Hydrophobic bioactives can be more easily incorporated into food and have their bioavailability enhanced if nanostructured lipid carriers (NLC) are used as carriers. In the present study, ...beta-carotene-loaded NLC were produced by low emulsification using murumuru butter and a mixture of Span 80 and Cremophor RH40 as surfactants. Their average diameter was 35 nm and alpha-tocopherol was required to protect the encapsulated β-carotene. Besides the evaluation of their physicochemical stability, NLC were submitted to dynamic in vitro digestion and cell viability assays with Caco-2 and HEPG cells. The bioaccessibility of beta-carotene in the dynamic system was about 42%. Regarding cell viability, results indicated NLC were toxic to the cell cultures tested. Such high toxicity is probably related to the type of surfactant used and to the extremely reduced particle size, which may have led to an intense and fast permeation of the NLC through the cells.
The accuracy of cup drawing simulation of aluminum alloy sheets strongly depends on the hardening and yield functions used in the simulation. In this study, stress-strain curves for an AA3104 ...aluminum alloy sheet were measured by uniaxial tensile and rolling tensile tests to identify parameters for various hardening functions. In addition, in-plane uniaxial tensile, biaxial tensile, and combined tension-compression tests were carried out to measure the successive contours of plastic work in the stress space and the directions of plastic strain rates under linear stress paths. The anisotropic coefficients, α1-α8, and exponent M of the Yld2000-2d yield function were determined so that the mean square error of the analytical yield locus from the measured plastic work contour was minimized. Finite element simulations of cup drawing of the aluminum alloy sheet were carried out using the determined parameters for hardening functions and the Yld2000-2d yield function implemented in LS-DYNA. By comparing the simulation results with the results of cup drawing experiments, the effects of the hardening functions on the accuracy of earing profile prediction were investigated.
This study aimed to characterize the endometrial transcriptome and functional pathways overrepresented in the endometrium of cows treated to ovulate larger (≥13 mm) versus smaller (≤12 mm) follicles. ...Nelore cows were presynchronized prior to receiving cloprostenol (large follicle LF group) or not (small follicle SF group), along with a progesterone (P4) device on Day (D) -10. Devices were withdrawn and cloprostenol administered 42-60 h (LF) or 30-36 h (SF) before GnRH agonist treatment (D0). Tissues were collected on D4 (experiment Exp. 1; n = 24) or D7 (Exp. 2; n = 60). Endometrial transcriptome was obtained by RNA-Seq, whereas proliferation and apoptosis were assessed by immunohistochemistry. Overall, LF cows developed larger follicles and corpora lutea, and produced greater amounts of estradiol (D-1, Exp. 1, SF: 0.7 ± 0.2; LF: 2.4 ± 0.2 pg/ml; D-1, Exp. 2, SF: 0.5 ± 0.1; LF: 2.3 ± 0.6 pg/ml) and P4 (D4, Exp. 1, SF: 0.8 ± 0.1; LF: 1.4 ± 0.2 ng/ml; D7, Exp. 2, SF: 2.5 ± 0.4; LF: 3.7 ± 0.4 ng/ml). Functional enrichment indicated that biosynthetic and metabolic processes were enriched in LF endometrium, whereas SF endometrium transcriptome was biased toward cell proliferation. Data also suggested reorganization of the extracellular matrix toward a proliferation-permissive phenotype in SF endometrium. LF endometrium showed an earlier onset of proliferative activity, whereas SF endometrium expressed a delayed increase in glandular epithelium proliferation. In conclusion, the periovulatory endocrine milieu regulates bovine endometrial transcriptome and seems to determine the transition from a proliferation-permissive to a biosynthetic and metabolically active endometrial phenotype, which may be associated with the preparation of an optimally receptive uterine environment.
Osteoarthritis (OA) is one of the main locomotor disorders in horses. Although nonsteroidal anti-inflammatory drugs are the first-line treatment for OA, opioids could also be used. In previous ...studies, opioids showed promising anti-inflammatory and analgesic effects. In this study, we aimed to investigate the effects of two opioids (morphine and methadone) against inflammation in lipopolysaccharide (LPS)-stimulated synoviocytes by analyzing microsomal prostaglandin E synthase-1 (mPGES-1) and prostaglandin-endoperoxide synthase 2 (PTGS2) expression. Synoviocytes were obtained from the joints at the distal limbs of dead animals. The cytotoxic effects of LPS, morphine, and methadone were investigated by using a cell viability assay with crystal violet dye. Synoviocytes were treated with LPS, LPS plus morphine, or LPS plus methadone for 3, 6, and 12 h, and mPGES-1 and PTGS2 expression was measured using real-time polymerase chain reaction. LPS, and morphine did not affect the viability of synoviocytes, even at high concentrations. LPS treatment increased mPGES-1 and PTGS2 expression, whereas morphine inhibited the increase in mPGES-1 and PTGS2 expression in LPS-stimulated synoviocytes. Methadone did not inhibit mPGES-1 or PTGS2 expression. These results suggest that morphine may exhibit anti-inflammatory effect; therefore, it might be beneficial for the treatment of OA.
•LPS and morphine did not affect the viability of synoviocytes, even at high concentrations.•A concentration of 10−6 M of methadone showed some synoviocyte toxicity.•LPS-stimulated synoviocytes showed higher mPGES-1 and PTGS2 expression than the non-stimulated synoviocytes.•Morphine inhibited the increase of mPGES-1 and PTGS2 expression in LPS-stimulated synoviocytes.
•Global gene expression profiling in 12 canine mammary carcinomas revealed 1699 differentially expressed genes.•The HYAL-1 gene was identified as a potential marker associated with tumorigenesis and ...local recurrence.•Larger studies are needed to confirm the role of HYAL-1 in malignancy and disease progression.
Mammary gland tumors are a heterogeneous group of neoplastic diseases. Genetic studies make it possible to determine genetic profiles and identify new molecular markers. The aim of the study was to evaluate the gene expression profile of canine mammary carcinomas and identify potential prognostic markers. Twelve mammary cancer samples from bitches were collected for the evaluation of global gene expression. Microarray assays were performed using commercial kits. Statistical analysis of the microarray was done using moderate t-statistic and adjusted using the Benjamini and Hochberg procedure. Differential connectivity analysis was also performed. Enrichment analyses were conducted using WebGestalt. P-values were calculated using hypergeometric statistics and adjusted using the Benjamini and Hochberg procedure. The HYAL-1 gene was validated using quantitative PCR (qPCR).
There were 878 upregulated genes and 821 downregulated genes in the neoplasms studied. Enrichment analysis (individual analysis) identified the HYAL-1 gene as a potential marker of tumorigenesis and tumor recurrence. Differential connectivity analysis demonstrated 262 differentially connected genes.
Takahashi and Yamanaka established the first technique in which transcription factors related to pluripotency are incorporated into the genome of somatic cells to enable reprogramming of these cells. ...The expression of these transcription factors enables a differentiated somatic cell to reverse its phenotype to an embryonic state, generating induced pluripotent stem cells (iPSCs). iPSCs from canine fetal fibroblasts were produced through lentiviral polycistronic human and mouse vectors (hOSKM/mOSKM), aiming to obtain pluripotent stem cells with similar features to embryonic stem cells (ESC) in this animal model. The cell lines obtained in this study were independent of LIF or any other supplemental inhibitors, resistant to enzymatic procedure (TrypLE Express Enzyme), and dependent on bFGF. Clonal lines were obtained from slightly different protocols with maximum reprogramming efficiency of 0.001%. All colonies were positive for alkaline phosphatase, embryoid body formation, and spontaneous differentiation and expressed high levels of endogenous OCT4 and SOX2. Canine iPSCs developed tumors at 120 days post-injection in vivo. Preliminary chromosomal evaluations were performed by FISH hybridization, revealing no chromosomal abnormality. To the best of our knowledge, this report is the first to describe the ability to reprogram canine somatic cells via lentiviral vectors without supplementation and with resistance to enzymatic action, thereby demonstrating the pluripotency of these cell lines.
Overview of the steps for obtaining the iPS cells. From the somatic cell chosen, genic factors are added, and after the formation of colonies, they must be characterized as positive for alkaline phosphatase, embryoid body formation, gene markers for immunofluorescence and real-time PCR, in vivo tumor formation and the possible abnormalities in chromosomes, resulting from reprogramming process. Display omitted
•iPSCs cells were generated by canine fetal fibroblast.•IPSCs were generated without LIF and resistant to enzymatic passages.•The strains were positive for all characterization testes and capable to forming tumor mass after 120 days.•The evaluated strains showed no chromosomal changes.
In-plane biaxial tension and combined tension-compression tests are carried out for AA3104 aluminum alloy sheets. Linear stress paths are applied to cruciform specimens to measure the contours of ...plastic work in the stress space and the directions of plastic strain rates at each stress path. Coefficients α1- α8 and exponent M of the Yld2000-2d yield function are determined to minimize the mean square error of the analytical yield locus from a measured work contour. The values of the weighting coefficients in the evaluation of the error are varied to check the effect of a specific stress state on the earing behavior. The effects of the combinations of the weighting coefficients on the accuracy of earing prediction in the cup drawing process are discussed.
Nuclear receptor subfamily 1, group I, member 3 (NR1I3) is reported to be a possible novel therapeutic target for some cancers, including lung, brain and hematopoietic tumors. Here, we characterized ...expression of NR1I3 in a mouse model of lung carcinogenesis induced by 4-(methylnitrosamino)-4-(3-pyridyl)-1-butanone (NNK), the most potent tobacco carcinogen. Lung tumors were collected from mice treated with NNK (400 mg/kg) and euthanized after 52 weeks. Benign and malignant lesions were formalin-fixed and paraffin-embedded for histology and immunohistochemistry, with samples snap-frozen for mRNA analysis. Immunohistochemically, we found that most macrophages and type I and II pneumocytes expressed NR1I3, whereas fibroblasts and endothelial cells were NR1I3-. Compared with benign lesions, malignant lesions had less NR1I3+ tumor cells. Gene expression analysis also showed an inverse correlation between NR1I3 mRNA expression and tumor size (P=0.0061), suggesting that bigger tumors expressed less NR1I3 transcripts, in accordance with our immunohistochemical NR1I3 tests. Our results indicate that NR1I3 expression decreased during progression of malignant lung tumors induced by NNK in mice.
The aim of this study was to identify candidate genes and genomic regions associated with ultrasound‐derived measurements of the rib‐eye area (REA), backfat thickness (BFT) and rumpfat thickness ...(RFT) in Nellore cattle. Data from 640 Nellore steers and young bulls with genotypes for 290 863 single nucleotide polymorphisms (SNPs) were used for genomewide association mapping. Significant SNP associations were explored to find possible candidate genes related to physiological processes. Several of the significant markers detected were mapped onto functional candidate genes including ARFGAP3, CLSTN2 and DPYD for REA; OSBPL3 and SUDS3 for BFT; and RARRES1 and VEPH1 for RFT. The physiological pathway related to lipid metabolism (CLSTN2, OSBPL3, RARRES1 and VEPH1) was identified. The significant markers within previously reported QTLs reinforce the importance of the genomic regions, and the other loci offer candidate genes that have not been related to carcass traits in previous investigations.
This study aimed to characterize the endometrial transcriptome and functional pathways overrepresented in the endometrium of cows treated to ovulate larger (≥13 mm) versus smaller (≤12 mm) follicles. ...Nelore cows were presynchronized prior to receiving cloprostenol (large follicle LF group) or not (small follicle SF group), along with a progesterone (P4) device on Day (D) −10. Devices were withdrawn and cloprostenol administered 42–60 h (LF) or 30–36 h (SF) before GnRH agonist treatment (D0). Tissues were collected on D4 (experiment Exp. 1; n = 24) or D7 (Exp. 2; n = 60). Endometrial transcriptome was obtained by RNA-Seq, whereas proliferation and apoptosis were assessed by immunohistochemistry. Overall, LF cows developed larger follicles and corpora lutea, and produced greater amounts of estradiol (D−1, Exp. 1, SF: 0.7 ± 0.2; LF: 2.4 ± 0.2 pg/ml; D−1, Exp. 2, SF: 0.5 ± 0.1; LF: 2.3 ± 0.6 pg/ml) and P4 (D4, Exp. 1, SF: 0.8 ± 0.1; LF: 1.4 ± 0.2 ng/ml; D7, Exp. 2, SF: 2.5 ± 0.4; LF: 3.7 ± 0.4 ng/ml). Functional enrichment indicated that biosynthetic and metabolic processes were enriched in LF endometrium, whereas SF endometrium transcriptome was biased toward cell proliferation. Data also suggested reorganization of the extracellular matrix toward a proliferation-permissive phenotype in SF endometrium. LF endometrium showed an earlier onset of proliferative activity, whereas SF endometrium expressed a delayed increase in glandular epithelium proliferation. In conclusion, the periovulatory endocrine milieu regulates bovine endometrial transcriptome and seems to determine the transition from a proliferation-permissive to a biosynthetic and metabolically active endometrial phenotype, which may be associated with the preparation of an optimally receptive uterine environment.
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