Reagent Costs Research Articles

Lipidure-based micropattern fabrication for stereotyping cell geometry

Cell autonomous behaviors such as migration and orchestration of cell polarity programs are required for physiological tissue formation. Micropatterns are cell-adhesive shapes that confine cell(s) to a user defined geometry. This biophysical confinement allows researchers to standardize the cell shape, and in doing so, stereotype organelle and cytoskeletal systems that can have an arbitrary organization. Thus, micropatterning can be a powerful tool in interrogation of polarity programs by enforcing a homogenous cell shape and cytoskeletal organization. A major drawback of this approach is the equipment and reagent costs associated with fabrication. Here, we provide a characterization of a compound called Lipidure (2-Methacryloyloxy ethyl phosphorylcholine) that is up to 40X less expensive than other cell repulsive coating agents. We found that Lipidure is an effective cell-repulsive agent for photolithography-based micropattern fabrication. Our results demonstrate that Lipidure is sensitive to deep UV irradiation for photolithography masking, stable in both benchtop and aqueous environments, non-toxic in prolonged culture, and effective at constraining cell geometry for quantification of cytoskeletal systems.

An efficient and low-cost method for the determination of selenium in selenium-enriched tea by high-performance liquid chromatography coupled with 3,3′-Diaminobenzidine derivatization

In this study, a novel analytical method was developed and validated for the quantification of selenium in selenium-enriched tea. The various valence states of selenium present in tea were converted to Se(IV) through oxidation and reduction reactions. To chemically react with Se(IV), 3,3′-Diaminobenzidine (DAB) was employed to produce Se-DAB derivatives. These derivatives were then analyzed and quantified using high-performance liquid chromatography (HPLC) with a NH2 column specifically designed for the target compound. The peak area of Se-DAB exhibited excellent linearity within the concentration range of 0.01–50 μgmL−1 (R2 = 0.9999). The method demonstrated a limit of detection (LOD) of 0.01 mgkg−1 and a limit of quantification (LOQ) of 0.03 mgkg−1. Moreover, the recoveries of Se-DAB spiked at different concentrations ranged from 81.2 % to 96.7 %, with relative deviations (RSDs) of 0.72 %–4.23 % (n = 6). Additionally, the RSDs under different batches varied from 3.42 % to 8.57 % (n = 9). The method exhibits good stability and repeatability. The results obtained by applying the method to 24 samples of selenium-rich tea show high consistency with the AFS method. The difference range between the two methods is 0.017–0.027 mg/kg, with an average difference of 0.005 mg/kg. Furthermore, this method offers the advantage of flexibility in analyzing large quantities of green tea samples, while utilizing lower instrument and reagent costs. Therefore, it can be considered as a new and cost-effective method for detecting selenium content in selenium-rich tea.

Characterizing a Cost-Effective Hydrogel-Based Transparent Soil.

Transparent soil (TS) was specifically designed to support root growth in the presence of air, water, and nutrients and allowed the time-resolved phenotyping of roots in vivo. Nevertheless, it is imperative to further optimize the reagent cost of TS to enable its wider utilization. We substituted the costly Phytagel obtained from Sigma with two more economical alternatives, namely Biodee and Coolaber. TS beads from each brand were prepared using 12 different polymer concentrations and seven distinct crosslinker concentrations. A comprehensive assessment encompassing transparency, mechanical characteristics, particle size, porosity, and stability of TS was undertaken. Compared to the Sigma Phytagel brand, both Biodee and Coolaber significantly reduced the transparency and collapse stress of the TS they produced. Consequently, this led to a significant reduction in the allowable width and height of the growth box, although they could still simultaneously exceed 20 cm and 19 cm. There was no notable difference in porosity and stability among the TS samples prepared using the three Phytagel brands. Therefore, it is feasible to consider replacing the Phytagel brand to reduce TS production costs. This study quantified the differences in TS produced using three Phytagel brands at different prices that will better promote the application of TS to root phenotypes.

Instantaneous and reversible flocculation of Scenedesmus via Chitosan and Xanthan Gum complexation

An instantaneous and reversible flocculation method for Scenedesmus harvesting was developed, based on the complexation of Chitosan (CTS) and Xanthan Gum (XG). Under rapid stirring, Scenedesmus cells formed centimeter-sized flocs within 20 s using binary flocculants of 4 mg/L CTS and 16 mg/L XG. These flocs exhibited a remarkable harvest efficiency exceeding 95 % when filtered through 500-μm-pore-sized sieves. Furthermore, the flocs could be completely disintegrated by using alkaline or NaCl solutions (pH > 11 or NaCl concentration > 1.5 mol/L). Adjusting pH allowed recovery of 50 % CTS and 75 % XG, resulting in microalgae biomass with lower flocculant content and reducing reagent costs. Electrostatic interaction of –COO– of XG and –NH3+ of CTS deduced the formation of polyelectrolyte complexes (PECs), which shrink and wrap the coexisting algal cells to form the flocs under stirring. CTS and XG complexation was instantaneous and reversible, explaining quick flocculation and disintegration.

Application of microfluidic technologies in forensic analysis.

The application of microfluidic technology in forensic medicine has steadily expanded over the last two decades due to the favorable features of low cost, rapidity, high throughput, user-friendliness, contamination-free, and minimum sample and reagent consumption. In this context, bibliometric methods were adopted to visualize the literature information contained in the Science Citation Index Expanded from 1989 to 2022, focusing on the co-occurrence analysis of forensic and microfluidic topics. A deep interpretation of the literature was conducted based on co-occurrence results, in which microfluidic technologies and their applications in forensic medicine, particularly forensic genetics, were elaborated. The purpose of this review is to provide an impartial evaluation of the utilization of microfluidic technology in forensic medicine. Additionally, the challenges and future trends of implementing microfluidic technology in forensic genetics are also addressed.

Lab-on-a-chip – The advent of instantaneous diagnosis for a plethora of diseases

A lab-on-a-chip (LOC) is a device that facilitates the incorporation of a concatenation of various laboratory functions on a unique integrated circuit. The size of this instrument is merely a few millimeters to centimeters designed to attain automation and high-throughput screening. Microfluidic systems used in LOC devices allow the manufacture of millions of microchannels, each measuring mere micrometers. These microchannels enable control of fluids in infinitesimal quantities for a variety of diagnoses. Several labs on a chip have been commercialized in recent times for fundamental procedures, including glucose monitoring, human immunodeficiency virus, early tumor detection, and cardiac diagnostics. The LOC integrates microfluidics, nanosensors, micro-electrics, and biochemistry on one device. The advantages of the chip include its sustainability and cutback wastage. It expedites a decline in reagent costs and requires minimal sample volumes. The analysis and response are faster and the response is better controlled by equipping micro-channels. Countries with exiguous healthcare are in the face of adversity due to increased fatality rates from infectious diseases that are often curable in developed nations. In certain circumstances, impoverished healthcare clinics have the medications requisite to treat a specific condition but are in dearth of the diagnostic equipment needed to determine, in which individuals are in need of the medications. This is where the role of LOC as a potent novel diagnostic instrument would benefit humankind in the nearest future, according to eminent researchers. This article highlights the applications of LOC in a miscellany of fields, its advantages, feasible means to overcome the drawbacks, and the propitious prospects of this technology.

Bioethanol production from mixed sugars at a semi-pilot scale through two-step repeated sequential fermentation

Assessing the industrial suitability of sequential culture for efficient conversion of mixed sugars from plant biomass hydrolysate is essential. In the present study, a two-stage sequential fermentation of mixed sugar-rich sugar cane bagasse hydrolysate was performed to produce bioethanol. In the first stage, glucose fermentation using Z. mobilis was carried out, followed by xylose fermentation using S. stipitis in the second. However, xylose fermentation was done only after completely separating primary cells and residual ethanol. Sequential fermentation of mixed sugar-rich bagasse hydrolysate yielded an ethanol titer of 77.7 g.L−1 with an average ethanol yield of 0.48 g.g−1 and overall ethanol productivity of 1.38 g.L−1.h−1. Further, repeated sequential fermentation (RSF) of mixed sugars was performed for two cycles. RSF assessed the fermentation performance of high cell density sequential culture achieved through sequential cell recycling. The maximum ethanol productivity achieved from glucose and xylose was 6.92 g.L−1.h−1 and 2.28 g.L−1.h−1, respectively. In addition, a techno-economic analysis found that the minimum selling price of the ethanol (MSP) produced from sequential fermentation of mixed sugar-rich bagasse hydrolysate (scenario I) was up to 1.7 US$. L−1. On the other hand, the MSP calculated after omitting reagent costs of synthetic glucose and xylose of the first scenario was up to 0.52 US$. L−1.

B-242 Fast and Cost-effective RNA Extraction in Onco-hematology Laboratories

Abstract Background Chronic myeloid leukemia (CML) is a myeloproliferative neoplasm characterized by the presence of a BCR-ABL1 rearrangement; its detection and quantification by real-time quantitative PCR (RQ-PCR) plays a central role in CML diagnosis, therapy monitoring, and sequencing to identify resistance mutations in case of therapy failure. To evaluate the molecular response of CML, BCR-ABL1 quantification must be calculated on the international scale (IS%) for which RQ-PCR has the necessary sensitivity (up to 10−5). Currently, BCR-ABL1 quantifications are ubiquitous in onco-hematology laboratories (OHLs; ∼70% of tests performed in our OHL), whereby quality, cost, and turn-around-time are of critical importance. Today, the majority of OHLs in Brazil rely on laborious manual RNA-extraction methods (e.g., Trizol) that usually require 8 mL of peripheral blood (PB) to monitor CML. Here, we report a strategy that requires only 2 mL of PB in combination with the automated magnetic DNA/RNA-extraction kit Extracta-MPTA (Loccus do Brasil) for CML diagnosis and monitoring. Methods: BCR:ABL1 was quantified and its IS% determined using our developed kit, which was calibrated using ERM-AD263 and UK-NEQAS (external quality assessment). Total RNA was extracted from 40 PB samples, after erythrocyte lysis using two different methods: 1) Leukocytes from 8 mL of PB were extracted manually using Trizol; 2) leukocytes from 2 mL of PB were automatically extracted using Extracta-MPTA (Loccus do Brasil). BCR-ABL1 quantification, BCR-ABL1 IS%, and BCR quantification (internal control) were compared statistically (paired t-test). The reproducibility was evaluated by analyzing 20 replicates; the low limit of the leucocyte number to BCR:ABL1 quantification using Extracta-MPTA were also evaluated. Median and moving means of 135 unpaired samples were used to monitor BCR-ABL1 IS% between Trizol, Extracta-MPTA, while EQA from UK-NEQAS was also evaluated using both methods. In parallel, RNA was extracted by both methods from 20 samples for the diagnosis of acute lymphoblastic leukemia (ALL) (BCR-ABL p190 transcript) and acute promyelocytic leukemia (APL) (PML-RARA transcript). Results A comparison of BRC-ABL1, BCR, and %IS in 40 paired samples showed that the results obtained from Trizol or Extracta-MPTA are comparable (paired t-test: P = 0.4222, P = 0.9263, and P = 0.071, respectively). The reproducibility was evaluated by BCR quantification (24%). A comparison of the medians from 135 unpaired samples extracted using Trizol and Extracta-MPTA revealed that BCR-ABL1 IS% distribution among our patients is stable over time (P = 0.7704; Mann Whitney: r2 = 0.0012). EQA was extracted using both methods and the results were identical. A comparison between Trizol and EXTRACTA_MPTA for ALL and APL was 100% identical (Fischer; P = 1.000). Conclusion Both RNA extraction methods are suitable for OHLs for PML-RARA and BCR-ABL (p190 and p210) detection and CML diagnosis/monitoring by BCR-ABL1 detection/quantification. By using Extracta-MPTA the amount of PB used to obtain RNA can be reduced by 75%, test results can be obtained within 4 h, and direct reagent costs are reduced by 25% with a minimum of hands-on time.

A CRISPR-Cas12a-based platform facilitates the detection and serotyping of Streptococcus suis serotype 2

Streptococcus suis serotype 2 is an economically important zoonotic pathogen that causes septicemia, arthritis, and meningitis in pigs and humans. S. suis serotype 2 is responsible for substantial economic losses to the swine industry and poses a serious threat to public health, and accurate and rapid detection is important for the prevention and control of epidemic disease. In this study, we developed a high-fidelity detection and serotyping platform for S. suis serotype 2 based on recombinase polymerase amplification (RPA) and a clustered regularly interspaced short palindromic repeat (CRISPR)-Cas12a system called Cards-SSJ/K. Cards-SSJ had a detection limit of 10 CFU, takes <60 min, and no cross-reaction was found with other S. suis serotypes, closely related Streptococcus spp., or common pig pathogens, and Cards-SSK could differentiate serotype 2 from serotype 1/2. Results from Cards-SSJ and qPCR were equivalent in detecting S. suis serotype 2 in tissue samples. Analysis indicated that despite a relatively high reagent cost compared to PCR and qPCR, Cards-SSJ was less time-consuming and had low requirements for equipment and personnel. Thus, it is an excellent method for point-of-care detection for S. suis serotype 2.

Separation and preparation of level I standard lead concentrate from lead ore via S-HGMS technology: description of separation mechanism

ABSTRACT To eliminate the adverse effects of traditional lead ore flotation methods, such as high reagent cost, complex operation process, and a large amount of wastewater, a superconducting high gradient magnetic separation (S-HGMS) technology was proposed to prepare level I standard lead concentrate (IGC) from lead ore. Under optimal conditions including a magnetic flow ratio of 0.076 T·s/m, a pulp concentration of 1.5%, a slurry flow velocity of 500 mL/min, and a 90% mass fraction of minus 45 μm, the Pb grade of the IGC product increased from 45.36% to 68.79% with 55.43% of Pb recovery, while all Cu, Zn, As, Fe, MgO, and Al2O3 concentrations of IGC decreased. The obtained IGC product meets the standard of level I in YS/T319–2013. The characterization results showed that lead ore comprised galena, sphalerite, and chalcopyrite. After S-HGMS, galena was effectively concentrated in the IGC product, while sphalerite and chalcopyrite were separated into the tailings. The magnetic field intensity and mineral magnetization characteristics were vital factors affecting the Pb grade and recovery of the IGC product. The S-HGMS technology efficiently achieved the preparation of the IGC product from lead ore, without using flotation agents and generating wastewater, and provides a valuable guideline for separating Cu – Pb, Zn – Pb, and Cu – Zn – Pb minerals.

Polyaniline and polystyrene sulfonic acid modified on conductive biochar nanoparticle with strong signal enhancement for trace atrazine detection

A label and reagent free and cost effective immunosensing system was newly developed for detection of trace leveled atrazine in water. This was constructed by synthesis of polyaniline (PA) and polystyrene sulfonic acid (PSS) modified biochar (BC) nanoparticle composite (PA-PSS-BC) and coating them on screen printed electrode as conductive and electrochemically active layer, and covalently immobilizing anti-atrazine antibody on electrode for specific target binding. The low cost biochar from sugarcane was highly conductive and favorable for attaching PA and PSS due to large surface area and sp2 carbon structure. Crediting to dopant PSS, PA-PSS-BC achieved significant enhancement of redox current signal even in phosphate buffered saline of neutral pH compared to that of without PSS. And redox current of PA was extremely stable with negligible signal change even after twenty cycles repeated reactions owning to protection by BC. These essentially ensured sensing platform being sensitively performed in neutral solution with exemption of adding external redox reagent. As a result, fairly low concentrations of atrazine could be detected within 5 min, with limit of detection being 0.407 ng/L in a dynamic detection range varying from 1 to 100 ng/L. Moreover, the immunosensor exhibited reasonable signal recoveries for detecting trace leveled atrazine in real water samples, high selectivity, and good stability with dry and cold storage.

HPV prevalence and genotype distribution in 2,306 patients with cervical squamous cell carcinoma in central and eastern China.

To explore the positivity rate and genotype distribution of human papillomavirus (HPV) in cervical squamous cell carcinoma (CSCC) tissues in central and eastern China and to provide theoretical basis for cervical cancer screening and prophylactic HPV vaccine development in China. DNA was extracted from paraffin-embedded tissues of CSCC samples and exfoliated cervical cells of cervical cancer screening populations. 23 HPV genotypes were detected by combining polymerase chain reaction (PCR) and reverse dot hybridized gene chip detection technology in 2,306 CSCC tissues and 10,245 cervical cancer screening populations. The genotype distribution of HPV infection was analyzed. The overall infection rate of HPVs in 2,306 CSCC patients was 92.71%. The frequency of single-type HPV infection and multiple-type HPV infection were 86.48% and 13.51%, respectively. The most common HPV genotypes detected in Chinese CSCC tissues were HPV-16, HPV-18, HPV-31, HPV-33, HPV-45, HPV-52, HPV-58, and HPV-59. The overall positivity rate of these eight high-risk HPV (HR-HPV) genotypes in HPV-positive CSCC was as high as 96.91%. Of which the positivity rate of seven HR-HPV genotypes related to nine-valent HPV vaccines in HPV-positive CSCC was 95.09%. Meanwhile, the overall infection rates of HR-HPV and low-risk HPV (LR-HPV) in female aged 35-64 years who underwent cervical cancer screening were 13.16% and 1.32%, respectively. The high-frequency HR-HPV genotypes in cervical cancer screening women were HPV-52, HPV-58, HPV-16, HPV-53, HPV-68, HPV-39, HPV-51, and HPV-56, with positivity rates of 2.25%, 1.60%, 1.31%, 1.22%, 0.93%, 0.92%, 0.78%, and 0.74%, respectively. Among women screened for cervical cancer in China, detecting the 8 high-frequency HR-HPV genotypes can reduce technical difficulty and reagent costs, while also improving the efficiency and effectiveness of cervical cancer screening. HPV genotyping assists gynecologists in assessing the risk of HR-HPV-positive cervical intraepithelial neoplasia and guiding them in implementing appropriate interventions. Furthermore, HPV genotyping is helpful for doctors to follow up HR-HPV-positive women and to evaluate the protective effect of HPV vaccine.

A Universal Hydrogel Microneedle for on-Site Detection of Small Molecules, Proteins, and Ribonucleic Acids

Point-of-care testing (POCT) of clinical biomarkers is critical to health monitoring and timely treatment, yet biosensing assays capable of detecting biomarkers without the need for costly external equipment and reagents are limited. Blood-based assays are, specifically, challenging as blood collection is invasive, and several processing steps are required. A promising alternative to blood is interstitial fluid (ISF), which originates from blood and fills the extracellular space; thus, it has common biomarkers with plasma/serum while also containing biomarkers unique to the local cells. A wide range of metabolites, including amino acids, lipids, nucleotides, and protein biomarkers, can be detected in ISF, emphasizing the potential of ISF for health monitoring. Microneedle (MN)-based biosensors have recently emerged as a promising approach for investigating the ISF. They enable minimally invasive penetration through the skin for ISF access. Specifically, Hydrogel-based MNs have two main advantages; i) their fabrication is cost-effective and simple, ii) they are highly biocompatible. Therefore, they overcome the challenges with conventional solid MNs. However, Hydrogel Microneedles (HMN) capable of both extracting ISF and sensing an analyte on-microneedle (without any need for post-sampling processing) have been limited. Here, we report a versatile assay that employs HMNs to both extract (ISF) and perform on-MN sensing of a specific analyte/biomarker in a minimally invasive manner. Our assay incorporates graphene oxide-nucleic acid (GO.NA) optical sensors for sensing. The GO.NA optical sensor consists of GO nanosheets conjugated to fluorophore-modified nucleic acid (NA), in which GO acts as a quencher. Single-stranded NAs have high affinity for binding to GO, therefore, in the absence of the biomarker of interest the NAs are tightly bound to GO; in turn, their fluorophore tag is quenched. However, in the presence of a specific biomarker, the NAs bind to their target, inducing a conformational change that distances the fluorophore tag from GO, leading to fluorescence recovery and generation of an optical signal. GO also acts as a substrate for NA immobilization; therefore, it protects the NA from degradation and prevents any potential NA release from HMN-GO.NA. To enable microscope-free patch visualization and optical measurements for POCT, we developed a miniaturized smartphone-based system that captures fluorescence images of the HMN-GO.NA patches (Scheme 1A and 1B), which are then analyzed using freely available software (ImageJ). Our system could successfully measure six clinically important biomarkers (glucose, uric acid, insulin, and serotonin as well as microribonucleic acid 210 and 21) in-vitro (using phantom gel) and ex-vivo (using porcine skin). Further, we have been able to accurately detect glucose and uric acid in live diabetic animal models, proving the efficacy of our system. Figure 1

Silver nanocluster-based aptasensor for the label-free and enzyme-free detection of ochratoxin A

Mycotoxin contamination in cereal is a global concern, threatening food safety and human health, necessitating the development of rapid on-site methods. Here, a label- and enzyme-free biosensor was developed based on aptamer-regulated DNA-silver nanoclusters (AgNCs) for rapid detection of ochratoxin A (OTA). A novel DNA-templated AgNCs emitting strong red fluorescence was designed and synthesized in this study. The partial sequence of the DNA template was selected from the complementary OTA aptamer (Apt-OTA) sequence, which can quench fluorescence from the AgNCs via hybridization in the absence of OTA. In the presence of OTA, the high OTA-Aptamer affinity prevented the Apt-OTA from quenching the AgNCs, resulting in “turn on” of the fluorescence. This biosensor eliminated the use of costly reagents, complex pretreatments, and sophisticated equipment, which could realize the point-of-care testing (POCT) of OTA with a limit of detection (LOD) of 1.3 nM and a detection time of 45 min.

Bioenergy Production from Agro-Industrial Wastewater Using Advanced Oxidation Processes as Pre-Treatment

Agro-industrial activities generate large volumes of wastewater. When this wastewater is discharged to the environment without proper treatment, it represents a serious problem. Bioenergy production can be conducted using wastewater, but the presence of some recalcitrant compounds may require a pre-treatment step. Advanced oxidation processes (AOPs) were traditionally used to treat hazardous materials but have recently been applied in various bioenergy production processes. AOPs are highly competitive water/wastewater treatment technologies and their application in the bioenergy sector is increasing as a pre-treatment process. Despite the increasing interest in using AOPs to enhance biofuel production, there is a lack of comprehensive documentation on their integration into biofuel production operations. This critical review highlights the application of AOPs as pre-treatment for agro-industrial wastewater (AIW) to enhance bioenergy production. It was noted that AOP applications can reduce the COD, VS, TS and total polyphenols, resulting in an improvement in their biodegradability. Moreover, these processes help remove hemicellulose and lignin contents, increasing the production of biogas, biodiesel and bioethanol. Among the different AOPs presented in this work, wet air oxidation showed promise for pre-treating lignocellulosic biomass to produce various energy types, while sonolysis and ozonation proved effective as a biosolid pre-treatment. Ozonolysis, Fenton reagents and photocatalysis are commonly used to selectively remove phenolic compounds and colorants from organic effluents. The high energy requirements and chemicals reagents costs are identified as obstacles to the application of AOPs in bioenergy production. Further studies should investigate the integration of AOPs with other treatment processes to improve the cost-effectiveness.

Micro-Colony Forming Unit Assay for Efficacy Evaluation of Vaccines against Tuberculosis.

Tuberculosis (TB), the leading cause of death worldwide by an infectious agent, killed 1.6 million people in 2022, only being surpassed by COVID-19 during the 2019-2021 pandemic. The disease is caused by the bacterium Mycobacterium tuberculosis (M.tb). The Mycobacterium bovis strain Bacillus Calmette-Guérin (BCG), the only TB vaccine, is the oldest licensed vaccine in the world, still in use. Currently, there are 12 vaccines in clinical trials and dozens of vaccines under pre-clinical development. The method of choice used to assess the efficacy of TB vaccines in pre-clinical studies is the enumeration of bacterial colonies by the colony-forming units (CFU) assay. This time-consuming assay takes 4 to 6 weeks to conclude, requires substantial laboratory and incubator space, has high reagent costs, and is prone to contamination. Here we describe an optimized method for colony enumeration, the micro-CFU (mCFU), that offers a simple and rapid solution to analyze M.tb vaccine efficacy results. The mCFU assay requires tenfold fewer reagents, reduces the incubation period threefold, taking 1 to 2 weeks to conclude, reduces lab space and reagent cost, and minimizes the health and safety risks associated with working with large numbers of M.tb. Moreover, to evaluate the efficacy of a TB vaccine, samples may be obtained from a variety of sources, including tissues from vaccinated animals infected with Mycobacteria. We also describe an optimized method to produce a unicellular, uniform, and high-quality mycobacterial culture for infection studies. Finally, we propose that these methods should be universally adopted for pre-clinical studies of vaccine efficacy determination, ultimately leading to time reduction in the development of vaccines against TB.

Biosensors as early warning detection systems for waterborne Cryptosporidium.

Waterborne disease is a global health threat contributing to a high burden of diarrhoeal disease, and growing evidence indicates a prospective increase in incidence coinciding with the profound effects of climate change. A major causative agent of gastrointestinal disease is Cryptosporidium, a protozoan waterborne parasite identified in over 70 countries. Cryptosporidium is a cause of high disease morbidity in children and the immunocompromised with limited treatment options for patients at risk of severe illness. The hardy nature of the organism leads to its persistence in various water sources, with certain water treatment procedures proving inefficient for its complete removal. While diagnostic methods for Cryptosporidium are well-defined in the clinical sphere, detection of Cryptosporidium in water sources remains suboptimal due to low dispersion of organisms in large sample volumes, lengthy processing times and high costs of equipment and reagents. A need for improvement exists to identify the organism as an emerging threat in domestic water systems, and the technological advantages that biosensors offer over current analytical methods may provide a preventative approach to outbreaks of Cryptosporidium. Biosensors are innovative, versatile and adaptable analytical tools that could provide highly sensitive, rapid, on-site analysis needed for Cryptosporidium detection in low-resource settings.

Methods for removing chloride ions to manufacture zinc from arc melting dust

In this work, we review literature sources to identify an industrial-scale technology for manufacturing cathode zinc from technogenic chlorine-containing raw materials of a complex chemical composition, in particular, from arc melting dust. The conducted review of Russian and foreign publications was used to search for a method of purifying process solutions with a high concentration of chloride ions produced by hydrometallurgical processing of technogenic arc melting dusts. It was found that the existing purification methods are associated with substantial limitations, including strict requirements for the acidity of the treated solution, low efficiency of treatment, secondary contamination of the medium by released chloride ions, and the high cost of reagents or equipment. Approaches to reducing the chlorine content in the initial dust and dechlorination of process solutions based on principles of sedimentation, ion exchange, sorption, and oxidation are described. In addition, we analyzed information published on chlorine removal from process effluents and solutions of various nature. Industrially-implemented and recently-reported laboratory methods of solution dechlorination were compared in terms of their feasibility, economic efficiency, and extent of chloride ion removal. It was concluded that the disadvantages of existing methods for processing technogenic chlorine-containing raw materials of arc melting can be eliminated by developing large, stable, and cheap hydride technologies based on principles of extraction, ion exchange, and sedimentation.

Smartphone-readable RPA-LFA for the high-sensitivity detection of Leishmania kDNA using nanophosphor reporters.

Early diagnosis of infectious diseases improves outcomes by enabling earlier delivery of effective treatment, and helps prevent further transmission by undiagnosed persons. We demonstrated a proof-of-concept assay combining isothermal amplification and lateral flow assay (LFA) for early diagnosis of cutaneous leishmaniasis, a vector-borne infectious disease that affects ca. 700,000 to 1.2 million people annually. Conventional molecular diagnostic techniques based on polymerase chain reaction (PCR) require complex apparatus for temperature cycling. Recombinase polymerase amplification (RPA) is an isothermal DNA amplification method that has shown promise for use in low-resource settings. Combined with lateral flow assay as the readout, RPA-LFA can be used as a point-of-care diagnostic tool with high sensitivity and specificity, but reagent costs can be problematic. In this work, we developed a highly-sensitive smartphone-based RPA-LFA for the detection of Leishmania panamensis DNA using blue-emitting [(Sr0.625Ba0.375)1.96Eu0.01Dy0.03]MgSi2O7 (SBMSO) persistent luminescent nanophosphors as LFA reporters. The greater detectability of nanophosphors allows the use of a reduced volume of RPA reagents, potentially reducing the cost of RPA-LFA. The limit of detection (LOD) of RPA with gold nanoparticle-based LFA readout is estimated at 1 parasite per reaction, but LOD can be 100-fold better, 0.01 parasites per reaction, for LFA based on SBMSO. This approach may be useful for sensitive and cost-effective point-of-care diagnosis and contribute to improved clinical and economic outcomes, especially in resource-limited settings.

JOINT PROCESSING OF QUARTZ-LEUCOXENE CONCENTRATE AND BRUCITE CONTAINING WASTE FROM THE PRODUCTION OF REFRACTORY MATERIALS TO OBTAIN COMPLEX COAGULANTS

The volumes of large-tonnage mineral waste generation cause serious concern from the side of environmental supervision. Most of the mineral waste is a valuable secondary raw material, but the lack of integrated technologies for their processing significantly slows down their processing. As part of the work, an assessment was made of the possibility of joint processing of quartz-leucoxene concentrate (waste from the oil production process) and brucite-containing waste from the production of refractories. Joint pyrometallurgical processing of two waste products made it possible to obtain magnesium titanium, a valuable product for various industries. Hydrometallurgical processing of magnesium titanate made it possible to obtain a complex magnesium-titanium-containing coagulant, which showed extremely high efficiency in the process of treating wastewater with a high pH level (concrete mixing unit). The use of a complex reagent, in addition to increasing the cleaning efficiency (reducing residual concentrations of pollutants), made it possible to reduce reagent costs and intensify the processes of sedimentation and filtration of coagulation sludge (simplification of the instrumental scheme).