Platelet-rich plasma (PRP) is an autologous blood product that contains a variety of growth factors (GFs) that are released upon platelet activation. Despite some therapeutic potential of PRP in ...vitro, in vivo data are not convincing. Bolus injection of PRP is cleared rapidly from the body diminishing its therapeutic efficacy. This highlights a need for a delivery vehicle for a sustained release of PRP to improve its therapeutic effect. In this study, we used microfluidics to fabricate biodegradable PRP-loaded polyethylene glycol (PEG) microspheres. PRP was incorporated into the microspheres as a lyophilized PRP powder either as is (powder PRP) or first solubilized and pre-clotted to remove clots (liquid PRP). A high PRP loading of 10%
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was achieved for both PRP preparations. We characterized the properties of the resulting PRP-loaded PEG microspheres including swelling, modulus, degradation, and protein release as a function of PRP loading and preparation. Overall, loading powder PRP into the PEG microspheres significantly affected the properties of microspheres, with the most pronounced effect noted in degradation. We further determined that microsphere degradation in the presence of powder PRP was affected by platelet aggregation and clotting. Platelet aggregation did not prevent but prolonged sustained PRP release from the microspheres. The delivery system developed and characterized herein could be useful for the loading and releasing of PRP to promote tissue regeneration and wound healing or to suppress tissue degeneration in osteoarthritis, and intervertebral disc degeneration.
Abstract
We describe here a novel nanoplatform called the Protein-Like Polymer (PLP) with unique characteristics allowing for sustained delivery of tumor antigens in conjunction with STING agonists.
...Methods:
A library of PLP formulations were synthesized via ROMP and characterized. Cell uptake/functional assays were conducted with payload-specific T Cells. Ability of PLPs to co-deliver adjuvants was tested by electrostatically coupling 2′3′ cGAMP, forming stable nanostructures.
Results:
Conjugating antigens to the polymer backbone using a cleavable disulfide linkage (reduces in APCs) resulted in increased endosomal localization and T cell proliferation/activation. Incorporating OEG side chains reduced enzymatic degradation while increasing immunogenicity and APC uptake. Increasing the density of side chains further improved vaccine efficacy and resistance to proteolysis. Antigen-PLP conjugates is effective only when paired with cells from their cognate system, demonstrating payload-specificity. Mice bearing established B16F10 tumors treated with gp100-PLPs resulted in increased survival time and reduced tumor burden with corresponding changes in immune cell profiles. Notably, STING-PLP complexes showed significantly smaller tumors at day 14 and allowed for subcutaneous administration of 2′3′ cGAMP. Studies looking at multiplexing pools of neoantigens onto one PLP and generation of immunological memory are ongoing.
Conclusion:
The modularity of the PLP platform allows for complex nano-architectures including systems capable of delivering challenging compounds, ie small molecule STING agonists, subcutaneously through electrostatic coupling, highlighting its potential to revolutionize cancer vaccinology.
Supported by grants from the NIH (5F30CA257519-03)
The current study aimed to understand how older adults with mild cognitive impairment (MCI) perceived their condition following diagnosis and to explain the process of coping with concomitant changes ...in their lives. Using theoretical sampling methods, in-depth interviews were conducted between August 2015 and September 2017 in 20 older adults with MCI; the collected data were analyzed using the constant comparative method. The core category was living a daily life with self-awareness. The four stages of daily life experience among participants were accepting their diagnosis, strengthening their volition, taking care of their health, and maintaining a daily life with self-awareness. After being diagnosed with MCI, participants were more attentive to behaviors in their daily lives. Participants accepted their diagnosis as well as changes to their lifestyles to avoid causing inconvenience to others. Research in Gerontological Nursing, 13(1), 41-48..
한국어판 간호사의 위임에 대한 태도 및 준비성 측정도구의 타당도와 신뢰도 김미영(Kim, Miyoung); 박진화(Park, Jinhwa); 최미란(Choi, Miran)
Kanho Haengjŏng Hakhoe chi = The journal of Korean Nursing Administration Academic Society,
2016, Letnik:
22, Številka:
1
Journal Article
Recenzirano
Odprti dostop
Purpose: The purpose of this study was to develop and test the validity and reliability of the Korean version of nurses' attitudes toward delegation and preparedness to delegate (APD). Methods: The ...Korean version of APD was developed through forward-backward translation methods. Internal consistency reliability, criterion validity, and construct validity using exploratory and confirmatory factor analysis were conducted using IBM SPSS Statistics 19 and AMOS 20.0. Survey data were collected from 161 nurses working in 2 general hospitals. Results: The Korean version of APD showed Cronbach's alphas of .68 and .85. Factor loadings of the 8 attitude items on the 3 subscales ranged from .60 to .86 and the 15 preparedness items on the 4 subscales ranged from .47 to .90. The model of 3 subscales for the Korean nurses' attitude toward delegation and the model of 4 subscales for the Korean nurses' preparedness to delegate were both validated by confirmatory factor analysis(NC<3, CFI>.90, RMSEA<.10). Criterion validity compared to job satisfaction showed significant correlation. Conclusion: The findings of this study demonstrate that this modified Korean version of APD is applicable for measuring Korean nurses' attitude toward delegation and preparedness to delegate.
Abstract Background: Patient-specific cancer vaccines derived from tumor antigen have been explored as a promising therapeutic strategy, however, challenges delivering vaccine components in a ...coordinated fashion to elicit antitumor responses remain. To overcome these, we utilize a novel nanoplatform called the Protein-Like Polymer (PLP), which allows for sustained and targeted delivery of tumor antigens in conjunction with adjuvants. Methods: PLPs containing peptide antigens were synthesized via ring-opening metathesis polymerization (ROMP) and characterized. A library of compounds with different sidechain linkage chemistries, degrees of polymerization (DP), and inclusion/exclusion of Oligo-ethylene glycol (OEG) were made to determine design rules for immune activation. Cell uptake and functional assays using payload-specific T Cells were conducted. Immunization in three independent tumor models was done to show generalizability. Ability of PLPs to co-deliver adjuvants was tested by electrostatically coupling small molecule STING agonist, 2’3’ cGAMP. Results: Conjugating peptide antigens to the polymer via a cleavable disulfide linkage, which reduces intracellularly in APCs, resulted in increased endosomal localization, higher levels of induced T cell proliferation, cytokine production, and expression of activation markers in CTLs and APCs. Incorporating a diluent amount of OEG side chains reduced enzymatic degradation while increasing immunogenicity and uptake. Additionally, increasing the DP, and therefore the density of antigen side chains, further improved vaccine efficacy and resistance to proteolysis. Antigen-PLP conjugates enhanced dendritic cell activation and T-cell response only when paired with cells from their cognate system, with no activity in immune cells not expressing receptors for the payload demonstrating antigen-specificity. Mice bearing established B16F10, MC38, or TC-1 tumors treated with PLPs containing gp100, adpgk, or E7 respectively all showed increased survival times, reduced tumor burden with corresponding changes in immune cell profiles, and immunological memory upon rechallenge. Impressively, mice treated with STING-PLP complexes had significantly smaller tumors vs control at day 14 (0.038g vs 0.76g; p <0.0001) and allowed for subcutaneous administration of 2’3’ cGAMP, which traditionally requires intratumoral injection. Studies on the effects of vaccinating with pools of neoantigens multiplexed onto one PLP are ongoing. Conclusion: This work validates the ability of PLPs to overcome major limitations in cancer vaccine development. The modularity of the platform allows for complex nano-architectures including systems capable of delivering challenging compounds, ie small molecule STING agonists, subcutaneously through electrostatic coupling, highlighting its potential to revolutionize cancer vaccinology. Citation Format: Max Wang, Miran Choi, Bin Zhang, Nathan Gianneschi. Proteomimetic polymers limit tumor growth in multiple models via potent antigen-specific T cell activation abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 481.
Abstract
Introduction: Altered metabolism is a hallmark of cancer. We have reported that a set of lipid metabolism (LiMe) genes is over-expressed in the contralateral unaffected breasts (CUBs) of ...women with unilateral estrogen receptor negative (ER-) breast cancer, who are at high risk for future ER- tumors. We hypothesized that a potential mechanism connecting lipid metabolism to risk of ER- disease may involve epigenetic regulation of gene expression in breast cells by lipids, as shown by McDonnel et. al., Cell reports 2016 in hepatocytes.
Methods: Mass spectrometry was performed to profile the composition of lipid species in the sera of patients with ER+ versus ER- disease. MCF-10A cells and mammary organoids from reduction mammoplasty patients were treated with Octanoic (OA) and Linoleic acid (LA) followed by western blot analysis of acetylated histones and sequencing of RNA with qPCR confirmation of significant findings. Partial wave spectroscopic (PWS) microscopy was employed to study change in chromatin. Epithelial RNA from microdissected benign breast samples from CUBs of 84 patients (28 ER+, 28 ER- and 28 healthy controls) were subjected to qPCR.
Results: Sera from ER- patients showed higher levels of two triacylglycerides: TAG46:4-FA18:2 (1.25-fold, p=0.05) and TAG 47:2-FA18:2 (1.35-fold, p=0.07), both of which contain LA. MCF10A cells and human mammary organoids showed induction of H3 acetylation at lysine 9 & 14 after treatment with LA and OA treatment at physiological levels. This was accompanied by evidence of chromatin remodeling and 2-fold increase in DNA accessibility on PWS. RNA-Seq of OA treated MCF-10A cells revealed significant upregulation of LiMe genes together with pathways associated with breast malignancy: Notch, Wnt, EMT and adenylate cyclase. The adenylate cyclase pathway (known to be specifically associated with ER- breast cancer risk), was enriched 3.7-fold (p=0.007). In RNA from CUBs of 84 patients, we found overexpression of Notch pathway intermediates that responded to lipid exposure in MCF10A in the ER- group, specifically Notch1 (1.8-fold, p=0.02) and Hey1 (2.9-fold, p=0.05).
Conclusion: Specific classes of dietary lipids are the source of acetyl-CoA which acetylates histones and remodels chromatin to modulate gene transcription in ER- MCF10A cells. Thus revved-up fatty acid oxidation may play a role in ER- breast oncogenesis via Notch signaling. Our novel findings point to potential interception strategies that target connections between lipid metabolism and histone modifications.
Citation Format: Shivangi Yadav, MiRan Choi, David Van Derway, Greta Braur, Vadim Backman, Seema Ahsan Khan, Susan Clare. Fatty acid metabolism is associated with chromatin remodeling in mammary epithelial cells abstract. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P3-02-07.
αGal-deficient xenografts are protected from hyperacute rejection during xenotransplantation but are still rejected more rapidly than allografts. Despite studies showing the roles of non-Gal ...antibodies and αβ T cells in xenograft rejection, the involvement of γδ T cells in xenograft rejection has been limitedly investigated.
Six male cynomolgus monkeys were transplanted with porcine vessel xenografts from wild-type (n = 3) or GGTA1 knockout (n = 3) pigs. We measured the proportions and T cell receptor (TCR) repertoires of blood γδ T cells before and after xenotransplant. Grafted porcine vessel-infiltrating immune cells were visualized at the end of experiments.
Blood γδ T cells expanded and infiltrated into the graft vessel adventitia following xenotransplantation of α-Gal-deficient pig blood vessels. Pre- and post-transplant analysis of γδ TCR repertoire revealed a transition in δ chain usage post-transplantation, with the expansion of several clonotypes of δ1, δ3, or δ7 chains. Furthermore, the distinctions between pre- and post-transplant δ chain usages were more prominent than those observed for γ chain usages.
γδ TCR repertoire was significantly altered by xenotransplantation, suggesting the role of γδ T cells in sustained xenoreactive immune responses.
The gut microbiota has been known to modulate the immune responses in chronic liver diseases. Recent evidence suggests that effects of dietary foods on health care and human diseases are related to ...both the immune reaction and the microbiome. The gut-microbiome and intestinal immune system play a central role in the control of bacterial translocation-induced liver disease. Dysbiosis, small intestinal bacterial overgrowth, translocation, endotoxemia, and the direct effects of metabolites are the main events in the gut-liver axis, and immune responses act on every pathways of chronic liver disease. Microbiome-derived metabolites or bacteria themselves regulate immune cell functions such as recognition or activation of receptors, the control of gene expression by epigenetic change, activation of immune cells, and the integration of cellular metabolism. Here, we reviewed recent reports about the immunologic role of gut microbiotas in liver disease, highlighting the role of diet in chronic liver disease.
Abstract
Background: Triple negative breast cancer (TNBC) is over-represented in indigenous African women and in women of African descent. The majority of women diagnosed with breast cancer in ...Nigeria, Uganda and Kenya have TNBC. Studies of TNBC in the US consistently report the percentage of TNBC to be 10-24% greater in African-Americans (AA) than in Caucasians (C). The abundance of TNBCs in both African and African-American women has yet to be explained and the incidence rates suggest that there may be a genetic predisposition to this particularly aggressive form of breast cancer.
Methods: Epithelial cell lines were established from normal, healthy breast tissue donated to the Susan G. Komen for the Cure® Tissue Bank at the IU Simon Cancer Center. Six cell lines from AA donors were matched on the basis of age, menopausal status and Gail-risk score to six cell lines from C donors. RNA was extracted from the cells and gene expression assayed using the Illumina HumanWG6 v3 chip. Significant differences in gene expression were identified using PADE (PAirwise Differential Expression; Expression Analysis, Durham, NC).
Results: 1552 genes were differentially expressed (p≤ 0.05). The largest number of genes over-expressed in AAs mapped to the long arm of chromosome 1. Copy number gains have been observed in this region in multiple TNBC data sets. Many of these genes are in the 1q21 region, a region frequently rearranged in epithelial tumors as well as soft tissue and bone. Analysis of differentially expressed genes using MetaCore from Thomson Reuters revealed that the over-expressed genes are responsible for skin development, cell adhesion/cell junctions, and functions of the innate immune system. Pattern recognition is in the vanguard of the innate immune system. The pattern recognition pathways mediated by RAGE (receptor for advanced glycation end products) and the Toll-like receptors are activated in the normal AA epithelium. The expression of genes in these pathways, specifically S100A12, S100A8, S110A9, and TLR5, is known to increase in the presence of Salmonella and Shigella. MDA5 recognizes rotavirus infection. LYST and DUSP4 are active in the control of Leishmania infection. LCN2 disrupts iron acquisition, which protects against a number of pathogens including Salmonella, Klebsiella and mycobacteria.
Conclusions: The results of the above gene expression analysis are best understood in light of our recently published findings using these same cells (Sauder et al, BMC Cell Biology, 2014). Epithelial cells grown from mammary explant culture are basal-like and phenotypically plastic. They can assume a wide range of cell types depending on the cues provided by the microenvironment. When grown in basement membrane matrix or on human dermal fibroblasts, these cells produce a stratified squamous epithelium. Although the cells used for the gene expression analysis were grown in plastic tissue culture flasks and grew as a monotonous monolayer, the data suggest that they are, to some extent, transcriptionally “squamous”. For example, they express involucrin and the genes that code for the corneodesmosome proteins: corneodesmin, desmoglein and desmocollin.
The skin of humans living in sub-Saharan Africa has evolved to provide optimal barrier protection. It specifically protects against water loss and, pathogenic microorganisms and parasites. Mammary glands are ectodermal appendages of the epithelium and they may share similar immune defense mechanisms. The increased expression of genes involved in innate immunity in AA breast epithelial cells is of significant interest. We hypothesize that a subclinical, chronic inflammatory state is incited within basal breast cells in response to a pathogen that is endemic to Africa and overrepresented in low income communities in the US. This persistent and unresolved innate immunity predates the development of TNBC and can thus participate in its initiation and promotion.
Citation Format: Candice A.M. Sauder, Jillian E. Koziel, MiRan Choi, Brittney-Shea Herbert, Susan E. Clare. Clues to the causes of the abundance of triple-negative breast cancer in women of African descent. abstract. In: Proceedings of the Seventh AACR Conference on The Science of Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; Nov 9-12, 2014; San Antonio, TX. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2015;24(10 Suppl):Abstract nr B48.
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