Small molecule conjugation reduces macrophage uptake and increases in vivo blood circulation of polystyrene nanoparticles.

Nanomedicine often failed clinically to show therapeutic efficacy due to reduced particle circulation and enhanced capture by the reticuloendothelial system (RES), including the liver. Developing novel immunomodulatory surface coating can prevent macrophage capture and increase the particle circulation of the nanomedicine, resulting in higher therapeutic efficiency. Herein, we demonstrate the development of immunomodulatory small molecule (RZA15) with triazole functionality using copper-catalyzed click chemistry to conjugate onto spherical polystyrene nanoparticles using amide coupling reactions, achieving higher blood circulation and lesser macrophage uptake of the nanoconjugates. In this work, we evaluated the effectiveness of RZA15 coating for the enhanced circulation of polystyrene nanoparticles of 100 nm size, which is commonly utilized for various drug delivery applications, and compared with poly(ethylene)glycol (PEG) coatings. Several polystyrene nanoconjugate formulations were analyzed in vitro in normal and macrophage cells for cell viability and cellular uptake studies. In vitro studies demonstrated lesser macrophage uptake of the nanoconjugates following RZA15 coating. Finally, in vivo, blood-circulation, pharmacokinetics, and biodistribution studies were performed in the C57BL/6J mouse model that endorsed the substantial role of RZA15 in preventing liver and spleen capture and results in extended circulation. Coating immunomodulatory small molecules to nanoparticles can severely enhance the potential therapeutic effects of nanomedicine at lower doses.

Fibroblasts in metastatic lymph nodes confer cisplatin resistance to ESCC tumor cells via PI16.

Although many studies have compared tumor fibroblasts (T-Fbs) and nontumor fibroblasts (N-Fbs), less is understood about the stromal contribution of metastatic lymph node fibroblasts (LN-Fbs) to the evolving microenvironment. Here, we explored the characteristics of LN-Fbs in esophageal squamous cell carcinoma (ESCC) and the interactions between fibroblasts and ESCC tumor cells in metastatic lymph nodes. Fibroblasts were isolated from tumor, nontumor and metastatic lymph node tissues from different patients with ESCC. Transcriptome sequencing was performed on the fibroblasts. Tumor growth and drug-resistance assays were carried out, and characteristics of T-Fbs, N-Fbs and LN-Fbs were determined. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to assay the culture medium of fibroblasts. The results demonstrated that fibroblasts derived from different tissues had different characteristics. Coculture with LN-Fbs conditioned medium inhibited ESCC tumor cell growth and induced chemoresistance in ESCC cells. LN-Fbs induced chemoresistance to cisplatin in ESCC cells by secreting PI16. Coculture with LN-Fbs conditioned medium decreased cisplatin-induced apoptosis in ESCC cells by regulating the p38 and JNK cell signaling pathways. Survival analyses showed that patients with high PI16 expression in Fbs of lymph nodes exhibited worse overall survival. We also examined PI16 expression in interstitial tissues in ESCC tumor samples of patients receiving platinum-based therapy postsurgery and found that high PI16 expression in tumor interstitial tissues was an independent prognostic factor for ESCC patients. In addition, an in vivo assay demonstrated that PI16 knockdown increased the sensitivity of ESCC cells to cisplatin. Our results suggest that fibroblasts in metastatic lymph nodes decrease apoptosis of ESCC cells via PI16, thereby providing a cisplatin-resistance niche and supporting ESCC tumor cells to survive in metastatic lymph nodes. PI16 is also a potential target for effectively blocking the chemoresistance niche signaling circuit in response to cisplatin.

The influence of physiological and pathological perturbations on blood-brain barrier function.

The blood-brain barrier (BBB) is located at the interface between the vascular system and the brain parenchyma, and is responsible for communication with systemic circulation and peripheral tissues. During life, the BBB can be subjected to a wide range of perturbations or stresses that may be endogenous or exogenous, pathological or therapeutic, or intended or unintended. The risk factors for many diseases of the brain are multifactorial and involve perturbations that may occur simultaneously (e.g., two-hit model for Alzheimer's disease) and result in different outcomes. Therefore, it is important to understand the influence of individual perturbations on BBB function in isolation. Here we review the effects of eight perturbations: mechanical forces, temperature, electromagnetic radiation, hypoxia, endogenous factors, exogenous factors, chemical factors, and pathogens. While some perturbations may result in acute or chronic BBB disruption, many are also exploited for diagnostic or therapeutic purposes. The resultant outcome on BBB function depends on the dose (or magnitude) and duration of the perturbation. Homeostasis may be restored by self-repair, for example, via processes such as proliferation of affected cells or angiogenesis to create new vasculature. Transient or sustained BBB dysfunction may result in acute or pathological symptoms, for example, microhemorrhages or hypoperfusion. In more extreme cases, perturbations may lead to cytotoxicity and cell death, for example, through exposure to cytotoxic plaques.

Screening hydrogels for antifibrotic properties by implanting cellularly barcoded alginates in mice and a non-human primate.

Screening implantable biomaterials for antifibrotic properties is constrained by the need for in vivo testing. Here we show that the throughput of in vivo screening can be increased by cellularly barcoding a chemically modified combinatorial library of hydrogel formulations. The method involves the implantation of a mixture of alginate formulations, each barcoded with human umbilical vein endothelial cells from different donors, and the association of the identity and performance of each formulation by genotyping single nucleotide polymorphisms of the cells via next-generation sequencing. We used the method to screen 20 alginate formulations in a single mouse and 100 alginate formulations in a single non-human primate, and identified three lead hydrogel formulations with antifibrotic properties. Encapsulating human islets with one of the formulations led to long-term glycaemic control in a mouse model of diabetes, and coating medical-grade catheters with the other two formulations prevented fibrotic overgrowth. High-throughput screening of barcoded biomaterials in vivo may help identify formulations that enhance the long-term performance of medical devices and of biomaterial-encapsulated therapeutic cells.

Modeling angiogenesis in the human brain in a tissue-engineered post-capillary venule.

Angiogenesis plays an essential role in embryonic development, organ remodeling, wound healing, and is also associated with many human diseases. The process of angiogenesis in the brain during development is well characterized in animal models, but little is known about the process in the mature brain. Here, we use a tissue-engineered post-capillary venule (PCV) model incorporating stem cell derived induced brain microvascular endothelial-like cells (iBMECs) and pericyte-like cells (iPCs) to visualize the dynamics of angiogenesis. We compare angiogenesis under two conditions: in response to perfusion of growth factors and in the presence of an external concentration gradient. We show that both iBMECs and iPCs can serve as tip cells leading angiogenic sprouts. More importantly, the growth rate for iPC-led sprouts is about twofold higher than for iBMEC-led sprouts. Under a concentration gradient, angiogenic sprouts show a small directional bias toward the high growth factor concentration. Overall, pericytes exhibited a broad range of behavior, including maintaining quiescence, co-migrating with endothelial cells in sprouts, or leading sprout growth as tip cells.

Corrigendum: The influence of physiological and pathological perturbations on blood-brain barrier function.

[This corrects the article DOI: 10.3389/fnins.2023.1289894.].

Differing views regarding diet and physical activity: adolescents versus parents' perspectives.

BACKGROUND: Today, approximately one in five United States adolescents age 12 to 19 years is obese and just over a third are either overweight or obese. This study examines how parents and peers influence diet and physical activity behaviors of older adolescents (14-18 years) with overweight or obesity to inform weight management interventions. METHODS: Adolescent participants included 14 to 18-year-olds with a Body Mass Index (BMI) greater than the 85th percentile for their age and sex who were receiving care in a large healthcare system in Northern California. Adolescents and their parents participated in separate focus groups and interviews (if not able to attend focus groups) that were held at the same time in the same location. We used qualitative thematic analysis to identify common themes discussed in the adolescent and parent focus groups as well as paired analysis of adolescent-parent dyads. RESULTS: Participants included 26 adolescents and 27 parents. Adolescent participants were 14 to 18 years old. Half were female and the participants were almost evenly distributed across year in school. The majority self-identified as White (56%) and Asian (36%).Three themes were identified which included 1) parents overestimated how supportive they were compared to adolescents' perception 2) parents and adolescents had different views regarding parental influence on adolescent diet and physical activity behaviors 3) parents and adolescents held similar views on peers' influential role on lifestyle behaviors. CONCLUSION: Parents' and adolescents' differing views suggest that alignment of parent and adolescent expectations and behaviors for supporting effective weight management could be incorporated into interventions.

Recent Advancements of Magnetic Nanomaterials in Cancer Therapy.

Magnetic nanomaterials belong to a class of highly-functionalizable tools for cancer therapy owing to their intrinsic magnetic properties and multifunctional design that provides a multimodal theranostics platform for cancer diagnosis, monitoring, and therapy. In this review article, we have provided an overview of the various applications of magnetic nanomaterials and recent advances in the development of these nanomaterials as cancer therapeutics. Moreover, the cancer targeting, potential toxicity, and degradability of these nanomaterials has been briefly addressed. Finally, the challenges for clinical translation and the future scope of magnetic nanoparticles in cancer therapy are discussed.

Racial/ethnic differences in reporting versus rating of healthcare experiences.

Asians are reported to have poorer healthcare experience than non-Hispanic Whites (NHWs), but the sources of the differences are not understood. One explanation is Asian's reluctance to choose extreme responses in survey. We thus sought to compare NHW-Asian differences in responses to healthcare experience surveys when asked to report versus rate their experiences. Patients of an outpatient care system in 2013 to 2014 in the United States were studied. Patient experience surveys were sent after randomly selected clinic visits. Responses from 6 major Asian subgroups and NHWs were included (N = 61,115). The surveys used a combined questionnaire of Clinician and Group Consumer Assessment of Healthcare Providers and Systems (CG-CAHPS) and Press Ganey surveys. CG-CAHPS questions are framed as "reporting" and Press Ganey questions as "rating" of experiences. We compared the proportion of favorable (or top box) responses to 2 related questions, one from CG-CAHPS and another from Press Ganey, and assessed racial/ethnic differences when using each of the 2 related questions, using a Pearson chi-squared test for independence. All Asian subgroups were less likely to select top box than NHWs for all questions. The Asian-NHW differences in 'rating" questions were larger than the difference in related "reporting" questions. Of those who chose top box to CG-CAHPS questions (e.g., "Yes" on a question asking "Waited < 15 minutes"), their responses to related Press Ganey questions varied widely: 47% to 57% of Asian subgroups versus 67% of NHWs rated wait time as "Very good." The extent of racial/ethnic differences in patient-reported experiences varies based on how questions are framed. The observed poorer experiences by Asians are in part explained by their worse rating of similar objectively measurable experiences.

Promoting Policy and Environmental Change in Faith-Based Organizations: Outcome Evaluation of a Mini-Grants Program.

High rates of heart disease, cancer, and stroke exist in rural South Georgia where the Emory Prevention Research Center's Cancer Prevention and Control Research Network provided mini-grant funding to six churches to implement policy and environmental change to promote healthy eating and physical activity. This study sought to determine whether perceptions of the health promotion environment changed over time and whether perceived environmental change was associated with healthy behavior at church and in general. This study used a single-group pre-post design with 1-year follow-up. Parishioners (N = 258) completed self-administered questionnaires assessing perceptions of the church health promotion environment relative to healthy eating and physical activity, eating behavior and intention to use physical activity facilities at church, and eating and physical activity behaviors generally. Results indicate that perceived improvements in church nutrition environments were most strongly associated with decreases in unhealthy food consumed and stronger intentions to use physical activity resources at church (ps ≤ .05). Perceived changes in the physical activity environment were unrelated to church or general behavior. Findings suggest that church environments may play an important role in supporting healthy eating and physical activity at church; however, whether the influence of the church environment extends to other settings is unknown.

Colorectal Cancer Control Program grantees' use of evidence-based interventions.

BACKGROUND: Colorectal cancer (CRC) screening is recommended for adults aged 50-75 years, yet screening rates are low, especially among the uninsured. The CDC initiated the Colorectal Cancer Control Program (CRCCP) in 2009 with the goal of increasing CRC screening rates to 80% by 2014. A total of 29 grantees (states and tribal organizations) receive CRCCP funding to (1) screen uninsured adults and (2) promote CRC screening at the population level. PURPOSE: CRCCP encourages grantees to use one or more of five evidence-based interventions (EBIs) recommended by the Guide to Community Preventive Services. The purpose of the study was to evaluate grantees' EBI use. METHODS: A web-based survey was conducted in 2011 measuring grantees' use of CRC screening EBIs and identifying their implementation partners. Data were analyzed in 2012. RESULTS: Twenty-eight grantees (97%) completed the survey. Most respondents (96%) used small media. Fewer used client reminders (75%); reduction of structural barriers (50%); provider reminders (32%); or provider assessment and feedback (50%). Provider-oriented EBIs were rated as harder to implement than client-oriented EBIs. Grantees partnered with several types of organizations to implement EBIs, many with county- or state-wide reach. CONCLUSIONS: Almost all grantees implement EBIs to promote CRC screening, but the EBIs that may have the greatest impact with CRC screening are implemented by fewer grantees in the first 2 years of the CRCCP.

FM-GA and CM-GA for gene microarray analysis.

In this paper, we propose two new approaches, FM-GA and CM-GA, to identify significant genes from microarray datasets. FM-GA and CM-GA combine our innovative FM-test and CM-test with genetic algorithm (GA), respectively, and leverage the strengths of GA. The performance of FM-GA and CM-GA was evaluated by the classification accuracy of decision trees constructed with the selected genes. Experiments were conducted to demonstrate the superiority of the proposed method over other approaches.

Cationic lipid/DNA complexes (JVRS-100) combined with influenza vaccine (Fluzone) increases antibody response, cellular immunity, and antigenically drifted protection.

Safe and effective adjuvants for influenza vaccines that could increase both the levels of neutralizing antibody, including against drifted viral subtypes, and T-cell immunity would be a major advance in vaccine design. The JVRS-100 adjuvant, consisting of DOTIM/cholesterol cationic liposome-DNA complexes, is particularly promising for vaccines that require induction of high levels of antibody and T-cell immunity, including CD8(+) cytotoxic T lymphocytes (CTL). Inclusion of protein antigens with JVRS-100 results in the induction of enhanced humoral and cell-mediated (i.e., CD4(+) and CD8(+) T cells) immune responses. The JVRS-100 adjuvant combined with a split trivalent influenza vaccine (Fluzone-sanofi pasteur) elicited increased antibody and T-cell responses in mice and non-human primates compared to vaccination with Fluzone alone. Mice vaccinated with JVRS-100-Fluzone and challenged with antigenically drifted strains of H1N1 (PR/8/34) and influenza B (B/Lee/40) viruses had higher grade protection, as measured by attenuation of weight loss and increased survival, compared to recipients of unadjuvanted vaccine. The results indicate that the JVRS-100 adjuvant substantially increases immunogenicity and protection from drifted-strain challenge using an existing influenza vaccine.

A novel vaccine adjuvant for recombinant flu antigens.

Many new vaccines under development consist of rationally designed recombinant proteins that are relatively poor immunogens unless combined with potent adjuvants. There is only one adjuvant in common use in the U.S., aluminum phosphate or hydroxide (e.g. alum). This adjuvant, however, has significant limitations, particularly regarding the generation of strong cell-mediated (T-cell) immune responses. A novel adjuvant, JVRS-100, composed of cationic liposome-DNA complexes (CLDC) has been evaluated for immune enhancing activity. The JVRS-100 adjuvant has been shown to elicit robust immune responses compared to CpG oligonucleotides, alum, and MPL adjuvants, and efficiently enhances both humoral and cellular immune responses. Safety has been evaluated in preclinical studies, and the adjuvant is now in early-stage clinical development. One application of this novel adjuvant is to augment the immune responses to recombinant subunit antigens, which are often poorly immunogenic. The JVRS-100 adjuvant, when combined with a recombinant influenza hemagglutinin (H1), elicited increased specific antibody and T-cell responses in mice. Single-dose vaccination and prime/boost vaccinations with JVRS-100-H1 were both shown to be protective (i.e., survival, reduced weight loss) following H1N1 (PR/8/34) virus challenge. Enhanced immunological responses could be critically important for improved efficacy and dose-sparing of a recombinant influenza vaccine.

MCM-test: a fuzzy-set-theory-based approach to differential analysis of gene pathways.

BACKGROUND: Gene pathway can be defined as a group of genes that interact with each other to perform some biological processes. Along with the efforts to identify the individual genes that play vital roles in a particular disease, there is a growing interest in identifying the roles of gene pathways in such diseases. RESULTS: This paper proposes an innovative fuzzy-set-theory-based approach, Multi-dimensional Cluster Misclassification test (MCM-test), to measure the significance of gene pathways in a particular disease. Experiments have been conducted on both synthetic data and real world data. Results on published diabetes gene expression dataset and a list of predefined pathways from KEGG identified OXPHOS pathway involved in oxidative phosphorylation in mitochondria and other mitochondrial related pathways to be deregulated in diabetes patients. Our results support the previously supported notion that mitochondrial dysfunction is an important event in insulin resistance and type-2 diabetes. CONCLUSION: Our experiments results suggest that MCM-test can be successfully used in pathway level differential analysis of gene expression datasets. This approach also provides a new solution to the general problem of measuring the difference between two groups of data, which is one of the most essential problems in most areas of research.

Effect of beef and soy proteins on the absorption of non-heme iron and inorganic zinc in children.

BACKGROUND: Iron and zinc deficiency remain substantial problems in small children in both developed and developing nations. Optimizing mineral absorption is an important strategy in minimizing this problem. OBJECTIVES: To assess the effects of beef and soy proteins on the bioavailability of non-heme iron and zinc in children. METHODS: We measured iron (n = 26) and zinc (n = 36) absorption in 4-8 y old children from meals differing only in protein source (beef or a low-phytate soy protein concentrate). Iron and zinc absorption were measured using multi-tracer stable isotope techniques. Iron absorption was calculated from the red blood cell iron incorporation measured after 14 days and zinc absorption from the ratio of the oral and intravenous excretion of the zinc tracers 48 hours after dosing. RESULTS: Iron absorption from the beef meal was significantly greater (geometric mean, 7.6%) than from the soy meal (3.5%, p = 0.0015). Zinc absorption from the beef meal was greater (mean +/- SD, 13.7 +/- 6.0%) than from the soy meal (10.1 +/- 4.1%, p = 0.047). CONCLUSION: These findings indicate that beef protein increases both non-heme iron and zinc absorption compared to soy protein. The effect of protein source on non-heme iron and inorganic zinc absorption should be one of the factors taken into account when designing diets for children. The inhibitory effect of the soy based meal on iron and zinc absorption could be overcome by fortifying the soy protein with these minerals during the production process.

FM-test: a fuzzy-set-theory-based approach to differential gene expression data analysis.

BACKGROUND: Microarray techniques have revolutionized genomic research by making it possible to monitor the expression of thousands of genes in parallel. As the amount of microarray data being produced is increasing at an exponential rate, there is a great demand for efficient and effective expression data analysis tools. Comparison of gene expression profiles of patients against those of normal counterpart people will enhance our understanding of a disease and identify leads for therapeutic intervention. RESULTS: In this paper, we propose an innovative approach, fuzzy membership test (FM-test), based on fuzzy set theory to identify disease associated genes from microarray gene expression profiles. A new concept of FM d-value is defined to quantify the divergence of two sets of values. We further analyze the asymptotic property of FM-test, and then establish the relationship between FM d-value and p-value. We applied FM-test to a diabetes expression dataset and a lung cancer expression dataset, respectively. Within the 10 significant genes identified in diabetes dataset, six of them have been confirmed to be associated with diabetes in the literature and one has been suggested by other researchers. Within the 10 significantly overexpressed genes identified in lung cancer data, most (eight) of them have been confirmed by the literatures which are related to the lung cancer. CONCLUSION: Our experiments on synthetic datasets show that FM-test is effective and robust. The results in diabetes and lung cancer datasets validated the effectiveness of FM-test. FM-test is implemented as a Web-based application and is available for free at http://database.cs.wayne.edu/bioinformatics.

A combination of prebiotic short- and long-chain inulin-type fructans enhances calcium absorption and bone mineralization in young adolescents.

BACKGROUND: Short-term studies in adolescents have generally shown an enhancement of calcium absorption by inulin-type fructans (prebiotics). Results have been inconsistent; however, and no studies have been conducted to determine whether this effect persists with long-term use. OBJECTIVE: The objective was to assess the effects on calcium absorption and bone mineral accretion after 8 wk and 1 y of supplementation with an inulin-type fructan. DESIGN: Pubertal adolescents were randomly assigned to receive 8 g/d of a mixed short and long degree of polymerization inulin-type fructan product (fructan group) or maltodextrin placebo (control group). Bone mineral content and bone mineral density were measured before randomization and after 1 y. Calcium absorption was measured with the use of stable isotopes at baseline and 8 wk and 1 y after supplementation. Polymorphisms of the Fok1 vitamin D receptor gene were determined. RESULTS: Calcium absorption was significantly greater in the fructan group than in the control group at 8 wk (difference: 8.5 +/- 1.6%; P < 0.001) and at 1 y (difference: 5.9 +/- 2.8%; P = 0.04). An interaction with Fok1 genotype was present such that subjects with an ff genotype had the least initial response to fructan. After 1 y, the fructan group had a greater increment in both whole-body bone mineral content (difference: 35 +/- 16 g; P = 0.03) and whole-body bone mineral density (difference: 0.015 +/- 0.004 g/cm(2); P = 0.01) than did the control group. CONCLUSION: Daily consumption of a combination of prebiotic short- and long-chain inulin-type fructans significantly increases calcium absorption and enhances bone mineralization during pubertal growth. Effects of dietary factors on calcium absorption may be modulated by genetic factors, including specific vitamin D receptor gene polymorphisms.

Height and height Z-score are related to calcium absorption in five- to fifteen-year-old girls.

CONTEXT: Understanding the relationship between calcium absorption and growth has been limited. We have developed a database of calcium absorption measurements in 315 girls aged 5.0-15.0 yr. DESIGN: We have used this database to assess the relationship between height, its age- and gender-normalized value (height Z-score), and calcium absorptive efficiency. RESULTS: Overall, height was significantly related to calcium absorption (corrected for calcium intake, age, Tanner, stage, and ethnicity) (P = 0.001). Similarly, height Z-score was significantly related to calcium absorption (P < 0.007). About 3-3.5% of the variability (eta2) of absorption was associated with height or height Z-score. We found that calcium absorption was significantly lower in girls with height Z-score equal to or less than zero compared with those with a height Z-score more than zero (difference, 3.9 +/- 1.4%, mean +/- sem; P = 0.007). Limiting the analysis to those girls in which Tanner staging was performed or those age 9 yr or older did not substantively affect these relationships. CONCLUSION: These results indicate that a small but significant component of the variability in calcium absorption is due to height. Identifying genetic risk factors for lowered calcium absorption during growth could lead to individual approaches for prevention of inadequate bone mass.

Metabolic adaptations to low zinc intakes in premenarcheal girls.

BACKGROUND: Zinc deficiency is increasingly recognized as an important cause of mortality and morbidity. Children in developing countries are at especially high risk because of relatively low zinc intakes and poor bioavailability. OBJECTIVE: We assessed the effect of 2-wk adaptation to low zinc intake (4 mg/d) on fractional zinc absorption, endogenous fecal zinc excretion, and urinary zinc excretion. DESIGN: Sixteen healthy 9-14-y-old girls were studied twice in random order after 2-wk adaptation to diets providing either 12 mg/d (high) or 4 mg/d (low) zinc. Fractional zinc absorption and endogenous fecal zinc excretion were measured with use of established stable isotope techniques. RESULTS: Plasma zinc was not significantly lower during the low dietary intake period (1.06 +/- 0.18 mg/L) than during the high dietary intake period (1.14 +/- 0.23 mg/L, P = 0.30). Endogenous fecal zinc excretion was significantly lower during the low intake period (1.08 +/- 0.62 mg/d) than during the high intake period (1.82 +/- 0.95 mg/d, P < 0.026), but there was no significant change in fractional zinc absorption (30.6% +/- 12.4% compared with 26.6% +/- 9.0%, P = 0.32) or urinary zinc excretion (0.68 +/- 0.35 mg/d compared with 0.59 +/- 0.24 mg/d, P = 0.30). Approximate zinc balance was significantly lower during the low-intake period than during the high-intake period (P = 0.007) and significantly (P < 0.0001) less than zero. CONCLUSION: Short-term zinc restriction in premenarcheal girls leads to a significant decrease in endogenous fecal zinc excretion, which was inadequate to restore normal zinc balance.