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This study assessed the exposure of free-ranging Amazon River dolphins (Inia geoffrensis) to phthalate esters (PAE) in a remote area of the Brazilian Amazon. Blubber samples were analyzed for four PAEs - dimethyl phthalate, diethyl phthalate, dibutyl phthalate, and di(2-ethylhexyl) phthalate - and cholesterol contents to evaluate potential metabolic disturbances. All dolphins were contaminated with at least two PAEs, with DEHP (242.16 ng/μL) and DBP (191.62 ng/μL) being the most frequent, detected in 93 % and 79 % of the sample, respectively. Significant positive correlations were found between DBP and DEP (r = 0.857), DEHP and DEP (r = 0.794), and DBP and body length (r = 0.642), suggesting bioaccumulation. Despite the cholesterol data not showing a correlation with the other findings, these results highlight PAE pollution in a supposedly pristine environment and their potential impact on the health and conservation of Amazon River dolphins' health and Amazonian ecosystem., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The present study investigated the effect of adding allopathic doxorubicin (DOX 0.3 µg/mL), the vehicle of ultradiluted/dynamized doxorubicin (0.2 % ethanol), different dynamizations of ultradiluted/dynamized doxorubicin (DOX 6CH, DOX 12CH and DOX 30CH), both in the absence or presence of chemical stress induced by doxorubicin at 0.3 µg/mL on follicular survival and activation, antioxidant capacity of the medium, Catalase activity (CAT), production of reactive protein thiol, maintenance of type I and III collagen fibers and accumulation of lipofuscin in porcine ovarian tissue cultured in vitro for 48 hours. To do this, part of the ovarian tissue fragments was fixed for the uncultured control and the rest were cultured in: MEM (cultured control), DOX 0.3 µg/mL, Ethanol, DOX 6CH, DOX 12CH, DOX 30CH, DOX (0.3 µg/mL) + DOX 6CH, DOX (0.3 µg/mL) + DOX 12CH, DOX (0.3 µg/mL) + DOX 30CH treatments. The results showed that, in general, ultradiluted/dynamized doxorubicin (DOX 6CH, DOX 12CH and DOX 30CH) mitigated the toxic effect of allopathic doxorubicin (0.3 µg/mL) on the morphology of preantral follicles, the content of type I and III collagen fibers, and the production of lipofuscin in the tissue. However, only DOX (0.3 µg/mL) + DOX 6CH attenuated the oxidative stress induced by DOX (0.3 µg/mL), maintaining adequate CAT activity that was similar to the uncultured control. Additionally, when the three isolated ultradiluted/dynamized doxorubicin were considered, only DOX 12CH increased the reduced thiol levels compared to the uncultured control and MEM. In conclusion, supplementing the culture medium with ultradiluted/dynamized DOX (DOX 6CH, DOX 12CH and DOX 30CH) attenuated the toxicity induced by allopathic doxorubicin during the in vitro culture of pig preantral follicles enclosed in ovarian tissue., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

This brief review delves into the topic of in vitro follicle culture for in vitro embryo production, with a particular emphasis on goat models. Specifically, we examine the main findings from LAMOFOPA-Brazil over the last 20 years, highlighting the challenges posed by oxidative stress and epigenetic changes. Our focus is on strategies to improve follicular development and oocyte maturation. Furthermore, we underscore the valuable role of the antioxidant anethole in optimizing the efficacy of in vitro follicle culture and improving outcomes in in vitro embryo production., Competing Interests: Conflicts of interest: The authors have no conflict of interest to declare., (Copyright © The Author(s).)

A wearable glove-based sensor is a portable and practical approach for onsite detection/monitoring of a variety of chemical threats. Herein, we report a flexible and sensitive wearable sensor fabricated on the nitrile glove fingertips by stencil-printing technique. The working electrodes were modified with multiwalled carbon nanotubes (MWCNTs)/poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) for sensitive and real-time analyses of hazardous or chemical treats, as picric acid (PA) explosive, diazepam (DZ) as drug-facilitated crimes and the emerging pollutant 4-nitrophenol (4-NP). The multi-sensing platform towards PA, 4-NP, and DZ offers the ability of in-situ qualitative and quantitative analyses of powder and liquid samples. A simple sampling by touching or swiping the fingertip sensor on the sample or surface under investigation using an ionic hydrogel combined with fast voltammetry measurement provides timely point-of-need analyses. The wearable glove-based sensor uses the square wave voltammetry (SWV) technique and exhibited excellent performance to detect PA, 4-NP, and DZ, resulting in limits of detection (LOD) of 0.24 μM, 0.35 μM, 0.06 μM, respectively, in a wide concentration range (from 0.5 μM to 100 μM). Also, we obtained excellent manufacturing reproducibility with relative standard deviations (RSD) in the range of 3.65%-4.61% using 7 different wearable devices (n = 7) and stability in the range of 4.86%-6.61% using different electrodes stored for 10 days at room temperature (n = 10), demonstrating the excellent sensor-to-sensor reproducibility and stability for reliable in-field measurements. The stretchable sensor presented great mechanical robustness, supporting up to 80 bending or stretching deformation cycles without significant voltammetric changes. Collectively, our wearable glove-based sensor may be employed for analyses of chemical contaminants of concern, such as explosives (PA), drugs (DZ), and emerging pollutants (4-NP), helping in environmental and public safety control., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The concerning of plastic pollution in different ecosystems has been worsened by the widespread presence. Phthalate esters (PAEs), plasticizers found in everyday products, can migrate into the environment, especially into the oceans. Researches on their effects on cetaceans are still rare. Metabolomics helps assess perturbations induced by exposure to PAEs, which act as persistent endocrine disruptors. Four PAEs (dimethyl phthalate - DMP, diethyl phthalate - DEP, dibutyl phthalate - DBP, and di(2-ethylhexyl phthalate - DEHP) were analyzed, along with cholesterol and fatty acid profiles of P. blainvillei's blubber samples collected in southern Brazil. The study reveals pervasive contamination by PAEs - especially DEHP, present in all samples - with positive correlations between DEP content and animal size and weight, as well as between the DEHP amount and the C17:1 fatty acid. These findings will be relevant to conservation efforts aimed at this threatened species and overall marine ecosystems., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study aimed to investigate the effect of including mouse feed with different concentrations (5, 10, or 20%) of Pereskia aculeata Miller (PAM) leaves on the morphology and development of preantral ovarian follicles and ovarian stromal cell density. The oral toxicity was performed using repeated dose toxicity assays subdivided into experiments of 30 days and 90 days of treatment. After the experiments, the ovaries of each animal were collected and submitted to classical histology. At 30 and 90 days, there was an equivalent percentage of normal, primordial, and developing follicles (P > 0.05) between PAM treatments compared to the control. Regarding the different stages of follicular development, after 90 days, there was a higher percentage (P < 0.05) of developing follicles only in the control group compared to day 30. The PAM 5% treatment was the only one that affected the cell density in the stroma after 90 days of treatment. Thus, we observed that supplementing the diet with P. aculeata did not pose any risk concerning animal consumption; specifically, there were no toxic reproductive effects observed from adding Pereskia aculeata Miller to the mouse diet., Competing Interests: Conflicts of interest: The authors have no conflict of interest to declare., (Copyright © The Author(s).)

Epidural anesthesia in dogs is a locoregional anesthesia technique used in veterinary medicine, becoming an important integrated application in the anesthetic protocol to provide safer and more effective analgesia to patients. For this, professionals must adhere to rigorous guidelines and possess technical skills. In this context, in veterinary education, the development of practical clinical skills represents a crucial aspect in the training of these professionals. However, traditional teaching methods have proven insufficient to ensure a consistent level of competence among recent graduates. The introduction of non-animal alternatives for educational purposes has contributed to the development of simulation-based teaching, an innovative and accessible field capable of enhancing pre-clinical proficiency in students and reducing the use of live animals and cadavers. Despite its application in various areas of veterinary education, there are no conclusive results regarding the development of accessible simulators capable of effectively enhancing training in epidural anesthesia in dogs. Therefore, this article represents a pioneering study aimed at sharing a method for creating SimuVet, a realistic simulator for training epidural anesthesia in dogs. The simulator was fully developed by veterinary researchers with limited experience in 3D printing and, after preliminary analysis, demonstrated excellent performance and ultrasonographic anatomy. Future work will focus on the formal validation of this simulator with the aim of improving the teaching and learning process for students and experts in performing epidural anesthesia in companion animals., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Lobo Moraes, Ghisi, Paes de Barros, de Carvalho Peixoto, Brandini Népoli, Moleta Colodel, Souza de Lima and Lopes de Souza.)

The present study highlights the integration of lignin with graphene oxide (GO) and its reduced form (rGO) as a significant advancement within the bio-based products industry. Lignin-phenol-formaldehyde (LPF) resin is used as a carbon source in polyurethane foams, with the addition of 1 %, 2 %, and 4 % of GO and rGO to produce carbon structures thus producing carbon foams (CFs). Two conversion routes are assessed: (i) direct addition with rGO solution, and (ii) GO reduction by heat treatment. Carbon foams are characterized by thermal, structural, and morphological analysis, alongside an assessment of their electrochemical behavior. The thermal decomposition of samples with GO is like those having rGO, indicating the effective removal of oxygen groups in GO by carbonization. The addition of GO and rGO significantly improved the electrochemical properties of CF, with the GO2% sensors displaying 39 % and 62 % larger electroactive area than control and rGO2% sensors, respectively. Furthermore, there is a significant electron transfer improvement in GO sensors, demonstrating a promising potential for ammonia detection. Detailed structural and performance analysis highlights the significant enhancement in electrochemical properties, paving the way for the development of advanced sensors for gas detection, particularly ammonia, with the prospective market demands for durable, simple, cost-effective, and efficient devices., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Monkeypox virus (MPXV) infection was classified as a public health emergency of international concern by the World Health Organization (WHO) in 2022, being transmitted between humans by large respiratory droplets, in contact with skin lesions, fomites, and sexually. Currently, there are no available accessible and simple-to-use diagnostic tests that accurately detect MPXV antigens for decentralized and frequent testing. Here, we report an electrochemical biosensor to detect MPXV antigens in saliva and plasma samples within 15 min using accessible materials. The electrochemical system was manufactured onto a paper substrate engraved by a CO 2 laser machine, modified with gold nanostructures (AuNS) and a monoclonal antibody, enabling sensitive detection of A29 viral protein. The diagnostic test is based on the use of electrochemical impedance spectroscopy (EIS) and can be run by a miniaturized potentiostat connected to a smartphone. The impedimetric biosensing method presented excellent analytical parameters, enabling the detection of A29 glycoprotein in the concentration ranging from 1 × 10 -14 to 1 × 10 -7 g mL -1 , with a limit of detection (LOD) of 3.0 × 10 -16 g mL -1 . Furthermore, it enabled the detection of MPXV antigens in the concentration ranging from 1 × 10 -1 to 1 × 10 4 PFU mL -1 , with an LOD of 7.8 × 10 -3 PFU mL -1 . Importantly, no cross-reactivity was observed when our device was tested in the presence of other poxvirus and nonpoxvirus strains, indicating the adequate selectivity of our nanobiosensor for MPXV detection. Collectively, the nanobiosensor presents high greenness metrics associated with the use of a reproducible and large-scale fabrication method, an accessible and sustainable paper substrate, and a low volume of sample (2.5 μL), which could facilitate frequent testing of MPXV at point-of-care (POC).

The increasing antifungal resistance rates against conventional drugs reveal the urgent need to search for new therapeutic alternatives. In this context, natural bioactive compounds have a critical role in antifungal drug development. Since evidence demonstrates that abietic acid, a diterpene found in Pinus species, has significant antimicrobial properties, this study aimed to evaluate the antifungal activity of abietic acid against Candida spp and its ability to potentiate the activity of fluconazole. Abietic acid was tested both individually and in combination with fluconazole against Candida albicans (CA INCQS 40006), Candida krusei (CK INCQS 40095), and Candida tropicalis (CT INCQS 40042). The microdilution method was used to determine the IC 50 and the cell viability curve. Minimum Fungicidal Concentration (MFC) was determined by subculture in a solid medium. The plasma membrane permeability was measured using a fluorescent SYTOX Green probe. While the IC 50 of the drugs alone ranged between 1065 and 3255 μg/mL, the IC 50 resulting from the combination of abietic acid and fluconazole ranged between 7563 and 160.1 μg/mL. Whether used in combination with fluconazole or isolated, abietic acid exhibited Minimum Fungicidal Concentration (MFC) values exceeding 1024 μg/mL against Candida albicans , Candida krusei and Candida tropicalis . However, it was observed that the antifungal effect of fluconazole was enhanced when used in combination with abietic acid against Candida albicans and Candida tropicalis . These findings suggest that while abietic acid alone has limited inherent antifungal activity, it can enhance the effectiveness of fluconazole, thereby reducing antifungal resistance.

Herpes simplex virus type 2 (HSV-2) infection, which is almost exclusively sexually transmitted, causes genital herpes. Although this lifelong and incurable infection is extremely widespread, currently there is no readily available diagnostic device that accurately detects HSV-2 antigens to a satisfactory degree. Here, we report an ultrasensitive electrochemical device that detects HSV-2 antigens within 9 min and costs just $1 (USD) to manufacture. The electrochemical biosensor is biofunctionalized with the human cellular receptor nectin-1 and detects the glycoprotein gD2, which is present within the HSV-2 viral envelope. The performance of the device is tested in a guinea pig model that mimics human biofluids, yielding 88.9% sensitivity, 100.0% specificity, and 95.0% accuracy under these conditions, with a limit of detection of 0.019 fg mL -1 for gD2 protein and 0.057 PFU mL -1 for titered viral samples. Importantly, no cross-reactions with other viruses were detected, indicating the adequate robustness and selectivity of the sensor. Our low-cost technology could facilitate more frequent testing for HSV-2., Competing Interests: DECLARATION OF INTERESTS A non-provisional patent application has been filed on the de la Fuente Lab’s related work (ID number 23–10377). C.F.N. provides consulting services to Invaio Sciences and is a member of the Scientific Advisory Boards of Nowture S.L. and Phare Bio. The de la Fuente Lab has received research funding or in-kind donations from United Therapeutics, Strata Manufacturing PJSC, and Procter & Gamble, none of which were used in support of this work. C.F.N. is on the Advisory Board of Cell Reports Physical Science.

Acepromazine (ACP) is a phenothiazine derivative drug commonly used as a tranquilizer veterinary medication due to its sedative properties. Benefiting from sedative properties, ACP has emerged as a drug of abuse and has been associated with drug-facilitated sexual assaults. Herein, we report, for the first time, the electrochemical behavior of ACP using a miniaturized and environmentally friendly laser-scribed graphene-based (LSG) sensor fabricated on a polyetherimide (PEI) substrate. The LSG device presented high porosity, as demonstrated by scanning electron microscopy (SEM). Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) measurements of the PEI-LSG electrode confirmed the enhanced electroactive area (3.1-fold increase) caused by the rough surface and revealed a low charge transfer resistance of the electrode material, with a heterogeneous electron transfer rate constant ( k 0 ) of 8.66 × 10 -3 cm s -1 for potassium ferricyanide redox probe. A simple and accurate method was applied to quantify ACP by using square wave voltammetry (SWV) under optimized experimental conditions, which exhibited high sensitivity (0.686 ± 0.008 A L mol -1 cm -2 ) and a low limit of detection (LOD) of 7.43 × 10 -8 mol L -1 , with a linear concentration ranging from 0.5 to 100 μmol L -1 ACP. Aiming for on-site analysis, the PEI-LSG sensor was integrated with a miniaturized potentiostat controlled by using a smartphone and applied as proof of applicability to ACP detection in commercial beverage and synthetic urine samples. These studies demonstrated adequate recoveries, ranging from 95.1% to 115.8%. The analytical parameters highlight the robustness and reliability of the proposed method for analyses of ACP directly at a potential crime scene.

COVID-19 has led to over 6.8 million deaths worldwide and continues to affect millions of people, primarily in low-income countries and communities with low vaccination coverage. Low-cost and rapid response technologies that enable accurate, frequent testing of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants are crucial for outbreak prevention and infectious disease control. Here we produce and characterize cellulose fibers naturally generated by the bacterium Gluconacetobacter hansenii as an alternative biodegradable substrate for manufacturing an eco-friendly diagnostic test for COVID-19. Using this green technology, we describe a novel and label-free potentiometric diagnostic test that can detect SARS-CoV-2 within 10 min and costs US$3.50 per unit. The test has bacterial cellulose (BC) as its substrate and a carbon-based electrode modified with graphene oxide and the human angiotensin-converting enzyme-2 (ACE2) as its receptor. Our device accurately and precisely detects emerging SARS-CoV-2 variants and demonstrates exceptional sensitivity, specificity, and accuracy for tested clinical nasopharyngeal/oropharyngeal (NP/OP) samples., Competing Interests: DECLARATION OF INTERESTS A non-provisional patent application has been filed on the de la Fuente Lab’s related work (ID number 23-10301). C.F.N. provides consulting services to Invaio Sciences and is a member of the Scientific Advisory Boards of Nowture S.L. and Phare Bio. The de la Fuente Lab has received research funding or in-kind donations from United Therapeutics, Strata Manufacturing PJSC, and Procter & Gamble, none of which were used in support of this work. C.F.N. is on the Advisory Board of Cell Reports Physical Science.

This work investigated the effect of zinc oxide nanoparticles functionalized with curcumin (ZnO (np)  + CUR) supplementation during the in vitro embryo culture (IVC) on the bovine in vitro embryo production, and the cellular antioxidant response. The cumulus-oocyte complexes (COCs) were matured, fertilized and then the presumptive zygotes were cultured in the medium in the absence (0 μM-control) or presence of different concentrations of ZnO (np)  + CUR (3, 6 or 12 μM). After IVC, the embryos were destined either to assay intracellular ROS levels and mitochondrial membrane potential. The results demonstrated that only the addition of 12 μM ZnO (np)  + CUR during IVC decreased intracellular ROS production and the rate of blastocyst production when compared to the control (p < .05). In conclusion, ZnO (np)  + CUR addition during the IVC impaired in concentration-dependent-manner bovine in vitro embryo production., (© 2023 Wiley-VCH GmbH. Published by John Wiley & Sons Ltd.)

Acute myocardial infarction (AMI) is the main cause of death worldwide, and the time of diagnosis is decisive for the effectiveness of the treatment of patients with AMI. Creatine kinase-myocardial band (CK-MB) has a predominance and high affinity with myocardial tissue, making it considered one of the main biomarkers for the diagnosis of AMI. In this work, we report a novel biodegradable composite material based on a polymer blend of Poly(3-hydroxybutyrate- co -3-hydroxyvalerate) and Poly(butylene adipate- co -terephthalate) (PHBV:Ecoflex) and graphite microparticles for sensitive and selective electrochemical detection of CK-MB. The morphological and physicochemical characterizations of the thermoplastic composite material revealed a homogeneous and synergistic distribution of the graphite microparticles through the blend structure, providing low defects and high electrical conductivity with high electron transfer kinetics ( k 0 = 3.54 × 10 -3 cm s -1 ) features with adequate flexibility for point-of-care applications. The portable and disposable devices were applied to detect CK-MB using the electrochemical impedance spectroscopy (EIS) technique in a relevant clinical concentration ranging from 5.0 ng mL -1 to 100.0 ng mL -1 and presented a limit of detection of 0.26 ng mL -1 CK-MB. The selectivity of the sensor was confirmed by testing the potential interference of major biomolecules found in biofluids and other relevant macromolecules. The accuracy and robustness were assessed by addition and recovery protocol in urine and saliva samples without sample pretreatment and demonstrated the potential of our method for rapid and decentralized tests of AMI. In addition, the study of the thermal, biological, and photodegradation of the devices after being used was also carried out, aiming at the disposal of the material more sustainably.

A portable and disposable laser-scribed graphene (LSG) device was fabricated on polyetherimide (PEI) substrate for electrochemical detection of benzodiazepines (BZ) drugs such as diazepam (DZ) and midazolam (MZ) in commercial beverage samples. Morphological characterizations of the LSG material recorded by scanning electron microscopy (SEM) revealed the porous nature of the proposed electrochemical device, which contributed to the enhancement of the electroactive area. Besides, the structural and electrochemical characterizations performed by Raman spectroscopy, cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS) measurements revealed that the PEI-LSG material presents highly disordered graphene-like structures and high electron transfer features, respectively. The electrochemical detection of DZ and MZ was carried out by Square Wave Voltammetry (SWV), whose analytical curves exhibited two linear intervals in concentrations ranging from 2.5 μmol L -1 to 25.0 μmol L -1 and from 25.0 μmol L -1 to 100.0 μmol L -1 for both BZ. We obtained limits of detection (LOD) and quantification (LOQ) of 0.66 and 2.18 μmol L -1 for DZ and 0.61 μmol L -1 and 2.01 μmol L -1 for MZ, respectively. The developed sensor was applied to detect DZ and MZ in commercial beverages such as juice, whisky, and sugarcane spirit samples to mimic potential forensic evidence of drug-facilitated crimes. The recoveries ranged from 97.1% to 117.2% for DZ and from 92.2% to 114.3% for MZ. In addition, the proposed method presented high manufacturing reproducibility (relative standard deviation (RSD) = 2.18% for DZ and RSD = 3.82% for MZ, n = 8 sensors) and adequate selectivity, highlighting the potential of PEI-LSG sensor as an excellent alternative method for forensic detection of crime facilitating drugs in commercial beverage samples., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Although cryopreservation of ovarian tissue has advanced greatly, it remains a challenge, and protocols should be optimized to handle the heterogeneous nature of ovarian samples. In an effort to address this factor, the present study evaluated the effects of corpus luteum (CL) and side of ovaries (right versus left) on cellular morphology and viability of vitrified bovine ovarian fragments in a closed system. The ovaries were categorized according to whether they had a CL and which side they were on, and then divided into six groups: 1) CL+ (with CL) group; 2) CL- (without CL) group; 3) right ovaries group; 4) left ovaries group; 5) fresh control group (ovaries without vitrification or culture that were not selected for CL or ovarian side) and 6) In vitro culture medium control group (non-vitrified ovaries that were not selected for the presence or absence of CL or side of the ovaries). The current study shows that the CL- and right groups had the greatest percentage of follicles with normal morphology compared to other vitrified-warmed groups. Furthermore, the levels of necrosis and tissue damage of the right cultured group were the lowest compared to other groups. It was shown that bovine ovarian tissues derived from right ovaries and ovaries without a corpus luteum can be functionally and morphologically preserved after vitrification. For the first time, the present study suggests that bovine ovarian tissue vitrification can be improved by considering the origin of the ovaries., Competing Interests: The author declares that there is no conflict of interest., (© 2023 Urmia University. All rights reserved.)

A laser-scribed graphene (LSG) device fabricated on polymeric polyetherimide (PEI) substrate is reported for sensitive electrochemical detection of xylazine (XLZ), a veterinary drug that has been associated with drug-facilitated crimes. Morphological characterization was made by scanning electron microscopy (SEM), demonstrating that the electrochemical device presents a highly porous carbonaceous structure. In addition, Raman spectroscopy, Fourier-transform infrared spectroscopy (FTIR), wettability, electrochemical impedance spectroscopy (EIS), and cyclic voltammetry (CV) measurements were used to characterize the PEI-LSG material, which showed superior electroanalytical performance after a simple anodic treatment in an alkaline medium (applying 2 V for 120 s in 0.1 mol L -1 NaOH solution). The electrochemical determination of XLZ was carried out using square wave voltammetry (SWV), which showed sensitivity and coefficient of determination (R 2 ) equal to 0.206 ± 0.008 A L mol -1  cm -2 and 0.991, respectively. In addition, we obtained a low limit of detection (LOD) of 1.39 × 10 -7  mol L -1 with a linear response in the concentration range 5.0 to 200.0 × 10 -6  mol L -1 . The PEI-LSG electrochemical sensor was applied to XLZ detection in commercial beverage and synthetic urine samples, providing recoveries between 96.0 and 114.8%. Collectively, our electrochemical sensor presents an easy method to manufacture reproducible (relative standard deviation (RSD) of 2.56%), low-cost ($0.12), and single-use (disposable) devices, which is a promising way for in-field determination of drug-facilitated sexual assaults and other relevant applications., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Lectins participate in the defense against microorganisms and in signaling the damage caused by pathogens to the cell surface and/or intracellular in plants. This study aims to analyze the antifungal potential of lectins extracted from seeds of Canavalia ensiformis (L.) DC and Canavalia rosea (Sw.) DC, against Candida albicans and Candida tropicalis. The antimicrobial tests were performed by microdilution against Candida spp. The test to verify the combined lectin/fluconazole effect was performed using subinhibitory concentrations of lectins and with antifungal ranging from 0.5 to 512 µg/mL. The ability to inhibit the morphological transition of Candida spp. was evaluated by microcultivation in a moist chamber. The results of the minimum inhibitory concentration revealed no antifungal activity against the tested strains. However, lectins modified the action of fluconazole, reducing the IC 50 of the drug against C. albicans. Lectins were also able to discretely modulate the morphological transition of the tested strains., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

COVID-19 has killed over 6 million people worldwide. Currently available methods to detect SARS-CoV-2 are limited by their cost and need for multistep sample preparation and trained personnel. Therefore, there is an urgent need to develop fast, inexpensive, and scalable point-of-care diagnostics that can be used for mass testing. Between January and March 2021, we obtained 321 anterior nare swab samples from individuals in Philadelphia (PA, USA). For the Real-time Accurate Portable Impedimetric Detection prototype 1.0 (RAPID) test, anterior nare samples were tested via an electrochemical impedance spectroscopy (EIS) approach. The overall sensitivity, specificity, and accuracy of RAPID in this cohort study were 80.6%, 89.0%, and 88.2%, respectively. We present a rapid, accurate, inexpensive (<$5.00 per unit), and scalable test for diagnosing COVID-19 at the point-of-care. We anticipate that further iterations of this approach will enable widespread deployment, large-scale testing, and population-level surveillance., Competing Interests: A non-provisional patent application has been filed on related work., (© 2022 The Authors.)

(1) Background: Candida is a genus of yeasts with notable pathogenicity and significant ability to develop antimicrobial resistance. Gossypium hirsutum L., a medicinal plant that is traditionally used due to its antimicrobial properties, has demonstrated significant antifungal activity. Therefore, this study investigated the chemical composition and anti- Candida effects of aqueous (AELG) and hydroethanolic (HELG) extracts obtained from the leaves of this plant. (2) Methods: The extracts were chemically characterized by UPLC-QTOF-MS/MS, and their anti- Candida activities were investigated by analyzing cell viability, biofilm production, morphological transition, and enhancement of antifungal resistance. (3) Results: The UPLC-QTOF-MS/MS analysis revealed the presence of twenty-one compounds in both AELG and HELG, highlighting the predominance of flavonoids. The combination of the extracts with fluconazole significantly reduced its IC 50 values against Candida albicans INCQS 40006, Candida tropicalis INCQS 40042, and C. tropicalis URM 4262 strains, indicating enhanced antifungal activity. About biofilm production, significant inhibition was observed only for the AELG-treated C. tropicalis URM 4262 strain in comparison with the untreated control. Accordingly, this extract showed more significant inhibitory effects on the morphological transition of the INCQS 40006 and URM 4387 strains of C. albicans (4) Conclusions: Gossypium hirsutum L. presents promising antifungal effects, that may be potentially linked to the combined activity of chemical constituents identified in its extracts.

Several single nucleotide polymorphisms (SNPs) could indirectly, as well directly, influence metabolic parameters related to health effects in response to selenium (Se) supplementation. This study aimed to investigate whether the selenoprotein SNPs were associated with the response of Se status biomarkers to the Brazil nut consumption in patients using statins and if the variation in Se homoeostasis could affect antioxidant protection, lipid profile, muscle homoeostasis and selenoproteins mRNA. The study was performed in the Ribeirão Preto Medical School University Hospital. Thirty-two patients using statins received one unit of Brazil nut daily for 3 months. Body composition, blood Se concentrations, erythrocyte glutathione peroxidase (GPX) activity, total cholesterol, low-density lipoprotein (LDL), high-density lipoprotein (HDL), triacylglycerol (TAG), creatine kinase (CK) activity and gene expression of GPX1 and selenoprotein P (SELENOP) were evaluated before and after Brazil nut consumption. The volunteers were genotyped for SNP in GPX1 (rs1050450) and SELENOP (rs3877899 and rs7579). SNPs in selenoproteins were not associated with plasma and erythrocyte Se, but SNPs in SELENOP influenced the response of erythrocyte GPX activity and CK activity, TAG and LDL after Brazil nut consumption. Also, Brazil nut consumption increased GPX1 mRNA expression only in subjects with rs1050450 CC genotype. SELENOP mRNA expression was significantly lower in subjects with rs7579 GG genotype before and after the intervention. Thus, SNP in SELENOP could be associated with interindividual differences in Se homeostasis after Brazil nut consumption, emphasising the involvement of genetic variability in response to Se consumption towards health maintenance and disease prevention.

This article aims to evaluate the predictive factors of morbidity and mortality in pediatric patients who suffered gunshot wounds to the head. We reviewed a series of 43 patients who were admitted to a referential neurosurgical hospital between 2010 and 2019. Data from 43 patients who underwent a surgical treatment in our institution were collected, and the following parameters were considered in the analysis: the initial Glasgow Coma Scale (GCS), age, sex, bullet entry site, and bullet trajectory. Computed tomography (CT) scans at admission, complications, midline crossing, and Glasgow score scale at the time of discharge (Glasgow Outcome Scale; GOS) were also factored in. Male sex corresponded to 90.7% of cases ( N  = 39), and 16-17 years of age was the most common age (60.5%). The frontal region was the most common entry site (41.9%), followed by the parietal wall and occipital entry. Penetrating trajectory was shown in 48.8% of cases, perforation/transfixing in 39.5%, and tangential in 11.6%. CT showed that sinking is the most common alteration (74.4%), followed by cerebral contusion (44.2%). According to the GOS, 23.3% died, 23.3% were classified by an unfavorable outcome (GOS, 2-3), and 53.5% a favorable outcome (GOS, 4 and 5). In our study, there was a significant association between the low GCS scores on admission and low GOS (1-3; p  = 0.001) at time of discharge. Patients with wounds that crossed the midline also had a significant association with low GOS ( p  = 0.014) in our clinical experience. We concluded that low GCS scores at admission and children with a wound that crosses the midline are predictive factors of high mortality and morbidity, in our clinical experience., Competing Interests: No competing financial interests exist., (© Luiz Severo Bem Junior et al., 2021; Published by Mary Ann Liebert, Inc.)

The COVID-19 pandemic has exacerbated our society's tremendous health equity gap. Disadvantaged populations have been disproportionally affected by COVID-19, lacking access to affordable testing, a known effective tool for preventing viral spread, hospitalizations, and deaths. Here, we describe COVID-19 Low-cost Optodiagnostic for Rapid testing (COLOR), a colorimetric biosensor fabricated on cotton swabs using gold nanoparticles modified with human angiotensin-converting enzyme 2 (ACE2), which costs 15¢ to produce and detects SARS-CoV-2 within 5 min. COLOR detected very low viral particle loads (limit of detection: 0.154 pg mL -1 of SARS-CoV-2 spike protein), and its color intensity correlated with the cycle threshold (Ct) values obtained using reverse transcription polymerase chain reaction (RT-PCR). The performance of COLOR was assessed using 100 nasopharyngeal/oropharyngeal (NP/OP) clinical samples, yielding sensitivity, specificity, and accuracy values of 96%, 84%, and 90%, respectively. In summary, each COLOR test can be manufactured for 15¢ and presents rapid minute-time scale detection of SARS-CoV-2, thus providing a solution to enable high-frequency testing, particularly in low-resource communities.

Background: Variable resistance training has recently become a component of strength and conditioning programs., Objective: This randomized counterbalanced cross-over study aimed to investigate the use of elastic bands (EB) and the traditional method (TRAD) and force indicators in a training session., Methods: 12 Paralympic athletes (age: 28.60 ± 7.60 years) participated in this three-week study. In the first week, the participants were familiarized with EB and TRAD and were tested for maximal repetition (1-RM). The research occurred in weeks 2 and 3, which included the pre-post training, during which the following measures were extracted: maximum isometric force (MIF), the peak torque (PT), rate of force development (RFD), fatigue index (FI), and time to MIF (Time). The athletes performed two tests, EB and TRAD, separated by a one-week interval., Results: Significant differences were found between the pre- and post-test for 1RM ( p = 0.018, η2p = 0.412), MIF ( p = 0.011, η2p = 0.415), PT ( p = 0.012, η2p = 0.413), and RFD ( p = 0.0002, η2p = 0.761). With the use of EB, there was a difference in RFD between TRAD before and EB after ( p = 0.016, η2p = 0.761). There were significant differences in the before and after for FI between TRAD and EB ( p < 0.001) and for Time ( p < 0.001), indicating that training with the use of elastic bands promotes overload, characterized by increased fatigue and decreased strength., Conclusions: Training with EB did not decrease 1RM, PT, MIF or RFD, however, there was an increase in fatigue and time to reach MIF when compared to the method with fixed resistance.

COVID-19 has led to over 3.47 million deaths worldwide and continues to devastate primarily middle- and low-income countries. High-frequency testing has been proposed as a potential solution to prevent outbreaks. However, current tests are not sufficiently low-cost, rapid, or scalable to enable broad COVID-19 testing. Here, we describe LEAD (Low-cost Electrochemical Advanced Diagnostic), a diagnostic test that detects severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) within 6.5 min and costs $1.50 per unit to produce using easily accessible and commercially available materials. LEAD is highly sensitive toward SARS-CoV-2 spike protein (limit of detection = 229 fg⋅mL -1 ) and displays an excellent performance profile using clinical saliva (100.0% sensitivity, 100.0% specificity, and 100.0% accuracy) and nasopharyngeal/oropharyngeal (88.7% sensitivity, 86.0% specificity, and 87.4% accuracy) samples. No cross-reactivity was detected with other coronavirus or influenza strains. Importantly, LEAD also successfully diagnosed the highly contagious SARS-CoV-2 B.1.1.7 UK variant. The device presents high reproducibility under all conditions tested and preserves its original sensitivity for 5 d when stored at 4 °C in phosphate-buffered saline. Our low-cost and do-it-yourself technology opens new avenues to facilitate high-frequency testing and access to much-needed diagnostic tests in resource-limited settings and low-income communities., Competing Interests: The authors declare no competing interest., (Copyright © 2021 the Author(s). Published by PNAS.)

SARS-CoV-2, the virus that causes COVID-19, has killed over 3 million people worldwide. Despite the urgency of the current pandemic, most available diagnostic methods for COVID-19 use RT-PCR to detect nucleic acid sequences specific to SARS-CoV-2. These tests are limited by their requirement of a large laboratory space, high reagent costs, multistep sample preparation, and the potential for cross-contamination. Moreover, results usually take hours to days to become available. Therefore, fast, reliable, inexpensive, and scalable point-of-care diagnostics are urgently needed. Here, we describe RAPID 1.0, a simple, handheld, and highly sensitive miniaturized biosensor modified with human receptor angiotensin-converting enzyme-2. RAPID 1.0 can detect SARS-CoV-2 using 10 μL of sample within 4 min through its increased resistance to charge transfer of a redox probe measured by electrochemical impedance spectroscopy. The sensitivity and specificity of RAPID for nasopharyngeal/oropharyngeal swab and saliva samples are 85.3% and 100% and 100% and 86.5%, respectively., Competing Interests: The authors declare no competing interests., (© 2021 Elsevier Inc.)

A new method to manufacture electrochemical devices based on the graphite and colorless nail polish (N-grap) film was developed for tartrazine (Tz) detection. Scanning Electron Microscopy (SEM) demonstrates that the composite material presents a high porous carbon structure. Cyclic voltammetry (CV) and Electrochemical Impedance Spectroscopy (EIS) were employed to electrochemically characterize the electrode material, which corroborates the porous structure of the N-graph due to the enhanced electroactive area (5.4-fold increase) and presented a heterogeneous electron transfer rate constant (k 0 ) of 5.82 × 10 -3  cm s -1 for potassium ferricyanide. The electrochemical determination of the Tz was carried out using square-wave voltammetry (SWV), under the optimized experimental conditions, which showed high sensitivity (0.793 A L mol -1 ) and a lower limit of detection (LOD) of 2.10 × 10 -8  mol L -1 with a linear concentration ranging from 2.0 to 50.0 μmol L -1 . The developed sensor was applied for the analysis of Tz in sports drink samples and the result obtained by N-grap device was statistically compared with a spectrophotometric method demonstrating good accordance and the accuracy of the proposed method. Based on these results, we believe that this new fabrication method to produce disposable and low-cost electrochemical devices can be an alternative method for in-field analysis of dye in commercial sport drink samples and other relevant applications., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The α-adrenergic agents are known to influence erection and ejaculation, and ejaculatory reflex is a primarily α-adrenergically mediated event. This paper aims to describe the urethral catheterization technique after pharmacological induction for semen collection in domestic dogs. For dose determination, semen collection was performed using increasing doses of dexmedetomidine (5 μg/kg, n = 2; 10 μg/kg, n = 2 and 15 μg/kg, n = 2) associated to 3 mg/kg ketamine in six animals. Once the dose was established, additional semen collection using association of 15 μg/kg dexmedetomidine and 3 mg/kg ketamine was carried out in eight dogs. After the procedure, sperm volume (0.092 ± 0.03 mL), total motility (58.33 ± 8.7%), vigor (1.89 ± 0.309), sperm concentration (1186.67 ± 304.667 × 10 6 sperm/mL), sperm plasma membrane integrity (83.22 ± 4.82%), acrosomal integrity (73 ± 6.187%) and normal sperm morphology (53.22 ± 5.6%) were assessed. Thus, this study demonstrates that urethral catheterization after pharmacological ejaculation using dexmedetomidine-ketamine association can be performed in dogs and it may be possible to obtain good semen quality, despite urine contamination., (Copyright © 2020 Elsevier Inc. All rights reserved.)

This study investigated the effect of Folliculinum 6 cH on the oocyte meiosis resumption and viability rates, progesterone production and mitochondrial activity after in vitro maturation of cumulus-oocyte complexes (COCs) in sheep. Sheep ovaries were collected at a local slaughterhouse and COCs were recovered by slicing technique. The selected COCs were maturated in TCM199 (Control treatment), or control medium supplemented with 0.05% ethanol (v/v) (the vehicle of the homeopathic preparation - Ethanol treatment) or with Folliculinum 6 cH . After 24 h of in vitro maturation (IVM), oocytes were mechanically denuded and incubated with Hoechst 33342 and MitoTracker (0.5 μM) Orange CMTMRos for analysis of viability and chromatin configuration, and mitochondrial activity, respectively. The results showed that Folliculinum 6 cH addition increased oocyte degeneration and reduced meiotic resumption compared to the control (P < 0.05). Interestingly, the percentages meiotic resumption and oocyte maturation were lower in the Folliculinum 6 cH treatment compared to its vehicle (Ethanol treatment) (P < 0.05). On the other hand, when the treatments were compared, higher mitochondrial activity was observed in the Ethanol treatment (P < 0.05). In conclusion, contrary to its vehicle, the addition of Folliculinum 6 cH to the IVM medium promoted oocyte degeneration and affected negatively the mitochondrial distribution, impairing meiosis resumption., Competing Interests: Conflicts of interest: The authors have no conflict of interest to declare., (Copyright © The Author(s).)

Type and concentration of cryoprotective agents (CPAs) are important factors which influence the likelihood of a successful ovarian tissue vitrification outcome. In an attempt to address this factor, the present study was conducted to evaluate the impacts of different synthetic polymers (Supercool X-1000, Supercool Z-1000 and PVP K-12) on vitrification of bovine ovarian tissue. From each ovarian pair, fragments were recovered and immediately fixed for analysis (fresh control) or submitted to vitrification, either or not followed by in vitro culture for one or five days. Vitrification was performed using the ovarian tissue cryosystem (OTC) system. The ovarian tissues were intended for histological and viability analysis [Reactive oxygen species (ROS) production and degenerate cells assay (Ethidium homodimer-1)], as well as immunolocalization of AQP3 and AQP9 were measured. The results showed that during almost all the periods after warming, in treatment groups which contain polymer (X-1000, Z-1000 and PVP), the percentage of morphologically normal follicles was the highest in the X-1000 samples. Furthermore, post-thawed X-1000 group revealed stronger labeling for AQP9 in primordial and transitional follicles, when compared with others. However, morphology after cryopreservation did not correlate with follicle viability and function where the levels of degeneration and tissue damage of PVP K-12 group were lower in comparison with X-1000 group and only in PVP K-12 group, ROS level was similar to that of the fresh control group. We believe that in addition to permeating CPAs, the addition of one (Supercool X-1000) or maybe a combination (Supercool X-1000 and PVP K-12) of non-permeating polymers could be useful to improve the outcome for vitrified bovine ovarian tissue., Competing Interests: Declaration of competing interest There is no competing interest., (Copyright © 2020 Elsevier Inc. All rights reserved.)

The in vitro follicle culture (IVFC) represents an outstanding tool to enhance our understanding of the control of folliculogenesis and to allow the future use of a large number of immature oocytes enclosed in preantral follicles (PFs) in assisted reproductive techniques in humans as well as in others mammalian species including the ruminants. So far, the best results of IVFC were reported from mice with the production of live offspring from primordial follicles cultured in vitro. Live birth has been obtained after the in vitro culture of bovine early antral follicles. However, in other ruminant species, these results have been limited to the production of a variable number of mature oocytes and low percentages of embryos after in vitro culture of goat, buffalo and sheep isolated secondary preantral follicles. The present review presents and discusses the main findings, limitations, and prospects of in vitro folliculogenesis in ruminants focusing on bovine, caprine, and ovine species., (Copyright © The Author(s). Published by CBRA.)

The culture of preantral follicles as an in vitro model to evaluate the toxicity of new anticancer drug has being established. Therefore, the aim of this study was to evaluate the effect of quinoxaline derivative the 2 2- (XYZC 6H 3 -CH=N-NH)-quinoxaline, 1 (QX) on caprine preantral follicles. We evaluate the follicular morphology and activation, proliferation and apoptosis of granulosa cells and finally the protein (ABCB1) and genes expression (cyclin/Cdks), respectively involved in multidrug resistance and cell cycle progression. Ovarian fragments containing primordial and developing follicles were exposed (in vitro culture) to different concentrations of QX (QX1.5, QX3.0 or QX6.0 μM/mL) during 6 days. To evaluate the effect of QX, the ovarian tissue was exposed to Paclitaxel 0.1 μg/mL (PTX - negative control) or in culture media without QX (MEM). At the end of exposure time, we realized that the QX (all concentrations) increased (P < .05) the normal morphology of preantral follicles compared to control (not treated ovarian tissue) or MEM. However, QX6.0 showed a enhanced (P < .05) on follicular activation (burnout) and apoptosis than QX1.5 and QX3.0. Expression of ABCB1 was similar between QX1.5 and QX6.0 and both were lower than control, MEM and PTX. Interestingly, the apoptosis rate in QX3.0 was similar to control and MEM and lower then QX1.5; QX6.0 and PTX. We conclude that quinoxaline may be a promising chemotherapeutic agent, however, other concentrations within a defined range (2-5.5 μM) could be widely investigated., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The Piper mikanianum species were investigated by the antimicrobial potential and chemical composition. Chemical analysis was performed by gas chromatography coupled to mass spectrometry (GC/MS). The Minimum Inhibitory Concentration (MIC) as well as the 50% Inhibitory Concentration against Candida strains were determined by microdilution. The effect of the drug-oil combination was also evaluated to verify possible synergism. The Minimum Fungicidal Concentration (MFC) was evaluated by subculturing the microdilution in Petri dishes and the anti-pleomorphism potential of the oil was tested in humid chambers. Chemical analysis revealed safrol as the major compound. The results from the intrinsic activity evaluation of the oil did not reveal a clinical importance, however, it presented a synergistic effect when associated with gentamicin against the multidrug resistant E. coli strain and when associated with fluconazole against fungal strains. Moreover, the oil possessed a fungistatic effect. Total inhibition of filamentous structures occurred in both Candida species in the anti-virulence test. The P. mikanianum essential oil showed a potentiating activity of drugs for which resistance exists and an inhibitory effect of one of the main virulence factors of the Candida genus, morphological transition, which has been previously shown to be responsible for causing invasive infections in human tissues., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Early nutritional aggressions promote epigenetic adjustments that culminate in the loss of phenotype plasticity (with permanent long-term modifications). Maternal diet and inadequate neonatal nutrition can result in fetal programming that presents susceptibility to infections in adult life. Thus, it becomes essential to verify the impacts of neonatal malnutrition (even following nutritional replacement) on the immunological response to methicillin resistant Staphylococcus aureus (MRSA) infections. Male rats were divided into two distinct groups: Nourished and Malnourished. After isolation of mononuclear cells, four systems were established: negative control, positive control and two testing systems, (MSSA and MRSA). Tests were performed to analyze expression of TLR-9, NF-kB, IL-1β, IL-18 and IL-33. For statistical analysis, we used the Student t and ANOVA tests p < 0.05. Even after nutritional replacement, malnutrition in the neonatal period compromised the animals' weight gains p < 0.05. There was a reduction in the expression of the immunological response in the positive control, however deregulation was observed in the gene expression of MRSA-infected macrophages, with a reduction in TLR-9 expression, and overexpression in NF-kB and cytokines p < 0.05. Puppies inflicted with protein-calorie malnutrition were compromised; (long-term) body growth and immune response. In the infectious scenario, immune collapse is reflected in inflammatory response exacerbation with a likely histolytic character. Immune disabling (resulting from gene expression deregulation) causes susceptibility to infections due to ineffective recognition, intense pro-inflammatory mediation, and cell death. It is suggested that neonatal malnutrition can program susceptibility to multiresistant bacterial infections, and generally favors a triggering of more intense confrontations with fatal outcomes., (Copyright © 2019. Published by Elsevier Ltd.)

The Withanolide D is a chemotherapeutic potential against the human tumor cell. However, there is no report on the effect of this compound on ovarian function, especially on preantral folliculogenesis. The aim of this study was to evaluate the toxicity of a new candidate to anticancer drug, Withanolide D (WD) on morphologic integrity, development (activation and granulosa cell proliferation) and gene expression of ABCB1 protein of caprine preantral follicles. Ovarian fragments were cultured in vitro for 2 or 6 days in α-MEM or α-MEM added with paclitaxel (PTX -0.1 μg/mL; negative control) and different concentrations of WD (WD1.5, WD3.0 or WD6.0). The higher dose of WD showed a toxic effect similar to PTX and higher (P < 0.05) than other treatments after 2 and 6 days. In addition, WD6.0 reduced the cell proliferating compared to PTX or mild dose. The expression of ABCB1 remained unchanged in the presence of the chemotherapeutic agents (PTX and WD) throughout the culture period. In conclusion, WD exerted a toxic effect observed by decreasing follicular survival and cell proliferation, on the preantral caprine follicles similar to PTX, whose negative effect on folliculogenesis is already widely known., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Aldehydes are relevant analytes in a wide range of samples, in particular, food and beverages but also body fluids. Hydrazines can undergo nucleophilic addition with aldehydes or ketones giving origin to hydrazones (a group of stable imines) that can be suitably used in the identification of aldehydes. Herein, 4-hydrazinobenzoic acid (HBA) was, for the first time, used as the derivatizing agent in analytical methodologies using liquid chromatography aiming the determination of low-molecular aldehydes. The derivatization reaction was simultaneously performed along with the extraction process, using gas-diffusion microextraction (GDME), which resulted in a clean extract containing the HBA-aldehyde derivates. The corresponding formed imines were determined by both high-performance liquid chromatography (LC) with UV spectrophotometric detection (HPLC-UV) and capillary electrophoresis with diode array detection (CE-DAD). HBA showed to be a rather advantageous derivatization reagent due to its stability, relatively high solubility in water and other solvents, high selectivity and sensibility, reduced impurities, simple preparation steps and applicability to different separation and/or different detection techniques. Limits of detections (LODs) of the optimized methodologies (in terms of time and pH among other experimental variables) were all below 0.5 mg L -1 , using both instrumental techniques. Furthermore, for the first time, the HBA-aldehyde derivatives were analyzed by LC with mass spectrometry (LC-MS), demonstrating the possibility of identification by MS of each compound. The developed methodologies were also successfully applied in the analysis of formaldehyde and acetaldehyde in several alcoholic beverages. This was also the first time GDME was combined with CE, showing that it can be a valuable sample preparation tool for electrophoresis, in particular by eliminating the interference of ions and inorganic constituents present in the samples., (Copyright © 2018. Published by Elsevier B.V.)

The regulation of folliculogenesis involves a complex interaction among endocrine, paracrine and autocrine factors. The mechanisms involved in the initiation of the growth of the primordial follicle, i.e., follicular activation and the further growth of primary follicles up to the pre-ovulatory stage, are not well understood at this time. The present review focuses on the regulation and development of early stage (primordial, primary, and secondary) folliculogenesis highlighting the mechanisms of primordial follicle activation, growth of primary and secondary follicles and finally transition from secondary to tertiary follicles. We also discuss the importance of in vitro follicle culture for the understanding of folliculogenesis during the preantral phase. Studies suggest that follicular development from primordial to early antral stages is primarily controlled by intra-ovarian ligands but it can also be influenced by many extra-ovarian factors. The control of early folliculogenesis is, therefore, extremely complex because several ligands act through distinct signaling pathways that form sophisticated information networks responding to multiple, often opposing, stimuli. The balance among different stimuli determines follicular survival or death as well as quiescence or activation (growth). The distribution of the ligands and their corresponding receptors varies among follicular compartments and species, and significant changes in gene expression pattern among follicular categories have been reported. Knowing that follicular requirements during early folliculogenesis can be stage-specific and species-specific, in vitro culture studies offer an alternative to evaluate single and combined factors during a specific period of follicular development. Herewith we summarize the main findings obtained in vitro together with the mechanisms regulating folliculogenesis.

The multidrug resistance proteins ABCB1, ABCC2 and ABCG2 are an energy-dependent efflux pump that functions in systemic detoxification processes. Physiologically expressed in a variety of tissues, most abundantly in the liver and intestinal epithelia, placenta, blood-brain barrier and various stem cells, until now, these pumps were not identified in goat ovarian tissue. Therefore, the aim of this study is to analyze ABCB1, ABCC2, and ABCG2 mRNA and protein expression in goat preantral follicles. Fragments (3 × 3 × 1 mm) from five pairs of ovary (n = 10) obtained from five goat were collected and immediately submitted to qPCR, Western blot, and immunofluorescence assay for mRNA detection and identification and localization of the ABC transporters, respectively. mRNA for ABCB1, ABCC2, and ABCG2 and the presence of their proteins were observed on ovarian tissue samples. Positive marks were observed for the three transport proteins in all follicular categories studied. However, the marks were primarily localized in the oocyte of primordial, transition and primary follicle categories. In conclusion, goat ovarian tissue expresses mRNA for the ABCB1, ABCC2 and ABCG2 transporters and the expression of these proteins in the preantral follicles is a follicle-dependent stage.

Objective: This study investigated the effect of two different follicle stimulating hormone (FSH) preparations (diluted/dynamised and diluted) on the in vitro development and steroid production (estradiol, progesterone and testosterone) of isolated porcine preantral follicle after in vitro culture., Methods: Secondary follicles were cultured in Alpha Minimum Essential Medium (α-MEM + ) supplemented with grain ethanol (AL - 0.2%, v/v), diluted/dynamised FSH (rFSH 6cH - 0.05 fg/mL) or diluted-only FSH (1.5 ng/mL) for 4 days. Follicle development was evaluated on the basis of follicular growth, morphology and hormone production., Results: The percentage of follicular integrity and extrusion were not affected by the treatments after culture. For all treatments, follicular diameter increased significantly from Day 0 to Day 4. On Day 2 of culture, the estradiol production was significantly higher in AL and diluted-only FSH treatments compared with diluted/dynamised FSH. However, diluted/dynamised FSH showed a significantly higher progesterone production on Day 2. Only on Day 4, the testosterone production was higher in the AL than diluted-only FSH treatments, but similar to diluted/dynamised FSH treatment. Except for diluted/dynamised FSH treatment, progesterone production increased (P < 0.05) from Day 2 to Day 4; only for AL treatment, a significant increase of testosterone production was observed during culture., Conclusion: Compared to control the diluted/dynamised FSH addition increased progesterone production but decreased the estradiol production after in vitro culture of isolated porcine preantral follicles. Taken together the results suggest that at least for progesterone production the mechanism of action of diluted/dynamised FSH differs from its vehicle., (Copyright © 2017 The Faculty of Homeopathy. Published by Elsevier Ltd. All rights reserved.)

Despite the advances made in monitoring and treatment of sepsis and septic shock, many septic patients ultimately develop multiple organ dysfunction (MODS) and die, suggesting that other players are involved in the pathophysiology of this syndrome. Mitochondrial dysfunction occurs early in sepsis and has a central role in MODS development. MODS severity and recovery of mitochondrial function have been associated with survival. In recent clinical and experimental investigations, mitochondrion-target therapy for sepsis and septic shock has been suggested to reduce MODS severity and mortality. This intervention, which might be named "metabolic resuscitation", would lead to improved mitochondrial activity afforded by pharmacological and nutritional agents. Of particular interest in this therapeutic strategy is thiamine, a water-soluble vitamin that plays an essential role in cellular energy metabolism. Critical illness associated with hypermetabolic states may predispose susceptible individuals to the development of thiamine deficiency, which is not usually identified by clinicians as a source of lactic acidosis. The protective effects of thiamine on mitochondrial function may justify supplementation in septic patients at risk of deficiency. Perspectives of supplementation with other micronutrients (ascorbic acid, tocopherol, selenium and zinc) and potential metabolic resuscitators [coenzyme Q10 (CoQ10), cytochrome oxidase (CytOx), L-carnitine, melatonin] to target sepsis-induced mitochondrial dysfunction are also emerging. Metabolic resuscitation may probably be a safe and effective strategy in the treatment of septic shock in the future. However, until then, preliminary investigations should be replicated in further researches for confirmation. Better identification of groups of patients presumed to benefit clinically by a certain intervention directed to "mitochondrial resuscitation" are expected to increase driven by genomics and metabolomics.

Cryoinjuries caused by vitrification of tissues and organs lead to the loss of membrane proteins that mediate intercellular communications, such as connexins 37 (Cx37) and 43 (Cx43). Thus, the present study aimed to evaluate ovine Cx37 and Cx43 gene and protein expressions and developmental competence by in vitro-cultured secondary follicles retrieved from vitrified ovarian tissue. Ovarian fragments for the same ovary pair were distributed into six treatments: (1) fresh ovarian tissue (FOT); (2) vitrified ovarian tissue (VOT); (3) isolated follicles from fresh ovarian tissue (FIF); (4) isolated follicles from vitrified ovarian tissue; (5) isolated follicles from fresh ovarian tissue followed by in vitro culture (CFIF); (6) isolated follicles from vitrified ovarian tissue followed by in vitro culture (CVIF). In all treatments, Cx37 and Cx43 gene and protein expression patterns were evaluated by reverse transcription polymerase chain reaction and immunocytochemistry. In addition, secondary follicles were analyzed according to follicular integrity and growth, apoptosis, and cell proliferation. In vitro-cultured secondary follicles (CFIF and CVIF) were evaluated based on morphology (extruded follicles), antrum formation, and viability. The percentage of intact follicles was higher, whereas antrum formation, oocyte extrusion rate, and follicle viability were lower in CVIF than in CFIF treatment (P < 0.05). Terminal deoxynucleotidyl transferase-mediated biotinylated deoxyuridine triphosphates nick end-labeling assay demonstrated that apoptosis was absent in FIF, whereas follicles from all other treatments showed positive labeling. Cell proliferation index was higher in isolated follicles from vitrified ovarian tissue and CVIF treatments than in follicles from FIF. Expression of Cx43 messenger RNA was lower in CVIF treatment when compared with follicles from all other treatments (P < 0.05). Follicle Cx37 messenger RNA levels did not show alterations in any treatment (P > 0.05). Cx37 and Cx43 immunolabeling was localized mainly on granulosa cells and oocytes, respectively. In conclusion, isolation of ovine secondary follicles could be done successfully after vitrification of ovarian tissue, and the basement membrane integrity remained intact after in vitro culture. Although the gene and protein expression of Cx37 did not change after vitrification of ovarian tissue, Cx43 turned out to be altered in secondary follicles after vitrification and in vitro culture., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Cryopreservation of preantral follicles is a promising technique to preserve female fertility. The aim of this study was to evaluate the effect of vitrification on the development of secondary follicles included in ovarian tissue or isolated after microdissection. An important end point included is the capacity of grown oocytes to resume meiosis. Sheep ovarian cortexes were cut into fragments and split into three different groups: (1) fresh (control): secondary follicles isolated without any previous vitrification; (2) follicle-vitrification (follicle-vit): secondary follicles vitrified in isolated form; and (3) tissue-vitrification (tissue-vit): secondary follicles vitrified within fragments of ovarian tissue (in situ former) and subsequently subjected to isolation. From the three groups, isolated secondary follicles were submitted to IVC for 18 days. After IVC, cumulus-oocyte complexes (COCs) were harvested from follicles. As an additional control group, in vivo grown, in vivo-grown COCs were collected from antral ovarian follicles. All, recovered COCs were matured and the chromatin configuration was evaluated. Data were analyzed by ANOVA, and the means were compared by Student-Newman-Keuls test, and by chi-square. Differences were considered to be significant when P < 0.05. Isolated preantral follicles from all treatments had normal morphology, antrum formation, and low follicle degeneration after IVC. The growth rate between control and follicle-vit did not differ (P > 0.05), and was higher (P < 0.05) than for tissue-vit. The percentage of follicles that decreased diameter during IVC was significantly higher in tissue-vit than the in follicle-vit. Recovery rate of oocytes from normal follicles was higher in follicle-vit than in tissue-vit. Furthermore, oocyte viability was lower in tissue-vit than other treatments, and follicle-vit did not differ from control and in vivo grown. The percentage of oocytes meiosis resuming was not different between treatments except for in vivo grown. After vitrification, only follicle-vit showed metaphase I oocyte. We conclude that secondary follicles vitrified after isolation displayed a better follicular growth rate, oocyte viability, percentage of oocytes reaching the metaphase I stage, and fewer follicles with decreased diameter after IVC., (Copyright © 2016 Elsevier Inc. All rights reserved.)

The risk of reintroducing malignant cells after ovarian graft into patients following post-cancer treatment is an obstacle for clinical applications (autotransplantation). In this context, in vitro follicle culture would be an alternative to transplantation in order to minimize such risks. Therefore, the aim of this study was to compare the development of secondary follicles after vitrification in isolated form (without stroma) with vitrification in in situ form (within fragments of ovarian tissue). Follicles were first isolated from ovarian fragments from mixed-breed ewes and then vitrified; these comprised the Follicle-Vitrification group (Follicle-Vit), or fragments of ovarian tissue were first vitrified, followed by isolation of the follicles, resulting in the Tissue-Vitrification group (Tissue-Vit). Control and vitrified groups were submitted to in vitro culture (6 days) and follicular morphology, viability, antrum formation, follicle and oocyte diameter, growth rate, ultrastructural characteristics and cell proliferation were evaluated. The percentages of morphologically normal follicles and antrum formation were similar among groups. Follicular viability and oocyte diameter were similar between Follicle-Vit and Tissue-Vit. The follicular diameter and growth rate of Follicle-Vit were similar to the Control, while those of Tissue-Vit were significantly lower compared to the Control. Both vitrified groups had an augmented rate of granulosa cellular proliferation compared to Control. Secondary follicles can be successfully vitrified before or after isolation from the ovarian tissue without impairing their ability to survive and grow during in vitro culture.

The mRNA expression and localization of Aquaporin 3 (AQP3) were investigated in the ovarian follicles of ewes at different stages of development (primordial, primary, secondary, small, and large antral). The gene expression was quantified by qPCR, while the protein identification and localization were determined by Western blot and immunohistochemistry, respectively. Analysis revealed that AQP3 mRNA was detected only in the antral follicles, whereas the protein expression was detected in the oocyte and granulosa cells in all stages of follicular development. The latter observation suggests that the presence of AQP3 in follicles of all categories, especially in the antral follicles, provides novel insights on the mechanisms that regulate the flow of water between cells during the formation of antral follicles in sheep., (Copyright © 2014 Elsevier GmbH. All rights reserved.)

Objective: Iodine is an essential micro nutrient, and a deficiency or excessive intake of this mineral is related to changes in thyroid function. In Brazil, both deficiency and excessive intake of iodine are common; however, excessive intakes have recently been observed. Thus, the objective of the present study was to assess the iodine concentration in maternal milk, taking into account the salt iodine concentration of the participating households and in the infants' urine., Method: Urine samples from 33 infants (less than 6 months of age), maternal milk samples and samples of the kitchen salt used by the mothers were collected. The iodine levels in the urine and maternal milk were assessed by ICP-MS; the iodine levels in the salt were assessed by titration., Result: The median iodinuria value in the infants was 293 μg/L; the mean iodine concentration was 206 μg/L in the maternal milk and 39.9 mg I/kg in the salt. There was a positive correlation between the iodine concentration in the maternal milk and the infant iodinuria value., Conclusion: The median infant iodinuria was elevated due to the high iodine concentration present in the maternal milk. High iodine values were caused by high salt iodine levels, which should be reduced., (Copyright © 2013 Elsevier GmbH. All rights reserved.)

Objectives: To determine whether hyperglycemia and hypoglycemia are associated with higher mortality, longer length of intensive care unit (ICU) stay, and fewer ventilator-free days in critically ill children while taking into account the clinical severity and nutrition status., Patients and Methods: A prospective observational cohort study was conducted on 221 children admitted to the ICU. Blood glucose levels were analyzed in the first 72 hours. Potential exposure variables for adverse prognosis included hyperglycemia (blood glucose >150 mg/dL), hypoglycemia (blood glucose ≤60 mg/dL), age <1 year, sex, nutrition status, the revised Pediatric Index of Mortality (PIM 2), and the Pediatric Logistic Organ Dysfunction (PELOD)., Results: Of the patients, 47.1% were malnourished. Controlling for nutrition status, both hyperglycemia and hypoglycemia increased the risk of mortality in the malnourished patients compared with the well-nourished ones. Adjusting for clinical severity, the odds ratio of mortality was higher in malnourished patients with hyperglycemia (odds ratio [OR], 3.98; 95% confidence interval [CI], 1.14-13.94; P = .03), whereas no significant associations were detected in the well-nourished patients. After controlling for nutrition status, hypoglycemia was associated with longer length of ICU stay (OR, 6.5; 95% CI, 1.30-32.57; P < .01) and fewer ventilator-free days (OR, 4.11; 95% CI, 1.26-13.40; P < .01) only in the malnourished group of patients., Conclusions: Compared with the well nourished, malnourished patients with hyperglycemia are at a greater risk of mortality, independent of clinical severity. Hypoglycemia was shown to be associated with mortality, longer length of ICU stay, and fewer ventilator-free days only in malnourished patients.

Cytological smear is widely employed to analyse specimens obtained from endosonography-guided fine-needle aspiration (EUS-FNA), but false-negative or inconclusive results may occur. A better diagnostic yield can be obtained from processing cell blocks. We compared the effectiveness of the cell block technique and cytological smear in the diagnosis of pancreatic neoplasms. From January 1997 to December 2006, 611 patients with pancreatic tumors were evaluated by EUS-FNA. Surgery was performed in 356 cases, and the other 255 patients were followed clinically for an average of 12.8 months. In total, 282 (46.2%) patients were evaluated with cytological smears, and 329 (53.8%) were evaluated using only cell blocks. Malignant disease was detected in 352 (57.6%) cases, in which adenocarcinoma accounted for 236 (67%) cases. A benign disease was found in the other 259 cases, including 35.1% focal chronic pancreatitis and 32.4% pseudocysts. Aspiration samples were satisfactory in 595 (97.4%) patients after an average of 2.2 (1-4) passes of the needle. Regardless of the cytopathological examination technique, EUS-FNA confirmed malignancy in 269 of 352 (76.4%) cases, and a benign disease in 257 of 259 (99.2%) cases. For patients who received surgery with histologically confirmed lesions, the sensitivity specificity, positive and negative predictive values, and accuracy of the smears versus cell blocks in diagnosing pancreatic tumors were 61% versus 85.2% (P<0.001), 100% versus 93.1%, 100% versus 98.4%, 36% versus 55.1% (P=0.046) and 68% versus 86.5% (P<0.001), respectively The cell block technique demonstrated a hig her sensitivity, negative predictive value and accuracy than cytological smears.