Cholinergic cells in the nucleus basalis of mice express the N-methyl-D-aspartate-receptor subunit NR2C and its replacement by the NR2B subunit enhances frontal and amygdaloid acetylcholine levels

It is known that glutamatergic and cholinergic systems interact functionally at the level of the cholinergic basal forebrain. The N-methyl-D-aspartate receptor (NMDA-R) is a multiprotein complex composed of NR1, NR2 and/or NR3 subunits. The subunit composition of NMDA-R of cholinergic cells in the nucleus basalis has not yet been investigated. Here, by means of choline acetyl transferase and NR2B or NR2C double staining, we demonstrate that mice express both the NR2C and NR2B subunits in nucleus basalis cholinergic cells.We generated NR2C-2B mutant mice in which an insertion of NR2B cDNA into the gene locus of the NR2C gene replaced NR2C by NR2B expression throughout the brain. This NR2C-2B mutant was used to examine whether a subunit exchange in cholinergic neurons would affect acetylcholine (ACh) content in several brain structures. We found increased ACh levels in the frontal cortex and amygdala in the brains of NR2C-2B mutant mice. Brain ACh has been implicated in neuroplasticity, novelty-induced arousal and encoding of novel stimuli. We therefore assessed behavioral habituation to novel environments and objects as well as object recognition in NR2C-2B subunit exchange mice. The behavioral analysis did not indicate any gross behavioral alteration in the mutant mice compared with the wildtype mice. Our results show that the NR2C by NR2B subunit exchange in mice affects ACh content in two target areas of the nucleus basalis.

Medicaid Expenditures for Cancer: Evidence from Medicaid-only Beneficiaries in Four States

This study estimates the cost burden of 6 prevalent invasive cancers—breast, cervical, colorectal, lung, melanoma, and prostate—on Medicaid programs in 4 states. The analyses use Medicaid claims and enrollment data for all Medicaid-only beneficiaries over age 18 in Georgia, Illinois, Louisiana, and Maine with at least 1 month of enrollment in fee-for-service Medicaid from 2000 to 2003. We applied ordinary least squares regression analysis to a data set created from Medicaid claims and enrollment data to estimate annual expenditures attributable to each cancer after controlling for age, race, gender, and comorbid conditions. Cancers and comorbid conditions were identified on the basis of claims with an appropriate diagnosis code. Cancers include both incident and prevalent cases. In 2003 dollars, annualized Medicaid expenditures attributable to the 6 cancers combined in the Medicaid-only population were $84.0 million in Georgia,$79.7 million in Illinois, $51.4 million in Louisiana, and$29.4 million in Maine. Attributable annualized per-capita Medicaid expenditures were highest for lung cancer, then colorectal cancer. After adjusting for sociodemographics and comorbidities, only 10% to 50% of medical expenditures among Medicaid-only beneficiaries with cancer were attributable to cancer. Estimates of the costs of care for Medicaid-eligible cancer patients are critical to understanding the implications of cancer for state and federal budgets. The Patient Protection and Affordable Care Act (ACA) of 2010 is expected to substantially expand the adult Medicaid population. These estimates provide important baseline information for assessing the potential effects of increased Medicaid enrollment on Medicaid expenditures for cancer

Exploring the Pathogenic and Drug Resistance Mechanisms of Staphylococcus aureus

We have previously identified σS, an ECF sigma factor that is important in the virulence and stress response of S. aureus. Transcriptional profiling of sigS revealed that it is differentially regulated in a variety of laboratory and clinical strains of S. aureus, suggesting that there exists a regulatory network that modulates its expression. In order to identify direct regulators of sigS expression, we performed a biotin pull down assay in tandem with mass spectrometry. We identified CymR as a direct regulator and observed that sigS expression is increased in cells lacking cymR. In addition, transposon mutagenesis was performed to identify regulators of sigS expression. We identified insertions in genes that are transcriptional regulators, and elements involved in amino acid biosynthesis and DNA replication, recombination and repair as influencing sigS expression. Finally, methyl nitro-nitrosoguanidine mutagenesis in conjunction with whole genome sequencing was employed and revealed mutations in the lactose repressor, lacR, and the membrane sensor histidine kinase, kdpD, as negatively effecting sigS expression. EMSAs revealed that LacR is an indirect regulator of sigS expression, while the response regulator KdpE is a direct repressor. These results indicate that a complex regulatory network is in place for sigS that modulates its expression. In a continuation of studies on σS regulation, we next explored interplay with the products of genes conserved within the sigS locus. We determined that this region is conserved amongst all the sequenced staphylococci, and includes four genes: SAUSA300_1721 (a conserved hypothetical protein), as well as sigS, ecfX, and ecfY. In order to investigate the relationship between EcfX and σS we performed protein pull down assays and observed that these two protein interact. Further to this, transcriptional analysis of sigS in an ecfX mutant reveal that expression of sigS is decreased, indicating that it is an activator. Architectural analysis of the sigS locus via RNAseq revealed that the majority of transcription in this region comes from ecfY, a gene that is downstream and divergent to sigS. We demonstrate that inactivation of ecfY leads to a significant increase in sigS expression, and that ecfY null strains are more resistant to DNA damaging agents such as UV, H2O2, MMS, and ethidium bromide, which we have previously demonstrated that a sigS mutant is highly sensitive to. Our studies also revealed that an ecfY null strain is better able to survive intracellularly following phagocytosis by RAW 264.7 cell and demonstrates increased survival in whole-human blood, which is again opposed to that previously observed for sigS deficient strains. Because the ecfY null strain overexpresses sigS, we investigated the regulon of this sigma factor using this mutant in conjunction with RNAseq analysis. We identified that genes putatively under the control of σS are involved in DNA damage and repair, virulence, amino acid starvation and nucleic acid biosynthesis. Collectively, our results indicate that σS is regulated via a unique mechanism: positively through an apparent need for an activator protein (EcfX) and negatively via RNA-RNA interaction (the 3’ UTR of ecfY). We suggest that the evidence presented here greatly adds not only to our understanding of the regulatory circuits extant within S. aureus, but also to alternative sigma factor biology in general. Finally, we evaluated the efficacy of a novel library of quinazoline-based compounds against a highly drug resistant strain of S. aureus. We performed structure activity and structure property relationship assays in order to identify lead compounds. These methods lead to the identification of N2,N4-disubstituted quinazoline-2,4-diamines that had low minimum inhibitory concentrations, along with favorable physiochemical properties. Evaluation of their biological activity demonstrated limited potential for resistance of to our quinazoline based compounds, low toxicity to human epithelial cells, and strong efficacy in vivo. Taken together, our findings support the use of quinazoline derivatives as potential new antimicrobials against multidrug resistant S. aureus

Human Neutrophil Elastase Proteolytic Activity in Ulcerative Colitis Favors the Loss of Function of Therapeutic Monoclonal Antibodies

Purpose: Proteases play an essential role in the pathophysiology of inflammatory bowel disease (IBD), contributing to the intestinal mucosal lesions through the degradation of the extracellular matrix and alteration of the barrier function. Ulcerative colitis (UC) is characterized by an extensive infiltrate of neutrophils into the mucosa and hence, increased proteolytic activity. Human neutrophil elastase (HNE) is a serine protease that has been reported to be increased in UC patients’ intestinal mucosa. Based on our previous studies, we hypothesized that HNE might induce proteolytic degradation and loss of function of therapeutic monoclonal antibodies in IBD patients. Patients and Methods: Elastase expression and elastinolytic activity were determined in mucosal explants from ulcerative colitis patients (n=6) and cultured ex vivo in the presence or absence of recombinant elafin. Enzymatic digestions of therapeutic monoclonal antibodies were performed using recombinant HNE and elafin. The integrity of the therapeutic antibodies was evaluated by immunoblotting and protein G binding assay, whereas their TNFneutralizing activity was assessed with a reporter cell line. Results: We found that HNE and its elastinolytic activity were increased in the gut mucosa of UC patients. We also demonstrated that HNE cleaved biological drugs, impairing the TNF-α neutralizing capacity of anti-TNF monoclonal antibodies. This proteolytic degradation was inhibited by the addition of the specific inhibitor, elafin. Conclusion: Our results suggest that the high level of proteolytic degradation by mucosal neutrophil elastase, along with a potential imbalance with elafin, contributes to the loss of function of biologic agents, which are currently used in patients with IBD. These findings might explain the non-responsiveness of UC patients to therapeutic monoclonal antibodies and suggest the potential beneficial concomitant use of elafin in this treatment.Facultad de Ciencias ExactasInstituto de Estudios Inmunológicos y Fisiopatológico

Exploring the Simultaneous Effect of Total Ion Concentration and K:Ca:Mg Ratio of the Nutrient Solution on the Growth and Nutritional Value of Hydroponically Grown Cichorium spinosum L.

Nutrient-efficient plants and agricultural systems could tackle issues resulting from conventional agriculture. Spiny chicory (Cichorium spinosum L.), a very adaptive, wild edible vegetable, is gaining commercial interest as a functional food. Floating-raft hydroponics is a method commonly used for the commercial cultivation of leafy vegetables due to numerous advantages compared to soil cultivation. In this paper, the simultaneous effects of different potassium, calcium and magnesium ratios and different electrical conductivity (EC) levels on the growth and mineral composition of hydroponically grown C. spinosum were investigated. Four nutrient solutions (NS) were compared, two NS with low EC (L, 2.4 dS/m) and two with high EC (H, 3.6 dS/m) with K:Ca:Mg ratios of either 50:40:10 or 40:50:10. The results showed no interactions between the two factors. No significant effects were observed on the fresh and dry weight, leaf number and leaf area. High EC levels increased the K content and decreased the Mn and Zn content in the leaf tissues. The 40:50:10 ratio led to increased Ca content in plant tissues. The Nitrate-N was only affected by the EC level and was increased under H conditions, whereas the total-N was not affected

Air pollution abatement performances of green infrastructure in open road and built-up street canyon environments – A review

Intensifying the proportion of urban green infrastructure has been considered as one of the remedies for air pollution levels in cities, yet the impact of numerous vegetation types deployed in different built environments has to be fully synthesised and quantified. This review examined published literature on neighbourhood air quality modifications by green interventions. Studies were evaluated that discussed personal exposure to local sources of air pollution under the presence of vegetation in open road and built-up street canyon environments. Further, we critically evaluated the available literature to provide a better understanding of the interactions between vegetation and surrounding built-up environments and ascertain means of reducing local air pollution exposure using green infrastructure. The net effects of vegetation in each built-up environment are also summarised and possible recommendations for the future design of green infrastructure are proposed. In a street canyon environment, high-level vegetation canopies (trees) led to a deterioration in air quality, while low-level green infrastructure (hedges) improved air quality conditions. For open road conditions, wide, low porosity and tall vegetation leads to downwind pollutant reductions while gaps and high porosity vegetation could lead to no improvement or even deteriorated air quality. The review considers that generic recommendations can be provided for vegetation barriers in open road conditions. Green walls and roofs on building envelopes can also be used as effective air pollution abatement measures. The critical evaluation of the fundamental concepts and the amalgamation of key technical features of past studies by this review could assist urban planners to design and implement green infrastructures in the built environment

05/18/1993 - Eastern\u27s 1993 All Student Show Winner Laura Brahos.pdf

This paper reports on the stages forming a model evaluation protocol for urban flow and dispersion models proposed within the COST Action 732 on "Quality Assurance and Improvement of Micro-Scale Meteorological Models". It discusses the different components forming model evaluation with emphasis on validation and implementation of the protocol for the test case Mock Urban Setting Test (MUST). The protocol was proposed with building-resolving models in mind, but integral models have also been included. The suggested approach can be used for further micro-scale model evaluation and for the standardisation of their applications

Protein-energy wasting and nutritional supplementation in patients with end-stage renal disease on hemodialysis

© 2016 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism Background & aims Protein-Energy Wasting (PEW) is the depletion of protein/energy stores observed in the most advanced stages of Chronic Kidney Disease (CKD). PEW is highly prevalent among patients on chronic dialysis, and is associated with adverse clinical outcomes, high morbidity/mortality rates and increased healthcare costs. This narrative review was aimed at exploring the pathophysiology of PEW in end-stage renal disease (ESRD) on hemodialysis. The main aspects of nutritional status evaluation, intervention and monitoring in this clinical setting were described, as well as the current approaches for the prevention and treatment of ESRD-related PEW. Methods An exhaustive literature search was performed, in order to identify the relevant studies describing the epidemiology, pathogenesis, nutritional intervention and outcome of PEW in ESRD on hemodialysis. Results and conclusion The pathogenesis of PEW is multifactorial. Loss of appetite, reduced intake of nutrients and altered lean body mass anabolism/catabolism play a key role. Nutritional approach to PEW should be based on a careful and periodic assessment of nutritional status and on timely dietary counseling. When protein and energy intakes are reduced, nutritional supplementation by means of specific oral formulations administered during the hemodialysis session may be the first-step intervention, and represents a valid nutritional approach to PEW prevention and treatment since it is easy, effective and safe. Omega-3 fatty acids and fibers, now included in commercially available preparations for renal patients, could lend relevant added value to macronutrient supplementation. When oral supplementation fails, intradialytic parenteral nutrition can be implemented in selected patients