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Bovine laminitis disorder results in animal welfare and economic concerns in dairy and beef farms worldwide. However, the affected metabolic pathways, pathophysiologic characteristics, and inflammatory mechanisms remain unclear, hampering the development of new diagnostics. Using cerumen (earwax) as a source of volatile metabolites (cerumenomic) that carry valuable biological information has interesting implications for veterinary medicine. Nonetheless, up to now, no applications of veterinary cerumenomic assays have been made to identify bovine laminitis. This work aims to develop a veterinary cerumenomic assay for bovine laminitis identification that is non-invasive, robust, accurate, and sensitive to detecting the metabolic disturbances in bovine volatile metabolome. Twenty earwax samples (10 from healthy/control calves and 10 from laminitis calves) were collected from Nellore cattle, followed by Headspace/Gas Chromatography-Mass Spectrometry (HS/GC-MS) analysis and biomarker selection in two multivariate approaches: semiquantitative (intensity data) and semiqualitative (binary data). Following the analysis, cerumen volatile metabolites were indicated as candidate biomarkers for identifying bovine laminitis by monitoring their intensity or occurrence. In the semiquantitative strategy, the p-cresol presented the highest diagnostic figures of merit (area under the curve: 0.845, sensitivity: 0.700, and specificity: 0.900). Regarding the binary approach, a panel combining eight variables/volatiles, with formamide being the most prominent one, showed an area under the curve, sensitivity, and specificity of 0.97, 0.81, and 0.90, respectively. In summary, this work describes the first veterinary cerumenomic assay for bovine laminitis that indicates new metabolites altered during the inflammatory condition, paving the way for developing laminitis early diagnosis by monitoring the cerumen metabolites.
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Enfermedades de los Bovinos , Dermatitis , Bovinos , Animales , Dermatitis/veterinaria , Cerumen/metabolismo , Enfermedades de los Bovinos/diagnóstico , BiomarcadoresRESUMEN
Facial filling is widespread in society, albeit associated with inherent risks. This review analyzes clinical studies using laser therapy for filler complications to assess its safety and efficacy as an alternative treatment. A literature search was conducted up until April 2023, encompassing five different databases: PubMed, Scopus, Embase, Web of Science, and Medline, to find clinical trials addressing patients who underwent laser treatment for adverse reactions to injectable facial filling. The outcome variables were the clinical assessment of the lesion and the occurrence of post-intervention complications/sequelae. The risk of bias was assessed using the ROBINS-I tool. In total, six studies were included, all classified as having a "moderate risk" of bias. A total of 533 patients underwent laser treatment for adverse reactions to injectable facial fillers. The diode laser was the most frequently utilized equipment, with positive results reported in five studies. Among all treated patients, 96.24% achieved partial or complete resolution, 0.22% experienced some sequelae or complications, and only 0.01% showed no improvement. Laser treatment can eliminate the necessity for surgical intervention for adverse reactions to injectable facial fillers, resulting in partial or complete improvement of the condition.
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Terapia por Láser , Terapia por Luz de Baja Intensidad , Humanos , Terapia por Láser/métodos , Terapia por Luz de Baja Intensidad/efectos adversos , Rayos LáserRESUMEN
Surface topographies of cell culture substrates can be used to generate in vitro cell culture environments similar to the in vivo cell niches. In vivo, the physical properties of the extracellular matrix (ECM), such as its topography, provide physical cues that play an important role in modulating cell function. Mimicking these properties remains a challenge to provide in vitro realistic environments for cells. Artificially generated substrates' topographies were used extensively to explore this important surface cue. More recently, the replication of natural surface topographies has been enabling to exploration of characteristics such as hierarchy and size scales relevant for cells as advanced biomimetic substrates. These substrates offer more realistic and mimetic environments regarding the topographies found in vivo. This review will highlight the use of natural surface topographies as a template to generate substrates for in-vitro cell culture. This review starts with an analysis of the main cell functions that can be regulated by the substrate's surface topography through cell-substrate interactions. Then, we will discuss research works wherein substrates for cell biology decorated with natural surface topographies were used and investigated regarding their influence on cellular performance. At the end of this review, we will highlight the advantages and challenges of the use of natural surface topographies as a template for the generation of advanced substrates for cell culture.
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The interaction between cells and biomaterials is essential for the success of biomedical applications in which the implantation of biomaterials in the human body is necessary. It has been demonstrated that material's chemical, mechanical, and structural properties can influence cell behaviour. The surface topography of biomaterials is a physical property that can have a major role in mediating cell-material interactions. This interaction can lead to different cell responses regarding cell motility, proliferation, migration, and even differentiation. The combination of biomaterials with mesenchymal stem cells (MSCs) for bone regeneration is a promising strategy to avoid the need for autologous transplant of bone. Surface topography was also associated with the capacity to control MSCs differentiation. Most of the topographies studied so far involve machine-generated surface topographies. Herein, our strategy differentiates from the above mentioned since we selected natural surface topographies that can modulate cell functions for regenerative medicine strategies.Rubus fruticosusleaf was the selected topography to be replicated in polycaprolactone (PCL) membranes through polydimethylsiloxane moulding and using soft lithography. Afterwards, rat bone marrow stem cells (rBMSCs) were seeded at the surface of the imprinted PCL membranes to characterize the bioactive potential of our biomimetic surface topography to drive rBMSCs differentiation into the osteogenic lineage. The selected surface topography in combination with the osteogenic inductive medium reveals having a synergistic effect promoting osteogenic differentiation.
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Biomimética , Osteogénesis , Ratas , Humanos , Animales , Diferenciación Celular , Materiales Biocompatibles/farmacología , HuesosRESUMEN
BRAF and TERT oncogenes hotspot mutations are associated with a more aggressive outcome in thyroid carcinomas (TC). TERT promoter (pTERT) mutations (C228T and C250T) are related to cancer growth and reduced overall- and disease-free survivals in TC. We report a patient followed up for 8 years with a poorly differentiated thyroid carcinoma (PDTC) presenting an extremely aggressive course, who developed a large volume of metastases in a short period. Molecular analysis of the primary tumor revealed two pTERT mutations (C228T and C250T), and no BRAF V600E mutation. pTERT mutations C228T and C250T have been described as mutually exclusive, indicating that one mutation is enough for telomerase activation and exerts its action in thyroid tumorigenesis. This report describes both pTERT hotspot mutations in the same PDTC patient presenting a very aggressive course, even for PDTC, suggesting a relationship between the two events. However, more studies are needed to prove this causality.
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Adenocarcinoma , Telomerasa , Neoplasias de la Tiroides , Humanos , Neoplasias de la Tiroides/genética , Neoplasias de la Tiroides/patología , Mutación , Regiones Promotoras Genéticas/genética , Adenocarcinoma/genética , Proteínas Proto-Oncogénicas B-raf/genética , Telomerasa/genéticaRESUMEN
In tumors, somatic mutations of the PTEN suppressor gene are associated with advanced disease, chemotherapy resistance, and poor survival. PTEN loss of function may occur by inactivating mutation, by deletion, either affecting one copy (hemizygous loss) leading to reduced gene expression or loss of both copies (homozygous) with expression absent. Various murine models have shown that minor reductions in PTEN protein levels strongly influence tumorigenesis. Most PTEN biomarker assays dichotomize PTEN (i.e. presence vs. absence) ignoring the role of one copy loss. We performed a PTEN copy number analysis of 9793 TCGA cases from 30 different tumor types. There were 419 (4.28%) homozygous and 2484 (25.37%) hemizygous PTEN losses. Hemizygous deletions led to reduced PTEN gene expression, accompanied by increased levels of instability and aneuploidy across tumor genomes. Outcome analysis of the pan-cancer cohort showed that losing one copy of PTEN reduced survival to comparable levels as complete loss, and was associated with transcriptomic changes controlling immune response and the tumor microenvironment. Immune cell abundances were significantly altered for PTEN loss, with changes in head and neck, cervix, stomach, prostate, brain, and colon more evident in hemizygous loss tumors. These data suggest that reduced expression of PTEN in tumors with hemizygous loss leads to tumor progression and influences anticancer immune response pathways.
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Evasión Inmune , Neoplasias de la Próstata , Masculino , Humanos , Animales , Ratones , Fosfohidrolasa PTEN/genética , Genoma , Neoplasias de la Próstata/patología , Próstata/patología , Genómica , Microambiente TumoralRESUMEN
INTRODUCTION/OBJECTIVES: To evaluate regurgitant fraction (RF) using Simpson's method of discs to estimate total stroke volume (RFSMOD_TSV) and using Motion-mode to estimate total stroke volume (RFM-modeTSV) in dogs with subclinical myxomatous mitral valve disease (MMVD). We also sought to evaluate the effects of pimobendan on RF, and to determine the reproducibility of RFSMOD_TSV and RFM-modeTSV. ANIMALS, MATERIALS, AND METHODS: Echocardiography was performed on 57 dogs with MMVD (30 stage B1 and 27 stage B2). Ten dogs received pimobendan for 7-10 days and had a second echocardiogram. Nine dogs underwent six repeated echocardiographic examinations by two operators on three nonconsecutive days within one week for reproducibility analysis. RESULTS: Both RFSMOD_TSV and RFM-modeTSV exhibited a curvilinear relationship with left atrium-to-aortic root ratio. Both RFSMOD_TSV and RFM-modeTSV varied considerably within stage B1 (minimum-maximum: -9.1%-58.2% and -35.7%-66.2%, respectively) and B2 (13.6%-76.2% and 20.1%-85.7%, respectively). Method comparison showed RFSMOD_TSV and RFM-modeTSV were not interchangeable with proportional bias. Pimobendan significantly reduced RFSMOD_TSV (-32.0% ± 23.3%) and RFM-modeTSV (-19.2% ± 10.9%) within the same dog and relative to controls. Good inter-day and between-operator reproducibility was observed for RFSMOD_TSV and RFM-modeTSV based on intraclass correlation coefficients 0.86-0.90 and 0.83-0.90, respectively. Reproducibility coefficients were 19.6%-24.1% and 24.1%-27.0%, respectively. CONCLUSIONS: Use of RF using the total stroke volume method to aid the assessment of dogs with subclinical MMVD might be of clinical value. However, further study is warranted. Based on response to pimobendan and reproducibility analysis, RF SMOD_TSV might be a more reliable technique to quantify RF.
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Enfermedades de los Perros , Enfermedades de las Válvulas Cardíacas , Perros , Animales , Válvula Mitral/diagnóstico por imagen , Reproducibilidad de los Resultados , Enfermedades de los Perros/diagnóstico por imagen , Enfermedades de los Perros/tratamiento farmacológico , Enfermedades de las Válvulas Cardíacas/veterinaria , Ecocardiografía/veterinariaRESUMEN
The aim of this study was to verify the relationship between quantitative T2 relaxation measurements of lumbar intervertebral discs (IVDs) and spinopelvic parameters in patients with chronic low back pain. The study was approved by the Clinical Hospital of the Ribeirao Preto Medical School (USP) Ethics Committee, and written consent was obtained from all patients. A total of 455 IVDs from 91 consecutive patients with chronic low back pain were included in this prospective study. All subjects were assessed using the Oswestry Disability Index and visual analogue scale questionnaires and were confirmed to have no other spine diseases except disc degeneration. Spinopelvic parameters including the pelvic incidence (PI), pelvic tilt (PT), sacral slope (SS), sagittal vertical axis (SVA), global tilt (GT), T1 pelvic angle (TPA), lumbar lordosis (LL), thoracic kyphosis (TK), pelvic incidence minus lumbar lordosis mismatch (PI-LL), and lack of lumbar lordosis (LLL) were measured. The study group was categorized according to the Roussouly classification. Sagittal T2 maps were acquired to extract the IVD relaxation times, and the complete manual segmentation of IVDs at all levels was performed using Display® software. Lumbar IVD T2 relaxation times showed significant correlation with PT (P<0.01), GT (P<0.01), TPA (P<0.01), PI-LL (P=0.01), and LLL (P=0.01). No difference was noted between Roussouly subtypes regarding T2 relaxation times at any disc level. Data from questionnaires showed no correlation with T2 relaxation times. Global tilt and T1 pelvic angle were correlated with IVD composition changes (T2 relaxometry). There was no correlation between clinical symptoms and IVD T2 relaxation times.
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Disco Intervertebral , Lordosis , Dolor de la Región Lumbar , Humanos , Lordosis/diagnóstico por imagen , Vértebras Lumbares/diagnóstico por imagen , Dolor de la Región Lumbar/diagnóstico por imagen , Estudios Prospectivos , Estudios Retrospectivos , Disco Intervertebral/diagnóstico por imagenRESUMEN
The implantation of biomaterial devices can negatively impact the local microenvironment through several processes including the injury incurred during the implantation process and the associated host inflammatory response. Immune cell responses to implantable biomaterial devices mediate host-material interactions. Indeed, the immune system plays a central role in several biological processes required for the integration of biomaterials such as wound healing, tissue integration, inflammation, and foreign body reactions. The implant physicochemical properties such as size, shape, surface area, topography, and chemistry have been shown to provide cues to the immune system. Its induced immune-modulatory responses towards inflammatory or wound healing phenotypes can determine the success of the implant. In this work, we aim to evaluate the impact of some biomimetic surface topographies on macrophages' acute inflammatory response. For that, we selected 4 different biological surfaces to replicate through soft lithography on spin casting PCL membranes. Those topographies were: the surface of E. coli, S.eppidermidis and L929 cells cultured in polystyrene tissue culture disks, and an Eggshell membrane. We selected a model based on THP-1-derived macrophages to study the analysis of the expression of both pro-inflammatory and anti-inflammatory markers. Our results revealed that depending on the surface where these cells are seeded, they present different phenotypes. Macrophages present a M1-like phenotype when they are cultured on top of PCL membranes with the surface topography of E. coli and S. epidermidis. When cultured on membranes with L929 monolayers or Eggshell membrane surface topography, the macrophages present a M2-like phenotype. These results can be a significant advance in the development of new implantable biomaterial devices since they can help to modulate the inflammatory responses to implanted biomaterials by controlling their surface topography.
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Materiales Biocompatibles , Poliestirenos , Antiinflamatorios/química , Materiales Biocompatibles/efectos adversos , Biomimética , Escherichia coli , Humanos , Inflamación/metabolismo , Macrófagos , Poliestirenos/químicaRESUMEN
Bone healing after a tumor removal can be promoted by biomaterials that enhance the bone regeneration and prevent the tumor relapse. Herein, we obtained several nanopatterns by self-assembly of polystyrene-block-poly-(2-vinylpyridine) (PS-b-P2VP) with different molecular weights and investigated the adhesion and morphology of human bone marrow mesenchymal stem cells (BMMSC) and osteosarcoma cell line (SaOS-2) on these patterns aiming to identify topography and chemistry that promote bone healing. We analyzed > 2000 cells per experimental condition using imaging software and different morphometric descriptors, namely area, perimeter, aspect ratio, circularity, surface/area, and fractal dimension of cellular contour (FDC). The obtained data were used as inputs for principal component analysis, which showed distinct response of BMMSC and SaOS-2 to the surface topography and chemistry. Among the studied substrates, micellar nanopatterns assembled from the copolymer with high molecular weight promote the adhesion and spreading of BMMSC and have an opposite effect on SaOS-2. This nanopattern is thus beneficial for bone regeneration after injury or pathology, e.g. bone fracture or tumor removal.
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BACKGROUND: Accurate diagnosis of TB infection (TBI) is challenging due to the lack of a gold standard. Tuberculin skin test (TST) and interferon-gamma release assay (IGRA) are currently useful in TBI diagnosis, but both have several limitations. This study aims to evaluate inter-operator variability in TST measurements and determine its impact on TBI diagnosis and treatment.METHODS: This was a retrospective analysis of patients screened for TBI using at least TST at a public outpatient clinic specialised in TB from January 2019 to August 2021. TST readings performed by five experienced nurses were compared.RESULTS: A total of 671 screenings were analysed. TST positivity rate (P < 0.001) and mean TST measurements obtained by our nurses were significantly different (P < 0.001). Concordance of TST and IGRA results was of 83.4% in the overall population (κ = 0.479). However, TST/IGRA agreement was significantly different among nurses (P = 0.003).CONCLUSION: Our analysis of TST measurements by experienced nurses shows significant differences in TST positivity rate, mean measured values and overall concordance with IGRA. This led to significant different outcomes in TBI diagnosis and subsequent treatment. TST measurement differences could potentially be more pronounced if we considered untrained operators or those with occasional reading experience.
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Tuberculosis Latente , Humanos , Ensayos de Liberación de Interferón gamma/métodos , Tuberculosis Latente/diagnóstico , Tamizaje Masivo/métodos , Estudios Retrospectivos , Prueba de Tuberculina/métodosRESUMEN
Glioblastoma is the most prevalent and malignant brain tumor identified in adults. Surgical resection followed by radiotherapy and chemotherapy, mainly with temozolomide (TMZ), is the chosen treatment for this type of tumor. However, the average survival of patients is around 15 months. Novel approaches to glioblastoma treatment are greatly needed. Here, we aimed to investigate the anti-glioblastoma effect of the combination of matteucinol (Mat) (dihydroxyflavanone derived from Miconia chamissois Naudin) with the chemotherapeutic TMZ in vitro using tumor (U-251MG) and normal astrocyte (NHA) cell lines and in vivo using the chick embryo chorioallantoic membrane (CAM) assay. The combination was cytotoxic and selective for tumor cells (28 µg/mL Mat and 9.71 µg/mL TMZ). Additionally, the combination did not alter cell adhesion but caused morphological changes characteristic of apoptosis in vitro. Notably, the combination was also able to reduce tumor growth in the chick embryo model (CAM assay). The docking results showed that Mat was the best ligand to the cell death membrane receptor TNFR1 and to TNFR1/TMZ complex, suggesting that these two molecules may be working together increasing their potential. In conclusion, Mat-TMZ can be a good candidate for pharmacokinetic studies in view of clinical use for the treatment of glioblastoma.
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Glioblastoma , Receptores Tipo I de Factores de Necrosis Tumoral , Animales , Apoptosis , Línea Celular Tumoral , Proliferación Celular , Embrión de Pollo , Cromonas , Biología Computacional , Glioblastoma/tratamiento farmacológico , Humanos , Temozolomida/farmacología , Temozolomida/uso terapéuticoRESUMEN
Sustainable intensification of tropical grasslands has been identified by researchers and stakeholders as a solution to decrease greenhouse gas emissions and deforestation. However, there are concerns about food security and the role of livestock in feed-food competition between animals and humans involving land and other resources. We aimed to determine the net protein contribution (NPC), a feed-food competitiveness index, of tropical beef cattle raised on extensive systems or finished in pastures or conventional feedlots, under different levels of intensification. We modelled five scenarios, from cow-calf to slaughter, based on common beef cattle practices in Brazil, whose main production system is grazing. Scenario 1 represented the lowest level of intensification and the most extensive system. Scenario 2 represented a moderately extensive system. Scenarios 3, 4, and 5 represented different degrees and practices of intensification, with animals in cow-calf and stocker phases raised solely on well-managed permanent pastures. In Scenario 3, the animals were finished in a feedlot. In Scenarios 4 and 5, all animals in the stocker phase received a protein-energy supplement, but in Scenario 4, animals were finished in a permanent pasture with high-concentrate intake. In Scenario 5, animals were finished in a feedlot. The human-edible protein (heP) conversion efficiency (hePCE) was calculated as the ratio of heP produced (meat) to heP consumed as feed, and the NPC was the product of hePCE using the protein quality ratio, accounting for the digestible indispensable amino acid score content. An hePCEâ¯>â¯1 indicated that meat production did not compete with humans for food, and an NPCâ¯>â¯1 indicated that it contributed positively to meet human requirements. Meat production and heP intake consistently increased with intensification. The greatest hePCE values were from Scenarios 1 (9.2), 2 (2.2), and 3 (1.2), which were essentially pasture-fed systems, compared to Scenarios 4 and 5 (average of 1.0). The NPC varied from 24.1 (Scenario 1) to 2.6 (Scenario 5). The area required to produce 1â¯kg of carcass decreased from 147 to 45â¯m2, and the slaughter age decreased from 36 to 21â¯months from the most extensive to intensive systems. Brazilian beef cattle production contributes positively to the protein requirements of humans without limiting human food supplies. The intensification of tropical grazing beef systems is a key strategy to save land and produce more meat without limiting food for humans, playing an important role in the food security agenda.
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Alimentación Animal , Gases de Efecto Invernadero , Alimentación Animal/análisis , Crianza de Animales Domésticos , Animales , Bovinos , Suplementos Dietéticos , Femenino , Humanos , CarneRESUMEN
The tumor microenvironment (TME) is like the Referee of a soccer match who has constant eyes on the activity of all players, such as cells, acellular stroma components, and signaling molecules for the successful completion of the game, that is, tumorigenesis. The cooperation among all the "team members" determines the characteristics of tumor, such as the hypoxic and acidic niche, stiffer mechanical properties, or dilated vasculature. Like in soccer, each TME is different. This heterogeneity makes it challenging to fully understand the intratumor dynamics, particularly among different tumor subpopulations and their role in therapeutic response or resistance. Further, during metastasis, tumor cells can disseminate to a secondary organ, a critical event responsible for approximately 90% of the deaths in cancer patients. The recapitulation of the rapidly changing TME in the laboratory is crucial to improve patients' prognosis for unraveling key mechanisms of tumorigenesis and developing better drugs. Hence, in this chapter, we provide an overview of the characteristic features of the TME and how to model them, followed by a brief description of the limitations of existing in vitro platforms. Finally, various attempts at simulating the TME using microfluidic platforms are highlighted. The chapter ends with the concerns that need to be addressed for designing more realistic and predictive tumor-on-a-chip platforms.
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Dispositivos Laboratorio en un Chip , Neoplasias , Carcinogénesis , Humanos , Microfluídica , Neoplasias/patología , Microambiente TumoralRESUMEN
This study investigated the osteogenic performance of new brushite cements obtained from Li+-doped ß-tricalcium phosphate as a promising strategy for bone regeneration. Lithium (Li+) is a promising trace element to encourage the migration and proliferation of adipose-derived stem cells (hASCs) and the osteogenic differentiation-related gene expression, essential for osteogenesis. In-situ X-ray diffraction (XRD) and in-situ 1H nuclear magnetic resonance (1H NMR) measurements proved the precipitation of brushite, as main phase, and monetite, indicating that Li+ favored the formation of monetite under certain conditions. Li+ was detected in the remaining pore solution in significant amounts after the completion of hydration. Isothermal calorimetry results showed an accelerating effect of Li+, especially for low concentration of the setting retarder (phytic acid). A decrease of initial and final setting times with increasing amount of Li+ was detected and setting times could be well adjusted by varying the setting retarder concentration. The cements presented compressive mechanical strength within the ranges reported for cancellous bone. In vitro assays using hASCs showed normal metabolic and proliferative levels. The immunodetection and gene expression profile of osteogenic-related markers highlight the incorporation of Li+ for increasing the in vivo bone density. The osteogenic potential of Li-doped brushite cements may be recommended for further research on bone defect repair strategies.
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Effective and safe therapies to counteract persistent inflammation are necessary. We developed erythrocyte-derived liposomes (EDLs) with intrinsic anti-inflammatory activity. The EDLs were prepared using lipids extracted from erythrocyte membranes, which are rich in omega-3 fatty acids with several health benefits. Diclofenac, a widely used anti-inflammatory drug, was incorporated into EDLs in relevant therapeutic concentrations. The EDLs were also functionalised with folic acid to allow their active targeting of M1 macrophages, which are key players in inflammatory processes. In the presence of lipopolysaccharide (LPS)-stimulated macrophages, empty EDLs and EDLs incorporating diclofenac were able to reduce the levels of important pro-inflammatory cytokines, namely interleukin-6 (IL-6; ≈85% and 77%, respectively) and tumour necrosis factor-alpha (TNF-α; ≈64% and 72%, respectively). Strikingly, cytocompatible concentrations of EDLs presented similar effects to dexamethasone, a potent anti-inflammatory drug, in reducing IL-6 and TNF-α concentrations, demonstrating the EDLs potential to be used as bioactive carriers in the treatment of inflammatory diseases.
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Liposomas , Factor de Necrosis Tumoral alfa , Antiinflamatorios/farmacología , Antiinflamatorios/uso terapéutico , Citocinas , Diclofenaco/farmacología , Diclofenaco/uso terapéutico , Eritrocitos , Humanos , Inflamación/tratamiento farmacológico , Interleucina-6RESUMEN
The objective of these studies was to determine the effects of feeding a novel rumen-protected Lys (RP-Lys) product on plasma AA, lactational performance, and Lys bioavailability. To evaluate RP-Lys on lactation performance a corn-based diet (42.5% of corn silage and 21.9% of corn and corn by-products, on DM basis) was formulated to be Lys deficient but adequate in Met, energy, and metabolizable protein. Thirty-six lactating Holstein cows were fed either a Lys-deficient control diet (CON) with no added RP-Lys, or diets containing 0.3% of RP-Lys (0.3RP-Lys) or 0.6% of RP-Lys (0.6RP-Lys) for 8 wk. There were no effects on dry matter intake (mean ± SD; 26.1 ± 0.58 kg/d), milk yield (37.9 ± 0.72 kg/d), or milk composition to the RP-Lys supplementation. No effect was observed on plasma AA concentrations except for His. Plasma His was linearly reduced by Lys feeding (42.6, 41.2, 30.0 ± 4.09 µM, for CON, 0.3RP-Lys, and 0.6RP-Lys, respectively). Calculated efficiency of Lys utilization decreased linearly with RP-Lys supplementation. In the companion study, 3 rumen-cannulated lactating dairy cows were used in a 3 × 3 Latin square design to assess the bioavailability of the RP-Lys. Free Lys (HCl-Lys), RP-Lys, and water were administered separately by postruminal bolus dosing. The Lys bioavailability was assessed by the ratio of area under the curve of Lys plasma concentration for RP-Lys compared with HCl-Lys and discounted for the area under the curve for water bolus dose. The estimated bioavailability of the RP-Lys was 24.4% ± 4.61. In summary, increased supplemental doses of Lys had no effect on Lys plasma concentration and lactational performance when fed to dairy cows on a corn-based diet, although altered Lys as % of essential AA was observed. However, the lack of effects should be considered in light of the lower-than-expected bioavailability of the RP-Lys.
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Lisina , Rumen , Aminoácidos/metabolismo , Animales , Bovinos , Dieta/veterinaria , Femenino , Lactancia , Leche/química , Proteínas de la Leche/análisis , Rumen/metabolismo , Ensilaje , Zea mays/metabolismoRESUMEN
The host immunologic response to a specific material is a critical aspect when considering it for clinical implementation. Collagen and gelatin extracted from marine sources have been proposed as biomaterials for tissue engineering applications, but there is a lack of information in the literature about their immunogenicity. In this work, we evaluated the immune response to collagen and/or gelatin from blue shark and codfish, previously extracted and characterized. After endotoxin evaluation, bone marrow-derived macrophages were exposed to the materials and a panel of pro- and anti-inflammatory cytokines were evaluated both for protein quantification and gene expression. Then, the impact of those materials in the host was evaluated through peritoneal injection in C57BL/6 mice. The results suggested shark collagen as the less immunogenic material, inducing low expression of pro-inflammatory cytokines as well as inducible nitric oxide synthase (encoded by Nos2) and high expression of Arginase 1 (encoded by Arg1). Although shark gelatin appeared to be the material with higher pro-inflammatory expression, it also presents a high expression of IL-10 (anti-inflammatory cytokine) and Arginase (both markers for M2-like macrophages). When injected in the peritoneal cavity of mice, our materials demonstrated a transient recruitment of neutrophil, being almost non-existent after 24 hours of injection. Based on these findings, the studied collagenous materials can be considered interesting biomaterial candidates for regenerative medicine as they may induce an activation of the M2-like macrophage population, which is involved in suppressing the inflammatory processes promoting tissue remodeling. STATEMENT OF SIGNIFICANCE: Marine-origin biomaterials are emerging in the biomedical arena, namely the ones based in marine-derived collagen/gelatin proposed as cell templates for tissue regeneration. Nevertheless, although the major cause of implant rejection in clinical practice is the host's negative immune response, there is a lack of information in the literature about the immunological impact of these marine collagenous materials. This work aims to contribute with knowledge about the immunologic response to collagen/gelatin extracted from blue shark and codfish skins. The results demonstrated that despite some differences observed, all the materials can induce a macrophage phenotype related with anti-inflammation resolution and then act as immuno-modulators and anti-inflammatory inducible materials.
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Gelatina , Ingeniería de Tejidos , Animales , Antiinflamatorios , Arginasa , Materiales Biocompatibles/farmacología , Colágeno , Citocinas/metabolismo , Gelatina/farmacología , Ratones , Ratones Endogámicos C57BLRESUMEN
We describe the antithrombotic properties of nanopatterned coatings created by self-assembly of poly(styrene-block-2-vinylpyridine) (PS-b-P2VP) with different molecular weights. By changing the assembly conditions, we obtained nanopatterns that differ by their morphology (size and shape of the nanopattern) and chemistry. The surface exposition of P2VP block allowed quaternization, i.e. introduction of positive surface charge and following electrostatic deposition of heparin. Proteins (albumin and fibrinogen) adsorption, platelet adhesion and activation, cytocompatibility, and reendothelization capacity of the coatings were assessed and discussed in a function of the nanopattern morphology and chemistry. We found that quaternization results in excellent antithrombotic and hemocompatible properties comparable to heparinization by hampering the fibrinogen adhesion and platelet activation. In the case of quaternization, this effect depends on the size of the polymer blocks, while all heparinized patterns had similar performance showing that heparin surface coverage of 40 % is enough to improve substantially the hemocompatibility.