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Purpose: Treatment for metastatic castration-resistant prostate cancer (mCRPC) includes chemotherapy and inhibition of the androgen receptor pathway. However, the optimal treatment sequence in this scenario is not yet fully understood. Therefore, we conducted a systematic review and meta-analysis comparing cabazitaxel versus abiraterone or enzalutamide for efficacy and safety outcomes as second-line therapy in mCRPC patients after docetaxel failure., Methods: We searched PubMed, Embase, and Cochrane databases for interventional studies comparing cabazitaxel versus abiraterone or enzalutamide for patients with mCRPC who have experienced treatment failure with docetaxel as their first-line therapy. We computed hazard ratios (HRs) or odds ratios (ORs) with 95% confidence intervals (CIs)., Results: Eight studies, comprising 1,897 patients were included, of whom 548 (28.8%) received cabazitaxel. Mean follow-up time ranged from 3 to 16.4 months. Median age ranged from 68.1 to 73.9 years in the cabazitaxel group, and 68.0 to 73.1 years in the abiraterone or enzalutamide group. In our meta-analysis, cabazitaxel significantly improved progression-free survival (PFS) rates (HR 0.60; 95% CI 0.47-0.78; p < 0.001) compared to abiraterone or enzalutamide. There were no differences between groups in overall survival (HR 0.76; 95% CI 0.46-1.24; p = 0.27), therapy-related grade ≥ 3 adverse events (AEs) (OR 3.00; 95% CI 0.72-12.40; p = 0.12), and PSA decline ≥ 50% (OR 1.20; 95% CI 0.51-2.80; p = 0.67)., Conclusions: In this systematic review and meta-analysis of men with mCRPC after docetaxel failure, second-line therapy with cabazitaxel was associated with a longer PFS compared with abiraterone or enzalutamide, though without a significant difference in OS., Competing Interests: Declarations. Conflict of interests: All authors report no relationships that could be construed as a conflict of interest. All authors take responsibility for all aspects of the reliability and freedom from bias of the data presented and their discussed interpretation. Ethical approval: No ethical approval was required for this systematic review with meta-analysis, as all data were already published in peer-reviewed journals. Furthermore, no patients were involved in the design, conduct, or interpretation of our study. Informed consent: For this type of study, no informed consent is required., (© 2025. The Author(s), under exclusive licence to Federación de Sociedades Españolas de Oncología (FESEO).)

Cows produce a substantial amount of immunoglobulin in the colostrum, and nutraceutical products derived from these antibodies are gaining attention for their potential role in human viral disease prevention. The objective of our study was to develop an immunization schedule for pregnant cows to produce hyperimmune colostrum with antibodies presenting high avidity and neutralizing activity against SARS-CoV-2. The recombinant spike receptor-binding domain (RBD) from SARS-CoV-2, expressed using the Expi293F system and purified via Ni-affinity chromatography, was solubilized in (1) saponin (QuilA) or (2) a suspension of potassium and aluminum hydroxide (Alum). Vaccination of pregnant cows and serum sample collection were performed 45, 30, and 15 d before the expected calving date. Serum and colostrum were also collected on the day of parturition. Anti-RBD IgG, IgG1, and IgG2 production, viral neutralization, and antibody avidity were evaluated by ELISA. Cows immunized with recombinant RBD with the QuilA adjuvant produced higher amounts of all antibody subclasses than cows in the Alum group. The viral neutralization index from serum samples was also higher in the QuilA group. Significant differences were not observed in the avidity of antibodies, except for that of IgG2, which was higher in the serum of cows receiving the Alum formulation. As the IgG1 antibody subclass and its avidity are crucial for SARS-CoV-2 neutralization, QuilA might be the optimal adjuvant for producing hyperimmune colostrum in cows. These findings support the use of cows as biofactories of neutralizing antibodies against SARS-CoV-2 or any future emerging and re-emerging viral diseases, with the possibility of simply substituting the subunit antigen in the vaccine formulation. Further tests must be done to evaluate the efficacy of using hyperimmune colostrum as a nutraceutical or purified bovine antibodies as a pharmacological approach for COVID-19 prevention., (© 2025, The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Chitosan (CS) is a promising polymeric biomaterial for use in scaffolds for in vitro skin models and wound dressings, owing to its non-antigenic and antimicrobial properties. However, CS often exhibits insufficient physicochemical properties, mechanical strength, and bioactivity, limiting its efficacy in demanding applications. To address these challenges, cotton cellulose nanofibers (CNFs) represent a promising nanomaterial for enhancing CS-based scaffolds in tissue engineering. CNF offers superior stiffness, and mechanical properties that enhance cellular adhesion and proliferation, both crucial for effective tissue regeneration and healing. This study aimed to develop and characterize a scaffold combining cotton CNF and CS, focusing on its cytocompatibility with human fibroblasts and keratinocytes. The cotton CNF/CS scaffold was fabricated using the casting technique, and its physicochemical properties and cellular compatibility were assessed in vitro . The results demonstrated that incorporating cotton CNF significantly enhanced the stability of the CS matrix. The CS scaffold with 1000 μg ml -1 of cotton CNF exhibited increased roughness and reduced rupture strain compared to the pure CS scaffold. The cotton CNF/CS scaffold effectively promoted the adhesion, viability, proliferation, migration, and collagen synthesis of skin cells. Notably, increased cell viability was observed in human fibroblasts cultured on scaffolds with higher concentrations of cotton CNF (100 and 1000 μg ml -1 ). Based on the findings, the cotton CNF/CS scaffold demonstrates enhanced physicochemical properties and bioactivity, making it a promising candidate for the development of in vitro human skin models and wound healing dressings., (© 2024 IOP Publishing Ltd. All rights, including for text and data mining, AI training, and similar technologies, are reserved.)

Zinc oxide nanoparticles (ZnO-NPs) have garnered significant attention in biological applications due to their known antioxidant properties. However, their potential impact on assisted reproduction techniques remains largely unexplored, particularly in the context of oocyte quality maintenance within in vitro culture systems, where free radicals can exert detrimental effects. This study investigated the effects of incorporating ZnO-NPs to in vitro maturation (IVM) media on the developmental, cryosurvival, and metabolic profiles of bovine embryos. Three concentrations of ZnO-NPs (0, 1.0, and 1.5 μg/mL) were evaluated. We observed, for the first time, that the inclusion of ZnO-NPs at a concentration of 1.0 μg/mL led to a significant increase in the number of embryonic cells (p < 0.05) accompanied by a reduction in reactive oxygen species production (p < 0.05). Notably, ZnO-NPs did not alter embryonic development, cryosurvival rates, or mitochondrial viability. These findings suggested that ZnO-NPs has antioxidant properties and are compatible with bovine oocytes. Consequently, they may serve as promising supplements to the IVM media, potentially enhancing the efficiency of assisted reproduction techniques., 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.)

Natural polymeric nanobiocomposites hold promise in repairing damaged bone tissue in tissue engineering. These materials create an extracellular matrix (ECM)-like microenvironment that induces stem cell differentiation. In this study, we investigated a new cytocompatible nanobiocomposite made from cotton cellulose nanofibers (CNFs) combined with chitosan polymer to induce osteogenic stem cell differentiation. First, we characterized the chemical composition, nanotopography, swelling properties, and mechanical properties of the cotton CNF/chitosan nanobiocomposite scaffold. Then, we examined the biological characteristics of the nanocomposites to evaluate their cytocompatibility and osteogenic differentiation potential using human mesenchymal stem cells derived from exfoliated deciduous teeth. The results showed that the nanobiocomposite exhibited favorable cytocompatibility and promoted osteogenic differentiation of cells without the need for chemical inducers, as demonstrated by the increase in alkaline phosphatase activity and ECM mineralization. Therefore, the cotton CNF/chitosan nanobiocomposite scaffold holds great promise for bone tissue engineering applications., (© 2023 IOP Publishing Ltd.)

We report a nanoarchitectonic electronic tongue made with flexible electrodes coated with curcumin carbon dots and zein electrospun nanofibers, which could detect Staphylococcus aureus ( S. aureus ) in milk using electrical impedance spectroscopy. Electronic tongues are based on the global selectivity concept in which the electrical responses of distinct sensing units are combined to provide a unique pattern, which in this case allowed the detection of S. aureus through non-specific interactions. The electronic tongue used here comprised 3 sensors with electrodes coated with zein nanofibers, carbon dots, and carbon dots with zein nanofibers. The capacitance data obtained with the three sensors were processed with a multidimensional projection technique referred to as interactive document mapping (IDMAP) and analyzed using the machine learning-based concept of multidimensional calibration space (MCS). The concentration of S. aureus could be determined with the sensing units, especially with the one containing zein as the limit of detection was 0.83 CFU/mL (CFU stands for colony-forming unit). This high sensitivity is attributed to molecular-level interactions between the protein zein and C-H groups in S. aureus according to polarization-modulated infrared reflection-absorption spectroscopy (PM-IRRAS) data. Using machine learning and IDMAP, we demonstrated the selectivity of the electronic tongue in distinguishing milk samples from mastitis-infected cows from milk collected from healthy cows, and from milk spiked with possible interferents. Calibration of the electronic tongue can also be reached with the MCS concept employing decision tree algorithms, with an 80.1% accuracy in the diagnosis of mastitis. The low-cost electronic tongue presented here may be exploited in diagnosing mastitis at early stages, with tests performed in the farms without requiring specialized laboratories or personnel., Competing Interests: The authors declare no competing financial interest., (© 2023 The Authors. Published by American Chemical Society.)

Cellulose nanofibers (CNFs) are natural polymers with physical-chemical properties that make them very attractive for modulating stem cell differentiation, a crucial step in tissue engineering and regenerative medicine. Although cellulose is cytocompatible, when materials are in nanoscale, they become more reactive, needing to evaluate its potential toxic effect to ensure safe application. This study aimed to investigate the cytocompatibility of cotton CNF and its differentiation capacity induction on stem cells from human exfoliated deciduous teeth. First, the cotton CNF was characterized. Then, the cytocompatibility and the osteogenic differentiation induced by cotton CNF were examined. The results revealed that cotton CNFs have about 6-18 nm diameters, and the zeta potential was -10 mV. Despite gene expression alteration, the cotton CNF shows good cytocompatibility. The cotton CNF induced an increase in phosphatase alkaline activity and extracellular matrix mineralization. The results indicate that cotton CNF has good cytocompatibility and can promote cell differentiation without using chemical inducers, showing great potential as a new differentiation inductor for tissue engineering and regenerative medicine applications.

The delivery of nucleic acids to cells is considered a crucial step for the success of genetic modifications aimed at therapeutic purposes or production of genetically modified animals. In this context, nanotechnology is one of the most promising fields of science, with the potential to solve several existing problems. Nanostructures have desirable characteristics to be used as carriers, such as nanometric size, large surface area, cell internalization capacity, prolonged and controlled release, among others. Genetically modified animals can contribute to the production of biopharmaceuticals, through the expression of high-associated-value molecules. The production of these animals, also known as biofactories, further enhances Brazilian agribusiness, since it allows adding value to the final product, and favors the integration between the agricultural market and the pharmaceutical sector. However, there is a growing concern about the safety and possible harmful effects of nanostructures, since data on the safe use of these materials are still insufficient. The objective of this review was to address aspects of the use of nanostructures, mainly carbon nanotubes as nucleic acid carriers, aiming at the production of genetically modified animals, with the certainty that progress in this field of knowledge depends on more information on the mechanisms of interaction between nanostructures, cells and embryos, as well as on its toxicity.

Antimicrobial peptides (AMP) are promising novel antibiotics but exhibit low stability and can be toxic. The AMP encapsulation can be used to protect the drug and control its release rates. The Lr-AMP1f encapsulated into chitosan nanoparticle (NP) by ionic gelation method reached 90% efficiency. The results indicated that the hydrodynamic particle size of NPs increased from 196.1 ± 3.14 nm (free NP) to 228.1 ± 12.22 nm (nanoencapsulated Lr-AMP1f), while the atomic force microscope showed the spherical shape. The Zeta potential of the nanoencapsulated Lr-AMP1f was high (+ 35 mV). These AMP-loaded NPs exhibited stability for up to 21 days of storage. The minimum inhibitory concentration (MIC) of free Lr-AMP1f was 8 µg/mL for E. coli and S. epidermidis. However, the nanoencapsulated Lr-AMP1f produced a bacteriostatic effect against both bacteria at 8 µg/mL. The MIC of nanoencapsulated Lr-AMP1f was 16 µg/mL for E. coli and 32 for S. epidermidis. Nanoencapsulated Lr-AMP1f was nontoxic to HEK293 cells. Promisingly, chitosan NP can be used as a vehicle for the antibacterial application of new AMP (Lr-AMP1f)., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Carboxylated multi-wall carbon nanotube (MWCNT-COOH) presents unique properties due to nanoscale dimensions and permits a broad range of applications in different fields, such as bone tissue engineering and regenerative medicine. However, the cytocompatibility of MWCNT-COOH with human stem cells is poorly understood. Thus, studies elucidating how MWCNT-COOH affects human stem cell viability are essential to a safer application of nanotechnologies. Using stem cells from the human exfoliated deciduous teeth model, we have evaluated the effects of MWCNT-COOH on cell viability, oxidative cell stress, and DNA integrity. Results demonstrated that despite the decreased metabolism of mitochondria, MWCNT-COOH had no toxicity against stem cells. Cells maintained viability after MWCNT-COOH exposure. MWCNT-COOH did not alter the superoxide dismutase activity and did not cause genotoxic effects. The present findings are relevant to the potential application of MWCNT-COOH in the tissue engineering and regenerative medicine fields., (© 2021 IOP Publishing Ltd.)

The titanium dioxide nanoparticles (NPs) have been applied to biomedical, pharmaceutical, and food additive fields. However, the effect on health and the environment are conflicting; thus, it has been reviewing several times. In this context, establishing standard robust protocols for detecting cytotoxicity and genotoxicity of nanomaterials became essential for nanotechnology development. The cell type and the intrinsic characteristics of titanium dioxide NPs can influence nanotoxicity. In this work, the cyto- and genotoxicity effects of standard reference material titanium dioxide NPs in primary bovine fibroblasts and immortalized Chinese hamster ovary epithelial (CHO) cells were determined and compared for the first time. Titanium dioxide NPs exposure revealed no cytotoxicity for primary bovine fibroblasts, while only higher concentrations tested (10 μg/ml) induce genotoxic effects in this cell model. In contrast, the lower concentrations of the titanium dioxide NPs cause the cyto- and genotoxic effects in CHO cells. Therefore, our finding indicates that the CHO line was more sensitive toward the effects of titanium dioxide NPs than the primary bovine fibroblast, which should be valuable for their environmental risk assessment., (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Many types of nanocomposites employed in food packaging are based on silver nanoparticles (AgNP) because of their antibacterial properties, which can increase food shelf-life. As the commercialization of AgNP products has been expanding, the released of such nanoparticles in the environment has caused enormous concern, once they can pose potential risks to the environment and human beings. For instance, exposure of the maternal environment to nanomaterials during pregnancy may impact the health of the dam, fetus and offspring. In this context, here we investigated the effects of prenatal exposure of AgNP on the pregnancy outcomes of dams and postnatal development of their offspring. Pregnant Wistar rats were exposed to distinct AgNP concentrations (0, 1, 3 and 5 μg/kg/day) from beginning to the end of pregnancy. At parturition, newborns were observed regarding clinical signs of toxicity and survival rate. The offspring was examined by evaluating developmental endpoints. A delay in time for vaginal opening and testes descent were detected in the offspring exposed to AgNP during embryonic development. Our results indicate that prenatal exposure to AgNP can compromise neonatal rats' postnatal development, especially the reproductive features., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Macrophages are classified upon activation as classical activated M1 and M2 anti-inflammatory regulatory populations. This macrophage polarization is well characterized in humans and mice, but M1/M2 profile in cattle has been far less explored. Bos primigenius taurus (taurine) and Bos primigenius indicus (indicine) cattle display contrasting levels of resistance to infection and parasitic diseases such as C57BL/6J and Balb/c murine experimental models of parasite infection outcomes based on genetic background. Thus, we investigated the differential gene expression profile of unstimulated and LPS stimulated monocyte-derived macrophages (MDMs) from Holstein (taurine) and Gir (indicine) breeds using RNA sequencing methodology. For unstimulated MDMs, the contrast between Holstein and Gir breeds identified 163 Differentially Expressed Genes (DEGs) highlighting the higher expression of C-C chemokine receptor type five (CCR5) and BOLA-DQ genes in Gir animals. LPS-stimulated MDMs from Gir and Holstein animals displayed 1,257 DEGs enriched for cell adhesion and inflammatory responses. Gir MDMs cells displayed a higher expression of M1 related genes like Nitric Oxide Synthase 2 (NOS2), Toll like receptor 4 (TLR4), Nuclear factor NF-kappa-B 2 (NFKB2) in addition to higher levels of transcripts for proinflammatory cytokines, chemokines, complement factors and the acute phase protein Serum Amyloid A (SAA). We also showed that gene expression of inflammatory M1 population markers, complement and SAA genes was higher in Gir in buffy coat peripheral cells in addition to nitric oxide concentration in MDMs supernatant and animal serum. Co-expression analyses revealed that Holstein and Gir animals showed different transcriptional signatures in the MDMs response to LPS that impact on cell cycle regulation, leukocyte migration and extracellular matrix organization biological processes. Overall, the results suggest that Gir animals show a natural propensity to generate a more pronounced M1 inflammatory response than Holstein, which might account for a faster immune response favouring resistance to many infection diseases., Competing Interests: The authors have declared that no competing interests exist.

Cellulose nanofibers have mechanical properties that make them very attractive in a myriad of fields such as biomedicine, tissue engineering, biosensors, cosmetics and food packet products. To evaluate the potential health risks of airborne cellulose nanofibers, the cellulose nanofiber was prepared and characterized and then its pulmonary potential toxicity to a mouse model was studied. Cellulose nanofiber has been prepared by acid hydrolysis of cotton cellulose and characterized by transmission electron microscopy, zeta potential and X-ray diffraction analysis. Then, using a short-term inhalation test, the pulmonary biocompatibility of cotton cellulose nanofibers at different concentrations (0.5 mg/mL, 1 mg/mL and 2 mg/mL) were evaluated. Transmission electron images showed needle-shaped particle with a diameter of about 6-18 nm and a length of 85-225 μ m. Zeta potential was -25.3±7.80 mV and the X-ray diffraction patterns indicate that cotton cellulose nanofiber has pure structural characteristics. The In Vivo results revealed that the exposure to cotton cellulose nanofiber did not alter the number of inflammatory cells or cytokine secretion by lung cells ( p > 0.05). The results demonstrate that the cotton cellulose nanofiber is biocompatible and it is an environment-friendly nanomaterial with promise in various industrial sectors.

Infectious bovine keratoconjunctivitis (IBK) is a widespread, contagious ocular disease that affects cattle, especially dairy breeds. The disease is caused by Gram-negative bacteria mainly Moraxella bovis, and its treatment consists of parenteral or topic antibiotic therapy. The topic treatment approach is used more commonly in lactating cows, to avoid milk disposal. However, treatment failures are common, because the antibiotic is removed during lacrimation. This study aimed to evaluate the susceptibility of commercial cloxacillin and evaluate the efficacy of nanostructured cloxacillin in clinical cases of IBK by Moraxella . The minimum inhibitory concentration (MIC) of nanoparticle cloxacillin nanocoated, the nanoparticle without the antibiotic and the commercial cloxacillin were determined in vitro with field samples of Moraxella ovis (5) and Moraxella bovis (5). The efficiency of nanoparticles was tested in three cows naturally infected that were treated with 1.0 mL (with 0.32 mg of nanostructured cloxacillin) for the ocular route. Moraxella bovis was isolated and identified by biochemical and molecular methods before the treatment. The animals were treated every 12 h for six days. The cure was considered by the absence of clinical symptoms and bacteria after treatment. The mucoadhesive nanoparticle-based formulation promoted clinical cure with a low number of doses of antibiotics, probably due to the maintenance of the MIC in the ocular mucosa for longer due to the mucoadhesive characteristics of the nanoparticle. The results indicate that the use of nanocoated cloxacillin is possible to control infectious bovine keratoconjunctivitis., (© 2020 The Authors.)

The present work shows the development and evaluation of the veterinary antibiotic cloxacillin benzathine (CLOXB) loaded into poly-ε-caprolactone (PCL) nanocapsules (NC), as a potential new treatment strategy to manage bovine intramammary infections, such as mastitis. Staphylococcus aureus-induced mastitis is often a recurrent disease due to the persistence of bacteria within infected cells. CLOXB-PCL NC were prepared by interfacial deposition of preformed biodegradable polymer followed by solvent displacement method. The mean diameter of NC varied from 241 to 428 nm and from 326 to 375 nm, when determined by dynamic light scattering and by atomic force microscopy, respectively. The zeta potential of NC was negative and varied from -28 to -51 mV. In vitro release studies from the NC were performed in two media under sink conditions: PBS with 1% polyethylene glycol or milk. A reversed-phase HPLC method was developed to determine the NC entrapment efficiency and kinetics of CLOXB release from the NC. Free CLOXB dissolution occurred very fast in both media, while drug release from the NC was slower and incomplete (below 50%) after 9 h. CLOXB release kinetics from polymeric NC was fitted with the Korsmeyer-Peppas model indicating that CLOXB release is governed by diffusion following Fick's law. The fluorescence confocal microscopy images of macrophage-like J774A.1 cells reveal NC uptake and internalization in vitro. In addition, antimicrobial effect of the intramammary administration of CLOXB-PCL NC in cows with mastitis resulted in no clinical signs of toxicity and allowed complete pathogen elimination after treatment. The in vivo results obtained in this work suggest that CLOXB-PCL NC could be a promising formulation for the treatment of intramammary infections in cattle, considering their physicochemical properties, release profiles and effects on bovine mastitis control., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Cellulose is a renewable polymer quite abundant on the Earth and very attractive for applications in the construction of eco-friendly biomedical products. The aim of this study was to investigate the chemical-physical characteristics of cotton cellulose nanofiber (CCN)/chitosan nanocomposite and its cytocompatibility with human embryonic kidney cells. First, the chemical composition, swelling ratio and surface topography of the nanocomposite were evaluated. Cytocompatibility was then assessed through spreading, proliferation and viability of cells. The experimental results showed that the CCN was an effective nanomaterial agent for increasing the roughness surface of chitosan film. Cell proliferation and changes in cell morphology indicated that the nanocomposite led to improved cell spreading and growth. Cell viability did not decrease after 24 h. However, the cell survival on the nanocomposite was affected at 72 h. The results indicate that CCN/chitosan nanocomposite could be a promising biocompatible biomaterial for biomedical applications.

This study aimed to evaluate the transcriptional changes occurring in isolated perfused mammary alveolar tissue in response to inoculation with S. agalactiae and to identify the most affected biological functions and pathways after 3 h. Four udders taken at slaughter from cows with healthy mammary gland were perfused ex situ with warmed and gassed Tyrode's solution. Mammary alveolar tissue samples were taken from the left fore and rear quarters (IQ-inoculated quarters) before inoculation (hour 0) and at 3 h post inoculation (hpi) and at the same times from control right fore and rear quarters (not inoculated: NIQ). A total of 1756 differentially expressed genes (DEGs) were identified between IQ and NIQ at 3 hpi using edgeR package. Within this set of DEGs, 952 were up regulated and mainly involved with innate immune response and inflammatory response, e.g., CD14, CCL5, TLR2, IL-8, SAA3, as well as in transcriptional regulation such as FOS, STAT3 and NFKBIA. Genes down-regulated (804) included those involved with lipid synthesis e.g., APOC2, SCD, FABP3 and FABP4. The most affected pathways were chemokine signaling, Wnt signaling and complement and coagulation cascades, which likely reflects the early stage response of mammary tissue to S. agalactiae infection. No significant gene expression changes were detected by RNA-Seq in the others contrasts. Real time-PCR confirmed the increase in mRNA abundance of immune-related genes: TLR2, TLR4, IL-1β, and IL-10 at 3 hpi between IQ and NIQ. The expression profiles of Casp1 and Bax for any contrasts were unaffected whereas Bcl2 was increased in IQ, which suggests no induction of apoptosis during the first hours after infection. Results provided novel information regarding the early functional pathways and gene network that orchestrate innate immune responses to S. agalactiae infection. This knowledge could contribute to new strategies to enhance resistance to this disease, such as genomic selection.

The aims of this study were to determine the antimicrobial susceptibility profile and genetic diversity of Staphylococcus spp. isolated from dairy cows in Minas Gerais, Brazil, and to assess the relationship among the isolates' susceptibility profiles and pulsed-field gel electrophoresis (PFGE) genotypes. Seventy-nine isolates were used, including S. aureus (n = 71) and coagulase-negative staphylococci (CoNS) (n = 8). Susceptibility to 12 antimicrobial agents was performed. All Staphylococcus spp. were subjected to PFGE. Staphylococcus aureus and CoNS isolates exhibited full susceptibility only to cephalothin. The greatest percentages of resistance among Staphylococcus spp. were observed to penicillins, folate pathway inhibitors, and tetracyclines. Twelve S. aureus and four CoNS were classified as multidrug resistance strains. Percentage of MRSA was also higher among CoNS (75%), compared to S. aureus isolates (2.81%). Adopting 100% of similarity, 34 different genotypes were identified. Association of minimum-spanning tree (MST) analysis with data from municipalities, herds, methicillin-resistant S. aureus (MRSA), and resistance patterns for all isolates did not show any clustering. However, a clustering pattern of bacterial species was observed. Results from this study indicate a high frequency of antimicrobial resistance, especially among CoNS, and a high genetic diversity among Staphylococcus spp. isolated from dairy cows with mastitis in Minas Gerais, Brazil., (© 2018 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.)

Nanomaterials can mimic properties of extracellular matrix molecules, promising great potential for scaffold composition in tissue engineering. In the present study, we investigated whether barium titanate nanoparticles (BT NP) combined with alginate polymer would provide a new cytocompatible three-dimensional (3D) scaffold to induce osteogenic stem cell differentiation. In vitro cytocompatibility and osteogenic differentiation potential were investigated using human mesenchymal stem cells (MSC). Firstly, we studied the cell viability and oxidative stress by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) thiazolyl blue tetrazolium bromide (MTT) and superoxide dismutase (SOD) assays. Overall, neither pure BT NP or BT NP/alginate 3D scaffold induced cytotoxicity. The scanning electron and atomic force microscopy revealed that BT NP/alginate 3D scaffold produced exhibited highly interconnected pores and surface nanotopography that were favorable for MSC differentiation. Von Kossa staining showed mineralization nodules and MSCs morphology changed from spindle to cuboid shape after 21 d. Finally, BMP-2 and ALP mRNA were significantly upregulated on cells grown into the BT NP/alginate 3D scaffold. Thus, the BT NP/alginate 3D scaffold showed an osteogenic differentiation induction potential, without the addition of osteogenic supplements. These results indicate that the BT NP/alginate 3D scaffold provides a cytocompatible and bioactive microenvironment for osteogenic human MSC differentiation.

Multi-walled carbon nanotubes (MWCNTs) have potential applications in the industrial, agricultural, pharmaceutical, medical, and environmental remediation fields. However, many uncertainties exist regarding the environmental implications of engineered nanomaterials. This study examined the effect of the MWCNTs on metabolic status and morphology of filamentous green microalgae Klebsormidium flaccidum. Appropriate concentrations of MWCNT (1, 50, and 100 μg mL -1 ) were added to a microalgal culture in the exponential growth phase and incubated for 24, 48, 72, and 96 h. Exposure to MWCNT led to reductions in algal growth after 48 h and decreased on cell viability for all experimental endpoints except for 1 µg mL -1 at 24 h and 100 µg mL -1 after 72 h. At 100 µg mL -1 , MWCNTs induced reactive oxygen species (ROS) production and had an effect on intracellular adenosine triphosphate (ATP) content depending on concentration and time. No photosynthetic activity variation was observed. Observations by scanning transmission electron microscopy showed cell damage. In conclusion, we have demonstrated that exposure to MWCNTs affects cell metabolism and microalgal cell morphology. To our best knowledge, this is the first case in which MWCNTs exhibit adverse effects on filamentous green microalgae K. flaccidum. These results contribute to elucidate the mechanism of MWCNT nanotoxicity in the bioindicator organism of terrestrial and freshwater habitats.

Intramammary infections are one of the main causes of productivity loss in dairy cows. To better understand the immune system response and to avoid the use of live animals, we validated the use of isolated bovine udder as an ex situ model. Six mammary glands were collected from cows ready for culling. Three udders were perfused with Tyrode's solution and three were not-perfused. During six hours, we collected perfusate samples for biochemical analysis. We also collected alveolar and teat canal tissue to evaluate gene expression. The biochemical parameters indicated that the perfused udders remained viable for the entire period of the experiment. A real-time polymerase chain reaction showed an increase in 18S rRNA gene expression in the alveolar tissue at 3 and 4 h after perfusion. There was also an increase in the Ubiquitin gene in the teat canal from not-perfused udders at 1, 3, and 4 h after slaughter. In general, gene expression was stable during the experiment. Our results indicated that the isolated perfused bovine udder model is appropriate for genetic studies, opening a new perspective in animal experimentation methods.

The pellucid zone (PZ) is a protective embryonic cells barrier against chemical, physical or biological substances. This put, usual transfection methods are not efficient for mammal oocytes and embryos as they are exclusively for somatic cells. Carbon nanotubes have emerged as a new method for gene delivery, and they can be an alternative for embryos transfection, however its ability to cross the PZ and mediated gene transfer is unknown. Our data confirm that multiwall carbon nanotubes (MWNTs) can cross the PZ and delivery of pDNA into in vitro-fertilized bovine embryos. The degeneration rate and the expression of genes associated to cell viability were not affected in embryos exposed to MWNTs. Those embryos, however, had lower cell number and higher apoptotic cell index, but this did not impair the embryonic development. This study shows the potential utility of the MWNT for the development of new method for delivery of DNA into bovine embryos.

Currently there is a growing interest in the use of nanotechnology in reproductive medicine and reproductive biology. However, their toxic effects on mammalian embryos remain poorly understood. In this work, we evaluate the biocompatibility of two fibrous nanomaterials (NMs): cotton cellulose nanofibers (CNF) and carboxylated multiwalled carbon nanotubes (MWCNT-COOH), by performing an investigation of the embryonic development, gene expression (biomarkers focused on cell stress, apoptosis and totipotency) and in situ apoptosis in bovine embryos. Exposure to NMs did not interfere in preimplantation development or in the incidence of apoptosis in the bovine embryo, but they did affect the gene expression. The results presented are important for an understanding of the toxicity of cotton CNF and MWCNT-COOH on mammalian embryos. To our knowledge, we report the first evaluation of biocompatibility between these NMs on preimplantation embryos, which may open a new window for reproductive biomedical applications., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Recently, cellulose nanofibers (CNFs) have attracted considerable attention as natural, abundant polymers with excellent mechanical properties and biodegradability. CNFs provide a new materials platform for the sustainable production of high-performance nano-enable products for various applications. Given the increasing rates of CNF production, the potential for their release to the environment and the subsequent impact on ecosystem is becoming an increasing concern that needs to be addressed. Here, we used the Klebsormidium flaccidum as a bioindicator organism of terrestrial and freshwater habitats pollution using a battery of biomarkers. Our results show that cotton CNFs inhibit the proliferation of algae and induce morphological changes in them. The two main toxicity mechanisms induced by cotton CNFs are: (i) a direct contact of CNFs with the cell wall and cellular membrane and (ii) an indirect effect through the generation of reactive oxygen species (ROS)., (Copyright © 2015 Elsevier Inc. All rights reserved.)

This study aimed to evaluate protection induced by the vaccine candidate B. ovis ΔabcBA against experimental challenge with wild type B. ovis in rams. Rams were subcutaneously immunized with B. ovis ΔabcBA encapsulated with sterile alginate or with the non encapsulated vaccine strain. Serum, urine, and semen samples were collected during two months after immunization. The rams were then challenged with wild type B. ovis (ATCC25840), and the results were compared to non immunized and experimentally challenged rams. Immunization, particularly with encapsulated B. ovis ΔabcBA, prevented infection, secretion of wild type B. ovis in the semen and urine, shedding of neutrophils in the semen, and the development of clinical changes, gross and microscopic lesions induced by the wild type B. ovis reference strain. Collectively, our data indicates that the B. ovis ΔabcBA strain is an exceptionally good vaccine strain for preventing brucellosis caused by B. ovis infection in rams.

This study aimed to evaluate the Brucella ovis ΔabcBA strain as a vaccine candidate in the murine model. BALB/c mice were subcutaneously or intraperitoneally immunized with a single dose or three doses of the B. ovis ΔabcBA strain and then were challenged with wild-type B. ovis. Single or multiple immunizations provided only mild protection, with significantly smaller numbers of wild-type B. ovis CFU in the livers of immunized mice but not in the spleens. Encapsulation of B. ovis ΔabcBA significantly improved protection against experimental challenges in both BALB/c and C57BL/6 mice. Furthermore, immunization with encapsulated B. ovis ΔabcBA markedly prevented lesions in the spleens and livers of experimentally challenged mice. These results demonstrated that the encapsulated B. ovis ΔabcBA strain confers protection to mice; therefore, this strain has potential as a vaccine candidate for rams., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

Studies have been demonstrating that smaller particles can lead to unexpected and diverse ecotoxicological effects when compared to those caused by the bulk material. In this study, the chemical composition, size and shape, state of dispersion, and surface's charge, area and physicochemistry of micro (BT MP) and nano barium titanate (BT NP) were determined. Green algae Chlorella vulgaris grown in Bold's Basal (BB) medium or Seine River water (SRW) was used as biological indicator to assess their aquatic toxicology. Responses such as growth inhibition, cell viability, superoxide dismutase (SOD) activity, adenosine-5-triphosphate (ATP) content and photosynthetic activity were evaluated. Tetragonal BT (~170 nm, 3.24 m(2) g(-1) surface area) and cubic BT (~60 nm, 16.60 m(2) g(-1)) particles were negative, poorly dispersed, and readily aggregated. BT has a statistically significant effect on C. vulgaris growth since the lower concentration tested (1 ppm), what seems to be mediated by induced oxidative stress caused by the particles (increased SOD activity and decreased photosynthetic efficiency and intracellular ATP content). The toxic effects were more pronounced when the algae was grown in SRW. Size does not seem to be an issue influencing the toxicity in BT particles toxicity since micro- and nano-particles produced significant effects on algae growth.

The interaction between live organisms and micro- or nanosized materials has become a current focus in toxicology. As nanosized barium titanate has gained momentum lately in the medical field, the aims of the present work are: (i) to assess BT toxicity and its mechanisms on the aquatic environment, using two photosynthetic organisms (Anabaena flos-aquae, a colonial cyanobacteria, and Euglena gracilis, a flagellated euglenoid); (ii) to study and correlate the physicochemical properties of BT with its toxic profile; (iii) to compare the BT behavior (and Ba(2+) released ions) and the toxic profile in synthetic (Bold's Basal, BB, or Mineral Medium, MM) and natural culture media (Seine River Water, SRW); and (iv) to address whether size (micro, BT MP, or nano, BT NP) is an issue in BT particles toxicity. Responses such as growth inhibition, cell viability, superoxide dismutase (SOD) activity, adenosine-5-triphosphate (ATP) content and photosynthetic efficiency were evaluated. The main conclusions are: (i) BT have statistically significant toxic effects on E. gracilis growth and viability even in small concentrations (1μgmL(-1)), for both media and since the first 24 h; on the contrary of on A. flos-aquae, to whom the effects were noticeable only for the higher concentrations (after 96 h: ≥75 μg mL(-1) for BT NP and =100 μg mL(-1) for BT MP, in BB; and ≥75 μg mL(-1) for both materials in SRW), in spite of the viability being affected in all concentrations; (ii) the BT behaviors in synthetic and natural culture media were slightly different, being the toxic effects more pronounced when grown in SRW - in this case, a worse physiological state of the organisms in SRW can occur and account for the lower resistance, probably linked to a paucity of nutrients or even a synergistic effect with a contaminant from the river; and (iii) the effects seem to be mediated by induced stress without a direct contact in A. flos-aquae and by direct endocytosis in E. gracilis, but in both organisms the contact with both BT MP and BT NP increased SOD activity and decreased photosynthetic efficiency and intracellular ATP content; and (iv) size does not seem to be an issue in BT particles toxicity since micro- and nano-particles produced significant toxic for the model-organisms., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Background: MWCNT and CNF are interesting NPs that possess great potential for applications in various fields such as water treatment, reinforcement materials and medical devices. However, the rapid dissemination of NPs can impact the environment and in the human health. Thus, the aim of this study was to evaluate the MWCNT and cotton CNF toxicological effects on freshwater green microalgae Chlorella vulgaris., Results: Exposure to MWCNT and cotton CNF led to reductions on algal growth and cell viability. NP exposure induced reactive oxygen species (ROS) production and a decreased of intracellular ATP levels. Addition of NPs further induced ultrastructural cell damage. MWCNTs penetrate the cell membrane and individual MWCNTs are seen in the cytoplasm while no evidence of cotton CNFs was found inside the cells. Cellular uptake of MWCNT was observed in algae cells cultured in BB medium, but cells cultured in Seine river water did not internalize MWCNTs., Conclusions: Under the conditions tested, such results confirmed that exposure to MWCNTs and to cotton CNFs affects cell viability and algal growth.

Cellulose nanofibers (CNF) have mechanical properties that make them very attractive for applications in the construction of polymeric matrices, drug delivery and tissue engineering. However, little is known about their impact on mammalian cells. The objective of this study was to evaluate the cytotoxicity of CNF and their effect on gene expression of fibroblasts cultured in vitro. The morphology of CNF was analyzed by transmission electron microscopy and the surface charge by Zeta potential. Cell viability was analyzed by flow cytometry assay and gene expression of biomarkers focused on cell stress response such as Heat shock protein 70.1 (HSP70.1) and Peroxiredoxin 1 (PRDX1) and apoptosis as B-cell leukemia (BCL-2) and BCL-2 associated X protein (BAX) by RT-PCR assay. Low concentrations of CNF (0.02-100 μg ml(-1)) did not cause cell death; however, at concentrations above 200 μg ml(-1), the nanofibers significantly decreased cell viability (86.41 ± 5.37%). The exposure to high concentrations of CNF (2000 and 5000 μg ml(-1)) resulted in increased HSP70.1, PRDX1 and BAX gene expression. The current study concludes that, under the conditions tested, high concentrations (2000 and 5000 μg ml(-1)) of CNF cause decreased cell viability and affect the expression of stress- and apoptosis-associated molecular markers.