relative percentages of the major chiral compounds and their prevailing enantiomeric form in crude essential oils (EOs). In a first step the infrared (IR) and vibrational circular dichroism (VCD) spectra of the crude essential oils were recorded and in a second step they were modelized as a linear weighted combination of the IR and VCD spectra of the individual spectra of pure enantiomer of the major chiral compounds present in the EOs. The VCD spectra of enantiomer of known enantiomeric excess shall be recorded if they are not yet available in a library of VCD spectra. For IR, the spectra of pure enantiomer or racemic mixture can be used. The full spectra modelizations were performed using a well known and powerful mathematical model (least square estimation: LSE) which resulted in a weighting of each contributing compound. For VCD modelization, the absolute value of each weighting represented the percentage of the associate compound while the attached sign addressed the correctness of the enantiomeric form used to build the model. As an example, a model built with the non-prevailing enantiomer will show a negative sign of the weighting value. For IR spectra modelization, the absolute value of each weighting represented the percentage of the compounds without of course accounting for the chirality of the prevailing enantiomers. Comparison of the weighting values issuing from IR and VCD spectra modelizations is a valuable source of information: if they are identical, the EOs are composed of nearly pure enantiomers, if they are different the chiral compounds of the EOs are not in an optically pure form. The method was applied on four samples of essential oil of Artemisia herba-alba in which the three major compounds namely (-)-α-thujone, (+)-ß-thujone and (-)-camphor were found in different proportions as determined by GC-MS and chiral HPLC using polarimetric detector. In order to validate the methodology, the modelization of the VCD spectra was performed on purpose using the individual VCD spectra of (-)-α-thujone, (+)-ß-thujone and (+)-camphor instead of (-)-camphor. During this work, the absolute configurations of (-)-α-thujone and (+)-ß-thujone were confirmed by comparison of experimental and calculated VCD spectra as being (1S,4R,5R) and (1S,4S,5R) respectively.

Attempts to separate (-)-α-thujone, (+)-ß-thujone epimers from camphor enantiomers by enantioselective HPLC with polarimetric detection.

In a first step, 26 chiral stationary phases (CSPs) have been screened for the separation of (-)-α-thujone, (+)-ß-thujone epimers and camphor enantiomers by LC. The separations were monitored by a polarimeter detector. None of these CSPs provided a noticeable resolution for camphor enantiomers. The three components of a test mixture were clearly baseline separated on Chiralpak AS-H, Chiralpak AZ-H and TCI-MBS (poly(N-alpha-(S)-methylbenzylmaleimide) coated on silica gel) in a mobile phase composed of hexane/2-PrOH (99:1 v/v). Interestingly, for a preparative application, the three CSPs produced different elution orders for the three constituents of the mixture. In a second step, it is shown that the use of online polarimetric detection constitutes an unprecedented method to reveal the occurrence and the relative content of thujone epimers and the chirality of the major camphor enantiomer in crude essential oils. A proof of concept is illustrated on crude essential oils from Rosmarinus tournefortii, Artemisia herba alba and A. arborescens, which grow in Morocco and have several traditional uses there. In a third step, pure (+)-ß-thujone was quantitatively collected from A. arborescens crude oil by semi-preparative HPLC on Chiralpak AZ-H monitored by a polarimeter.