Health of Holochilus chacarius (Rodentia: Cricetidae) in rice agroecosystem in a neotropical wetland assessed by histopathology.

Small mammals have a short lifetime and are strictly associated with their environment. This work aimed to use histopathology to assess the health of Holochilus chacarius in a rice agroecosystem in the Pantanal of Mato Grosso do Sul. During necropsy, fragments of the lung, kidney, skin, liver, and reproductive system of 33 animals were collected and submitted to histological processing. Tissue damages were evaluated as mild, moderate, and severe and arranged in a matrix for further statistical analysis. Furthermore, we used generalized linear models to verify the influence of tissue changes on the body condition, obtained by a regression between body mass and length. In the lungs, we found an intense inflammatory infiltrate associated with anthracosis that had a negative influence on the body's condition. Also, we observed degenerative and inflammatory changes in the liver, kidneys, skin, and reproductive system that ranged from mild to moderate. The histopathological lesions observed in this study may be associated with environmental alterations of anthropic origin such as the exposure to soot from wildfires and heavy metals, evidenced by lesions in the lung, kidney, and liver. The present study provided a histopathological matrix as a new approach that allows to classify and quantify the tissue alterations. Tissue changes when associated with body condition demonstrated to be an effective tool to assess the health of small free-living mammals, showing that these animals can be used as bioindicators of environmental condition.

Employing electrochemically derived pH gradients for Lab-on-PCB protein preconcentration devices.

Protein preconcentration is an essential sample preparation step for analysis in which the targeted proteins exist in low concentrations, such as bodily fluids, water, or wastewater. Nonetheless, very few practical implementations of miniaturized protein preconcentration devices have been demonstrated in practice, and even fewer have been integrated with other microanalytical steps. Existing approaches rely heavily on additional chemicals and reagents and introduce complexity to the overall assay. In this paper, we propose a novel miniaturized isoelectric focusing-based protein preconcentration screening device based on electrochemically derived pH gradients rather than existing chemical reagent approaches. In this way, we reduce the need for additional chemical reagents to zero while enabling device incorporation in a seamlessly integrated full protein analysis microsystem via Lab-on-PCB technology. We apply our previously presented Lab-on-PCB approach to quantitatively control the pH of a solution in the vicinity of planar electrodes using electrochemical acid generation through redox-active self-assembled monolayers. The presented device comprises a printed circuit board with an array of gold electrodes that were functionalized with 4-aminothiophenol; this formed a self-assembled monolayer that was electropolymerized to improve its electrochemical reversibility. Protein preconcentration was performed in two configurations. The first was open and needed the use of a holder to suspend a well of fluid above the electrodes; the second used microfluidic channels to enclose small volumes of fluid. Reported here are the resulting data for protein preconcentration in both these forms, with a quantitative concentration factor shown for the open form and qualitative proof shown for the microfluidic.

Cranial morphological variation of Ctenomys lami (Rodentia: Ctenomyidae) in a restricted geographical distribution.

The relationship between chromosomal and morphological variation in mammals is poorly understood. We analyzed the cranial size and shape variation in Ctenomys lami concerning to the geographic variation in their chromosome numbers. This subterranean rodent occurs in a narrow range of sand-dunes in the Coastal Plain of southern Brazil. This species presents a high karyotypic variation with diploid numbers varying from 2n = 54 to 2n = 58, involving the fission and fusion of chromosome pairs 1 and 2. Due to different chromosome rearrangement frequencies along their geographic distribution, four karyotypic blocks were proposed. This study, explored cranium shape and size variation in geographical, chromosomal polymorphism, and chromosome rearrangements contexts to test whether the four karyotypic blocks reflect morphologically distinct units. For this, we measured 89 craniums using geometric morphometrics and used uni and multivariate statistics to discriminate the predicted groups and test for an association among chromosomal and morphological variation. Our results show the size and shape of sexual dimorphism, with males larger than females, and support the existence of four karyotypic blocks for Ctenomys lami based on morphological variation. However, our results do not support a direct relationship between chromosomal and cranial morphological variation in C. lami.

Phylogenetic relationships, distribution, and conservation of Roosmalens' dwarf porcupine, Coendouroosmalenorum Voss & da Silva, 2001 (Rodentia, Erethizontidae).

The New World porcupines of the genus Coendou comprise 16 species of arboreal nocturnal rodents. Some of these species are poorly known and have not been included in phylogenetic analyses. Based on recently collected specimens with associated tissue from the Brazilian Amazonia, we investigate the distribution and phylogenetic relationships of Roosmalens' dwarf porcupine, Coendouroosmalenorum, using an integrative approach using mitochondrial gene sequences and morphological data from new specimens and localities. Our results recovered C.roosmalenorum in the subgenus Caaporamys. However, analyses of our molecular and combined datasets produced different topologies. The new record shows the presence of C.roosmalenorum 480 km to the southeast of the Rio Madeira and 95 km away from Rio Juruena in Mato Grosso state, indicating a wider distribution in southern Amazonia than suspected. All known records of C.roosmalenorum are in the Madeira biogeographical province, to which it might be endemic.

Silicon microfabrication technologies for biology integrated advance devices and interfaces.

Miniaturization is the trend to manufacture ever smaller devices and this process requires knowledge, experience, understanding of materials, manufacturing techniques and scaling laws. The fabrication techniques used in semiconductor industry deliver an exceptionally high yield of devices and provide a well-established platform. Today, these miniaturized devices are manufactured with high reproducibility, design flexibility, scalability and multiplexed features to be used in several applications including micro-, nano-fluidics, implantable chips, diagnostics/biosensors and neural probes. We here provide a review on the microfabricated devices used for biology driven science. We will describe the ubiquity of the use of micro-nanofabrication techniques in biology and biotechnology through the fabrication of high-aspect-ratio devices for cell sensing applications, intracellular devices, probes developed for neuroscience-neurotechnology and biosensing of the certain biomarkers. Recently, the research on micro and nanodevices for biology has been progressing rapidly. While the understanding of the unknown biological fields -such as human brain- has been requiring more research with advanced materials and devices, the development protocols of desired devices has been advancing in parallel, which finally meets with some of the requirements of biological sciences. This is a very exciting field and we aim to highlight the impact of micro-nanotechnologies that can shed light on complex biological questions and needs.

A Lateral Flow Assay for the Detection of Leptospira lipL32 Gene Using CRISPR Technology.

The clinical manifestation of leptospirosis is often misdiagnosed as other febrile illnesses such as dengue. Therefore, there is an urgent need for a precise diagnostic tool at the field level to detect the pathogenic Leptospira lipL32 gene at the molecular level for prompt therapeutic decisions. Quantitative polymerase chain reaction (qPCR) is widely used as the primary diagnostic tool, but its applicability is limited by high equipment cost and the lack of availability in every hospital, especially in rural areas where leptospirosis mainly occurs. Here, we report the development of a CRISPR dFnCas9-based quantitative lateral flow immunoassay to detect the lipL32 gene. The developed assay showed superior performance regarding the lowest detectable limit of 1 fg/mL. The test is highly sensitive and selective, showing that leptospirosis diagnosis can be achieved with a low-cost lateral flow device.

Antimicrobial releasing hydrogel forming microneedles.

Hydrogel-forming microneedle arrays as a technique for transdermal drug delivery show promise as an alternative to traditional drug delivery methods. In this work, hydrogel-forming microneedles have been created with effective, controlled delivery of amoxicillin and vancomycin within comparable therapeutic ranges to that of oral delivered antibiotics. Fabrication using reusable 3D printed master templates enabled quick and low-cost hydrogel microneedle manufacturing through micro-molding. By 3D printing at a tilt angle of 45° the resolution of the microneedle tip was improved by double (from ca. 64 µm down to 23 µm). Amoxicillin and vancomycin were encapsulated within the hydrogel's polymeric network through a unique room temperature swell/deswell drug loading method within minutes, eliminating the need for an external drug reservoir. The hydrogel-forming microneedle mechanical strength was maintained, and successful penetration of porcine skin grafts observed with negligible damage to the needles or surrounding skin morphology. Hydrogel swell rate was tailored by altering the crosslinking density, resulting in controlled antimicrobial release for an applicable delivered dosage. The potent antimicrobial properties of the antibiotic-loaded hydrogel-forming microneedles against both Escherichia coli and Staphylococcus aureus, highlights the beneficial use of hydrogel-forming microneedles towards the minimally invasive transdermal drug delivery of antibiotics.

Nanotechnology-based electrochemical biosensors for monitoring breast cancer biomarkers.

Breast cancer is categorized as the most widespread cancer type among women globally. On-time diagnosis can decrease the mortality rate by making the right decision in the therapy procedure. These features lead to a reduction in medication time and socioeconomic burden. The current review article provides a comprehensive assessment for breast cancer diagnosis using nanomaterials and related technologies. Growing use of the nano/biotechnology domain in terms of electrochemical nanobiosensor designing was discussed in detail. In this regard, recent advances in nanomaterial applied for amplified biosensing methodologies were assessed for breast cancer diagnosis by focusing on the advantages and disadvantages of these approaches. We also monitored designing methods, advantages, and the necessity of suitable (nano) materials from a statistical standpoint. The main objective of this review is to classify the applicable biosensors based on breast cancer biomarkers. With numerous nano-sized platforms published for breast cancer diagnosis, this review tried to collect the most suitable methodologies for detecting biomarkers and certain breast cancer cell types.

Population genetic structure and morphological diversity of Cruzia tentaculata (Nematoda: Ascaridida), a parasite of marsupials (Didelphinae), along the Atlantic Forest on the eastern coast of South America.

Cruzia tentaculata is a helminth parasite of marsupials and has a wide geographic distribution from Mexico to Argentina. The aim of this study was to analyse the genetic population structure of this nematode along the Atlantic Forest biome. Cruzia tentaculata specimens were recovered from Didelphis aurita, Didelphis albiventris and Philander quica in 9 localities. Morphological and morphometric data were investigated for phenotypic diversity among localities and hosts using multivariate discriminant analysis of principal components. Phylogenetic relationships of C. tentaculata were determined using maximum likelihood and Bayesian inference. The population structure was analysed by fixation indices, molecular variance analysis, Tajima's D and Fu's Fs neutrality tests, Mantel tests and Bayesian clustering analysis. A higher significant morphometric difference for males was observed between localities. In the haplogroup networks, 2 groups were recovered, separating locations from the north and from the south/southeast. The morphometric variation in C. tentaculata between different localities was compatible with this north and southeast/south pattern, suggesting adaptation to different ecological conditions. Population genetic analyses suggested a pattern of evolutionary processes driven by Pleistocene glacial refugia in the northeast and southeast of the Atlantic Forest based on the distribution of genetic diversity.

A lab-on-a-chip for the concurrent electrochemical detection of SARS-CoV-2 RNA and anti-SARS-CoV-2 antibodies in saliva and plasma.

Rapid, accurate and frequent detection of the RNA of SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) and of serological host antibodies to the virus would facilitate the determination of the immune status of individuals who have Coronavirus disease 2019 (COVID-19), were previously infected by the virus, or were vaccinated against the disease. Here we describe the development and application of a 3D-printed lab-on-a-chip that concurrently detects, via multiplexed electrochemical outputs and within 2 h, SARS-CoV-2 RNA in saliva as well as anti-SARS-CoV-2 immunoglobulins in saliva spiked with blood plasma. The device automatedly extracts, concentrates and amplifies SARS-CoV-2 RNA from unprocessed saliva, and integrates the Cas12a-based enzymatic detection of SARS-CoV-2 RNA via isothermal nucleic acid amplification with a sandwich-based enzyme-linked immunosorbent assay on electrodes functionalized with the Spike S1, nucleocapsid and receptor-binding-domain antigens of SARS-CoV-2. Inexpensive microfluidic electrochemical sensors for performing multiplexed diagnostics at the point of care may facilitate the widespread monitoring of COVID-19 infection and immunity.

Electrochemical sensors based on metal nanoparticles with biocatalytic activity.

Biosensors have attracted a great deal of attention, as they allow for the translation of the standard laboratory-based methods into small, portable devices. The field of biosensors has been growing, introducing innovations into their design to improve their sensing characteristics and reduce sample volume and user intervention. Enzymes are commonly used for determination purposes providing a high selectivity and sensitivity; however, their poor shelf-life is a limiting factor. Researchers have been studying the possibility of substituting enzymes with other materials with an enzyme-like activity and improved long-term stability and suitability for point-of-care biosensors. Extra attention is paid to metal and metal oxide nanoparticles, which are essential components of numerous enzyme-less catalytic sensors. The bottleneck of utilising metal-containing nanoparticles in sensing devices is achieving high selectivity and sensitivity. This review demonstrates similarities and differences between numerous metal nanoparticle-based sensors described in the literature to pinpoint the crucial factors determining their catalytic performance. Unlike other reviews, sensors are categorised by the type of metal to study their catalytic activity dependency on the environmental conditions. The results are based on studies on nanoparticle properties to narrow the gap between fundamental and applied research. The analysis shows that the catalytic activity of nanozymes is strongly dependent on their intrinsic properties (e.g. composition, size, shape) and external conditions (e.g. pH, type of electrolyte, and its chemical composition). Understanding the mechanisms behind the metal catalytic activity and how it can be improved helps designing a nanozyme-based sensor with the performance matching those of an enzyme-based device.

Pre-concentration of microRNAs by LNA-modified magnetic beads for enhancement of electrochemical detection.

MicroRNAs are extremely promising candidates for early cancer diagnosis and prognosis. The levels of circulating microRNAs provide valuable information about cancer disease at its early stages. However, the levels of microRNAs that need to be detected are extremely low and difficult to discriminate from a large pool of oligonucleotides. There is the need for accurate, rapid and sensitive detection methodologies for detection of microRNAs. We developed electrochemical impedance spectroscopy peptide nucleic acid (PNA)-based sensors that can detect miRNAs in diluted serum with a limit of detection of 0.38 fM. In order to further improve the accuracy and reliability of the sensors, we developed an assay using magnetic beads for simple and rapid fishing of target microRNAs from solution and its pre-concentration prior to electrochemical detection. Our methodology utilizes magnetic beads for the capture of the target microRNA from solution and brings the concentrated sample to the sensor surface. We modify the magnetic beads with locked nucleic acids (LNA), which have high affinity and specificity to their complementary microRNA sequence. The separated and concentrated microRNA is then detected using the PNA-based sensors. By exposing the sensing electrodes only to the captured microRNAs, interferences from other nucleotides or biomolecules from the sample are eliminated.

Utilising Commercially Fabricated Printed Circuit Boards as an Electrochemical Biosensing Platform.

Printed circuit boards (PCBs) offer a promising platform for the development of electronics-assisted biomedical diagnostic sensors and microsystems. The long-standing industrial basis offers distinctive advantages for cost-effective, reproducible, and easily integrated sample-in-answer-out diagnostic microsystems. Nonetheless, the commercial techniques used in the fabrication of PCBs produce various contaminants potentially degrading severely their stability and repeatability in electrochemical sensing applications. Herein, we analyse for the first time such critical technological considerations, allowing the exploitation of commercial PCB platforms as reliable electrochemical sensing platforms. The presented electrochemical and physical characterisation data reveal clear evidence of both organic and inorganic sensing electrode surface contaminants, which can be removed using various pre-cleaning techniques. We demonstrate that, following such pre-treatment rules, PCB-based electrodes can be reliably fabricated for sensitive electrochemical biosensors. Herein, we demonstrate the applicability of the methodology both for labelled protein (procalcitonin) and label-free nucleic acid (E. coli-specific DNA) biomarker quantification, with observed limits of detection (LoD) of 2 pM and 110 pM, respectively. The proposed optimisation of surface pre-treatment is critical in the development of robust and sensitive PCB-based electrochemical sensors for both clinical and environmental diagnostics and monitoring applications.

Multiplexed Prostate Cancer Companion Diagnostic Devices.

Prostate cancer (PCa) remains one of the most prominent forms of cancer for men. Since the early 1990s, Prostate-Specific Antigen (PSA) has been a commonly recognized PCa-associated protein biomarker. However, PSA testing has been shown to lack in specificity and sensitivity when needed to diagnose, monitor and/or treat PCa patients successfully. One enhancement could include the simultaneous detection of multiple PCa-associated protein biomarkers alongside PSA, also known as multiplexing. If conventional methods such as the enzyme-linked immunosorbent assay (ELISA) are used, multiplexed detection of such protein biomarkers can result in an increase in the required sample volume, in the complexity of the analytical procedures, and in adding to the cost. Using companion diagnostic devices such as biosensors, which can be portable and cost-effective with multiplexing capacities, may address these limitations. This review explores recent research for multiplexed PCa protein biomarker detection using optical and electrochemical biosensor platforms. Some of the novel and potential serum-based PCa protein biomarkers will be discussed in this review. In addition, this review discusses the importance of converting research protocols into multiplex point-of-care testing (xPOCT) devices to be used in near-patient settings, providing a more personalized approach to PCa patients' diagnostic, surveillance and treatment management.

The role and impact of Zootaxa in mammalogy in its first 20 years.

Zootaxa came as a new and innovative publication medium for taxonomy, amidst a scenario of devaluation of this important biological science. After 20 years, it has ascertained itself as one of the main journals in animal taxonomy. However, the contribution of the journal to the taxonomy of Mammalia (mammals), one of the most studied groups of animals with a long-standing, dedicated spectrum of specialized journals (mammalogy), could have been expected as minor. All the current and former editors of the Mammalia section of Zootaxa analyzed the relative contribution of the journal to the description of new species of mammals since 2001. We also analyzed the contribution of Zootaxa by taxon, geographic origin of taxa, and geographic origin of first authors. The taxonomic methodology of authors in species description is described as well as the temporal trends in publications and publication subjects. We highlight the editors' picks and eventually, the challenges for the future. We found that Zootaxa has had a significant contribution to mammalogy, being the second journal (the first being Journal of Mammalogy) in terms of number of new species described (76; 10.6% of the new mammalian species described between 2001 and 2020). The majority of the new species were described following an integrative taxonomic approach with at least two sources of data (86%). The analysis of published taxa, their geographic origin, and the country of origin of first authors shows a wide coverage and exhaustive representation, except for the species from the Nearctic. We conclude that Zootaxa has likely responded to a repressed demand for an additional taxonomic journal in mammalogy, with as possible appeals the absence of publication fees and an established publication speed. With 246 articles published in the past 20 years, the Mammalia section of Zootaxa embraces a large spectrum of systematic subjects going beyond alpha taxonomy. The challenges for the future are to encourage publications of authors from the African continent, still poorly represented, and from the palaeontology community, as the journal has been open to palaeontology since its early days.

Printable graphene BioFETs for DNA quantification in Lab-on-PCB microsystems.

Lab-on-Chip is a technology that aims to transform the Point-of-Care (PoC) diagnostics field; nonetheless a commercial production compatible technology is yet to be established. Lab-on-Printed Circuit Board (Lab-on-PCB) is currently considered as a promising candidate technology for cost-aware but simultaneously high specification applications, requiring multi-component microsystem implementations, due to its inherent compatibility with electronics and the long-standing industrial manufacturing basis. In this work, we demonstrate the first electrolyte gated field-effect transistor (FET) DNA biosensor implemented on commercially fabricated PCB in a planar layout. Graphene ink was drop-casted to form the transistor channel and PNA probes were immobilized on the graphene channel, enabling label-free DNA detection. It is shown that the sensor can selectively detect the complementary DNA sequence, following a fully inkjet-printing compatible manufacturing process. The results demonstrate the potential for the effortless integration of FET sensors into Lab-on-PCB diagnostic platforms, paving the way for even higher sensitivity quantification than the current Lab-on-PCB state-of-the-art of passive electrode electrochemical sensing. The substitution of such biosensors with our presented FET structures, promises further reduction of the time-to-result in microsystems combining sequential DNA amplification and detection modules to few minutes, since much fewer amplification cycles are required even for low-abundance nucleic acid targets.

Electrochemical Biosensors for Cytokine Profiling: Recent Advancements and Possibilities in the Near Future.

Cytokines are soluble proteins secreted by immune cells that act as molecular messengers relaying instructions and mediating various functions performed by the cellular counterparts of the immune system, by means of a synchronized cascade of signaling pathways. Aberrant expression of cytokines can be indicative of anomalous behavior of the immunoregulatory system, as seen in various illnesses and conditions, such as cancer, autoimmunity, neurodegeneration and other physiological disorders. Cancer and autoimmune diseases are particularly adept at developing mechanisms to escape and modulate the immune system checkpoints, reflected by an altered cytokine profile. Cytokine profiling can provide valuable information for diagnosing such diseases and monitoring their progression, as well as assessing the efficacy of immunotherapeutic regiments. Toward this goal, there has been immense interest in the development of ultrasensitive quantitative detection techniques for cytokines, which involves technologies from various scientific disciplines, such as immunology, electrochemistry, photometry, nanotechnology and electronics. This review focusses on one aspect of this collective effort: electrochemical biosensors. Among the various types of biosensors available, electrochemical biosensors are one of the most reliable, user-friendly, easy to manufacture, cost-effective and versatile technologies that can yield results within a short period of time, making it extremely promising for routine clinical testing.

Hydrogel-Forming Microneedles: Current Advancements and Future Trends.

In this focused progress review, the recent developments and trends of hydrogel-forming microneedles (HFMs) and potential future directions are presented. Previously, microneedles (solid, hollow, coated, and dissolving microneedles) have primarily been used to enhance the effectiveness of transdermal drug delivery to facilitate a wide range of applications such as vaccinations and antibiotic delivery. However, the recent trend in microneedle development has resulted in microneedles formed from hydrogels which have the ability to offer transdermal drug delivery and, due to the hydrogel swelling nature, passively extract interstitial fluid from the skin, meaning they have the potential to be used for biocompatible minimally invasive monitoring devices. Thus, in this review, these recent trends are highlighted, which consolidate microneedle design considerations, hydrogel formulations, fabrication processes, applications of HFMs and the potential future opportunities for utilizing HFMs for personalized healthcare monitoring and treatment.

Impedimetric aptamer-based glycan PSA score for discrimination of prostate cancer from other prostate diseases.

Prostate specific antigen (PSA) is the common biomarker for prostate cancer (PCa). However, its lack of specificity to differentiate PCa from benign prostate disorders stimulates the search for alternative cancer biomarkers to improve the clinical management of the patients. Different studies have described changes in the core-fucosylation level of PSA between PCa patients and healthy controls. To exploit these findings, we have adapted an impedimetric aptamer-based sensor to the dual recognition of PSA. Two different aptamers, PSAG-1 and anti-PSA, are immobilized onto two adjacent nanostructured gold electrodes. The direct binding from diluted serum samples of specific glycosylated-PSA to the first sensor and total PSA to the second one leads to changes in the charge transfer resistance, which correlate to the amount of glycosylated and total PSA in the sample. The sensors are able to measure PSA in serum with a dynamic range between 0.26 and 62.5 ng/mL (PSAG-1) and from 0.64 to 62.5 ng/mL (anti-PSA), with a reproducibility of 5.4 %. The final output of the proposed platform is the ratio between PSAG-1 reactive PSA and total PSA, defined as the glycan score. The glycan score was tested in serum samples from patients with different pathologies, showing excellent correlation between the measured score and the known diagnosis of the patients. Hence this dual aptamer-based impedimetric biosensor could be used as a minimally invasive method for the diagnosis of prostate cancer.

Label-Free DNA Biosensor Using Modified Reduced Graphene Oxide Platform as a DNA Methylation Assay.

This work reports the use of modified reduced graphene oxide (rGO) as a platform for a label-free DNA-based electrochemical biosensor as a possible diagnostic tool for a DNA methylation assay. The biosensor sensitivity was enhanced by variously modified rGO. The rGO decorated with three nanoparticles (NPs)-gold (AuNPs), silver (AgNPs), and copper (CuNPs)-was implemented to increase the electrode surface area. Subsequently, the thiolated DNA probe (single-stranded DNA, ssDNA-1) was hybridized with the target DNA sequence (ssDNA-2). After the hybridization, the double-stranded DNA (dsDNA) was methylated by M.SssI methyltransferase (MTase) and then digested via a HpaII endonuclease specific site sequence of CpG (5'-CCGG-3') islands. For monitoring the MTase activity, differential pulse voltammetry (DPV) was used, whereas the best results were obtained by rGO-AuNPs. This assay is rapid, cost-effective, sensitive, selective, highly specific, and displays a low limit of detection (LOD) of 0.06 U·mL-1. Lastly, this study was enriched with the real serum sample, where a 0.19 U·mL-1 LOD was achieved. Moreover, the developed biosensor offers excellent potential in future applications in clinical diagnostics, as this approach can be used in the design of other biosensors.