Variable Intrinsic Expression of Immunoregulatory Biomarkers in Breast Cancer Cell Lines, Mammospheres, and Co-Cultures.

Advances in immunotherapy have increased interest in knowing the role of the immune system in breast cancer (BC) pathogenesis. Therefore, immune checkpoints (IC) and other pathways related to immune regulation, such as JAK2 and FoXO1, have emerged as potential targets for BC treatment. However, their intrinsic gene expression in vitro has not been extensively studied in this neoplasia. Thus, we evaluated the mRNA expression of tumor-cell-intrinsic CTLA-4, PDCD1 (PD1), CD274 (PD-L1), PDCD1LG2 (PD-L2), CD276 (B7-H3), JAK2, and FoXO1 in different BC cell lines, derived mammospheres, and co-cultures with peripheral blood mononuclear cells (PBMCs) by real-time quantitative polymerase chain reaction (qRT-PCR). Our results showed that intrinsic CTLA-4, CD274 (PD-L1), and PDCD1LG2 (PD-L2) were highly expressed in triple-negative cell lines, while CD276 was predominantly overexpressed in luminal cell lines. In contrast, JAK2 and FoXO1 were under-expressed. Moreover, high levels of CTLA-4, PDCD1 (PD1), CD274 (PD-L1), PDCD1LG2 (PD-L2), and JAK2 were found after mammosphere formation. Finally, the interaction between BC cell lines and peripheral blood mononuclear cells (PBMCs) stimulates the intrinsic expression of CTLA-4, PCDC1 (PD1), CD274 (PD-L1), and PDCD1LG2 (PD-L2). In conclusion, the intrinsic expression of immunoregulatory genes seems very dynamic, depending on BC phenotype, culture conditions, and tumor-immune cell interactions.

ID1 and ID4 Are Biomarkers of Tumor Aggressiveness and Poor Outcome in Immunophenotypes of Breast Cancer.

Inhibitor of differentiation (ID) proteins are a family of transcription factors that contribute to maintaining proliferation during embryogenesis as they avoid cell differentiation. Afterward, their expression is mainly silenced, but their reactivation and contribution to tumor development have been suggested. In breast cancer (BC), the overexpression of ID1 has been previously described. However, whether the remaining ID genes have a specific role in this neoplasia is still unclear. We studied the mRNA expression of all ID genes by q RT-PCR in BC cell lines and 307 breast carcinomas, including all BC subtypes. Our results showed that ID genes are highly expressed in all cell lines tested. However, ID4 presented higher expression in BC cell lines compared to a healthy breast epithelium cell line. In accordance, ID1 and ID4 were predominantly overexpressed in Triple-Negative and HER2-enriched samples. Moreover, high levels of both genes were associated with larger tumor size, histological grade 3, necrosis and vascular invasion, and poorer patients' outcomes. In conclusion, ID1 and ID4 may act as biomarkers of tumor aggressiveness and worse prognosis in breast cancer, and they could be used as potential targets for new treatments discover.

Free clinic oversight and outcomes in the United States: a scoping review protocol.

OBJECTIVE: The objective of this review is to explore existing literature related to free clinic oversight, and patient and health outcomes reported with the use of free clinics in the United States, to examine and conceptually map the evidence, and to identify any gaps. INTRODUCTION: Free clinics serve a unique role in American health care; however, little is known about clinic oversight and the outcomes seen in uninsured and under-insured patients who are treated at free health clinics. INCLUSION CRITERIA: The review will consider studies that include hospital or privately owned and operated community free clinics, and adults and children who utilize free clinics in the United States. In addition to information about clinic oversight, patient outcomes to be considered include disease course management metrics and patient satisfaction. Health outcomes to be considered include hospitalization rates and emergency room visit rates. METHODS: This scoping review will consider both experimental and quasi-experimental study designs in addition to analytical observational studies. Qualitative studies, systematic reviews that meet the inclusion criteria, and text and opinion papers will also be included. The search will be limited to those studies published in English since 1967, the year the first free clinic opened in the United States. The proposed review will be conducted in accordance with the JBI methodology for scoping reviews. Data will be extracted from papers by two independent reviewers using an extraction tool developed by the reviewers.

Achieving balance between implementing effective infection prevention and control practices and maintaining a home-like setting in U.S. Department of Veterans Affairs nursing homes.

Nursing homes present a unique challenge for implementing infection prevention and control practices while striving to maintain a home-like environment. Medical devices such as urinary catheters and central venous catheters have become a part of nursing home care but can predispose residents to associated infections. Because evidence-based prevention bundles were implemented, catheter-associated urinary tract infections (CAUTIs) and central line-associated bloodstream infections (CLABSIs) were monitored in all U.S. Department of Veterans Affairs (VA) nursing homes, and outcomes were evaluated. Bundle components for CLABSIs focused on insertion technique, site selection, and routine assessment of central line necessity, while the CAUTI bundle focused on insertion technique, appropriate indication, and routine assessment of urinary catheter necessity. From October 2010 through September 2016, VA nursing homes reported nationwide reductions of CAUTIs (51.2%; P < .0001) and CLABSIs (25.0%; P = .0009).

Assessing a National Collaborative Program To Prevent Catheter-Associated Urinary Tract Infection in a Veterans Health Administration Nursing Home Cohort.

OBJECTIVECollaborative programs have helped reduce catheter-associated urinary tract infection (CAUTI) rates in community-based nursing homes. We assessed whether collaborative participation produced similar benefits among Veterans Health Administration (VHA) nursing homes, which are part of an integrated system.SETTINGThis study included 63 VHA nursing homes enrolled in the "AHRQ Safety Program for Long-Term Care," which focused on practices to reduce CAUTI.METHODSChanges in CAUTI rates, catheter utilization, and urine culture orders were assessed from June 2015 through May 2016. Multilevel mixed-effects negative binomial regression was used to derive incidence rate ratios (IRRs) representing changes over the 12-month program period.RESULTSThere was no significant change in CAUTI among VHA sites, with a CAUTI rate of 2.26 per 1,000 catheter days at month 1 and a rate of 3.19 at month 12 (incidence rate ratio [IRR], 0.99; 95% confidence interval [CI], 0.67-1.44). Results were similar for catheter utilization rates, which were 11.02% at month 1 and 11.30% at month 12 (IRR, 1.02; 95% CI, 0.95-1.09). The numbers of urine cultures per 1,000 residents were 5.27 in month 1 and 5.31 in month 12 (IRR, 0.93; 95% CI, 0.82-1.05).CONCLUSIONSNo changes in CAUTI rates, catheter use, or urine culture orders were found during the program period. One potential reason was the relatively low baseline CAUTI rate, as compared with a cohort of community-based nursing homes. This low baseline rate is likely related to the VHA's prior CAUTI prevention efforts. While broad-scale collaborative approaches may be effective in some settings, targeting higher-prevalence safety issues may be warranted at sites already engaged in extensive infection prevention efforts.Infect Control Hosp Epidemiol 2018;820-825.

Phytochemicals and cognitive/behavioral function in patients with Alzheimer's disease: a scoping review protocol.

REVIEW QUESTION/OBJECTIVE: The objective of this review is to explore the existing literature related to phytochemicals and cognitive/behavioral function in patients with Alzheimer's disease, to examine and conceptually map the evidence, and to identify any gaps.The question of this review is: what is the evidence on phytochemicals and cognitive/behavioral function in patients with Alzheimer's disease?

Effectiveness of droxidopa compared to midodrine in standing blood pressure and orthostatic tolerance in adults with neurogenic orthostatic hypotension: a systematic review protocol.

REVIEW QUESTION: The question of this review is: what is the effectiveness of droxidopa compared to midodrine on standing blood pressure and orthostatic intolerance symptoms in adults with neurogenic orthostatic hypotension?

Fat4-Dchs1 signalling controls cell proliferation in developing vertebrae.

The protocadherins Fat4 and Dchs1 act as a receptor-ligand pair to regulate many developmental processes in mice and humans, including development of the vertebrae. Based on conservation of function between Drosophila and mammals, Fat4-Dchs1 signalling has been proposed to regulate planar cell polarity (PCP) and activity of the Hippo effectors Yap and Taz, which regulate cell proliferation, survival and differentiation. There is strong evidence for Fat regulation of PCP in mammals but the link with the Hippo pathway is unclear. In Fat4(-/-) and Dchs1(-/-) mice, many vertebrae are split along the midline and fused across the anterior-posterior axis, suggesting that these defects might arise due to altered cell polarity and/or changes in cell proliferation/differentiation. We show that the somite and sclerotome are specified appropriately, the transcriptional network that drives early chondrogenesis is intact, and that cell polarity within the sclerotome is unperturbed. We find that the key defect in Fat4 and Dchs1 mutant mice is decreased proliferation in the early sclerotome. This results in fewer chondrogenic cells within the developing vertebral body, which fail to condense appropriately along the midline. Analysis of Fat4;Yap and Fat4;Taz double mutants, and expression of their transcriptional target Ctgf, indicates that Fat4-Dchs1 regulates vertebral development independently of Yap and Taz. Thus, we have identified a new pathway crucial for the development of the vertebrae and our data indicate that novel mechanisms of Fat4-Dchs1 signalling have evolved to control cell proliferation within the developing vertebrae.

Dchs1-Fat4 regulation of polarized cell behaviours during skeletal morphogenesis.

Skeletal shape varies widely across species as adaptation to specialized modes of feeding and locomotion, but how skeletal shape is established is unknown. An example of extreme diversity in the shape of a skeletal structure can be seen in the sternum, which varies considerably across species. Here we show that the Dchs1-Fat4 planar cell polarity pathway controls cell orientation in the early skeletal condensation to define the shape and relative dimensions of the mouse sternum. These changes fit a model of cell intercalation along differential Dchs1-Fat4 activity that drives a simultaneous narrowing, thickening and elongation of the sternum. Our results identify the regulation of cellular polarity within the early pre-chondrogenic mesenchyme, when skeletal shape is established, and provide the first demonstration that Fat4 and Dchs1 establish polarized cell behaviour intrinsically within the mesenchyme. Our data also reveal the first indication that cell intercalation processes occur during ventral body wall elongation and closure.

Lipid analysis of developing Camelina sativa seeds and cultured embryos.

Camelina sativa is a cultivated oilseed rich in triacylglycerols containing oleic, linoleic, α-linolenic and eicosenoic acids. As it holds promise as a model species, its lipid synthesis was characterized in vivo and in culture. Lipid accumulates at a maximum rate of about 26 µg/day/seed (11.5 mg lipid/day/g fresh seed weight), a rate comparable with other oilseeds. Noteworthy is a late stage surge in α-linolenic acid accumulation. Small amounts of unusual epoxy and hydroxy fatty acids are also present in the triacylglycerols. These include 15,16-epoxy- and 15-hydroxy-octadecadienoic acids and homologous series of ω7-hydroxy-alk-ω9-enoic and ω9/10-hydroxy-alkanoic acids. Mid-maturation embryos cultured in vitro have growth and lipid deposition rates and fatty acid compositions that closely match that of seeds, but extended culture periods allow these rates to rise and surpass those observed in planta. Optimized thin layer chromatography systems for characterization of labeled products from acetate or glycerol labeling are described. Glycerol label is only found in acylglycerols, largely as the intact glyceryl backbone, but acetate can label acyl groups and sterols, the latter to a much higher relative specific activity. This presumably occurs because mevalonic acid precursor is derived from the non-plastid pool of acetyl-CoA that is also the source for malonyl-CoA to drive FAE1-dependent chain elongation. Particular attention has been paid to the separation of sterols and diacylglycerols, and to hydrogenation of triacylglycerols to simplify their analysis. These improved methods will allow more accurate analyses of the fluxes of lipid metabolism in cultured plant embryos.

Lipid labeling from acetate or glycerol in cultured embryos of Camelina sativa seeds: A tale of two substrates.

Studies on the metabolism of lipids in seeds frequently use radiolabeled acetate and glycerol supplied to excised developing seeds to track the biosynthesis of acyl and lipid head groups, respectively. Such experiments are generally restricted to shorter time periods and the results may not quantitatively reflect in planta rates. These limitations can be removed by using cultured embryos, provided they mimic growth and lipid deposition observed for embryos in planta. Mid-maturation embryos from Camelina sativa were cultured in vitro to assess the use of sufficient acetate or glycerol concentrations and labeling periods for stable isotope labeling and mass spectrometric detection. Maximum incorporation of exogenous acetate into fatty acids occurred at 1mM and above. This provides about 5% of the total carbon flux entering fatty acids, enough for (13)C isotopomer analysis while maintaining normal biosynthetic rates for over 24h. Labeling analysis indicates that acetate reports lipid metabolism uniformly across the embryo. At higher acetate concentrations with longer incubations, the rate of fatty acid synthesis is reduced and the composition of newly synthesized fatty acids changes. While the mole fractions of oleate that undergo Δ12-desaturation or elongation are independent of biosynthetic flux, Δ15-desaturation shows a bimodal dependence. These observations are consistent with changes occurring in planta over seed development. Incorporation rates of the glyceryl moiety into lipids saturates at about 0.5mM exogenous glycerol. At saturation, the exogenous glycerol almost completely replaces the endogenous supply of glycerol-3-phosphate without affecting net lipid accumulation or fatty acid composition. It is concluded that acetate and glycerol labeling of cultured C. sativa embryos can provide an accurate representation of lipid metabolism in embryos in vivo, and that in Camelina embryos glycerol-3-phosphate levels do not co-limit triacylglycerol synthesis.

Abnormal high-density lipoprotein induces endothelial dysfunction via activation of Toll-like receptor-2.

Endothelial injury and dysfunction (ED) represent a link between cardiovascular risk factors promoting hypertension and atherosclerosis, the leading cause of death in Western populations. High-density lipoprotein (HDL) is considered antiatherogenic and known to prevent ED. Using HDL from children and adults with chronic kidney dysfunction (HDL(CKD)), a population with high cardiovascular risk, we have demonstrated that HDL(CKD) in contrast to HDL(Healthy) promoted endothelial superoxide production, substantially reduced nitric oxide (NO) bioavailability, and subsequently increased arterial blood pressure (ABP). We have identified symmetric dimethylarginine (SDMA) in HDL(CKD) that causes transformation from physiological HDL into an abnormal lipoprotein inducing ED. Furthermore, we report that HDL(CKD) reduced endothelial NO availability via toll-like receptor-2 (TLR-2), leading to impaired endothelial repair, increased proinflammatory activation, and ABP. These data demonstrate how SDMA can modify the HDL particle to mimic a damage-associated molecular pattern that activates TLR-2 via a TLR-1- or TLR-6-coreceptor-independent pathway, linking abnormal HDL to innate immunity, ED, and hypertension.

Integrated community education model: breast health awareness to impact late-stage breast cancer.

Race and ethnicity play roles in breast cancer mortality, particularly for African American women. As a result, a three-pronged integrated community education model (i.e., faith-based, community, and state agencies) was generated and tested in a medically underserved area with high mortality rates from breast cancer to increase participation in breast health education, provide early screening and detection practices, and provide access to annual mammograms and referral sources. The model provided three women with life-saving early diagnoses, in addition to providing potentially hundreds of women with a network of breast health, self-monitoring, and referral sources for future issues.

Effects of student participation in hand hygiene monitoring on knowledge and perception of infection control practices.

Studies report that students in health care professions do not retain knowledge of infection control and demonstrate poor hand hygiene compliance. This study describes the effect of a multifaceted approach (education, skills training, and monitoring) on nursing students' knowledge of infection control principles, opinions, hand hygiene practices, and value of nursing research in evidence-based practice. Students participated in hand hygiene monitoring of health care workers with 900 observations. Students demonstrated strong knowledge of hand hygiene principles: 63% reported that hand hygiene monitoring positively influenced their own compliance. Although posters have been identified as effective prompts, students did not perceive poster reminders as effective in prompting handwashing. Students reported that hand hygiene activities helped them value the role of the nurse in research and evidence-based practice. This study may help educators clarify misconceptions that result in student noncompliance in hand hygiene practices.

Comprehensive compositional analysis of plant cell walls (Lignocellulosic biomass) part I: lignin.

The need for renewable, carbon neutral, and sustainable raw materials for industry and society has become one of the most pressing issues for the 21st century. This has rekindled interest in the use of plant products as industrial raw materials for the production of liquid fuels for transportation(1) and other products such as biocomposite materials(7). Plant biomass remains one of the greatest untapped reserves on the planet(4). It is mostly comprised of cell walls that are composed of energy rich polymers including cellulose, various hemicelluloses (matrix polysaccharides, and the polyphenol lignin(6) and thus sometimes termed lignocellulosics. However, plant cell walls have evolved to be recalcitrant to degradation as walls provide tensile strength to cells and the entire plants, ward off pathogens, and allow water to be transported throughout the plant; in the case of trees up to more the 100 m above ground level. Due to the various functions of walls, there is an immense structural diversity within the walls of different plant species and cell types within a single plant(4). Hence, depending of what crop species, crop variety, or plant tissue is used for a biorefinery, the processing steps for depolymerization by chemical/enzymatic processes and subsequent fermentation of the various sugars to liquid biofuels need to be adjusted and optimized. This fact underpins the need for a thorough characterization of plant biomass feedstocks. Here we describe a comprehensive analytical methodology that enables the determination of the composition of lignocellulosics and is amenable to a medium to high-throughput analysis. In this first part we focus on the analysis of the polyphenol lignin (Figure 1). The method starts of with preparing destarched cell wall material. The resulting lignocellulosics are then split up to determine its lignin content by acetylbromide solubilization(3), and its lignin composition in terms of its syringyl, guaiacyl- and p-hydroxyphenyl units(5). The protocol for analyzing the carbohydrates in lignocellulosic biomass including cellulose content and matrix polysaccharide composition is discussed in Part II(2).

Comprehensive compositional analysis of plant cell walls (lignocellulosic biomass) part II: carbohydrates.

The need for renewable, carbon neutral, and sustainable raw materials for industry and society has become one of the most pressing issues for the 21st century. This has rekindled interest in the use of plant products as industrial raw materials for the production of liquid fuels for transportation(2) and other products such as biocomposite materials(6). Plant biomass remains one of the greatest untapped reserves on the planet(4). It is mostly comprised of cell walls that are composed of energy rich polymers including cellulose, various hemicelluloses, and the polyphenol lignin(5) and thus sometimes termed lignocellulosics. However, plant cell walls have evolved to be recalcitrant to degradation as walls contribute extensively to the strength and structural integrity of the entire plant. Despite its necessary rigidity, the cell wall is a highly dynamic entity that is metabolically active and plays crucial roles in numerous cell activities such as plant growth and differentiation(5). Due to the various functions of walls, there is an immense structural diversity within the walls of different plant species and cell types within a single plant(4). Hence, depending of what crop species, crop variety, or plant tissue is used for a biorefinery, the processing steps for depolymerisation by chemical/enzymatic processes and subsequent fermentation of the various sugars to liquid biofuels need to be adjusted and optimized. This fact underpins the need for a thorough characterization of plant biomass feedstocks. Here we describe a comprehensive analytical methodology that enables the determination of the composition of lignocellulosics and is amenable to a medium to high-throughput analysis (Figure 1). The method starts of with preparing destarched cell wall material. The resulting lignocellulosics are then split up to determine its monosaccharide composition of the hemicelluloses and other matrix polysaccharides1, and its content of crystalline cellulose(7). The protocol for analyzing the lignin components in lignocellulosic biomass is discussed in Part I(3).

Preferences for teaching methods in a baccalaureate nursing program: how second-degree and traditional students differ.

Accelerated baccalaureate nursing programs are in great demand in the United States. Currently there are 197 such programs, but little research has been conducted on student characteristics and program outcomes. This quantitative study explores preferences of second-degree students and traditional generic students with regard to teaching methods and relationships with faculty. The results indicate that statistically significant differences exist between the two groups of students. Three areas of significance are ability for self-directed learning, expectations of faculty and classroom structure, and obtaining a grade that really matters.

Generational (age) differences in nursing students' preferences for teaching methods.

A generational age transformation is occurring in nursing classrooms across the United States. Nurse educators need to prepare for the different values and expectations of students from Generation X and the newly emerging Generation Y in the educational environment. This quantitative, descriptive research begins to examine the preferences and expectations of these generations regarding teaching methods.

The effect of conventional and sustained delivery of thymoquinone and levodopa on SH-SY5Y human neuroblastoma cells.

The objective of this study was to determine the effects of thymoquinone on the viability and metabolic activity of SH-SY5Y human neuroblastoma cells alone or challenged with levodopa (L-dopa) using conventional and sustained drug delivery routes. The findings suggest that thymoquinone may protect SH-SY5Y cells against L-dopa toxicity to some degree although the exact mechanism for this is unknown. Thymoquinone may be a viable option to prevent quinone formation as a result of L-dopa auto-oxidation and could be investigated as a neuroprotective agent given the fact that inflammatory etiologies are strongly implemented in the pathogenesis of Parkinson's disease.