Plasma and serum concentrations of cytarabine administered via continuous intravenous infusion to dogs with meningoencephalomyelitis of unknown etiology.

The objective of this study was to evaluate the plasma and serum concentrations of cytarabine (CA) administered via constant rate infusion (CRI) in dogs with meningoencephalomyelitis of unknown etiology (MUE). Nineteen client-owned dogs received a CRI of CA at a dose of 25 mg/m2 /h for 8 h as treatment for MUE. Dogs were divided into four groups, those receiving CA alone and those receiving CA in conjunction with other drugs. Blood samples were collected at 0, 1, 8, and 12 h after initiating the CRI. Plasma (n = 13) and serum (n = 11) cytarabine concentrations were measured by high-pressure liquid chromatography. The mean peak concentration (CMAX ) and area under the curve (AUC) after CRI administration were 1.70 ± 0.66 µg/mL and 11.39 ± 3.37 h·µg/mL, respectively, for dogs receiving cytarabine alone, 2.36 ± 0.35 µg/mL and 16.91 + 3.60 h·µg/mL for dogs administered cytarabine and concurrently on other drugs. Mean concentrations for all dogs were above 1.0 µg/mL at both the 1- and 8-h time points. The steady-state achieved with cytarabine CRI produces a consistent and prolonged exposure in plasma and serum, which is likely to produce equilibrium between blood and the central nervous system in dogs with a clinical diagnosis of MUE. Other medications commonly used to treat MUE do not appear to alter CA concentrations in serum and plasma.

Biosafety testing of genetically modified ryegrass (Lolium perenne) using a model for the optimum selection of test invertebrates.

Selection of test species for use in biosafety evaluation of genetically modified plants is challenging but important, as regulators in many jurisdictions require tests to determine the potential for adverse environmental impacts before the release of plants into the environment. This contribution provides an example of an evidence-based process whereby species from the receiving environment can be ranked in order of susceptibility to potential impact, and guide test species selection. The case study used for this example was ryegrass, a forage plant, which had been modified to produce elevated levels of the lipid triacylglyceride. The previously described priority ranking of nontarget invertebrates model (PRONTI), designed to rank invertebrates for biosafety testing, has been adapted for use with these plants, which could, potentially, be beneficial to invertebrate populations, and applied to data on 246 known pasture invertebrate species. The output from the model for the top 20 ranked pasture invertebrate species is discussed, the attributes of these are considered along with the level of uncertainty in the information used. Consideration is given to how the model output can be interpreted and used in a biosafety risk assessment. While some subjectivity is involved in establishing the scores, all invertebrate species are subjected to the same analysis, and treated equally. In this way, regulators have a method of a risk assessment that is evidence-based, and transparent in its assumptions thereby avoiding potential for bias.

The pharmacokinetics of cytarabine in dogs when administered via subcutaneous and continuous intravenous infusion routes.

This crossover study compared the pharmacokinetics of cytarabine in six healthy dogs following intravenous constant rate infusion (CRI) and subcutaneous (SC) administrations, as these are two routes of administration commonly employed in the treatment of meningoencephalitis of unknown etiology. Each dog received a SC cytarabine injection of 50 mg/m(2) or an 8 h CRI of 25 mg/m(2) per hour, with a 7-day washout before receiving the alternative treatment. Blood samples were collected for 16 h after CRI initiation and for 8 h after SC injection. Plasma concentrations were measured by high-pressure liquid chromatography (HPLC). Pharmacokinetic parameters were estimated using the best-fit compartmental analysis for both CRI and SC routes. Terminal half-life (T(1/2) ) of cytarabine was 1.35 ± 0.3 and 1.15 ± 0.13 h after SC administration and CRI, respectively. Mean peak concentration (Cmax ) was 2.88 and 2.80 µg/mL for SC and CRI administration, respectively. Volume of distribution was 0.66 ± 0.07 l/kg. The 8-h CRI produced steady-state plasma concentrations as determined by consecutive measurement that did not decline until the end of the infusion. The SC administration did not achieve steady-state concentrations because cytarabine administered by this route was rapidly absorbed and eliminated quickly. The steady state achieved with the cytarabine CRI may produce a more prolonged exposure of cytarabine at cytotoxic levels in plasma compared to the concentrations after SC administration.

Apparent absence of Parahaemoproteus lophortyx and other hematozoa in North Florida populations of bobwhite quail (Colinus virginianus).

Parahaemoproteus lophortyx (formerly Haemoproteus lophortyx) is known to infect populations of bobwhite quail (Colinus virginianus) in California and to lead to considerable mortalities in these birds. Populations of bobwhite quail in Florida have never been surveyed for the presence of this parasite. The goal of this study was to determine whether P. lophortyx is present in populations of bobwhite quail in north Florida. To achieve this goal, blood was drawn from 294 bobwhite quail from 4 study sites in north Florida from 19 June 2007 to 1 August 2007. Blood smears were made, stained with Giemsa, and examined under x1,000 magnification for the presence of Parahaemoproteus lophortyx. No gametocytes were noted in any of the blood smears. Thirty randomly chosen samples were examined via polymerase chain reaction (PCR). This procedure may detect parasitemias too low to detect by microscopy. No PCR-positive samples were detected, however, adding support to the absence of hematozoa in Florida populations of bobwhite quail.

Field parasitism of nontarget weevil species (Coleoptera: Curculionidae) by the introduced biological control agent Microctonus aethiopoides Loan (Hymenoptera: Braconidae) over an altitude gradient.

The parasitoid, Microctonus aethiopoides Loan (Hymenoptera: Braconidae) was introduced into New Zealand in 1982 to control the alfalfa pest, Sitona discoideus Gyllenhal (Coleoptera: Curculionidae). Studies have shown that a number of nontarget weevil species are attacked in the field by this parasitoid. A field study was carried out to investigate nontarget parasitism by M. aethiopoides over an altitudinal sequence from the target host habitat (alfalfa) into native grassland. Three locations were selected for the study, and at each, the alfalfa growing in the valley floor was sampled annually for parasitism of the target pest that ranged between 17 and 78%. At progressively higher altitudes, three further grassland sites at each location were sampled monthly during spring to autumn for up to 6 yr. Weevil densities were estimated, species identified, and dissections carried out to determine reproductive status and parasitism. Almost 12,000 weevils were collected during the study, which were identified as 36 species in total from the three locations. Eight weevil species were found to be parasitized, including S. discoideus, the target host that was found at all sites. Parasitism of nontarget species was approximately 2% overall but varied with location, site, and season. Substantial nontarget parasitism was found at only one of the locations, with up to 24% parasitism of a native weevil, Nicaeana fraudator Broun (Coleoptera: Curculionidae), recorded. Another species, Irenimus egens (Broun) (Coleoptera: Curculionidae), was also found at this location at similar population densities but was attacked far less by M. aethiopoides. Results are discussed in relation to weevil phenology.

State scholarship, loan forgiveness, and related programs: the unheralded safety net.

CONTEXT: In the mid-1980s, states expanded their initiatives of scholarships, loan repayment programs, and similar incentives to recruit primary care practitioners into underserved areas. With no national coordination or mandate to publicize these efforts, little is known about these state programs and their recent growth. OBJECTIVES: To identify and describe state programs that provide financial support to physicians and midlevel practitioners in exchange for a period of service in underserved areas, and to begin to assess the magnitude of the contributions of these programs to the US health care safety net. DESIGN: Cross-sectional, descriptive study of data collected by telephone, mail questionnaires, and through other available documents, (eg, program brochures, Web sites). SETTING AND PARTICIPANTS: All state programs operating in 1996 that provided financial support in exchange for service in defined underserved areas to student, resident, and practicing physicians; nurse practitioners; physician assistants; and nurse midwives. We excluded local community initiatives and programs that received federal support, including that from the National Health Service Corps. MAIN OUTCOME MEASURES: Number and types of state support-for-service programs in 1996; trends in program types and numbers since 1990; distribution of programs across states; numbers of participating physicians and other practitioners in 1996; numbers in state programs relative to federal programs; and basic features of state programs. RESULTS: In 1996, there were 82 eligible programs operating in 41 states, including 29 loan repayment programs, 29 scholarship programs, 11 loan programs, 8 direct financial incentive programs, and 5 resident support programs. Programs more than doubled in number between 1990 (n = 39) and 1996 (n = 82). In 1996, an estimated 1306 physicians and 370 midlevel practitioners were serving obligations to these state programs, a number comparable with those in federal programs. Common features of state programs were a mission to influence the distribution of the health care workforce within their states' borders, an emphasis on primary care, and reliance on annual state appropriations and other public funding mechanisms. CONCLUSIONS: In 1996, states fielded an obligated primary care workforce comparable in size to the better-known federal programs. These state programs constitute a major portion of the US health care safety net, and their activities should be monitored, coordinated, and evaluated. State programs should not be omitted from listings of safety-net initiatives or overlooked in future plans to further improve health care access. JAMA. 2000;284:2084-2092.

Mouse beta defensin-1 is a functional homolog of human beta defensin-1.

Defensin are 3-4 kDa antimicrobial peptides of which three distinct families have been identified; alpha-defensin, beta-defensins, and insect defensins. Recent investigations have shown that beta-defensins are present in the human airways and may be relevant to the pathogenesis of cystic fibrosis (CF) lung disease. We report here the further characterization of a recently identified mouse beta-defensin gene, Defb1, sometimes referred to as mBD-1, which is homologous to the human airway beta defensin hBD-1. We report that Defb1 is expressed in a variety of tissues including the airways and, similar to hBD-1, is not upregulated by lipopolysaccharide (LPS). Defb1 was found to consist of two small exons separated by a 16-kb intron and cytogenetic, and physical mapping linked it to the alpha defensin gene cluster on mouse Chromosome (Chr) 8. Functional studies demonstrate that, like hBD-1, Defb1 demonstrates a salt-sensitive antimicrobial activity against Pseudomonas aeruginosa. Of relevance to CF lung disease is the fact that neither the hBD-1 nor the mBD-1 peptides are active against Burkholderia cepacia.

Inclusion of cholesterol in DOTAP transfection complexes increases the delivery of DNA to cells in vitro in the presence of serum.

The contrast between the relative efficiency of transfection by cationic lipid reagents in vitro and that in vivo is well recognised. One suggested reason for this is the presence of competing polyanionic surfaces in blood and other biological fluids, and even in vitro transfections have to be performed in low-serum medium. In this study we have shown that by preparing cationic lipid reagents based on DOTAP with cholesterol as a second constituent of the bilayer we can achieve significant levels of in vitro transfection in serum concentrations of up to 80% (DOTAP alone did not transfect at all in these conditions). In an effort to explain the behaviour of DOTAP/cholesterol mixes under these conditions, we examined the effect of serum on the transfection complex. We could detect protein bound to each type of cationic lipid complex, but there was no difference in the amount nor in the type of protein bound. DNA within either type of complex which were incubated with increasing amounts of serum remained resistant to digestion with DNase I, and there was no reduction in the condensation of the DNA as measured by ethidium bromide fluorescence. Finally, we measured the attachment and uptake into cells by the different complexes in the presence of serum and showed that more DOTAP-cholesterol than DOTAP complexes attach to and are taken up by cells in the presence of serum. We suggest that improved cell binding and uptake may be the main mechanism by which cholesterol acts to maintain transfection in the presence of serum.

Plasmid DNA molecules complexed with cationic liposomes are protected from degradation by nucleases and shearing by aerosolisation.

The cationic liposome DOTAP was complexed with plasmid DNA encoding beta-galactosidase in various ratios. As the concentration of DOTAP increased, the DNA became increasingly refractory to staining with ethidium bromide, presumably because the DNA was becoming condensed and being encapsulated by the liposomes. Transfection by DNA-DOTAP complexes at all ratios tested was unaffected by treatment of the complexes with DNase I. This finding has relevance to clinical trials for gene therapy of cystic fibrosis, in which patients are normally removed from treatment with DNase before receiving administration of DNA. We additionally tested the effect of aerosolisation of the liposome-DNA complex and of the DNA alone on the efficiency of in vitro transfection. Aerosolised DNA complexed with fresh DOTAP led to much lower reporter gene expression in Cos 7 cells than non-aerosolised complex, since aerosolisation appeared to destroy almost all of the plasmid. However, complexing the plasmid before passage through the nebuliser did protect most of the DNA from degradation, as reflected in the levels of transfection obtained. These findings contribute towards an overall understanding of both how DNA-cationic liposome complexes are formed and their fate following administration in vivo.

Enrichment of early fetal-liver hemopoietic stem cells of the rat using monoclonal antibodies against the transferrin receptor, Thy-1, and MRC-OX82.

Fetal livers from inbred rat fetuses at 14 days' gestation were dispersed into a single-cell suspension by physical disruption and collagenase digestion. Pluripotent stem cells were characterized and partially purified by a combination of monoclonal antibodies. These included CD71 (anti-transferrin receptor, MRC-OX26, used for rosetting), Cdw90 (anti-Thy-1, MRC-OX7), and the newly described MRC-OX82 (reacting with myeloiid cells in peritoneal exudate), employed in FACS sorting. Enrichment was monitored by long-term reconstitution of lethally irradiated congenic rats genetically distinguishable from the donor by an allelomorphic variant of the CD45 cell-surface antigen. At intervals from 3 months to 1 year, lymph-node cells and peritoneal exudate cells were biopsied for analysis by two-color flow cytometry--one color to determine donor origin, the other to identify Th cell (CD4+), Tc cell (CD8+), B cell (sIg+ or CD45RC+), neutrophil (OX82+ or OX43-), and macrophage (OX43+) compartments. The degree of chimaerism was taken as the read out of stem-cell activity. No significant differentials between lymph-node and peritoneal exudate chimaerisms were detected in any of the recipients; therefore, the enrichment procedure revealed only pluripotent cells, not stem cells of restricted potency. All recovered stem-cell activity was in the OX26(CD71)-negative, OX7(CDw90)-positive, OX82-positive fraction. In the optimum case, an enrichment of very roughly 200-fold in cell-for-cell activity was obtained. Rat bone-marrow colony-forming units in the spleen (CFUs-12) were found to lack the surface antigens recognized by the monoclonal antibodies CD53 (MRC-OX44), MRC-OX39, MRC-OX59, and 144.2.15. These would provide a strategy for their enrichment by depletion.

Endocytosis and presentation of liposome-associated antigens by B cells.

B cells have limited endocytic capacity and are reported to endocytose and present liposome-encapsulated antigens poorly. B cells also endocytose soluble antigens poorly, except those for which their surface immunoglobulin is specific, which are taken up and presented efficiently. We present results indicating that, in vitro, B cells endocytose small liposomes bearing antigen with affinity for their surface immunoglobulin. Antigen encapsulated in liposomes targeted by antibody specific for surface immunoglobulin is presented to T cells as efficiently as specific antigen in soluble form. These studies provide a rational basis for the design of liposomes optimized to stimulate T-dependent B-cell responses.

Induction of T lymphocyte subsets and levels of interleukin-2 and interleukin-3 after infection with Trichinella spiralis are similar in mice of high- and low-responder phenotypes.

Various aspects of T lymphocyte responses in mice with high- and low-responder phenotypes to infection with the nematode Trichinella spiralis were compared. Mesenteric lymph node cells (MLNC) from infected NIH strain mice (high-responder) showed a much higher level of proliferation in response to parasite antigen than MLNC from infected C57.B10 mice (low-responder). Levels of CD4+ and CD8+ T lymphocytes in the MLN remained relatively constant in both strains, ruling out the possibility that the more rapid worm expulsion in NIH mice (day 6 post-infection, as compared to day 12-15 in B10 mice) was due to quantitative differences in levels of CD4+ T helper cells. To examine any variation in the recruitment of Th1 v Th2 cell subsets, the production of the cytokines interleukins-2, -3, and -4 (IL-2, -3, and -4) by MLNC in response to parasite antigen was measured throughout the course of infection. Amounts of IL-2 and IL-3 were broadly similar in both strains, while almost background levels of IL-4 were detected. However, NIH strain mice produced IL-2 and IL-3 more rapidly than B10 mice. These data support the conclusion from other authors that the response to T. spiralis is not dependent on the absolute levels of cytokines produced, but rather on timing of production and rapidity of recruitment of effector cells at the level of bone marrow precursors.

Tarasoff in the Canadian context: Wenden and the duty to protect.

The precedent-setting 1976 US court decision of Tarasoff v. Regents of the University of California established a "duty to protect" whereby psychotherapists are held responsible to protect the potential victims of their clients' violent behaviour. The purpose of this article is to review and discuss this duty in the context of Canadian law and the 1991 Alberta court decision of Wenden v. Trikha, Royal Alexandra Hospital and Yaltho. The current Canadian interpretation of the duty to protect and guidelines for avoiding such liability are presented.

The facility master plan: a road map for your future.

Current and anticipated changes in the mental healthcare delivery system demand a comprehensive facility planning process. As reimbursement for acute care decreases and the influence of managed care increases, most private psychiatric hospitals will need to develop a facility master plan to best use their current buildings and property. The facility master plan will become a road map and provide a logical route for expanding, modifying, and using current hospital assets during the changes imminent in the years ahead.

Perceived family environments of depressed and anxious children: child's and maternal figure's perspectives.

This study examined perceived environment among families with a depressed, depressed and anxious, anxious, or normal child from the 4th to 7th grades. Fifty-one such children were classified according to criteria from the K-SADS and a set of self-ratings of depression and anxiety. Results showed that children in all three diagnostic groups, and to a lesser extent their mothers, experienced their families as more distressed on a host of dimensions relative to controls. In addition, significant differences were found between families with a depressed and anxious child and those with an anxious child. Discriminant function analyses revealed that 68.63% of the youngsters could be classified correctly into depressed and anxious groups on the basis of their family ratings alone.