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Abstract Recently, the interest in extracellular vesicles released by bacteria has rapidly increased. Bacterial extracellular vesicles (BEVs) have been involved in bacteria‐bacteria and bacteria‐host interactions, which strengthen health or bring about various pathologies. However, BEV separation, characterization, and functional studies require the establishment of guidelines and further optimization in order to stimulate the development of science in BEV research and a following successful transformation into clinical applications. This position paper is authored by the Microbial Vesicles Task Force of the Chinese Society for Extracellular Vesicles (CSEV) composed of experienced medical laboratory specialists, microbiologists, virologists, biologists and material biologists who are actively engaged in BEV research. Herein, we present a concise description of BEV research and discover challenges and critical gaps in current BEV‐based analyses for clinical applications. Finally, we also offer suggestions and considerations to improve experimental reproducibility and interoperability in BEV research to promote progress in the field.

The effective and cheap production of platform chemicals is a crucial step towards the transition to a bio-based economy. In this work, biotechnological methods using sustainable, cheap, and readily available raw materials bring bio-economy and industrial microbiology together: Microbial production of two platform chemicals is demonstrated [lactic (LA) and succinic acid (SA)] from a non-expensive side stream of pulp and paper industry (fibre sludge) proposing a sustainable way to valorize it towards economically important monomers for bioplastics formation. This work showed a promising new route for their microbial production which can pave the way for new market expectations within the circular economy principles. Fibre sludge was enzymatically hydrolysed for 72 h to generate a glucose rich hydrolysate (100 g·L−1 glucose content) to serve as fermentation medium for Bacillus coagulans A 541, A162 strains and Actinobacillus succinogenis B1, as well as Basfia succiniciproducens B2. All microorganisms were investigated in batch fermentations, showing the ability to produce either lactic or succinic acid, respectively. The highest yield and productivities for lactic production were 0.99 g·g−1 and 3.75 g·L−1·h−1 whereas the succinic acid production stabilized at 0.77 g·g−1 and 1.16 g·L−1·h−1.

Food allergies are adverse health effects that arise from specific immune responses, occurring upon exposure to given foods, even if present in traces. Egg allergy is one of the most common food allergies, mainly caused by egg white proteins, with ovalbumin being the most abundant. As allergens can also be present in foodstuff due to unintended contamination, there is a need for analytical tools that are able to rapidly detect allergens in food products at the point-of-use. Herein, we report an origami paper-based device for detecting ovalbumin in food samples, based on a competitive immunoassay with chemiluminescence detection. In this biosensor, magnetic microbeads have been employed for easy and efficient immobilization of ovalbumin on paper. Immobilized ovalbumin competes with the ovalbumin present in the sample for a limited amount of enzyme-labelled anti-ovalbumin antibody. By exploiting the origami approach, a multistep analytical procedure could be performed using reagents preloaded on paper layers, thus providing a ready-to-use immunosensing platform. The assay provided a limit of detection (LOD) of about 1 ng mL−1 for ovalbumin and, when tested on ovalbumin-spiked food matrices (chocolate chip cookies), demonstrated good assay specificity and accuracy, as compared with a commercial immunoassay kit.

As glucose biosensors play an important role in glycemic control, which can prevent the diabetic complications, the development of a glucose sensing platform is still in needed. Herein, the first proposal on the in-house fabricated paper-based screen-printed ionic liquid/graphene electrode (SPIL-GE) modified with MXene (Ti3C2Tx), prussian blue (PB), glucose oxidase (GOx), and Nafion is reported. The concentration of PB/Ti3C2Tx was optimized and the optimal detection potential of PB/Ti3C2Tx/GOx/Nafion/SPIL-GE is −0.05 V. The performance of PB/Ti3C2Tx/GOx/Nafion modified SPIL-GE was characterized by cyclic voltammetry and chronoamperometry technique. This paper-based platform integrated with nanomaterial composites were realized for glucose in the range of 0.0–15.0 mM with the correlation coefficient R2 = 0.9937. The limit of detection method and limit of quantification were 24.5 μM and 81.7 μM, respectively. In the method comparison, this PB/Ti3C2Tx/GOx/Nafion/SPIL-GE exhibits a good correlation with the reference hexokinase method. This novel glucose sensing platform can potentially be used for the good practice to enhance the sensitivity and open the opportunity to develop paper-based electroanalytical devices.

In this work, indoxyl-glucoside was used as the substrate to develop a cost-effective, paper-based analytical device for the fluorescent and colorimetric dual-mode detection of β-glucosidase activity through a smartphone. The β-glucosidase can hydrolyze the colorless substrate indoxyl-glucoside to release indoxyl, which will be self-oxidized to generate green products in the presence of oxygen. Meanwhile, the green products emit bright blue-green fluorescence under ultraviolet–visible light irradiation at 365 nm. Fluorescent or colorimetric images were obtained by a smartphone, and the red-green-blue channels were analyzed by the Adobe Photoshop to quantify the β-glucosidase activity. Under the optimum conditions, the relative fluorescent and colorimetric signals have a good linear relationship with the activity of β-glucosidase, in the range of 0.01–1.00 U/mL and 0.25–5.00 U/mL, and the limits of detection are 0.005 U/mL and 0.0668 U/mL, respectively. The activities of β-glucosidase in a crude almond sample measured by the fluorescent and colorimetric methods were 23.62 ± 0.53 U/mL and 23.86 ± 0.25 U/mL, respectively. In addition, the spiked recoveries of normal human serum and crude almond samples were between 87.5% and 118.0%. In short, the paper-based device, combined with a smartphone, can provide a simple, environmentally friendly, and low-cost method for the fluorescent and colorimetric dual-mode detection of β-glucosidase activity.

Herein, a paper-based electrochemical sensor based on PtNP/COFTFPB−DHzDS@rGO was developed for the sensitive detection of furazolidone. A cluster-like covalent organic framework (COFTFPB−DHzDS) was successfully grown on the surface of amino-functional reduced graphene oxide (rGO-NH2) to avoid serious self-aggregation, which was further loaded with platinum nanoparticles (PtNPs) with high catalytic activity as nanozyme to obtain PtNP/COFTFPB−DHzDS@rGO nanocomposites. The morphology of PtNP/COFTFPB−DHzDS@rGO nanocomposites was characterized, and the results showed that the smooth rGO surface became extremely rough after the modification of COFTFPB−DHzDS. Meanwhile, ultra-small PtNPs with sizes of around 1 nm were precisely anchored on COFTFPB−DHzDS to maintain their excellent catalytic activity. The conventional electrodes were used to detect furazolidone and showed a detection limit as low as 5 nM and a linear range from 15 nM to 110 μM. In contrast, the detection limit for the paper-based electrode was 0.23 μM, and the linear range was 0.69–110 μM. The results showed that the paper-based electrode can be used to detect furazolidone. This sensor is a potential candidate for the detection of furazolidone residue in human serum and fish samples.

Microfluidic paper-based analytical devices (μPADs) have been extensively proposed as ideal tools for point-of-care (POC) testing with minimal user training and technical requirements. However, most μPADs use dried bioreagents, which complicate production, reduce device reproducibility and stability, and require transport and storage under temperature and humidity-controlled conditions. In this work, we propose a μPAD produced using an affordable craft-cutter and stored at room temperature, which is used to partially automate a single-step colorimetric magneto-immunoassay. As a proof-of-concept, the μPAD has been applied to the quantitative detection of Plasmodium falciparum lactate dehydrogenase (Pf-LDH), a biomarker of malaria infection. In this system, detection is based on a single-step magneto-immunoassay that consists of a single 5-min incubation of the lysed blood sample with immuno-modified magnetic beads (MB), detection antibody, and an enzymatic signal amplifier (Poly-HRP). This mixture is then transferred to a single-piece paper device where, after on-chip MB magnetic concentration and washing, signal generation is achieved by adding a chromogenic enzyme substrate. The colorimetric readout is achieved by the naked eye or using a smartphone camera and free software for image analysis. This μPAD afforded quantitative Pf-LDH detection in −1 when the result was interpreted by the naked eye and 1.4 ng mL−1 when analysed using the smartphone imaging system. Moreover, the study of a battery of clinical samples revealed concentrations of Pf-LDH that correlated with those provided by the reference ELISA and with better sensitivity than a commercial rapid diagnostic test (RDT). These results demonstrate that magneto-immunoassays can be partly automated by employing a μPAD, achieving a level of handling that approaches the requirements of POC testing.

Paper-based analytical devices (PADs) are highly effective tools due to their low cost, portability, low reagent accumulation, and ease of use. Molecularly imprinted polymers (MIP) are also extensively used as biomimetic receptors and specific adsorption materials for capturing target analytes in various complex matrices due to their excellent recognition ability and structural stability. The integration of MIP and PADs (MIP-PADs) realizes the rapid, convenient, and low-cost application of molecular-imprinting analysis technology. This review introduces the characteristics of MIP-PAD technology and discusses its application in the fields of on-site environmental analysis, food-safety monitoring, point-of-care detection, biomarker detection, and exposure assessment. The problems and future development of MIP-PAD technology in practical application are also prospected.

The effectiveness of a biorefinery based on an HWE-Kraft-ECF bleaching process and the end use of pulp was systematically evaluated. Using a P-factor of 198, nearly 30% of xylan-based sugars were recovered. The resulting pulp and paper properties were found to be comparable with the control. A maximum xylan-based sugar recovery of nearly 50% was achieved at a P-factor of 738. Although the strength of this P-factor induced handsheet was lower than that of the control by about 20%, the corresponding pulp was sufficient for dissolving pulp application. However, once the P-factor rose above 1189, hemicellulose sugars were significantly degraded into furans; pulp and paper properties were also deteriorated due to cellulose degradation, lignin deposition and condensation. Thus, considering the different end use of pulps, the performance of an HWE-based biorefinery could be balanced by its HWE severity., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Monitoring ferroptosis-related miRNAs is crucial for the treatment and prognosis of patients with intracerebral hemorrhage. In this work, a novel hydrophobic paper (h-paper)-based plasmonic substrate was produced by dropping DS Au nanorods with a narrow range of sizes and morphologies onto h-paper. Raman reporter molecules were adsorbed to the array surface, and surface-enhanced Raman scattering spectra at randomly selected points reveal uniform and significant SERS enhancement. Hairpin DNAs labelled with Raman reporters and hybridized with placeholder DNAs were decorated on SERS substrate to fabricate SERS biosensor. Target miRNAs initiated the “inverse Molecular Sentinel” process. During the process, PHs were removed and the conformation of HPs changed toward the hairpin structure, thus eliciting the proximity of Raman reporter to substrate and a stronger SERS signal. The proposed SERS biosensor performs well in terms of stability, reproducibility, and selectivity. The limits of detection of miR-122-5p and miR-140-5p in serum were 4.17 aM and 4.49 aM, respectively. Finally, the fabricated SERS biosensor was applied to detect miR-122-5p and miR-140-5p in ICH patients and healthy subjects, and the results obtained by SERS were consistent with the results from quantitative real-time polymerase chain reaction, revealing the accuracy of the method. This simple, rapid approach offers great potential for the simultaneous detection of miRNAs in practical clinical applications.

A new N,O-rich covalent organic framework (COFDHNDA-BTH) was synthesized by an amine-aldehyde condensation reaction between 2,6-dialdehyde-1,5-dihydroxynaphthalene (DHNDA) and 1,3,5-phenyltriformylhydrazine (BTH) for carbaryl detection. The free NH, OH, and C=O groups of COFDHNDA-BTH not only covalently couples with acetylcholinesterase (AChE) into the pores of COFDHNDA-BTH, but also greatly improves the catalytic activity of AChE in the constrained environment of COFDHNDA-BTH’s pore. Under the catalysis of AChE, the acetylthiocholine (ATCl) was decomposed into positively charged thiocholine (TCl), which was captured on the COFDHNDA-BTH modified electrode. The positive charges of TCl can attract anionic probe [Fe(CN)6]3−/4− on the COFDHNDA-BTH-modified electrode to show a good oxidation peak at 0.25 V (versus a saturated calomel electrode). The carbaryl detection can inhibit the activity of AChE, resulting in the decrease in the oxidation peak. Therefore, a turn-off electrochemical carbaryl biosensor based on a flexible carbon paper electrode loaded with COFDHNDA-BTH and AChE was constructed using the oxidation peak of an anionic probe [Fe(CN)6]3−/4− as the detection signal. The detection limit was 0.16 μM (S/N = 3), and the linear range was 0.48~35.0 μM. The sensor has good selectivity, repeatability, and stability, and has a good application prospect in pesticide detection.

The spread and resurgence of the SARS-CoV-2 virus (COVID-19 disease) threatens human health and social relations. Prevention of COVID-19 disease partly relies on fabricating low-cost, point-of-care (POC) sensing technology that can rapidly and selectively detect the SARS-CoV-2 virus. We report a colorimetric, paper-based polydiacetylene (PDA) biosensor, designed to detect SARS-CoV-2 spike protein in artificial saliva. Analytical characterizations of the PDA sensor using NMR and FT-IR spectroscopy showed the correct structural elucidation of PCDA-NHS conjugation. The PDA sensor platform containing the N-Hydroxysuccinimide ester of 10, 12-pentacosadiynoic acid (PCDA-NHS) was divided into three experimental PCDA-NHS concentration groups of 10%, 20%, and 30% to optimize the performance of the sensor. The optimal PCDA-NHS molar concentration was determined to be 10%. The PDA sensor works by a color change from blue to red as its colorimetric output when the immobilized antibody binds to the SARS-CoV-2 spike protein in saliva samples. Our results showed that the PDA sensing platform was able to rapidly and qualitatively detect the SARS-CoV-2 spike protein within the concentration range of 1 to 100 ng/mL after four hours of incubation. Further investigation of pH and temperature showed minimal influence on the PDA sensor for the detection of COVID-19 disease. After exposure to the SARS-CoV-2 spike protein, smartphone images of the PDA sensor were used to assess the sensor output by using the red chromatic shift (RCS) of the signal response. These results indicate the potential and practical use of this PDA sensor design for the rapid, colorimetric detection of COVID-19 disease in developing countries with limited access to medical testing.

In vitro diagnosis (IVD) has become a hot topic in laboratory research and achievement transformation. However, due to the high cost, and time-consuming and complex operation of traditional technologies, some new technologies are being introduced into IVD, to solve the existing problems. As a result, IVD has begun to develop toward point-of-care testing (POCT), a subdivision field of IVD. The pandemic has made governments and health institutions realize the urgency of accelerating the development of POCT. Microfluidic paper-based analytical devices (μPADs), a low-cost, high-efficiency, and easy-to-operate detection platform, have played a significant role in advancing the development of IVD. μPADs are composed of paper as the core material, certain unique substances as reagents for processing the paper, and sensing devices, as auxiliary equipment. The published reviews on the same topic lack a comprehensive and systematic introduction to μPAD classification and research progress in IVD segmentation. In this paper, we first briefly introduce the origin of μPADs and their role in promoting IVD, in the introduction section. Then, processing and detection methods for μPADs are summarized, and the innovative achievements of μPADs in IVD are reviewed. Finally, we discuss and prospect the upgrade and improvement directions of μPADs, in terms of portability, sensitivity, and automation, to help researchers clarify the progress and overcome the difficulties in subsequent μPAD research.

In this study, we developed a rotatable paper device integrating loop-mediated isothermal amplification (RT-LAMP) and a novel naked-eye readout of the RT-LAMP results using a food additive, carmoisine, for infectious pathogen detection. Hydroxyl radicals created from the reaction between CuSO4 and H2O2 were used to decolor carmoisine, which is originally red. The decolorization of carmoisine can be interrupted in the presence of DNA amplicons produced by the RT-LAMP reaction due to how DNA competitively reacts with the hydroxyl radicals to maintain the red color of the solution. In the absence of the target DNA, carmoisine is decolored, owing to its reaction with hydroxyl radicals; thus, positive and negative samples can be easily differentiated based on the color change of the solution. A rotatable paper device was fabricated to integrate the RT-LAMP reaction with carmoisine-based colorimetric detection. The rotatable paper device was successfully used to detect SARS-CoV-2 and SARS-CoV within 70 min using the naked eye. Enterococcus faecium spiked in milk was detected using the rotatable paper device. The detection limits for the SARS-CoV-2 and SARS-CoV targets were both 103 copies/µL. The rotatable paper device provides a portable and low-cost tool for detecting infectious pathogens in a resource-limited environment.

The illegal use of β-adrenergic agonists during livestock growth poses a threat to public health; the long-term intake of this medication can cause serious physiological side effects and even death. Therefore, rapid detection methods for β-adrenergic agonist residues on-site are required. Traditional detection methods such as liquid chromatography have limitations in terms of expensive instruments and complex operations. In contrast, paper methods are low cost, ubiquitous, and portable, which has led to them becoming the preferred detection method in recent years. Various paper-based fluidic devices have been developed to detect β-adrenergic agonist residues, including lateral flow immunoassays (LFAs) and microfluidic paper-based analytical devices (μPADs). In this review, the application of LFAs for the detection of β-agonists is summarized comprehensively, focusing on the latest advances in novel labeling and detection strategies. The use of μPADs as an analytical platform has attracted interest over the past decade due to their unique advantages and application for detecting β-adrenergic agonists, which are introduced here. Vertical flow immunoassays are also discussed for their shorter assay time and stronger multiplexing capabilities compared with LFAs. Furthermore, the development direction and prospects for the commercialization of paper-based devices are considered, shedding light on the development of point-of-care testing devices for β-adrenergic agonist residue detection.

Sepsis is an immune response to a microbial invasion that causes organ injury and dysfunction due to a systemic inflammatory response. Sepsis is a serious, life-threatening condition and a widely recognized global health challenge. Given its high death rate, it is critical to diagnose sepsis and start treatment as early as possible. There is an urgent need for a sensitive and rapid screening method for detecting sepsis. In this study, we investigated the use of MMP-9 as a biomarker for sepsis. A colorimetric paper-based biosensor was used for the detection of MMP-9 utilizing peptide-magnetic nanoparticle conjugates. The method is based on the cleavage of the MMP-9-specific peptide by the protease leading to the detaching of the magnetic beads from the sensor surface and changing of color. A fecal intraperitoneal (FIP) challenge was used to induce sepsis in mice, and an MMP-9 secretion was measured by taking blood and Bronchoalveolar Lavage (BAL) fluid samples at 1 h, 2 h, 4 h, and 20 h (early sepsis) post-challenge intervals. The results of the paper-based sensor for the detection of MMP-9 levels in blood samples and BAL samples were compared with ELISA and Western Blot. We found that both blood and BAL levels of MMP-9 increased immediately and could be detected as early as 1 h in FIP mice post-challenge. Our work adds evidence to the assertion that MMP-9 is a reliable biomarker for the detection of sepsis at early stages.

This study investigated the autohydrolysis pre-treatment prior to chemi-thermomechanical pulping (CTMP) process including the effects of autohydrolysis pretreatment with a lower severity on characteristics of the autohydrolysis liquor and hydrolyzed chips. The intensity of autohydrolysis, characterized by the severity factor, increased from 1.76 to 3.54, the hydrolyzed chips yield decreased from 99.0% to 86.7% and the xylose extraction yield increased from 0.8% to 44.0%. The content of holocellulose, pentosan and acid soluble lignin remained in the hydrolyzed chips decreased dramatically with increasing the treatment severity. After the autohydrolysis pretreatment, the resultant poplar chemi-thermomechanical pulp with a severity factor of 2.37 could obtain a 15.3 Nm/g tensile index, 58.6% ISO brightness and a 2.39 cm(3)/g bulk., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

In this work, corn stover was refined by a pulp refining instrument (PFI refiner) after NaOH pretreatment under varied conditions. The quantitative characterization of the influence of PFI refining on enzymatic hydrolysis was studied, and it was proved that the enhancement of enzymatic saccharification by PFI refining of the pretreated corn stover was largely due to the significant increment of porosity of substrates and the reduction of cellulose crystallinity. Furthermore, a linear relationship between beating degree and final total sugar yields was found, and a simple way to predict the final total sugar yields by easily testing the beating degree of PFI refined corn stover was established. Therefore, this paper provided the possibility and feasibility for easily monitoring the fermentable sugar production by the simple test of beating degree, and this will be of significant importance for the monitoring and controlling of industrial production in the future., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Wheat straw was subjected to three different processes prior to saccharification, namely alkaline pulping, natural pulping and autohydrolysis, in order to study their effect on the rate of enzymatic hydrolysis. Parameters like medium concentration, temperature and time have been varied in order to optimize each method. Milling the raw material to a length of 4mm beforehand showed the best cost-value-ratio compared to other grinding methods studied. Before saccharification the pulp can be stored in dried form, leading to a high yield of glucose. Furthermore the relation of pulp properties (i.e. intrinsic viscosity, Klason-lignin and hemicelluloses content, crystallinity, morphology) to cellulose hydrolysis is discussed., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Lignin removal is essential for value-added utilization of hemicelluloses and acetic acid present in the prehydrolysis liquor (PHL) of a kraft-based hardwood dissolving pulp production. In this paper, a novel process concept, consisting of laccase-induced lignin polymerization, followed by filtration/flocculation, was developed to enhance the lignin removal. The results showed that the lignin removal increased from 11% to 46-61% at laccase concentration of 1-4 U mL(-1). The GPC results showed that the molecular weight of the lignin from the laccase treated PHL was increased by 160% in comparison with the original one. The subsequent flocculation using singular Poly-DADMAC system or dual polymer system of Poly-DADMAC/CPAM can further remove 10-15% lignin. The concentrations of hemicelluloses and acetic acid were negligibly affected during the laccase treatment, while flocculation caused 12-15% of total sugar loss. Additionally, the process incorporates this new concept into the kraft-based dissolving pulp production process was proposed., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Treating ground bagasse or Southern yellow pine in the biodegradable ionic liquid (IL), choline acetate ([Cho][OAc]), at 100°C for 24h led to dissolution of hemicellulose and lignin, while leaving the cellulose pulp undissolved, with a 54.3% (bagasse) or 34.3% (pine) reduction in lignin content. The IL solution of the dissolved biopolymers can be separated from the undissolved particles either by addition of water (20 wt% of IL) followed by filtration or by centrifugation. Hemicellulose (19.0 wt% of original bagasse, 10.2 wt% of original pine, containing 14-18 wt% lignin) and lignin (5.0 wt% of original bagasse, 6.0 wt% of original pine) could be subsequently precipitated. The pulp obtained from [Cho][OAc] treatment can be rapidly dissolved in 1-ethyl-3-methylimidazolium acetate (e.g., 17 h for raw bagasse vs. 7h for pulp), and precipitated as cellulose-rich material (CRM) with a lower lignin content (e.g., 23.6% for raw bagasse vs. 10.6% for CRM)., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The spent liquor (SL) of a neutral sulfite semichemical pulping process contains lignocelluloses that are currently treated in a waste water system. In this work, an adsorption process using activated carbon (AC) was considered for isolating the lignin and hemicelluloses from SL. The maximum adsorptions of 0.9 g/g lignin and 0.43 g/g of hemicelluloses on AC were achieved under the conditions of 30°C, pH 7 and 3h with SL/AC weight ratio of 90. The addition of polydiallyldimethylammonium chloride (PDADMAC) to the SL/AC system significantly improved the adsorption of lignin to 2.5 g/g on AC. The molecular weight of PDADMAC considerably affected the results in that the higher MW PDADMAC led to less lignin, but more hemicelluloses, turbidity and chemical oxygen demand removals from the SL. The thermal analysis also revealed that the higher MW PDADMAC generated precipitates with a lower incineration temperature and heating value., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Growing interest in alternative and renewable energy sources has brought increasing attention to the integration of a pulp mill into a forest biorefinery, where other products could be produced in addition to pulp. To achieve this goal, hemicelluloses were extracted, either by steam explosion or by steam treatment, from Eucalyptus globulus wood prior to pulping. The effects of both pre-treatments in the subsequent kraft pulping and paper strength were evaluated. Results showed a similar degree of hemicelluloses extraction with both options (32-67% of pentosans), which increased with the severity of the conditions applied. Although both pre-treatments increased delignification during pulping, steam explosion was significantly better: 12.9 kappa number vs 22.6 for similar steam unexploded pulps and 40.7 for control pulp. Finally, similar reductions in paper strength were found regardless of the type of treatment and conditions assayed, which is attributed to the increase of curled and kinked fibers., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The commercial pre-hydrolysis kraft-based dissolving pulp production process can be a typical example for the demonstration/implementation of the integrated forest biorefinery concept. In this study, the concept of cellulase treatment of this dissolving pulp for enhancement of accessibility/reactivity in terms of viscose rayon production was demonstrated. The cellulase treatment resulted in the formation of additional openings/surface areas in the fiber structure via the possible action of "etching". As a result, the pore volume of pulp fibers increased, which led to the increase in the accessibility to xanthation, and thus Fock reactivity. Results showed that the cellulase treatment was effective in increasing the Fock reactivity, at a cellulase dosage of 2u/g (based on the dry weight of pulp), the Fock reactivity increased from 47.67% to 79.9%. The adoption of cellulase treatment to hardwood kraft-based dissolving pulp can provide an efficient approach for enhancing its performance in the commercial viscose-rayon process., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Research background. Fish freshness and quality monitoring are of high importance for consumers, retailers and fishing industry. Therefore, developing novel approaches that are simple, fast, non-destructive and inexpensive to monitor fish freshness in real time is of great value. One alternative is using Intelligent or smart packaging to monitor the freshness or conditions of packaged fish. Experimental approach. On-package dual indicator label based on paper-based pH sensors was developed for real-time monitoring of the milkfish (Chanos chanos) freshness. The paper-based pH sensor was prepared using bromocresol purple (BCP) and bromothymol blue (BTB) that were immobilized onto a filter paper by dip coating. Herein, the fish degradation could be monitored visually by the dual indicator label, where the BCP changes from yellow to pink, then finally to purple, while the BTP changes from orange to green-yellow, and finally to green-blue to indicate fresh, medium fresh or spoiled product, respectively. Results and conclusion. The label responds to the pH change caused by the fish degradation and the colour of dual indicator changes to show the fish freshness at room temperature and chiller conditions. This pH change was followed by changes in the other parameters related to fish freshness, such as total volatile basic nitrogen (TVBN), total viable count (TVC), texture and odour. The threshold of fish spoilage at room temperature was observed at 8 h and under chiller conditions at 7 days when the deterioration time point was indicated by the colour changes. Thus, it can be concluded that the dual indicator label can be applied as a simple and low-cost on-package active label for fish freshness monitoring. Novelty and scientific contribution. Increasing consumer concerns about quality and safe food worldwide has boosted the search for a novel approach to food monitoring. In this work, a simple and practical on-package dual indicator label for real-time monitoring of fish freshness was developed. The colorimetric pH sensor was obtained simply by dip-coating of filter paper, yet it enables easy and accurate detection of fish spoilage with the naked eye. Similarly, the dual indicator label changes colour for other freshness parameters, such as TVBN, TVC, texture and odour.

A novel assay platform consisting of a microfluidic sliding double-track paper-based chip and a hand-held Raspberry Pi detection system is proposed for determining the albumin-to-creatine ratio (ACR) in human urine. It is a clinically important parameter and can be used for the early detection of related diseases, such as renal insufficiency. In the proposed method, the sliding layer of the microchip is applied and the sample diffuses through two parallel filtration channels to the reaction/detection areas of the microchip to complete the detection reaction, which is a simple method well suited for self-diagnosis of ACR index in human urine. The RGB (red, green, and blue) value intensity signals of the reaction complexes in these two reaction zones are analyzed by a Raspberry Pi computer to derive the ACR value (ALB and CRE concentrations). It is shown that the G + B value intensity signal is linearly related to the ALB and CRE concentrations with the correlation coefficients of R2 = 0.9919 and R2 = 0.9923, respectively. It is additionally shown that the ALB and CRE concentration results determined using the proposed method for 23 urine samples were collected from real suffering chronic kidney disease (CKD) patients are in fine agreement with those acquired operating a traditional high-reliability macroscale method. Overall, for point-of-care (POC) CKD diagnosis and monitoring in clinical applications, the results prove that the proposed method offers a convenient, real time, reliable, and low-spending solution for POC CKD diagnosis.

Bleached cellulose with good end-properties (≈ 90% ISO brightness and 62% cellulose preservation) was obtained by using a totally chlorine-free biobleaching process (TCF). Unbleached sulphite cellulose was treated with Trametes villosa laccase in combination with violuric acid. This enzymatic stage (L) was followed by a chelating stage (Q) and then by a hydrogen peroxide stage reinforced with pressurized oxygen (Po), resulting to an overall LQPo sequence. The use of violuric acid was dictated by the results of a preliminary study, where the bleaching efficiency of various natural (syringaldehyde and p-coumaric acid) and synthetic mediators (violuric acid and 1-hydroxybenzotriazole) were assessed. The outstanding results obtained with laccase-violuric acid system fulfil most of the characteristics of commercial dissolving pulp, totally acceptable for viscose manufacturing or CMC derivatives, with the added advantage that the enzymatic treatment saved 2h of reaction time and about 70% of hydrogen peroxide consumption, relative to a conventional sequence (Po)., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

While computational biology and bioinformatics became an inherent part of most life science degrees, it remains challenging to encourage students to employ these diverse skills in a practical research context. In this education paper, we present a method to enable students to acquire hands-on skills by the data-driven study of complex biomolecular datasets. The course starts with a series of introductory lectures, including a practical session on Cytoscape and associated computational network biology techniques. As we believe that enabling students to work on their own research questions will increase their motivation and interest, we introduced a recent hallmark virus-host protein-protein interaction article from which the database served as a starting point for the students to develop their research goal, which was presented and discussed in a first informal meeting. Over the next two months, about four interactive sessions were held, where students presented their problems, progress and challenges and exchanged ideas to help each other reach their goals. The role of the lecturers was to assist the students with independently collecting data and exploring new software tools for their research question. We present this concept and methods, and we share our experiences from an evaluation of the student learning outcomes.

Synthetic organic compounds (SOCs) are reported as xenobiotics compounds contaminating the environment from various sources including waste from the pulp and paper industries: Since the demand and production of paper is growing increasingly, the release of paper and pulp industrial waste consisting of SOCs is also increasing the SOCs’ pollution in natural reservoirs to create environmental pollution. In pulp and paper industries, the SOCs viz. phenol compounds, furans, dioxins, benzene compounds etc. are produced during bleaching phase of pulp treatment and they are principal components of industrial discharge. This review gives an overview of various biotechnological interventions for paper mill waste effluent management and elimination strategies. Further, the review also gives the insight overview of various ways to restrict SOCs release in natural reservoirs, its limitations and integrated approaches for SOCs bioremediation using engineered microbial approaches. Furthermore, it gives a brief overview of the sustainable remediation of SOCs via genetically modified biological agents, including bioengineering system innovation at industry level before waste discharge.

Paper is a popular platform material in all areas of sensor research due to its porosity, large surface area, and biodegradability, to name but a few. Many paper-based nanocomposites have been reported in the last decade as novel substrates for surface-enhanced Raman spectroscopy (SERS). However, there are still limiting factors, like the low density of hot spots or loss of wettability. Herein, we designed a process to fabricate a silver–chitosan nanocomposite layer on paper celluloses by a layer-by-layer method and pH-triggered chitosan assembly. Under microscopic observation, the resulting material showed a nanoporous structure, and silver nanoparticles were anchored evenly over the nanocomposite layer. In SERS measurement, the detection limit of 4-aminothiophenol was 5.13 ppb. Furthermore, its mechanical property and a strategy toward further biosensing approaches were investigated.

In order to facilitate monitoring of cannabidiol (CBD), we devised a gold immunochromatographic sensor based on a specific monoclonal antibody (mAb). To prepare the antigen, a novel hapten with CBD moiety and a linear carbon chain was employed. By utilizing hybridoma technology, a specific mAb was screened and identified that exhibited a 50% maximal inhibitory concentration against CBD ranging from 28.97 to 443.97 ng/mL. Extensive optimization led to the establishment of visual limits of detection for CBD, achieving a remarkable sensitivity of 8 μg/mL in the assay buffer. To showcase the accuracy and stability, an analysis of CBD-spiked wine, sparkling water, and sports drink was conducted. The recovery rates observed were as follows: 88.4–109.2% for wine, 89.9–107.8% for sparkling water, and 83.2–95.5% for sports drink. Furthermore, the coefficient of variation remained impressively low, less than 4.38% for wine, less than 2.07% for sparkling water, and less than 6.34% for sports drink. Importantly, the developed sensor exhibited no cross-reaction with tetrahydrocannabinol (THC). In conclusion, the proposed paper sensor, employing gold nanoparticles, offers a user-friendly and efficient approach for the precise, rapid, and dependable determination of CBD in products.

Abstract By way of broadening the use of diverse sustainable bioethanol feedstocks, the potentials of Paper mulberry fruit juice (PMFJ), as a non-food, sugar-based substrate, were evaluated for fuel ethanol production. The suitability of PMFJ was proven, as maximum ethanol concentration (56.4 g/L) and yield (0.39 g/g) were achieved within half a day of the start of fermentation, corresponding to very high ethanol productivity of 4.7 g/L/hr. The established potentials were further optimally maximized through the response surface methodology (RSM). At the optimal temperature of 30 °C, yeast concentration of 0.55 g/L, and pH of 5, ethanol concentration, productivity, and yield obtained were 73.69 g/L, 4.61 g/L/hr, and 0.48 g/g, respectively. Under these ideal conditions, diverse metal salts were afterward screened for their effects on PMFJ fermentation. Based on a two-level fractional factorial design, nutrient addition had no positive impact on ethanol production. Thus, under the optimal process conditions, and without any external nutrient supplementation, bioethanol from PMFJ compared favorably with typical sugar-based energy crops, highlighting its resourcefulness as a high-value biomass resource for fuel ethanol production. Graphical Abstract

DNA microarray technology is revolutionizing biological science. DNA microarrays (also called DNA chips) allow simultaneous screening of many genes for changes in expression between different cells. Now researchers can obtain information about genes in days or weeks that used to take months or years. The paper activity described in this article allows students to visualize how cells grown under different conditions would be analyzed using a DNA microarray. This simple microarray experiment involves looking for differences in gene expression of seedlings grown under two conditions: One set of seedlings receives full light and the other set germinates completely in the dark. This activity also reinforces concepts related to DNA structure, transcription, and experimental design. (Contains 5 figures.)

The pulp and paper industry is highly energy-intensive. In mills that use chemical pulping, roughly half of the higher heating value of the cellulosic material used to manufacture the product typically is incinerated to generate steam and electricity that is needed to run the processes. Additional energy, much of it non-renewable, needs to be purchased. This review considers publications describing steps that pulp and paper facilities can take to operate more efficiently. Savings can be achieved, for instance, by minimizing unnecessary losses in exergy, which can be defined as the energy content relative to a standard ambient condition. Throughout the long series of unit operations comprising the conversion of wood material to sheets of paper, there are large opportunities to more closely approach a hypothetical ideal performance by following established best-practices.

Abstract Aβ42 is one of the most extensively studied blood and Cerebrospinal fluid (CSF) biomarkers for the diagnosis of symptomatic and prodromal Alzheimer’s disease (AD). Because of the heterogeneity and transient nature of Aβ42 oligomers (Aβ42Os), the development of technologies for dynamically detecting changes in the blood or CSF levels of Aβ42 monomers (Aβ42Ms) and Aβ42Os is essential for the accurate diagnosis of AD. The currently commonly used Aβ42 ELISA test kits usually mis-detected the elevated Aβ42Os, leading to incomplete analysis and underestimation of soluble Aβ42, resulting in a comprised performance in AD diagnosis. Herein, we developed a dual-target lateral flow immunoassay (dLFI) using anti-Aβ42 monoclonal antibodies 1F12 and 2C6 for the rapid and point-of-care detection of Aβ42Ms and Aβ42Os in blood samples within 30 min for AD diagnosis. By naked eye observation, the visual detection limit of Aβ42Ms or/and Aβ42Os in dLFI was 154 pg/mL. The test results for dLFI were similar to those observed in the enzyme-linked immunosorbent assay (ELISA). Therefore, this paper-based dLFI provides a practical and rapid method for the on-site detection of two biomarkers in blood or CSF samples without the need for additional expertise or equipment. Graphical Abstract

Laccases are multicopper enzymes present in plants, fungi, bacteria, and insects, which catalyze oxidation reactions together with four electron reduction of oxygen to water. Plant, bacterial, and insect laccases have a polymerizing role in nature, implicated in biosynthesis of lignin, melanin formation, and cuticle hardening, respectively. On the other hand, fungal laccases carry out both polymerizing (melanin synthesis and fruit body formation) as well as depolymerizing roles (lignin degradation). This bifunctionality of fungal laccases can be attributed to the presence of multiple isoforms within the same as well as different genus and species. Interestingly, by manipulating culture conditions, these isoforms with their different induction patterns and unique biochemical characteristics can be expressed or over-expressed for a targeted biotechnological application. Consequently, laccases can be considered as one of the most important biocatalyst which can be exploited for divergent industrial applications viz. paper pulp bleaching, fiber modification, dye decolorization, bioremediation as well as organic synthesis. The present review spotlights the role of fungal laccases in various antagonistic applications, i.e., polymerizing and depolymerizing, and co-relating this dual role with potential industrial significance.

Kraft lignin (KL), is the major pollutant in pulp and paper effluent and due to its heterogeneous structure, it is resistant to the depolymerization process. It has drawn much attention from the researcher due to its challenging degradation process. In this study, a KL-degrading bacterium was isolated and screened from paper mill sludge. This bacterium was identified as ligninolytic Bacillus aryabhattai using biochemical and 16SrRNA gene analysis. B. aryabhattai showed maximum activities of lignin peroxidase-LiP (0.74 IU mL−1) and manganese peroxidase-MnP (9.2 IU mL−1) on the 4th day, and 5th day, respectively. A total 84% of KL (500 mg L−1) reduction was observed after 14 days. The KL bio-degradation was confirmed based on changes in chemical stracture of KL and new metabolites identification using FTIR and GC–MS, respectively. The study concluded that B. aryabhattai maybe becomes a potential biological agent in KL biodegradation and treatment of other lignin-containing industrial effluents.

Summary: Fifteen percent of couples of reproductive age suffer from infertility globally and the burden of infertility disproportionately impacts residents of developing countries. Assisted reproductive technologies (ARTs), including in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI), have been successful in overcoming various reasons for infertility including borderline and severe male factor infertility which consists of 20%–30% of all infertile cases. Approximately half of male infertility cases stem from suboptimal sperm parameters. Therefore, healthy/normal sperm enrichment and sorting remains crucial in advancing reproductive medicine. Microfluidic technologies have emerged as promising tools to develop in-home rapid fertility tests and point-of-care (POC) diagnostic tools. Here, we review advancements in fabrication methods for paper-based microfluidic devices and their emerging fertility testing applications assessing sperm concentration, sperm motility, sperm DNA analysis, and other sperm functionalities, and provide a glimpse into future directions for paper-based fertility microfluidic systems.

Xylanases have the potential to be used as bio-deinking and bio-bleaching materials and their application will decrease the consumption of the chlorine-based chemicals currently used for this purpose. However, xylanases with specific properties could act effectively, such as having significant thermostability and alkali resistance, etc. In this study, we found that TfXyl10A, a xylanase from Thermobifida fusca, was greatly induced to transcript by microcrystalline cellulose (MCC) substrate. Biochemical characterization showed that TfXyl10A is optimally effective at temperature of 80 °C and pH of 9.0. After removing the carbohydrate-binding module (CBM) and linker regions, the optimum temperature of TfXyl10A-CD was reduced by 10°C (to 70°C), at which the enzyme’s temperature tolerance was also weakened. While truncating only the CBM domain (TfXyl10AdC) had no significant effect on its thermostability. Importantly, polysaccharide-binding experiment showed that the auxiliary domain CBM2 could specifically bind to cellulose substrates, which endowed xylanase TfXyl10A with the ability to degrade xylan surrounding cellulose. These results indicated that TfXyl10A might be an excellent candidate in bio-bleaching processes of paper industry. In addition, the features of active-site architecture of TfXyl10A in GH10 family were further analyzed. By mutating each residue at the -2 and -1 subsites to alanine, the binding force and enzyme activity of mutants were observably decreased. Interestingly, the mutant E51A, locating at the distal -3 subsite, exhibited 90% increase in relative activity compared with wild-type (WT) enzyme TfXyl10A-CD (the catalytic domain of TfXyl110A). This study explored the function of a GH10 xylanase containing a CBM2 domain and the contribution of amino acids in active-site architecture to catalytic activity. The results obtained provide guidance for the rational design of xylanases for industrial applications under high heat and alkali-based operating conditions, such as paper bleaching.

This work validates a new strategy for complete utilization of palm residues by separating fibers and parenchyma for the respective purposes of pulping and biorefining. The parenchyma cells were fractionated from royal palm sheath (RPS) after neutral sulfite semichemical (NSSC) cooking for producing fermentable sugars, leaving vascular bundles for manufacturing pulp and paper. Parenchyma cells could be readily and completely screened out prior to defibration. They were more digestible by cellulase than vascular bundles or the pulp derived from them. Cellulose enzymatic digestibility (CED) of parenchyma cells rapidly reached 82% in 12-h hydrolysis and finally up to 92%. The CEDs of parenchyma were maintained around 90% at a medium solid consistency, 12% (w/w). The average length, retention and drainability of RPS pulp were all improved without loss of physical strength after removing parenchyma. This work may help establish a new platform for maximizing the utilization efficiency of parenchyma-rich biomass., (Copyright © 2013 Elsevier Ltd. All rights reserved.)