Aerosol delivery of synthetic DNA containing CpG motifs in broiler chicks at hatch under field conditions using a commercial-scale prototype nebulizer provided protection against lethal Escherichia coli septicemia.

Synthetic DNA containing CpG motifs (CpG-ODN) are potent innate immune stimulators in neonatal and adult broiler chickens against bacterial septicemia. We have recently demonstrated that intrapulmonary (IPL) delivery of CpG-ODN as microdroplets under laboratory conditions can protect neonatal chickens against lethal Escherichia coli septicemia. The objectives of this study were to develop a commercial-scale poultry nebulizer (CSPN) that can deliver CpG-ODN as microdroplets in neonatal broiler chicks in the hatcheries and study the efficacy of CSPN in inducing immune-protective effects under different environmental conditions in 2 geographical locations in Canada. Three field experiments were conducted in commercial poultry hatcheries during different seasons of the year in Saskatchewan and British Columbia, Canada. Neonatal broiler chicks (n = 8,000/experiment) received CpG-ODN by the IPL route in the CSPN chamber for 30 min, and control chicks received distilled water (DW) for 30 min. Broiler chicks (CpG-ODN-240 chicks/experiment and DW-40 chicks/experiment) were randomly sampled from all locations of the CSPN after nebulization and challenged with a lethal dose of E. coli to examine the CpG-ODN nebulization induced protection. We found a significant level (P < 0.05) of protection in broiler chicks against E. coli challenge, suggesting that the newly built CSPN successfully delivered CpG-ODN via the IPL route. We found that when the CSPN was maintained at humidex 28°C or below and relative humidity (RH) between 40 and 60%, neonatal birds were significantly (P < 0.05) protected against E. coli septicemia after IPL delivery of CpG-ODN. By contrast, protection in chicks was adversely affected when the CSPN was maintained at the humidex of 29°C or higher and RH of 70%. Overall, the present study successfully built a CSPN for CpG-ODN delivery in chicks at the hatchery and revealed that the temperature, humidity, and humidex were critical parameters in CSPN for efficient delivery of CpG-ODN.

Teriflunomide added to interferon-ß in relapsing multiple sclerosis: a randomized phase II trial.

OBJECTIVE: To evaluate teriflunomide as add-on therapy to ongoing stable-dosed interferon-ß (IFNß) in patients with relapsing forms of multiple sclerosis (RMS). METHODS: A total of 118 patients with RMS were randomly assigned 1:1:1 to receive oral placebo or teriflunomide, 7 or 14 mg, once daily for 24 weeks; 86 patients entered the 24-week extension. The primary objective was to evaluate safety; secondary objectives were to evaluate the effects of treatment on disease activity assessed by MRI and relapse rate. RESULTS: Teriflunomide was well tolerated with a low and similar incidence of treatment-emergent adverse events (TEAEs) across the 3 groups; TEAEs led to treatment discontinuation of 4.9%, 8.1%, and 7.9% of patients in the placebo, 7-mg, and 14-mg groups, respectively. The number of gadolinium-enhancing T1 (T1-Gd) lesions was reduced in both teriflunomide groups, with relative risk reductions (RRRs) of 84.6% (p = 0.0005) and 82.8% (p < 0.0001) for 7 and 14 mg, respectively, compared with IFNß alone at 48 weeks. T1-Gd lesion volume was also reduced in the 7-mg group (RRR 72.1%, p = 0.1104) and 14-mg group (RRR 70.6%, p = 0.0154). A trend toward dose-dependent reduction in annualized relapse rate was also noted (RRRs 32.6% [p = 0.4355] and 57.9% [p = 0.1005] for 7 and 14 mg, respectively). CONCLUSION: Teriflunomide as add-on therapy to IFNß had acceptable safety and tolerability and reduced MRI disease activity compared with IFNß alone. CLASSIFICATION OF EVIDENCE: This study provides Class II evidence that teriflunomide, 7 and 14 mg, added to IFNß, is safe. The T1-Gd lesion burden was significantly reduced with both teriflunomide doses.

The effect of structures on indoor humidity--possibility to improve comfort and perceived air quality.

The research presented in this paper shows that moisture transfer between indoor air and hygroscopic building structures can generally improve indoor humidity conditions. This is important because the literature shows that indoor humidity has a significant effect on occupant comfort, perceived air quality (PAQ), occupant health, building durability, material emissions, and energy consumption. Therefore, it appears possible to improve the quality of life of occupants when appropriately applying hygroscopic wood-based materials. The paper concentrates on the numerical investigation of a bedroom in a wooden building located in four European countries (Finland, Belgium, Germany, and Italy). The results show that moisture transfer between indoor air and the hygroscopic structure significantly reduces the peak indoor humidity. Based on correlations from the literature, which quantify the effect of temperature and humidity on comfort and PAQ for sedentary adults, hygroscopic structures can improve indoor comfort and air quality. In all the investigated climates, it is possible to improve the indoor conditions such that, as many as 10 more people of 100 are satisfied with the thermal comfort conditions (warm respiratory comfort) at the end of occupation. Similarly, the percent dissatisfied with PAQ can be 25% lower in the morning when permeable and hygroscopic structures are applied.

Processing of PDGF gene products determines interactions with glycosaminoglycans.

The platelet derived growth factor (PDGF), a mitogen for mesenchymal cells, may be bound to and inhibited by heparin and other glycosaminoglycans. PDGF is a homo- or heterodimer of A- and B-chains. They occur as short (A109 and B110) and long (A125 and B160) isoforms. The latter contain basic carboxyl-terminal extensions. Dimeric A125 binds to heparin through its basic extension in a two-step reaction. The mechanism involves a conformational change and is consistent with a Monod-Wyman-Changeux allosteric model. Previous indirect experiments suggested that three critical amino acids (basic R111, K116 and polar T125) might be involved. Here, direct binding experiments using dimeric full-length mutants in surface plasmon resonanse analysis showed that all three critical amino acids in an R(X)4K(X)8T-motif contributed in a concerted manner to the high affinity binding. Mutations of these amino acids to alanine resulted in large thermodynamic changes, loss of the allosteric mechanism and order(s) of magnitude lower binding affinity. The binding mechanism and affinity of long dimeric rB were similar to the mutants. Short dimeric rA109 and rB110 showed 100 times lower binding affinity than rA125. Consequently, interactions with glycosaminoglycans in tissues varies between PDGF isoforms and may influence their local accumulation and activity.

Effect of phenotype on the transcription of the genes for platelet-derived growth factor (PDGF) isoforms in human smooth muscle cells, monocyte-derived macrophages, and endothelial cells in vitro.

Proliferation of arterial smooth muscle cells (ASMCs) contributes considerably to enlargement of the arterial wall during atherosclerosis. The platelet-derived growth factor (PDGF) is a well-known mitogen and chemoattractant for ASMCs. Quantitative reverse transcription-polymerase chain reaction showed that cells appearing in atherosclerotic lesions, such as ASMCs, endothelial cells, and monocytes/macrophages, expressed mRNAs for both PDGF A and B chains in vitro, with the highest expression in endothelial cells. On proliferation, ASMCs and endothelial cells upregulated PDGF A mRNA. Differentiation of macrophages increased the amount of both mRNAs. Thus, the regulation of PDGF A- and B-chain expression depends on cell types and phenotypic states of the cells, which have also been found in vivo in human atherosclerotic lesions. PDGF A can be produced as short and long isoforms. The latter binds with high affinity to glycosaminoglycans. Irrespective of phenotype, only the minor part of total PDGF A mRNA consisted of the long variant in ASMCs, while endothelial cells produced 40% of total PDGF A as the long form. The differentiation of macrophages increased the production of the long PDGF A mRNA from 10% to 40%. Thus, increasing numbers of stimulated cells in the atherosclerotic lesion may increase the transcription of PDGF isoforms, and particularly of the long PDGF A isoform. Together with increasing amounts of ASMC-derived proteoglycans in developing lesions, this may contribute to accumulation of PDGF in the arterial wall matrix, resulting in prolonged stimulation of ASMCs.

Teaching caring to nursing students.

This article reports on phenomenological research designed to discover how caring was taught in a nursing education program. The basic questions were: 1) What is the meaning of caring to the faculty and students; 2) How do the faculty communicate this meaning to the students; and 3) How does this meaning shape the experience of the students? Data were collected from a small associate degree nursing program using: a) semi-structured interviews with all faculty and a selected group of students, b) classroom observations, and c) review of documents. Data were analyzed for and found to have content explaining the meaning of caring, how caring was being taught, and what students were learning about caring as the essence of nursing. Implications derived speak to the need for faculty and administrators to have caring as a way of being if they wish to communicate caring as the essence of nursing to students.

Cancer treatment in rural areas.

The inability to deliver cancer prevention and treatment to the rural population poses a significant barrier in the national effort to reduce cancer mortality. Since 25 percent of the U.S. population lives in rural areas and few rural areas are readily accessible to cancer centers or Community Clinical Oncology Programs (CCOPs), the prospects for accomplishing the National Cancer Institute (NCI) Goals for the Year 2000 are limited unless substantive changes occur in rural cancer care delivery. This article reviews the problem of cancer risk and care in rural areas and describes one effort to deliver state-of-the-art cancer treatment to rural patients in Virginia. It describes the needs and barriers to access in rural Virginia, the structural elements of the Rural Cancer Outreach Program, and the health policy issues that result when subspecialty care is exported to disadvantaged areas.

Teaching specialty cancer medicine in rural hospitals: the cancer outreach program as a model.

Cancer doctors and nurses are clustered in the metropolitan areas of Virginia. However, cancer patients are found throughout the state, and cancer mortality time trends are worse in the rural areas. Efforts to recruit cancer physicians and nurses to rural hospitals have been unsuccessful due to the practice characteristics, educational isolation, and economic disincentives. Our rural cancer education program involves the physicians and nurses currently in practice at two rural hospitals. We provide hands-on training in cancer care, continuing education, and intense week-long educational sessions. We have observed changes in physician and nurse practice styles that benefit the cancer patient including effective pain management, enrollment of patients on clinical trials, increased use of adjuvant therapy, and breast conservation. We are providing state-of-the-art cancer care at the rural hospitals in the Cancer Outreach Program. We have improved the educational opportunities and increased utilization of the resources of the academic career center. We can modify the practice characteristics by providing needed clinical programs and enhancing the rural hospital/academic medical center link. We have shown that rural cancer care can be revenue-neutral or positive, and we are seeking creative solutions to the financial disincentives of rural specialty practice. We can assist the rural hospital in the recruitment of oncology specialty nurses and physicians by providing cancer care services, and the patient caseload is available to teach prospective rural subspecialty practitioners at the rural hospitals.

Inducible UV repair potential of Pseudomonas aeruginosa PAO.

Pseudomonas aeruginosa PAO lacks UV-inducible Weigle reactivation and Weigle mutagenesis of UV-damaged bacteriophages. This lack of UV-inducible, error-prone DNA repair appears to be due to the absence of efficiently expressed umuDC-like genes in this species. When the P. aeruginosa recA gene is introduced into a recA(Def) mutant of Escherichia coli K12, the P. aeruginosa recA gene product is capable of mediating UV-induced mutagenesis, indicating that it could participate in a recA-lexA-like regulatory network and function in inducible DNA repair pathways if such existed in P. aeruginosa. The presence of the IncP9, UV-resistance plasmid R2 in RecA+ strains of P. aeruginosa PAO allows UV-inducible, mutagenic DNA repair of UV-irradiated bacteriophages. R2 also greatly stimulates the ability of UV radiation to induce mutagenesis of the bacterial chromosome. When R2 is introduced into P. aeruginosa strains containing either the recA908 or recA102 mutation, plasmid-mediated UV resistance and Weigle reactivation are not observed. These observations suggest that the increased protection afforded to P. aeruginosa by R2 is derived from a RecA-mediated, DNA-damage-inducible, error-prone DNA repair system which complements the lack of a chromosomally encoded umuDC-like operon.

Expression of a high-affinity mechanism for acquisition of transferrin iron by Neisseria meningitidis.

Iron-starved meningococci grown at either pH 7.2 or 6.6 were capable of removing and incorporating iron from human transferrin by a saturable, cell surface mechanism that specifically recognized transferrin rather than iron. The maximum expression of the iron uptake system occurred after 4 h of iron starvation. The uptake of the iron was dependent upon a functioning electron transport chain and was sensitive to 60 degrees C and trypsin. Cells grown under iron-sufficient conditions were incapable of accumulating iron from transferrin. No evidence was found for a primary role for cell-free soluble siderophores in the removal of iron from transferrin. The nonpathogenic neisseriae, Neisseria flava and N. sicca, were unable to utilize iron on transferrin.

Comparison of iron binding and uptake from FeCl3 and Fe-citrated by Neisseria meningitidis.

Iron-starve Neisseria meningitidis SD1C took up Fe added as FeCl3 by a two-step process: a rapid phase occurring during the 1st min after Fe addition and unaffected by 0.5 mM KCN and a slower secondary phase of uptake sensitive to various metabolic inhibitors. The rate and extent of energy-dependent Fe uptake from stage dicitrate and nitrilotriacetate Fe3+ complexes were essentially identical to FeCl3, indicating the presence of a high-affinity Fe acquisition system. The sulfonated siderophore Desferal (deferrioxamine beta-mesylate) effectively prevented any uptake of Fe from the citrate complex and was unable to remove that Fe already bound by either poisoned or active cells. The energy-independent system rapidly deferrated Fe3+ dicitrate and bound Fe to a finite number of cellular sites. Fe derived from FeCl3 was associated with these same sites as well as with a large number of apparently nonspecific sites. The extent of low-affinity, nonspecific binding was concentration dependent. Both energy-independent and energy-dependent systems involved in the uptake of Fe from the Fe3+ dicitrate were inactivated by 5 min at 60 degrees C, but not by 45 degrees C. The citrate carrier itself was recycled, being neither bound separately nor in concert with Fe3+ by these sites.

Energy-independent uptake of iron from citrate by isolated outer membranes of Neisseria meningitidis.

Cyanide-poisoned Neisseria meningitidis SD1C cells rapidly took up 55Fe from iron-citrate complexes during the first 2 min, after which no further iron was accumulated. [14C]citrate was not taken up concomitantly with 55Fe by these cells. The 55Fe taken up by the poisoned cells was found in the membrane fraction after cells were broken; 70% of the radioactivity was distributed in the outer membrane, and 30% was in the inner membrane. Isolated outer membranes from iron-starved cells were as capable of iron uptake from citrate as intact cells were. As with whole cells, [14C]citrate was not taken up by isolated outer membranes. A polyacrylamide gel electrophoresis analysis of the proteins from citrate-dialyzed outer membranes after the uptake of 55Fe revealed that the radioactivity was associated with a major band of 36,500 molecular weight.