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Microfluidic paper-based analytical devices (microPADs) are emerging as simple-to-use, low-cost point-of-care testing platforms. Such devices are mostly fabricated at present by creating hydrophobic barriers using wax or photoresist patterning on porous paper sheets. Even though devices fabricated using these methods are used and tested with a wide variety of analytes, still they pose many serious practical limitations for low-cost automated mass fabrication for their widespread applicability. We present an affordable and simple two-step process - cut and heat (CH-microPADs) - for the selective fabrication of hydrophilic channels and reservoirs on a wide variety of porous media such as tissue/printing/filter paper and cloth types, such as cotton and polyester, by a lamination process. The technique presents many advantages as compared to existing commonly used methods. The devices possess excellent mechanical strength against bending, folding and twisting, making them virtually unbreakable. They are structurally flexible and show good chemical resistance to various solvents, acids and bases, presenting widespread applicability in areas such as clinical diagnostics, biological sensing applications, food processing, and the chemical industry. Fabricated paper media 96 well-plate CH-microPAD configurations were tested for cell culture applications using mice embryonic fibroblasts and detection of proteins and enzymes using ELISA. With a simple two-step process and minimal human intervention, the technique presents a promising step towards mass fabrication of inexpensive disposable diagnostic devices for both resource-limited and developed regions.

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

Rabbit hemorrhagic disease virus 2 (RHDV2), a virulent and contagious viral pathogen that affects wild and domestic lagomorph populations, was identified in Wyoming, USA in December 2020. A surveillance program was developed involving full-carcass submission and liver analysis, although carcass quality as a result of predation and decomposition impeded analysis. To increase the number of submissions and provide flexibility to field staff, we evaluated 2 sample types: 77 dried blood on filter paper samples, 66 ear punch samples. At initial sampling, test specificity and sensitivity of the RT-rtPCR utilizing dried blood on filter paper and ear punch samples were both 100% compared to liver. Filter paper results were consistent over time; sensitivity stayed >96% through weeks 2, 4, and 6, with a maximum mean difference of 6.0 Ct from baseline liver Ct values (95% CI: 5.0–7.3) at 6 wk. Test sensitivity of the ear punch sample at 1, 3, 5, and 7 wk post-sampling remained at 100%, with a maximum mean difference of 5.6 Ct from baseline liver Ct values (95% CI: 4.3–6.9) at 5 wk. Filter paper and ear punch samples were suitable alternatives to liver for RHDV2 surveillance in wild lagomorph populations. Alternative sampling options provide more flexibility to surveillance programs, increase testable submissions, and decrease exposure of field personnel to zoonotic disease agents.

Allergy to Polistes dominula (European paper wasp) venom is of particular relevance in Southern Europe, potentially becoming a threat in other regions in the near future, and can be effectively cured by venom immunotherapy (VIT). As allergen content in extracts may vary and have an impact on diagnostic and therapeutic approaches, the aim was to compare five therapeutic preparations for VIT of P. dominula venom allergy available in Spain. Products from five different suppliers were analyzed by SDS-PAGE and LC-MS/MS and compared with a reference venom sample. Three products with P. dominula venom and one product with a venom mixture of American Polistes species showed a comparable band pattern in SDS-PAGE as the reference sample and the bands of the major allergens phospholipase A1 and antigen 5 were assignable. The other product, which consists of a mixture of American Polistes species, exhibited the typical band pattern in one, but not in another sample from a second batch. All annotated P. dominula allergens were detected at comparable levels in LC-MS/MS analysis of products containing P. dominula venom. Due to a lack of genomic information on the American Polistes species, the remaining products were not analyzed by this method. The major Polistes allergens were present in comparable amounts in the majority, but not in all investigated samples of venom preparations for VIT of P. dominula venom allergy.

Many studies have been conducted on measuring avian influenza viruses and their hemagglutinin (HA) antigens via electrochemical principles; most of these studies have used gold electrodes on ceramic, glass, or silicon substrates, and/or labeling for signal enhancement. Herein, we present a paper-based immunosensor for label-free measurement of multiple avian influenza virus (H5N1, H7N9, and H9N2) antigens using flexible screen-printed carbon nanotube-polydimethylsiloxane electrodes. These flexible electrodes on a paper substrate can complement the physical weakness of the paper-based sensors when wetted, without affecting flexibility. The relative standard deviation of the peak currents was 1.88% when the electrodes were repeatedly bent and unfolded twenty times with deionized water provided each cycle, showing the stability of the electrodes. For the detection of HA antigens, approximately 10-μl samples (concentration: 100 pg/ml–100 ng/ml) were needed to form the antigen–antibody complexes during 20–30 min incubation, and the immune responses were measured via differential pulse voltammetry. The limits of detections were 55.7 pg/ml (0.95 pM) for H5N1 HA, 99.6 pg/ml (1.69 pM) for H7N9 HA, and 54.0 pg/ml (0.72 pM) for H9N2 HA antigens in phosphate buffered saline, and the sensors showed good selectivity and reproducibility. Such paper-based sensors are economical, flexible, robust, and easy-to-manufacture, with the ability to detect several avian influenza viruses.

Rabies is a deadly viral disease present mainly in low-income countries of Africa and Asia. Dogs are the main reservoir and the source of human deaths. Mass vaccination campaigns of dogs are pivotal to achieve rabies elimination. The monitoring of the immune response of the dog population is necessary to evaluate the effectiveness of these campaigns, taking into account field conditions. This study explores the feasibility and the performance of a new tool using filter papers (FPs) to collect blood samples associated with an Enzyme-Linked ImmunoSorbent Assay (ELISA) titration of rabies antibodies in dogs. A total of 216 eluates from FP samples were collected from 111 dogs kept in experimental facilities in France and 29 dogs from the field in Tunisia. Sera were also analyzed using both the Fluorescence Antibody Virus Neutralization test (FAVNt) and ELISA. A high specificity (98.0%) was obtained by testing FP blood eluates from 51 unvaccinated dogs, with the results compared with those of FAVNt and ELISA on serum samples. The coefficients of concordance between FP eluates and tested sera were 88.9% for FAVNt and 88.0% for ELISA. Blood filter papers coupled with the titration of rabies antibodies by ELISA provide a reliable, simple, and effective solution to overcome the issues of the logistics and transport of samples, especially in low-income countries.

Paper spray ionization (PSI) is an extractive ambient ionization technique for mass spectrometry (MS), whereby a triangular paper tip serves as the sampling base and the electrospray tip. During PSI, analytes are extracted and transported to the edge of the paper tip by the applied spraying solvent. Analytes can be purified from a sample matrix and separated from each other by this transportation process. In this study, we investigated and utilized the analyte transportation process of PSI for the in situ separation and analysis of lipid mixtures. We found that differential transport of phosphatidylcholine (PC) and triacylglycerol (TAG), the two most abundant lipid classes in animals, occurred during PSI. We also found that the order in which these lipids moved strongly depended on how the spraying solvent was applied to the paper base. The more polar PC moved faster than the less polar TAG during PSI, when a polar solvent was slowly fed into a paper tip, whereas TAG was transported faster than PC when excess solvent was applied to the tip at once. In addition, we achieved a complete separation and detection of PC and TAG by slowly supplying a nonpolar solvent to a PSI tip.

Aristotle is often named as the first zoologist or biologist because of his writings on animals. Although Aristotle’s major intention in these books was to illustrate his ideas of how knowledge and understanding might advance, at least one modern biologist (C. Darwin) has recognized Aristotle's depth and breadth as being of surviving merit. Of greater surprise is the ongoing attention that his works continue to receive, including publications in contemporary scientific journals. This review identifies 38 peer-reviewed papers on various topics from Aristotle’s biological writings that have been published during the last 50 years. These papers are described according to content (genetics, population biology, anatomy, brain, movement), specific creatures (fish, scorpions, elephants, insects, birds), publication outlet, distribution over the fifty year period surveyed, and visible trends in the topics studied. It is concluded that, in the highly-competitive field of peer-reviewed scientific publication and citation, Aristotle's biology continues to excite the interest of scientists and remains salient to modern science itself.

With the appearance of plants and animals, new challenges emerged. These multicellular eukaryotes had to solve for example the difficulties of multifaceted communication between cells and adaptation to new habitats. In this paper, we are looking for one piece of the puzzle that made the development of complex multicellular eukaryotes possible with a focus on regulation of P2B autoinhibited Ca 2+-ATPases. P2B ATPases pump Ca 2+ out of the cytosol at the expense of ATP hydrolysis, and thereby maintain a steep gradient between the extra- and intracytosolic compartments which is utilized for Ca 2+-mediated rapid cell signaling. The activity of these enzymes is regulated by a calmodulin (CaM)-responsive autoinhibitory region, which can be located in either termini of the protein, at the C-terminus in animals and at the N-terminus in plants. When the cytoplasmic Ca 2+ level reaches a threshold, the CaM/Ca 2+ complex binds to a calmodulin-binding domain (CaMBD) in the autoinhibitor, which leads to the upregulation of pump activity. In animals, protein activity is also controlled by acidic phospholipids that bind to a cytosolic portion of the pump. Here, we analyze the appearance of CaMBDs and the phospholipid-activating sequence and show that their evolution in animals and plants was independent. Furthermore, we hypothesize that different causes may have initiated the appearance of these regulatory layers: in animals, it is linked to the appearance of multicellularity, while in plants it co-occurs with their water-to-land transition.

(1) Background: The medical practice of acupuncture involves the insertion of a specialized stainless needle into a specific body point, often called an acupoint, to initiate a perceived phenomenon of de-qi sensation. Therefore, the term “de-qi” describes bodily sensations experienced by the recipient during acupuncture, which may include feelings of soreness, heaviness, fullness, numbness, and migration. However, while acupuncture treatments reportedly result in acupoint activation and an increased release of neurotransmitters or cytokines, detecting these substances released into the acupoint microenvironment is often missed or delayed in clinical and basic practice. (2) Methods: To address this situation, we employed a paper-based enzyme-linked immunosorbent assay method to examine acupoint environmental changes using minute volumes of easily accessible acupoint fluids. (3) Results: Our results indicated that while levels of adenosine triphosphate (ATP), interleukin-1β, interleukin-6, glutamate, substance P, and histamine were all increased in the experimental group following electroacupuncture (EA) treatment, contrary results were observed in the sham EA and transient receptor potential vanilloid 1 (Trpv1−/−) groups. Subsequently, TRPV1 and its associated molecules were augmented in mouse dorsal root ganglion, spinal cord, thalamus, and the somatosensory cortex, then examined by Western blotting and immunofluorescence techniques. Investigations revealed that these elevations were still unobserved in the sham EA or EA in the Trpv1−/− groups. Furthermore, results showed that while administering ATP could mimic EA function, it could be reversed by the ATP P2 receptor antagonist, suramin. (4) Conclusions: Our data provide novel information, indicating that changes in neurotransmitter and cytokine levels can offer insight into acupuncture mechanisms and clinical targeting.

The study's goal was to develop a spore-based paper strip biosensor for detecting ß-lactam antibiotics in milk using the enzyme induction principle. A new spore-based paper strip biosensor has been developed after important operating parameters such as spore volume, substrate volume, exposure time and temperature, and incubation time and temperature were optimised. The limit of detection for various ß-lactam antibiotics, including amoxicillin, penicillin, ampicillin, carbenicillin, cloxacillin, nafcillin, oxacillin, cephalothin, cefalexin, cefoxitin, cefazolin, and cefuroxime, was determined in milk with detection sensitivity of 1 ppb, 2 ppb, 2 ppb, 10 ppb, 10 ppb, 10 ppb, 20 ppb, 10 ppb 1000 ppb, 10 ppb 300 ppb and 100 ppb, respectively. It was also tested with other contaminants such non-ß-lactam antibiotics, pesticides, aflatoxin, heavy metals, and other chemical contaminants, and no interference was found, indicating that the created biosensor had a low rate of false positive and negative results. In comparison to the AOAC-approved CHARM-ROSA ß-lactam strip test, which identified 7 raw milk and zero pasteurised milk samples positive for ß-lactam antibiotics, the sensor was further analysed and verified using 200 raw milk and 105 pasteurised milk samples. This indicates a perfect match between our biosensor and the AOAC-approved CHARM-ROSA ß-lactam strip test. The developed spore-based paper strip biosensors are expected to be useful in the rapid and cost-effective detection of ß-lactam antibiotic residues in milk samples at the dairy farm, reception dock, and production units, respectively.

Specification of germ cell fate depends on the asymmetric segregation of germ granules in early embryos. Now, a new paper in Development describes 'germline P-bodies', germ granules in Caenorhabditis elegans embryos, which function cooperatively with another condensate, P granules, in germline specification. To find out more, we caught up with first author Madeline Cassani and corresponding author Geraldine Seydoux, Professor at Johns Hopkins University School of Medicine.

Paper wasps are a model system for the study of social evolution due to a high degree of inter- and intraspecific variation in cooperation, aggression, and visual signals of social status. Increasing the taxonomic coverage of genomic resources for this diverse clade will aid comparative genomic approaches for testing predictions about the molecular basis of social evolution. Here, we provide draft genome assemblies for two well-studied species of paper wasps, Polistes exclamans and Mischocyttarus mexicanus. The P. exclamans genome assembly is 221.5 Mb in length with a scaffold N50 of 4.11 Mb. The M. mexicanus genome assembly is 227 Mb in length with a scaffold N50 of 1.1 Mb. Genomes have low repeat content (9.54–10.75%) and low GC content (32.06–32.4%), typical of other social hymenopteran genomes. The DNA methyltransferase gene, Dnmt3 , was lost early in the evolution of Polistinae. We identified a second independent loss of Dnmt3 within hornets (genus: Vespa).

The theory about the infectious causes of cancer was very popular at the turn of the nineteenth and twentieth centuries. The research methodology to confirm this hypothesis was based on diagnostic methods used by bacteriology. Various types of experiments were carried out on animals and humans. Attempts were made to "infect" cancer, as well as to transplant cancer cells into healthy animals. Disinfectants were studied looking for opportunities for anti-cancer prophylaxis. They were guided by analogy and empiricism, clinical images of diseases were compared. The aim of the article is to recall research on the infectious theory of cancer with a reminder of the achievements of Polish scientists in this field, and to try to determine the genesis of this theory.

Monoclonal antibodies (mAbs) deliver great benefits to patients with chronic and/or severe diseases thanks to their strong specificity to the therapeutic target. As a result of this specificity, non-human primates (NHP) are often the only preclinical species in which therapeutic antibodies cross-react with the target. Here, we highlight the value and limitations that NHP studies bring to the design of safe and efficient early clinical trials. Indeed, data generated in NHPs are integrated with in vitro information to predict the concentration/effect relationship in human, and therefore the doses to be tested in first-in-human trials. The similarities and differences in the systems defining the pharmacokinetics and pharmacodynamics (PKPD) of mAbs in NHP and human define the nature and the potential of the preclinical investigations performed in NHPs. Examples have been collated where the use of NHP was either pivotal to the design of the first-in-human trial or, inversely, led to the termination of a project prior to clinical development. The potential impact of immunogenicity on the results generated in NHPs is discussed. Strategies to optimize the use of NHPs for PKPD purposes include the addition of PD endpoints in safety assessment studies and the potential re-use of NHPs after non-terminal studies or cassette dosing several therapeutic agents of interest. Efforts are also made to reduce the use of NHPs in the industry through the use of in vitro systems, alternative in vivo models, and in silico approaches. In the case of prediction of ocular PK, the body of evidence gathered over the last two decades renders the use of NHPs obsolete. Expert perspectives, advantages, and pitfalls with these alternative approaches are shared in this review.

Over the years, conflicting data have meant that the role of Wnt in symmetry breaking and axis formation was unclear. Now, a new paper published in Development finds that maternal Wnt11b is required for robust cortical rotation inXenopus laevis, which, in turn, is required for axis induction. To find out more about this story, we caught up with two of the authors, Douglas Houston, Professor at The University of Iowa, and Marko Horb, Director of the National Xenopus Resource at the Marine Biological Laboratory in Chicago.

Milk adulteration is a common problem in developing countries, and it can lead to fatal diseases in humans. Despite several studies to identify different adulterants in milk samples, the effects of multiple adulterants remain unexplored. In this work, a three-dimensional (3D) paper-based microfluidic device is designed and fabricated to simultaneously detect multiple chemical adulterants in milk. This device comprises a top cover, a bottom cover, and a middle layer composed of transportation and a detection zone. By making cuts on the middle layer’s support, the device’s flow path is characterised by optimum and uniform velocity. For the first time, seven adulterants (urea, detergents, soap, starch, hydrogen peroxide, sodium-hydrogen-carbonate, and salt) are detected in the milk sample simultaneously with specificity evaluation and detailed color interference analysis. Only 1–2 mL of sample volume is required to detect 7 adulterants at one time. We have used only 10 $$\upmu$$ μ L of the reagent’s volume for the colorimetric reaction and found the results within a few seconds. Observation reveals that the limit of detection (LOD) of the adulterants lies in the range between $$0.05\%$$ 0.05 % (vol./vol.) to $$0.2\%$$ 0.2 % (vol./vol.) using the colorimetric detection technique. The unknown quantity of the added adulterants is measured using the calibration curves obtained from the experiments results. The repeatability and reproducibility of the process, sensitivity, and the linear range of detection of the calibration curves and the statistical study of the color intensity data are thoroughly analysed herein. In any resource-limited setting, this simple, portable, and user-friendly 3D microfluidic device is expected to be used for testing liquid foods before consumption.

Social insects are highly diverse in their social structures, aside from the consistent presence of reproductive castes. Among social insects, the Australian paper wasp Ropalidia plebeiana constructs extremely dense colony aggregations consisting of hundreds of colonies within a few square meters; however, little is known about the aggregation structures. We genetically analyzed the colony and population structure of R. plebeiana, and concomitant variations in colony sex ratios. In spring, the foundress (candidate queen) group started their colonies on a single old comb from the previous season, subsequently dividing these old combs via relatedness-based comb-cutting. Female philopatry, a prerequisite condition of Local Resource Competition (LRC), was confirmed. The colony sex ratio of reproductive individuals (male and female offspring for the next generation) became slightly male-biased in larger colonies, as predicted under LRC. However, the number of foundresses was positively associated with the number of reproductive individuals, suggesting that Local Resource Enhancement (LRE) also operates. Although the population structure appears to meet the prerequisites of LRC, the sex ratio appears to be modulated by factors other than LRC. Rather, through LRE, the availability of female helpers at the founding stage is likely to mitigate the sex ratios predicted under LRC.

Concept formation requires animals to learn and use abstract rules that transcend the characteristics of specific stimuli. Abstract concepts are often associated with high levels of cognitive sophistication, so there has been much interest in which species can form and use concepts. A key abstract concept is that of sameness and difference, where stimuli are classified as either the same as or different than an original stimulus. Here, we used a simultaneous two-item same-different task to test whether paper wasps ( Polistes fuscatus) can learn and apply a same-different concept. We trained wasps by simultaneously presenting pairs of same or different stimuli (e.g. colours). Then, we tested whether wasps could apply the concept to new stimuli of the same type (e.g. new colours) and to new stimulus types (e.g. odours). We show that wasps learned a general concept of sameness or difference and applied it to new samples and types of stimuli. Notably, wasps were able to transfer the learned rules to new stimuli in a different sensory modality. Therefore, P. fuscatus can classify stimuli based on their relationships and apply abstract concepts to novel stimulus types. These results indicate that abstract concept learning may be more widespread than previously thought.

Publishing a primary research article is typically the result of a collaborative effort between a variety of researchers across differing career stages. STAR Protocols can complement a research article and empower authors to share the expertise they contributed to the larger study. In this Backstory, we interview members of the Gennarino lab, who published a Cell paper and four protocols, covering bioinformatics, culturing of patient-derived cell lines, neuroimaging from mouse brain sections and primary neurons, and mouse seizure recordings. For more information on the protocols related to this backstory, please refer to (Gennarino et al., 2018).

Currently available European Alpine Altitude Climate Treatment (AACT) programs combine the physical characteristics of altitude with the avoidance of environmental triggers in the alpine climate and a personalized multidisciplinary pulmonary rehabilitation approach. The reduced barometric pressure, oxygen pressure, and air density, the relatively low temperature and humidity, and the increased UV radiation at moderate altitude induce several physiological and immunological adaptation responses. The environmental characteristics of the alpine climate include reduced aeroallergens such as house dust mites (HDM), pollen, fungi, and less air pollution. These combined factors seem to have immunomodulatory effects controlling pathogenic inflammatory responses and favoring less neuro-immune stress in patients with different asthma phenotypes. The extensive multidisciplinary treatment program may further contribute to the observed clinical improvement by AACT in asthma control and quality of life, fewer exacerbations and hospitalizations, reduced need for oral corticosteroids (OCS), improved lung function, decreased airway hyperresponsiveness (AHR), improved exercise tolerance, and improved sinonasal outcomes. Based on observational studies and expert opinion, AACT represents a valuable therapy for those patients irrespective of their asthma phenotype, who cannot achieve optimal control of their complex condition despite all the advances in medical science and treatment according to guidelines, and therefore run the risk of falling into a downward spiral of loss of physical and mental health. In the light of the observed rapid decrease in inflammation and immunomodulatory effects, AACT can be considered as a natural treatment that targets biological pathways. ispartof: ALLERGY vol:77 issue:7 pages:1991-2024 ispartof: location:Denmark status: published

Social interactions have large effects on individual physiology and fitness. In the immediate sense, social stimuli are often highly salient and engaging. Over longer time scales, competitive interactions often lead to distinct social ranks and differences in physiology and behavior. Understanding how initial responses lead to longer-term effects of social interactions requires examining the changes in responses over time. Here we examined the effects of social interactions on transcriptomic signatures at two times, at the end of a 45-minute interaction and 4 hours later, in female Polistes fuscatus paper wasp foundresses. Female P. fuscatus have variable facial patterns that are used for visual individual recognition, so we separately examined the transcriptional dynamics in the optic lobe and the non-visual brain. Results demonstrate much stronger transcriptional responses to social interactions in the non-visual brain compared to the optic lobe. Differentially regulated genes in response to social interactions are enriched for memory-related transcripts. Comparisons between winners and losers of the encounters revealed similar overall transcriptional profiles at the end of an interaction, which significantly diverged over the course of 4 hours, with losers showing changes in expression levels of genes associated with aggression and reproduction in paper wasps. On nests, subordinate foundresses are less aggressive, do more foraging and lay fewer eggs compared to dominant foundresses and we find losers shift expression of many genes in the non-visual brain, including vitellogenin, related to aggression, worker behavior, and reproduction within hours of losing an encounter. These results highlight the early neurogenomic changes that likely contribute to behavioral and physiological effects of social status changes in a social insect., Author summary Aggressive interactions often create inequalities–some individuals win while others lose. Winning versus losing can lead to large physiological differences between individuals, including different neurogenomic profiles between winners and losers. How this information about contest outcome leads to distinct neurogenomic profiles is poorly understood. Here we examine gene expression in response to aggressive social encounters in paper wasps, which naturally form dominance hierarchies on their nests in the wild. Shortly following encounters winners and losers have similar expression profiles, likely because similar mechanisms are engaged by social experiences. Four hours later, we find divergent neurogenomic profiles between winners and losers, with losers showing larger shifts in expression compared to winners. Many of the most dynamically expressed genes have been previously associated with dominance and caste differences in paper wasps showing how a single interaction can engage many of the same genomic networks that are involved in mediating more dramatic differences in queen-worker behavioral differences are also involved in responses shortly following social interactions.

To increase milk production, antibiotics are administered to animals to provide weight gain and to prevent or treat diseases. The inappropriate use of these substances can lead to antibiotic resistance and allergic reactions and toxic effects to milk consumers. We describe the development of a simple, fast, portable, and low-cost microfluidic paper-based analytical device (μPAD) to quantify sulfonamides in milk using the inhibition of the colorimetric reaction between carbonic anhydrase (CA) and 4-nitrophenyl acetate. The main advantages presented by the μPAD include reproducible batch production, simple application, and precise analysis without previous treatment. The µPAD displayed good linearity (R2 ≥ 0.986) in a wide range of sulfonamides in milk (2.5 to 40.0 µmol L−1), being selective for the drugs even in a highly complex matrix. We expect that this device allows in situ monitoring of milk quality, reducing the prejudicial conditions associated with high concentrations of sulfonamides in milk.

Viral-derived genes have had a huge impact during mammalian evolution, with many of the exapted genes being expressed in the placenta. Now, new research published in Development describes the importance of two genes with retroviral origins in microglia, the innate immune cells of the brain, which are derived from another extra-embryonic tissue, the yolk sac. We caught up with the first author, Masahito Irie, and the corresponding authors, Fumitoshi Ishino, Professor at Tokyo Medical and Dental University, and Tomoko Kaneko-Ishino, Professor at Tokai University, to hear about more about their research.

The 3D architecture of the uterus plays a key role in determine pregnancy outcome. A new paper in Development reveals the dynamic changes in luminal folding pattern that occur during the preimplantation period in mice. To hear more about the story, we caught up with first author Manoj Madhavan and corresponding author Ripla Arora, Assistant Professor at Michigan State University.

Allergic asthma (AA) is a common asthma phenotype, and its diagnosis requires both the demonstration of IgE-sensitization to aeroallergens and the causative role of this sensitization as a major driver of asthma symptoms. Therefore, a bronchial allergen challenge (BAC) would be occasionally required to identify AA patients among atopic asthmatics. Nevertheless, BAC is usually considered a research tool only, with existing protocols being tailored to mild asthmatics and research needs (eg long washout period for inhaled corticosteroids). Consequently, existing BAC protocols are not designed to be performed in moderate-to-severe asthmatics or in clinical practice. The correct diagnosis of AA might help select patients for immunomodulatory therapies. Allergen sublingual immunotherapy is now registered and recommended for controlled or partially controlled patients with house dust mite-driven AA and with FEV1 ≥ 70%. Allergen avoidance is costly and difficult to implement for the management of AA, so the proper selection of patients is also beneficial. In this position paper, the EAACI Task Force proposes a methodology for clinical BAC that would need to be validated in future studies. The clinical implementation of BAC could ultimately translate into a better phenotyping of asthmatics in real life, and into a more accurate selection of patients for long-term and costly management pathways.

The paper by K. L. Blaxter and J. L. Clapperton (1965) 'Prediction of the amount of methane produced by ruminants.

Carnosine (β-alanyl-L-histidine) is a naturally occurring endogenous dipeptide and an over-the-counter food supplement with a well-demonstrated multimodal mechanism of action that includes the detoxification of reactive oxygen and nitrogen species, the down-regulation of the production of pro-inflammatory mediators, the inhibition of aberrant protein formation, and the modulation of cells in the peripheral (macrophages) and brain (microglia) immune systems. Since its discovery more than 100 years ago, a plethora of in vivo preclinical studies have been carried out; however, there is still substantial heterogeneity regarding the route of administration, the dosage, the duration of the treatment, and the animal model selected, underlining the urgent need for “coordinated/aligned” preclinical studies laying the foundations for well-defined future clinical trials. The main aim of the present position paper is to critically and concisely consider these key points and open a discussion on the possible “alignment” for future studies, with the goal of validating the full therapeutic potential of this intriguing molecule.

Sustainability standards are increasingly used in the livestock sector.Insight into their actual contribution to sustainability is limited.This paper sets out a research agenda to study the role that standards play.We focus on the role of standard developers, consumers, producers and scientists.Standards need to be reliable and make an actual improvement to sustainability.

Although in a critical position in the economy, the paper industry releases a lot of wastewater that requires adequate treatment for sustainable development. This study presents an application of Life cycle assessment (LCA) with the ReCiPe tool on the wastewater treatment plant (WWTP) of a paper factory in Vietnam to evaluate the environmental effect of the individual techniques in WWTP, especially the internal circulation (IC) reactor, a pioneer and practical anaerobic technology. Both Midpoint and Endpoint categories results demonstrated that chemical use and electricity consumption mainly contributed to the environmental impact in the WWTP. The Dissolved air flotation (DAF) and Moving bed biofilm reactor (MBBR) are classified as effective techniques to reduce the impacts on the environment. Moreover, the comparison of LCA between IC and up-flow anaerobic sludge bed (UASB) shows that IC is the better practically green technique for the environment.

The deposition of trace elements around a pulp and paper industry in Morelia, Mexico, was evaluated using two lichen species as biomonitors. Samples of the foliose lichen Flavopunctelia praesignis and the fruticose lichen Usnea ceratina were collected in two remote areas and transplanted at different distances and directions from the pollution source. Lichen samples were exposed for 4 months (1) around the industrial area and (2) in their native habitats (control sites). We investigated the bioaccumulation of 11 trace elements in lichen thalli, and we compared the response of the two lichen species. To identify possible common sources, we evaluated the relationships between trace elements by correlations and cluster analyses. Our results showed that Cd was a good tracer for air pollution from the pulp and paper mills. In samples of Usnea ceratina exposed around the industrial area, Cd was significantly higher than in the remote area. Within the study area, trace element contents increase with the distance from the source, and they showed high depositions in the direction of prevailing winds. Moreover, we were able to detect groups of elements with similar behavior and common origins. Our results indicated that Flavopunctelia praesignis showed a higher capacity to accumulate trace elements than Usnea ceratina.

Specification of primordial germ cells requires a proportion of the cells in the posterior of the epiblast to reacquire pluripotency. A new paper in Development describes how OVOL2 is involved in regulating the balance between mesodermal fate and germ cell fate during gastrulation. We caught up with the first author, Yuki Naitou, and corresponding author, Katsuhiko Hayashi (Osaka University), to find out more about the paper and their future research.

We are glad to share with you our first Journal Club and to highlight some of the most interesting papers published recently. We hope we will tease your curiosity and encourage you to read full papers outside of your research area, which you may not have read otherwise. The Biomolecules Scientific Board wishes you an exciting read.

"Biological plausibility" is a concept frequently referred to in environmental and public health when researchers are evaluating how confident they are in the results and inferences of a study or evidence review. Biological plausibility is not, however, a domain of one of the most widely-used approaches for assessing the certainty of evidence (CoE) which underpins the findings of a systematic review, the Grading of Recommendations Assessment, Development and Evaluation (GRADE) CoE Framework. Whether the omission of biological plausibility is a potential limitation of the GRADE CoE Framework is a topic that is regularly discussed, especially in the context of environmental health systematic reviews. We analyse how the concept of "biological plausibility", as applied in the context of assessing certainty of the evidence that supports the findings of a systematic review, is accommodated under the processes of systematic review and the existing GRADE domains. We argue that "biological plausibility" is a concept which primarily comes into play when direct evidence about the effects of an exposure on a population of concern (usually humans) is absent, at high risk of bias, is inconsistent, or limited in other ways. In such circumstances, researchers look toward evidence from other study designs in order to draw conclusions. In this respect, we can consider experimental animal and in vitro evidence as "surrogates" for the target populations, exposures, comparators and outcomes of actual interest. Through discussion of 10 examples of experimental surrogates, we propose that the concept of biological plausibility consists of two principal aspects: a "generalisability aspect" and a "mechanistic aspect". The "generalisability aspect" concerns the validity of inferences from experimental models to human scenarios, and asks the same question as does the assessment of external validity or indirectness in systematic reviews. The "mechanistic aspect" concerns certainty in knowledge of biological mechanisms and would inform judgements of indirectness under GRADE, and thus the overall CoE. While both aspects are accommodated under the indirectness domain of the GRADE CoE Framework, further research is needed to determine how to use knowledge of biological mechanisms in the assessment of indirectness of the evidence in systematic reviews.

Zebrafish heart regeneration is dependent on the activation of a regenerative programme in the cells surrounding the heart, known as the epicardium. A new paper in Development uses genome-wide transcriptomics and chromatin accessibility profiles to identify and validate candidate enhancers linked to genes induced during regeneration in epicardial cells. To hear more about the story, we caught up with first author Yingxi Cao and senior authors Professor Ken Poss from Duke University and Jingli Cao, Assistant Professor at Weill Cornell Medicine.

We aimed to establish common guidelines for experimental studies with cattle.A book on “Methods in cattle physiology and behaviour research” was published.The book is designed as an open-access living handbook and is open to everyone.Citing the book saves space and avoids repetitions in scientific journals.Referencing guidelines reduces apparent plagiarism among authors.

Three-dimensional visualization of cellular and subcellular-structures in histological-tissues is essential for understanding the complexities of biological-phenomena, especially with regards structural and spatial relationships and pathologlical-diagnosis. Recent advancements in tissue-clearing technology, such as Magnified Analysis of Proteome (MAP), have significantly improved our ability to study biological-structures in three-dimensional space; however, their wide applicability to a variety of tissues is limited by long incubation-times and a need for advanced imaging-systems that are not readily available in most-laboratories. Here, we present optimized MAP-based method for paper-thin samples, Paper-MAP, which allow for rapid clearing and subsequent imaging of three-dimensional sections derived from various tissues using conventional confocal-microscopy. Paper-MAP successfully clear tissues within 1-day, compared to the original-MAP, without significant differences in achieved optical-transparency. As a proof-of-concept, we investigated the vasculature and neuronal-networks of a variety of human and rodent tissues processed via Paper-MAP, in both healthy and diseased contexts, including Alzheimer’s disease and glioma.

Since Nicolle, Manceaux and Splendore first described Toxoplasma gondii as a parasite of rodents and rabbits in the early 20th century, a diverse and vigorous research community has been built around studying this fascinating intracellular parasite. In addition to its importance as a pathogen of humans, livestock and wildlife, modern researchers are attracted to T. gondii as a facile experimental system to study many aspects of evolutionary biology, cellular biology, host-microbe interactions, and host immunity. For new researchers entering the field, the extensive literature describing the biology of the parasite, and the interactions with its host, can be daunting. In this review, we examine four foundational studies that describe various aspects of T. gondii biology, presenting a 'journal club'-style analysis of each. We have chosen a paper that established the beguiling life cycle of the parasite (Hutchison et al., 1971), a paper that described key features of its cellular biology that the parasite shares with related organisms (Gustafson et al., 1954), a paper that characterised the origin of the unique compartment in which the parasite resides within host cells (Jones and Hirsch, 1972), and a paper that established a key mechanism in the host immune response to parasite infection (Pfefferkorn, 1984). These interesting and far-reaching studies set the stage for subsequent research into numerous facets of parasite biology. As well as providing new researchers with an entry point into the literature surrounding the parasite, revisiting these studies can remind us of the roots of our discipline, how far we have come, and the new directions in which we might head.

On September 1, 1971, unknowingly to most, the world changed for the fields of behavioral genetics and circadian clocks. Ronald Konopka, a graduate student with Seymour Benzer at Caltech, published a paper (1) that I would argue is the most important discovery ultimately leading to our current molecular understanding of the circadian clock in animals. In this classic paper, Ron and Seymour reported the isolation of three single-gene mutants in Drosophila that dramatically altered circadian rhythms in pupal eclosion and locomotor activity. One mutant exhibited no rhythmicity, another had a short 19-h period, and a third had a long 28-h period. Remarkably, all three mutants mapped to the same locus on the X chromosome. They named this gene period . Seymour Benzer had recently joined the faculty at Caltech in 1967, after two previously successful careers in physics and molecular biology, and spurred on by Max Delbruck to do something more interesting, launched his third career in behavioral biology (2⇓⇓–5). Having done a sabbatical at Caltech with Roger Sperry, Seymour interacted with Ed Lewis, a giant in Drosophila genetics who trained with Alfred Sturtevant (descendent of Thomas Hunt Morgan) (6). Seymour chose Drosophila as a model system because its nervous system was intermediate in complexity “between a single neuron and the human brain” yet exhibited complex behavior and was amenable to genetic analysis (7). That same year, Seymour published his first paper (8) using mutagenesis and countercurrent technology to isolate phototaxis mutants in Drosophila . The opening sentence of this paper reads, “Complex as it is, much of the vast network of cellular functions has been successfully dissected, on a microscopic scale, by the use … [↵][1]1Email: joseph.takahashi{at}utsouthwestern.edu. [1]: #xref-corresp-1-1

The 2019 outbreak of coronavirus disease (COVID-19) in Wuhan (Hubei province of China) has given rise to a pandemic spread of virus, more than 240 million incidences and a death toll larger than 5 million people. COVID-19 has set off large efforts in research, therapy and patient care, as well as public and private debates in every imaginable form. A number of scientists used the publication platforms provided by the Federation of the European Biochemical Societies (FEBS) to present their research data, reviews, opinions and other contributions relating to COVID-19 and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Here, I highlight the recent COVID-19 papers which have been published and collected in a Virtual Issue in FEBS Letters, and discuss their implications towards understanding the molecular, biochemical and cellular mechanisms of SARS-CoV-2 infections, vaccine development and antiviral discovery strategies.

Polistes paper wasps are a widespread taxon inhabiting various climates. They build nests in the open without a protective outer layer, which makes them vulnerable to changing temperatures. To better understand the options they have to react to environmental variation and climate change, we here compare the thermoregulatory behavior of Polistes biglumis from cool Alpine climate with Polistes gallicus from warm Mediterranean climate. Behavioral plasticity helps both of them to withstand environmental variation. P. biglumis builds the nests oriented toward east-south-east to gain solar heat of the morning sun. This increases the brood temperature considerably above the ambience, which speeds up brood development. P. gallicus, by contrast, mostly avoids nesting sites with direct insolation, which protects their brood from heat stress on hot days. To keep the brood temperature below 40–42 °C on warm days, the adults of the two species show differential use of their common cooling behaviors. While P. biglumis prefers fanning of cool ambient air onto the nest heated by the sun and additionally cools with water drops, P. gallicus prefers cooling with water drops because fanning of warm ambient air onto a warm nest would not cool it, and restricts fanning to nests heated by the sun.