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Cocaine use causes significant cardiovascular morbidity from its hemodynamic effects. It is less clear whether cocaine promotes atherosclerosis. Vascular inflammation is one of the earliest steps in the pathophysiology of atherosclerosis. We hypothesized that cocaine results in an increase in inflammatory markers. Study objective was to measure the acute effects of intravenous cocaine on biomarkers of vascular inflammation. Eleven chronic cocaine users were enrolled. After a drug-free period, they received intravenous cocaine at 0.36 mg/kg dose in an in-hospital controlled environment. Serum levels of soluble CD40 ligand, monocyte chemoattractant protein-1, interleukin 6, and soluble intercellular adhesion molecule-1 were measured at baseline, 6 h, 24 h, and 6 days after cocaine challenge and at baseline for controls. After cocaine challenge, sCD40 ligand levels decreased in subjects and were significantly lower at 24 h. MCP-1 levels decreased and were significantly lower at the 6-day time point. No significant changes in IL-6 or sICAM-1 level were found. In conclusion, intravenous cocaine did not result in an increase in levels of inflammatory markers. Levels of MCP-1 and sCD40L decreased significantly. This unexpected finding suggests that chronic effects of cocaine on inflammation may be different from acute effects or that higher dosing may have differential effects as compared to lower dose used here.

Mast cells play an important role in immunomodulation and in the maintenance of vascular integrity. Interleukin-6 (IL-6) is one of the key biomarkers and therapeutic target in systemic vasculitis. The objective of the current study is to describe the role of mast cells in arterial IL-6 homeostasis. Eight- to ten-week-old male C57BL/6 (wild-type) mice were injected with either (a) saline, (b) compound 48/80 (a systemic mast cell degranulating agent), (c) lipopolysaccharide (LPS), or (d) a combination of C48/80 and LPS. Twenty-four hours after the injections, mice were sacrificed and serum samples and aortic tissues were analyzed for determining inflammatory response and cytokine expression profile. The results revealed that induction of mast cell degranulation significantly lowers serum IL-6 levels and aortic expression of IL-6 in LPS-treated mice. Significantly higher aortic expression of toll-like receptor-2 (TLR-2) and TNF-α was seen in the LPS and LPS+C48/80 groups of mice compared to controls. Aortic expression of TLR-4 was significantly decreased in LPS+C48/80 compared to C48/80 alone. LPS+C48/80-treated mice presented with a 3-fold higher aortic expression of suppressor of cytokine signaling (SOCS-1) compared to saline-injected groups. The inhibition of LPS-induced increase in serum IL-6 levels by mast cell degranulation was not seen in H1R knockout mice which suggests that mast cell-derived histamine acting through H1R may participate in the regulatory process. To examine whether the mast cell-mediated downregulation of LPS-induced IL-6 production is transient or cumulative in nature, wild-type mice were injected serially over a period of 10 days (5 injections) and serum cytokine levels were quantified. We found no significant differences in serum IL-6 levels between any of the groups. While mice injected with C48/80 or LPS had higher IL-10 compared to vehicle-injected mice, there was no difference between C48/80- and LPS+C48/80-injected mice. In conclusion, in an in vivo setting, mast cells appear to partially and transiently regulate systemic IL-6 homeostasis. This effect may be regulated through increased systemic IL-10 and/or aortic overexpression of SOCS-1.

Background We have previously demonstrated that mast cell degranulation acutely downregulates lipopolysaccharide induced aortic expression and serum levels of IL-6 in vivo. This is accompanied by aortic upregulation of suppressor of cytokine signaling-1 (SOCS-1) gene expression1. This effect is not seen in histamine H1 receptor-knockout mice suggesting that mast cell-derived histamine is a key mediator involved in IL-6 homoeostasis2. Mice injected with Candida albican water-soluble-fraction (CAWS) have been shown to develop coronary and aortic vasculitis3. Our long-term objective is to determine the pathogenic mechanism of large vessel vasculitis (LVV). Objectives The aim of this pilot study was to replicate and develop a working mouse model to determine the regulatory role of mast cells in LVV. Methods Eight to ten weeks-old male C57Bl6/J mice were randomly distributed into two groups [CAWS, N=8; and Control, N=8] and were injected i.p. daily for 5 days with either CAWS in normal saline (2 mg/day/mouse) or normal saline alone (controls). All mice were sacrificed 30 days after the 5th injection. We examined serum levels of IL-6 and TNF-α, as well as aortic tissue expressions of IL-6, TNF-a, IL-10 and SOCS-1 mRNA. Heart and aortic sections were evaluated for inflammation and mast cells after staining with H & E and toluidine blue, respectively. Results Treatment of mice with CAWS for 5-consecutive days led to overexpression of IL-6, TNF-a and IL-10 genes in the aortic tissue with modest upregulation of SOCS-1. At the root of the aorta, all animals in the CAWS group had intense inflammatory infiltrates composed of mixed acute and chronic inflammatory cells. There is also evidence of vasculitis in the coronary arteries. In contrast, none of the control mice had any evidence of aortic inflammation or vasculitis. Serum IL-6 concentrations were below detectable levels in both controls and CAWS-treated mice whereas TNF-a levels were elevated in 3 out of 8 mice in the CAWS group. There were no signs or increased presence of intact or degranulating mast cells in the area of inflammation. Conclusions These results suggest that CAWS-induced LVV involves acute and chronic inflammatory response and vascular tissue expression of both pro- and anti-inflammatory cytokines and SOC-1. Detailed kinetic studies are warranted to determine the optimum windows of peak inflammatory response and the expression of these genes to understand the pathobiology of CAWS-induced LVV. References Springer J, Raveendan V, Smith D, et al. [abstract] Ann Rheum Dis 2014;73:112–113. Raveendran V, Springer J, Smith D, et al. [abstract]. Arthritis Rheumatol 2015;67(suppl 10). Miura N, Shingo Y, Adachi Y, et al. Immunopharmacol Immunotoxicol 2004;26(4):527–43. Acknowledgements Supported by Basic Science Development Award from Department of Medicine at the University of Kansas. Disclosure of Interest None declared

We examined the effects of two over-the-counter H1-antihistamines on the progression of fatty liver disease in male C57Bl/6 wild-type and apolipoprotein E (ApoE)−/− mice. Mice were fed a high-fat diet (HFD) for 3 mo, together with administration of either cetirizine (4 mg/kg body wt) or fexofenadine (40 mg/kg body wt) in drinking water. Antihistamine treatments increased body weight gain, gonadal fat deposition, liver weight, and hepatic steatosis in wild-type mice but not in ApoE−/− mice. Lobular inflammation, acute inflammation, and necrosis were not affected by H1-antihistamines in either genotype. Serum biomarkers of liver injury tended to increase in antihistamine-treated wild-type mice. Serum level of glucose was increased by fexofenadine, whereas lipase was increased by cetirizine. H1-antihistamines reduced the mRNA expression of ApoE and carbohydrate response element-binding protein in wild-type mice, without altering the mRNA expression of sterol regulatory element-binding protein 1c, fatty acid synthase, or ApoB100, in either genotype. Fexofenadine increased both triglycerides and cholesterol ester, whereas cetirizine increased only cholesterol ester in liver, with a concomitant decrease in serum triglycerides by both antihistamines in wild-type mice. Antihistamines increased hepatic levels of conjugated bile acids in wild-type mice, with the effect being significant in fexofenadine-treated animals. The increase was associated with changes in the expression of organic anion transport polypeptide 1b2 and bile salt export pump. These results suggest that H1-antihistamines increase the progression of fatty liver disease in wild-type mice, and there seems to be an association between the severity of disease, presence of ApoE, and increase in hepatic bile acid levels.

Activation of histamine receptor 1 (H1R) reduces adiposity in mice, and chronic use of H1-antihistamines (H1-AH) increase weight gain in obese patients. Apolipoprotein E (ApoE) plays a major role in lipid transport. We examined the effects of chronic blockade of H1R by cetirizine or fexofenadine, two therapeutic H1-AH, on fatty liver disease, adiposity and weight gain in wild-type (WT) and ApoE-knockout (ApoE-/-) mice. Eight week old male WT or ApoE-/- mice were fed a high fat diet, and concurrently treated with equivalent therapeutic doses of either cetirizine or fexofenadine via drinking water for 3 months. H1-HA treatments increased weight gain, visceral adiposity, liver weight and macrovesicular hepatosteatosis in WT mice, but not in ApoE-/- mice. These changes were independent of food intake. Serum markers of liver injury tended to increase in H1-HA-treated WT mice in association with increased macrovesicular hepatosteatosis. Microvesicular hepatosteatosis and lobular inflammation were not affected b...

Mast cells are constituent cells of vascular tissue and their numbers are increased in atherosclerotic vessels. To gain insight into the role of mast cells in vascular inflammation, the effect of mast cell granules (MCG) on endothelial cell production of interleukin-6 (IL-6) was examined. Human umbilical vein endothelial cells (HUVEC) were cultured with lipopolysaccharide (LPS) in the presence or absence of rat peritoneal MCG and IL-6 production was assayed by enzymelinked immunosorbent assay. The interaction of MCG with HUVEC in culture was examined by electron microscopy (EM). The EM studies revealed that MCG are internalized by HUVEC and appear intact even after 24 h in culture. Unactivated HUVEC produced little or no IL-6 either in the presence or absence of MCG. Treatment of HUVEC with LPS stimulated IL-6 production in a dose- and time-dependent fashion. Addition of MCG to LPS-activated HUVEC resulted in the potentiation of IL-6 production at all LPS doses. MCG-induced enhancement of IL-6 production was evident even at a mast cell-to-endothelial cell ratio of 1:32. The enhancement of IL-6 production by MCG was also seen when tumor necrosis factor α was used as an activator. Although potentiation was evident when MCG were added 6 h before or after LPS stimulation, the maximum effect was noted when MCG and LPS were added simultaneously. MCG-mediated enhancement of IL-6 production was abrogated by pretreating MCG with protease inhibitors. Although MCG proteases potentiate IL-6 production by HUVEC, they do not degrade secreted IL-6. These results demonstrate that MCG interact with endothelial cells and modulate the production of an important inflammatory cytokine.

The cellular enzyme-linked immunosorbent assay (CELISA) permits assay of cell surface antigens on intact fixed cells. Using a monolayer of cells as the solid phase, the CELISA offers an inexpensive alternative to flow cytometry. In addition, this protocol has the decided advantages of miniaturization (small numbers of cells) and ease of replication (the 96-well cluster plate format). In efforts to optimize CELISA for detecting surface antigens on human fibroblastlike synoviocytes, the authors found that cell number, serum proteins, choice of culture plate, pipetting technique and fixatives may all impact the results of the CELISA.

We examined three disturbed, but distinctly different, sites on the historically forested eastern slope of the Sierra de Baoruco, Provincia de Barahona, Republica Dominicana, in order to evaluate the impact of habitat alteration on the composition of anoline lizard communities. Five species of Anolis (A. bahorucoensis, A. barahonae, A. coelestinus, A. cybotes, A. distichus) are known to occur in the immediate area of our study sites. Only three were found in the most substantially altered clear-cut site, whereas all five species occupied the most heterogeneous habitat, a coffee plantation characterized by a three-tiered canopy. Similarly, numbers of individuals of all but one species were greatest in the plot that most closely resembled the natural forests in this area. Calculated niche breadths, based on analysis of stomach contents, of the three species found in all sites increased with the degree of habitat alteration. The higher number of A. coelestinus in more heavily altered sites may be attributed to density compensation.

The societal and economic forces driving change in medical education are affecting communities as well as universities. Each of the four authors of this paper is deeply involved in one of the components of their locale's well-developed community-based medical educational system, and each describes how change is influencing his role in that system, whether the role be managing a community hospital, directing a local Area Health Education Center, participating as a family medicine faculty member, or being a community preceptor. They agree on some common themes: (1) that it is good that medical students' education is moving into the community (e.g., this validates the importance of the community hospital to medical education, is an acknowledgment of the importance of generalism, and provides students invaluable learning experiences); (2) that educating medical students in the community is expensive, and more funding and resources are needed so that the area's hospitals, community faculty, preceptors, and support services can be fairly compensated for their commitment; and (3) that their community-based education system can no longer absorb the costs of training more medical students. This is not a criticism of academic medical centers, which are under tremendous financial pressures themselves, but is simply to state the community perspective and to urge fairness in the distribution of resources for medical education. Community institutions and academic medical centers will work individually to create their own integrated health care systems but must work together to create a better, more cost-effective system for educating medical students.

Background Serum interleukin-6 (IL-6) has been shown to closely follow disease activity in large vessel vasculitis 1 . Prior research by our group has demonstrated that mast cell degranulation downregulates lipopolysaccharide (LPS) induced aortic expression of IL-6 in vivo. This is accompanied by aortic upregulation of SOCS-1 2 . This effect is lost in histamine 1 receptor knockout mice suggesting that histamine is a primary mediator of aortic IL-6 inhibition. Prior studies have also demonstrated that mice injected with Candida albicans water-soluble fraction (CAWS) develop coronary and aortic arteritis 3 . The mechanism through which arteritis is induced by CAWS remains poorly understood. Objectives The objective of this study is to determine the effects of histamine and CAWS on IL-6 production by human aortic endothelial cells (HAEC) in vitro. Methods HAEC were cultured with histamine (10μM), LPS (100ng/mL), CAWS (10μg/mL), CAWS (1μg/mL) or a combination of these. After 22 hours, levels of IL-6 and 6-ketoprostaglandin-F1α, a breakdown product of prostacyclin, were measured. Microculture tetrazolium (MTT) assays were used to determine cell viability. Results IL-6 levels were significantly lower in cells treated with CAWS 10μg/mL compared to controls (175.2±11.9 vs 270.0 pg/mL, p=0.006). Similarly, IL-6 levels were significantly lower with the addition 10μg/mL CAWS to LPS compared to LPS alone (359.8±37.74 vs 2906.5±318.02, p=0.0001). No significant differences were seen between the 10μg/mL and the 1μg/mL CAWS concentrations. Histamine synergistically enhanced IL-6 levels over LPS alone (3758.0±38.18 vs 2906.5±318.0, p=0.028). CAWS had no effect on prostacyclin levels compared to controls. MTT assays demonstrated no differences in cell viability in CAWS or histamine treated cells compared to controls. Conclusions Our experiments suggest that CAWS inhibits IL-6 production in HAEC. Since IL-6 is a key cytokine in the development of large vessel vasculitis we suspect that endothelial cells do not play a role in CAWS mediated arteritis. Similarly, the inhibitory effects of histamine do not appear to be mediated through endothelial cells. Future research will focus on the in vivo effects of histamine on the development of CAWS mediated vasculitis. References Weyand CM, Fulbright JW, Hunder GG, Evans JM, Goronzy JJ. Treatment of giant cell arteritis: interleukin-6 as a biologic marker of disease activity. Arthritis and rheumatism. May 2000;43(5):1041–1048. Springer J, Raveendran V., Smith D., Maz M., Dileepan K. Novel role of mast cells in the regulation of large vessel vasculitis. Paper presented at: European League Against Rheumatism 2014 meeting; June 12th, 2014, 2014; Paris, France. Nagi-Miura N, Shingo Y, Adachi Y, et al. Induction of coronary arteritis with administration of CAWS (Candida albicans water-soluble fraction) depending on mouse strains. Immunopharmacology and immunotoxicology. 2004;26(4):527–543. Acknowledgement Supported by Basic Science Development Award from Department of Medicine at the University of Kansas. Disclosure of Interest None declared

Sera from 273 wild mammals from Missouri and Kansas (USA), collected between December 1974 and December 1987, were tested for the presence of antibodies to Toxoplasma gondii using the Sabin-Feldman dye test. Sixty-five (24%) had antibodies at titers ofor = 1:8, including 38 (66%) of 58 carnivores, 14 (15%) of 94 omnivores, 13 (11%) of 117 herbivores, and none of four insectivores. The prevalence of antibodies in mice (Mus musculus and Peromyscus spp.) and rats (Rattus norvegicus and Sigmodon hispidus) was low (3%), while medium sized herbivores such as squirrels (Sciurus spp.), rabbits (Sylvilagus floridanus), and muskrats (Ondatra zibethicus) had prevalences of about 18%. Red foxes (Vulpes fulva) and mink (Mustela vison) had the highest prevalence of antibodies with frequencies of 90 and 66%, respectively. In 32 attempts to isolate Toxoplasma gondii from wild mammals with positive (or = 1:4) titers, only six (19%) were successful: a gray squirrel (Sciurus carolinensis), a beaver (Castor canadensis), an opossum (Didelphis marsupialis), a red fox and two mink. These findings are consistent with the hypothesis that the probability of infection with Toxoplasma gondii, and therefore prevalence of antibodies in wildlife, is greatest in carnivores.

Abstract Agonistic behaviour of male Anolis olssoni was elicited in natural habitats near Baraona, Barahona Province, Dominican Republic, through the introduction of tethered males. Videotaped responses consisted of various sequences and combinations of behaviours and modifiers and appeared to vary according to levels of arousal. Male A. olssoni assume a posture involving full head elevation and dewlap extension, which apparently signals the highest state of arousal to conspecifc males. This display was invariably accompanied by gaping with a protruded tongue, distinct extension of nuchal and dorsal crests, bulging eyes, and changes in body and tail colouration. Analysis of behaviours failed to identify a stereotypical display for this species. Males apparently monitor and defend their immediate surroundings, a strategy which may be adaptative in a densely occupied and complex microhabitat in which vision is restricted.

Mast cells are important cells of the immune system and are recognized as participants in the pathogenesis of atherosclerosis. In this study, we evaluated the role of mast cells on the progression of atherosclerosis and hepatic steatosis using the apolipoprotein E-deficient ( ApoE−/−) and ApoE−/−/mast cell-deficient ( KitW-sh/W-sh) mouse models maintained on a high-fat diet. The en face analyses of aortas showed a marked reduction in plaque coverage in ApoE−/−/KitW-sh/W-shcompared with ApoE−/−after a 6-mo regimen with no significant change noted after 3 mo. Quantification of intima/media thickness on hematoxylin and eosin-stained histological cross sections of the aortic arch revealed no significant difference between ApoE−/−and ApoE−/−/KitW-sh/W-shmice. The high-fat regimen did not induce atherosclerosis in either KitW-sh/W-shor wild-type mice. Mast cells with indications of degranulation were seen only in the aortic walls and heart of ApoE−/−mice. Compared with ApoE−/−mice, the serum levels of total cholesterol, low-density lipoprotein and high-density lipoprotein were decreased by 50% in ApoE−/−/KitW-sh/W-shmice, whereas no appreciable differences were noted in serum levels of triglycerides or very low density lipoprotein. ApoE−/−/KitW-sh/W-shmice developed significantly less hepatic steatosis than ApoE−/−mice after the 3-mo regimen. The analysis of Th1/Th2/Th17 cytokine profile in the sera revealed significant reduction of interleukin (IL)-6 and IL-10 in ApoE−/−/KitW-sh/W-shmice compared with ApoE−/−mice. The assessment of systemic generation of thromboxane A2(TXA2) and prostaglandin I2(PGI2) revealed significant decrease in the production of PGI2in ApoE−/−/KitW-sh/W-shmice with no change in TXA2. The decrease in PGI2production was found to be associated with reduced levels of cyclooxygenase-2 mRNA in the aortic tissues. A significant reduction in T-lymphocytes and macrophages was noted in the atheromas of the ApoE−/−/KitW-sh/W-shmice. These results demonstrate the direct involvement of mast cells in the progression of atherosclerosis and hepatic steatosis.

Although increased serum histamine levels and H1R expression in the plaque are seen in atherosclerosis, it is not known whether H1R activation is a causative factor in the development of the disease, or is a host defense response to atherogenic signals. In order to elucidate how pharmacological inhibition of histamine receptor 1 (H1R) signaling affects atherogenesis, we administered either cetirizine (1 and 4 mg/kg. b.w) or fexofenadine (10 and 40 mg/kg. b.w) to ApoE−/− mice maintained on a high fat diet for three months. Mice ingesting a low dose of cetirizine or fexofenadine had significantly higher plaque coverage in the aorta and cross-sectional lesion area at the aortic root. Surprisingly, the higher doses of cetirizine or fexofenadine did not enhance atherosclerotic lesion coverage over the controls. The low dose of fexofenadine, but not cetirizine, increased serum LDL cholesterol. Interestingly, the expression of iNOS and eNOS mRNA was increased in aortas of mice on high doses of cetirizine or fexofenadine. This may be a compensatory nitric oxide (NO)-mediated vasodilatory mechanism that accounts for the lack of increase in the progression of atherosclerosis. Although the administration of cetirizine did not alter blood pressure between the groups, there was a positive correlation between blood pressure and lesion/media ratio at the aortic root in mice receiving the low dose of cetirizine. However, this association was not observed in mice treated with the high dose of cetirizine or either doses of fexofenadine. The macrophages or T lymphocytes densities were not altered by low doses of H1-antihistamines, whereas, high doses decreased the number of macrophages but not T lymphocytes. The number of mast cells was decreased only in mice treated with low dose of fexofenadine. These results demonstrate that chronic ingestion of low therapeutic doses of cetirizine or fexofenadine enhance progression of atherosclerosis.

Background Our previous reports suggested that mast cell-mediated innate immune up-regulation plays a key role in vascular inflammation (1-2). The literature also document increased presence of mast cells in the temporal arteries of patients with giant cell arteritis, a type of large vessel vasculitis (LVV) (3). The activation of Toll-like receptor-4 (TLR4) has been implicated in the recruitment of dendritic cells and T-cells into the aortic wall in LVV. Endothelial cells, Th17 T-cells and macrophages produce interleukin 6 [IL-6] which is important in the pathogenesis of LVV. The mechanism by which mast cells modulate the pathogenesis of LVV is not known. Objectives The objective of this study was to test the hypothesis that mast cell degranulation will regulate lipopolysaccharide (LPS, a TLR4 ligand)-induced systemic production of IL6. Methods Two month old male C57BI6/J mice were randomized into 4 groups (N=4/group). They were injected intraperitoneally with saline (control), Compound 48/80 (mast cell degranulator, 1mg/kg), LPS (1mg/kg) or C48/80+LPS. Animals were sacrificed 24 h after injections, and serum IL-6 levels, aortic expressions of mRNAs encoding IL-6, TLR4 and suppressor of cytokine signaling-1 (Socs-1) were determined. Data were analyzed for statistical significance and p Results C48/80 injection did not increase serum levels of IL-6 or aortic expression of IL-6 mRNA compared to control. LPS injection significantly enhanced serum IL-6 (350±146 pg/ml vs 21.3±5.5 pg/ml) and aortic IL-6 gene expression (18.0±5.4 fold vs 1.03±0.025 fold). LPS-induced increase in serum IL-6 levels and IL-6 gene expression in the aorta were significantly reduced when mast cell degranulation was simultaneously induced by C48/80 (IL-6: 350±146 pg/ml vs 101±13 pg/ml; IL-6 mRNA: 18.0±5.4 vs 4.3±0.5 fold change). In comparison to controls, the aortic expression of Socs-1 mRNA was 2-fold and 3-fold higher in LPS-treated, and C48/80+LPS-treated mice, respectively. Aortic expression of TLR4 was not affected by any of the treatments. Conclusions The results demonstrate that mast cell degranulation decreases aortic expression of IL-6 mRNA as well as systemic production of IL-6. The decreased expression of IL-6 was associated with increased expression of Socs-1 in aortic tissues. One mechanism by which mast cells regulate LPS-induced systemic production of IL-6 may be by inhibiting the expression of IL-6 mRNA in the vasculature through Socs-1 activation. Since mast cells release many immunomodulatory substances further studies are warranted to identify the regulatory factors. References Talreja J, Kabir MH, Filla B, Stechschulte DJ, Dileepan KN. Histamine induces Toll-like receptor 2 and 4 expression in endothelial cells and enhances sensitivity to Gram-positive and Gram-negative bacterial cell wall components. Immunology. 113:224-33, 2004. Raveendran VV, Tan X, Sweeney ME, Levant B, Slusser J, Stechschulte DJ, Dileepan KN. Lipopolysaccharide induces H1 receptor expression and enhances histamine responsiveness in human coronary artery endothelial cells. Immunology. 132:578-88, 2011. Mayranpaa MI, Trosien JA, Nikkari ST, Kovanen PT. Mast cells associate with T-cells and neointimal microvessels in giant cell arteritis. Clin Exp Rheum. 26 (3 Suppl 49):S63-6, 2008. Acknowledgements Supported by NIH grants R01-HL070101 & 3R01-HL070101-04S1, and Joseph & Elizabeth Carey Arthritis Fund and Audrey Smith Fund from KU Endowment Association. Disclosure of Interest None declared DOI 10.1136/annrheumdis-2014-eular.2639

Background Synovial tissue from patients with osteoarthritis (OA) demonstrated synovial inflammation (Benito et al.). We hypothesized that a single intraarticular (IA) injection of an anti-TNF drug would result in decreased inflammatory cell infiltration and ultimately reduce articular injury. Objectives Evaluate the short-term benefit of IA anti-TNF therapy in knee osteoarthritis. Methods This study was a single center, 2:1:1 [INF:MP:P] randomization, double-blind, placebo-controlled treatment of knee OA with IA treatments of infliximab (INF) 100 mg, methylprednisolone (MP) 80 mg or saline (P) on Day 0. Subjects (n=16) had to have knee pain and show minimal to moderate osteoarthritic change on plain radiographs. Closed needle synovial biopsies (Bx) were obtained on Days 0 and 28. Total WOMAC was determined at Days 0, 14, 28 and 56. Bivariate associations were investigated using logistic and linear regression. Results Total WOMAC score improved significantly only for Group INF, comparing baseline to Day 56 (p Conclusions In this pilot study the Total WOMAC score showed significant improvement by Day 56 (p References Benito MJ, et al. Synovial tissue inflammation in early and late osteoarthritis. Ann Rheum Dis 2005;64:1263-7. Disclosure of Interest H. Lindsley Grant/Research support from: Janssen, J. Schue: None Declared, O. Tawfik: None Declared, R. Bolce: None Declared, D. Smith: None Declared, G. Hinson: None Declared, J. Wick: None Declared

The Western Commercial Space Center, Inc. (WCSC), a nonprofit corporation designated by California as the ‘‘California Spaceport Authority,’’ fosters the development of launch, manufacturing, education, and research operations related to space. After receiving recognition as the state’s authority on commercial space, WCSC oversees development of the California SpaceportTM on land leased from the Air Force at Vandenberg Air Force Base, California. When completed in 1997, the California SpaceportTM will support launches from nine different vehicle configurations on a common launch pad. This paper discusses WCSC efforts to establish the commercial space infrastructure in California.

We determined the ability of proinflammatory cytokines to enhance ICAM-1 (CD54) expression on, and PBMC adhesion to, human synoviocytes. Surface molecules were characterized by cell ELISA and by flow cytometry. Adhesion of PBMC to synoviocyte monolayers was measured by direct counting or by colorimetric staining. Most cytokines upregulated ICAM-1 expression (IL-1 betaTNF alphaIFN-gammaPDGF-bb, IL-6), but not GM-CSF or TGF beta. A similar concentration-dependent increase was observed for synoviocytes derived from patients with rheumatoid or osteoarthritis. Kinetic studies of ICAM-1 expression differed among several cytokines: an early rise with IL-1 beta or TNF alpha stimulation, a gradual increase with IFN-gamma, a transient increase with PDGF-bb, and a plateau with IL-6. Adhesion of PBMC to synoviocytes was increased by IL-1 beta or TNF alpha and reduced by MAb to CD54 or CD18. Increased synoviocyte adhesiveness may promote interactions with infiltrating inflammatory cells.

The capacity of synoviocytes to participate in inflammatory responses may be altered by the cytokine-enhanced expression of adhesion molecules such as intercellular adhesion molecule-1 (ICAM-1). To examine this possibility, the ability of selected cytokines to enhance ICAM-1 expression was examined. The data indicated that each of these cytokines (interleukin-1 beta greater than tumor necrosis factor-alpha, interferon-gamma much greater than interleukin-6) can up-regulate synoviocyte ICAM-1 expression. This can potentially increase the ability of these cells to interact with infiltrating inflammatory cells, thereby propagating immunologically mediated inflammation such as occurs in rheumatoid synovitis.

Human synoviocytes were stimulated for 48 hours or 72 hours with cytokines (recombinant interleukin-1 beta and/or recombinant tumor necrosis factor alpha) in the presence or absence of selected nonsteroidal antiinflammatory drugs. The drugs were tested at subtherapeutic and clinically therapeutic levels, as follows: piroxicam 0.3 pM to 3 X 10(6) pM, sodium salicylate 30 nM to 3 X 10(6) nM, and indomethacin 3 pM to 3 X 10(6) pM. At low concentrations, all 3 drugs showed significant enhancement of prostaglandin E2 (PGE2) secretion (piroxicam, salicylate greater than indomethacin). At high, clinically therapeutic concentrations, all drugs showed suppression of PGE2 secretion (indomethacin greater than piroxicam greater than salicylate). The enhancement of PGE2 secretion may be partially responsible for the flare of arthralgia or arthritis frequently seen after termination of drug therapy.

Mast cells (MC) are well known for their contribution to vascular inflammation and atherosclerosis. We have shown that MC proteases and histamine amplify lipopolysaccharide (LPS)-induced inflammatory responses in human coronary artery endothelial cells via stimulation of TLR4 expression. This study examined mRNA levels of COX1, COX2, TLR4 and eNOS in aortic tissues of MC-deficient (W/Wv) mice and wild type controls (WT) to gain insight into the role of MC in vascular pathophysiology. We also determined the effect of MC degranulation on gene expression in aortas 24 h after intraperitoneal injection of MC degranulation agent compound 48/80 (1 μg/g body wt). We found that, in comparison to WT, aortic tissues of W/Wv mice express lower levels of COX1, COX2, TLR4 and eNOS mRNA. Injection of compound 48/80 resulted in marked stimulation of expression of these genes in aortas of WT, but not in W/Wv mice, confirming the involvement of MC degranulation products in this process. To determine whether a deficiency of MC and/or decreased COX expression in W/Wv mice is associated with changes in prostanoid homoeostasis, the levels of prostacyclin and thromboxane A2 metabolites in urine samples were analyzed. The levels of 6-keto PGF1α and 11-dehydro TXB2 in 24 h urine of both genotypes were comparable when normalized for the volume and creatinine excretion. These results suggest that, although MC deficiency suppresses the expression of COX1 and COX2 genes in the vasculature, the systemic prostanoid homeostasis remains protected. (Supported by NIH R01-HL070101 and Carey Arthritis Fund)

Fecal samples from 25 Anolis armouri, 2 Anolis bahorucoensis, 48 Anolis cybotes, and 21 Anolis olssoni (Lacertilia: Polychrotidae) from southern Hispaniola were examined for coccidian oocysts. Two eimerians and 2 isosporans are herein described as new species. Sporulated oocysts of Eimeria schwartzi n. sp. from A. armouri are ellipsoidal, 22.7 (20.8-25.0) x 15.7 (14.6-17.7) microns, with spherical to subspherical sporocysts, 7.9 (6.2-9.4) x 7.4 (6.2-8.3) microns. Sporulated oocysts of Isospora reui n. sp. from A. bahorucoensis are spherical to subspherical, 18.2 (15.6-20.0) x 17.8 (15.6-19.8) microns, with ovoid sporocysts, 11.9 (10.4-12.7) x 8.5 (7.5-9.4) microns. Sporulated oocysts of Isospora hendersoni n. sp. from A. armouri and A. cybotes are spherical to subspherical, 23.2 (20.8-26.0) x 21.1 (18.4-23.9) microns, with ellipsoidal sporocysts, 14.7 (12.5-15.6) x 10.0 (9.2-11.4) microns. Sporulated oocysts of Eimeria avilae n. sp. from A. olssoni are cylindrical, 29.3 (26.0-33.3) x 15.9 (13.5-18.9) microns, with ellipsoidal sporocysts 10.2 (9.4-11.4) x 6.8 (5.2-8.0) microns.

WASSERSUG, R. J. 1984. The Pseudohemisus tadpole: a morphological link between microhylid (Orton Type 2) and Ranoid (Orton Type 4) larvae. Herpetologica 40(2):138-149. 1989. What, if anything is a microhylid (Orton type II) tadpole? Progr. Zool. 35:534-538. , AND W. F. PYBURN. 1987. The biology of the Pe-ret' toad, Otophryne robusta (Microhylidae), with special consideration on its fossorial larva and systematic relationships. Zool. J. Linn. Soc. 91:137169.

Estimates are obtained of the fraction of ingested or inhaled 239+240Pu transferred to blood and tissues of a reproducing herd of beef cattle, individuals of which grazed within fenced enclosures for up to 1064 d under natural conditions with no supplemental feeding at an arid site contaminated 16 y previously with Pu oxide. The estimated (geometric mean [GM]) fraction of Pu transferred from the gastrointestinal tract to blood serum was about 5 x 10(-6) (geometric standard error [GSE] = 1.4) with an approximate upper bound of about 2 x 10(-5). These results are in reasonable agreement with the value of 1 x 10(-5) recommended for human radiation protection purposes by the International Commission on Radiological Protection (ICRP) for insoluble Pu oxides that are free of very small particles. Also, results from a laboratory study by Stanley (St75), in which large doses of 238Pu were orally administered daily to dairy cattle for 19 consecutive days, suggest that aged 239+240Pu at this arid grazing site may not be more biologically available to blood serum than fresh 239+240Pu oxide. The estimated fractions of 239+240Pu transferred from blood serum to tissues of adult grazing cattle were: femur (3.2 X 10(-2), 1.8; GM, GSE), vertebra (1.4 X 10(-1), 1.6), liver (2.3 X 10(-1), 2.0), muscle (1.3 X 10(-1), 1.9), female gonads (7.9 X 10(-5), 1.5), and kidney (1.4 X 10(-3), 1.7). The blood-to-tissue fractional transfers for cattle initially exposed in utero were greater than those exposed only as adults by a factor of about 4 for femur (statistically significant) and of about 2 for other tissues (not significant). The estimated (GM) fraction of inhaled Pu initially deposited in the pulmonary lung was 0.34 (GSE = 1.3) for adults and 0.15 (GSE = 1.3) for cattle initially exposed in utero (a statistically significant difference), which may be compared with the expected fraction of 0.11 at the study site using the ICRP lung model for humans.

Opossums (Didelphis marsupialis), act as intermediate hosts for Besnoitia darlingi and could be infected orally with sporozoites (oocysts) and bradyzoites (tissue cysts), or intraperitoneally (i.p.) with tachyzoites. Infections could presumably be transmitted through cannibalism. Cats (Felis catus), the definitive host, could be infected only with bradyzoites but not sporozoites. Oocysts shed by cats measure about 12 × 12 μm, resemble similarly sized oocysts of Toxoplasma gondii and Hammondia hammondi, and must be differentiated by the appearance of tissue cysts after experimental infection of intermediate hosts. Cats did not form tissue cysts of B. darlingi. Tachyzoites from the related B. jellisoni could be used in the Sabin-Feldman dye test to determine the development of antibody to B. darlingi in opossums after infection.

Members of the family Sarcocystidae, as defined by Frenkel, have had a complicated history, principally due to the existence of both coccidial and cystic stages. This formerly clandestine relationship resulted in dual or partial designations of nomenclature for individual species. The problem was further compounded by the obligatory heteroxeny of several of the genera, making it impossible to transmit the parasites from one individual to another of a single host. As a result, oocysts similar in appearance, though from hosts separated taxonomically up to the familial level, were sometimes considered to be identical. Discoveries within the last decade have generated much interest and some understanding. Current studies of these and other coccidia should emphasize complete cyclical transmissions with cognizance of potential heteroxeny with the production of tissue cysts in intermediate hosts.

The role of the cat (Felis domestica) as a definitive host for Sarcocystis cuniculi of European rabbits (Oryctolagus cuniculus) was confirmed. It was shown that after dosing with sporocysts from cats, rabbits developed sarcocysts and these became infective for cats at not less than 93 days post-infection (p.i.). The earliest infection detected was at 142 days p.i. Infected muscle from an experimental rabbit did not transmit Sarcocystis when fed to other rabbits. Microscopically, sarcocysts in European rabbits (O. cuniculus) were morphologically indistinguishable from those in cottontail rabbits (Sylvilagus floridanus).

Follicle stimulating hormone (FSH) will induce red-orange pigmentation in non-ovariectomized female leopard lizards (Crotaphytus wislizenii) in 7-21 days depending upon the season. Progesterone is judged to be an important hormone in the process of color change, and will bring about pigmentation in the absence of ovarian tissue. Estrogen alone has no apparent effect on color change, but when administered before progesterone it appears to have a priming effect.

Arm-to-tongue circulation times in conjunction with a Valsalva maneuver at 40 mm Hg for 15 seconds were determined in 189 young men. In 112 subjects the 20% sodium dehydrocholate indicator was injected just as the Valsalva maneuver was begun. Circulation times ranged from 13 to 31 seconds. The frequency-distribution was tetramodal, means 14.6 ± 0.25 seconds, 18.8 ± 0.17 seconds, 24.2 ± 0.22 seconds and 29.5 ± 0.37 seconds; within the respective modes lay 9.8%, 42.9%, 35.7% and 11.6% of the sample. In 77 subjects the indicator was not injected until an expiratory pressure of 40 mm Hg had been attained. The range was 14–33 seconds and the frequency-distribution bimodal, means at 14.7 ± 0.18 seconds (9.1%) and 24.6 ± 0.40 seconds (90.9%). It is possible that the lowest mode in both cases may represent subjects with large patent foramina ovalia. The tetramodal nature of the first distribution is probably consequent upon differences in the manner in which the Valsalva maneuver was initiated. Submitted on November 7, 1957

A delayed-speech case was present in which the patient, a male with fair skin, blonde hair and blue eyes was given a histidine loading test. The patient was about two and a half when the test was administered. The test yielded results consistent with increased histidine. The patient was not producing any consonant sounds at this age and only limited vowel sounds. At the age of three, the patient was placed on a low histidine diet. After being on the diet for a month another test was administered and the results showed no appreciable level of histidine. During the next few months the patient appeared to develop more interest in producing words and more consonant sounds are now being used. More spontaneous words were produced and his overall language development has improved. He is enrolled in a language development program and his progress has been satisfactory.

Two of five opossums examined from the Kansas City area were found to be infected with Besnoitia darlingi. Inoculation of cysts resulted in acute lethal infections in mice and hamsters. Chronic infections with development of tissue cysts were obtained in mice by prophylaxis with sulfadiazine. Cysts were fed to cats and a dog which resulted in the shedding of isosporoid oocysts by cats. The prepatent period was 11 to 14 days. Unsporulated oocysts averaged 11.9 by 12.3 micrometer and sporulated in 48 to 72 hr. Mouse-to-mouse tramsmission was achieved by injecting triturated tissue containing cysts. This is the third species of Besonitia to be cyclically transmitted by cats.

ABSTRACr: Development of immunity to the shedding of oocysts was examined in 75 kittens that survived infection with the three stages of Toxoplasma gondii. Of 16 kittens fed bradyzoites in cysts, 94% were immune and did not shed oocysts. Of seven injected with tachyzoites 86% were immune. Of 18 fed sporozoites only 11% were immune, but following injection, 54% of 12 were immune. After the administration of either bradyzoites or tachyzoites from nonoocyst-producing strains, only 9% of 22 were immune. Considering all inocula, immunity was present in 93% of kittens that had previously shed oocysts, 25% of those that only develop antibody, and none that had neither shed nor developed an antibody titer. After a second challenge with a different isolate, a similar percentage of immunity was observed. Infection with killed tachyzoites, alone, or together with Freund's complete or incomplete adjuvants was followed by immunity in only one of 24 kittens. Eighty-five percent of 13 kittens were immune, after they had been treated prophylactically with 200 mg/kg monensin, or 60 mg/kg cat sulfadiazine combined with 1 mg/kg cat of pyrimethamine; oocyst shedding had been suppressed in all. It is concluded that cats can be immunized against oocyst shedding by infections where oocysts are produced, or where developmental stages are suppressed by chemoprophylaxis, but not if enteroepithelial stages are absent, as in the oocyst-less strain examined.

To assess the roles of the German cockroach (Blatella germanica) and the American cockroach (Periplaneta americana) in the transmission of Sarcocystis muris and of 3 other coccidia of cats-Toxoplasma gondii, Isospora felis, and Isospora rivolta, cockroaches exposed to feces containing these coccidia were periodically fed to mice, as was a portion of the fecal matter. Sarcocystis muris sporocysts, which in feces remained infectious for at least 20 days, were also transmitted to mice by P. americana for at least 20 days and by B. germanica for 5 days after exposure to infectious feces. Toxoplasma gondii oocysts were transmitted by P. americana intermittently up to 10 days, but by B. germanica only immediately after exposure to feces. Oocysts of 2 species of Isospora, when associated with fecal matter, remained infectious for 20 days. Those of I. rivolta were transmitted by both cockroach species for 10 days, but I. felis was transmitted only by by B. germanica, and for only 2 days.