Reduction In Severity Of Lesions Research Articles

SKIN NEEDLING THERAPY IS AN EFFECTIVE TREATMENT FOR ATROPHIC FACIAL ACNE SCARS

Background
 Acne is a common condition seen in up to 80% of people between 11 and 30 years of age. In some patients, the severe inflammatory response results in permanent scars. Atrophic facial scars are always a challenge to treat. Skin needling therapy is a new, simple and an effective treatment for such scars.
 Objective
 To assess the ef cacy of skin needling in the management of grade 2 and 3 atrophic facial acne scars.
 Patients and Methods
 Twenty five patients with facial acne scars were enrolled in the study (17 female and 8 male patients; age range 14–42 years). The severity of the lesions in each patient was scored on a 10-point scale (0 = no lesions; 10 = maximum severity) by the researcher involved in the study. Three groups of patients were identified and patients with Grade 2 and 3 atrophic facial acne scars were included. Patients were treated with skin needling procedure. Four photographs were taken for each patient by the researcher; one before, one immediately after the procedure done and another one after one month from the first session then one month after the second session of the treatment. At every follow-up, the same researcher evaluated the scars, scoring them using the same scale as previously, to assess any clinical improvement in the severity of the lesions. The last follow-up was conducted one month after the second treatment then the photographs were compared with the photographs taken before the first treatment and each patient was given a new severity score.
 Results
 One month after the rst session of skin needling; all patients had smoother facial skin and a slight reduction in lesion severity. One month after the second session of skin needling, the improvement in the acne rolling scars was evident as photographic comparison in each group of patients showed that the skin became thicker; and independent of the lesion grading the relative rolling scar depth was significantly reduced
 Conclusion
 Skin needling therapy is a simple, cheap, office method and an effective treatment for atrophic facial acne scars.

Ketorolac salt is a newly discovered DDX3 inhibitor to treat oral cancer

DDX3 belongs to DEAD box RNA helicase family and is involved in the progression of several types of cancer. In this work, we employed a High Throughput Virtual screening approach to identify bioactive compounds against DDX3 from ZINC natural database. Ketorolac salt was selected based on its binding free energy less than or equals to −5 Kcal/mol with reference to existing synthetic DDX3 inhibitors and strong hydrogen bond interactions as similar to crystallized DDX3 protein (2I4I). The anti-cancer activity of Ketorolac salt against DDX3 was tested using oral squamous cell carcinoma (OSCC) cell lines. This compound significantly down regulated the expression of DDX3 in human OSCC line (H357) and the half maximal growth inhibitory concentration (IC50) of Ketorolac salt in H357 cell line is 2.6 µM. Ketorolac salt also inhibited the ATP hydrolysis by directly interacting with DDX3. More importantly, we observed decreased number of neoplastic tongue lesions and reduced lesion severity in Ketorolac salt treated groups in a carcinogen induced tongue tumor mouse model. Taken together, our result demonstrates that Ketorolac salt is a newly discovered bioactive compound against DDX3 and this compound can be used as an ideal drug candidate to treat DDX3 associated oral cancer.

Therapeutic role of TLR9 agonist in the atherosclerosis

Background: Atherosclerosis is driven by inflammatory reactions that are shared with the innate immune system. Toll-like receptor-9 (TLR9) is an intracellular pattern recognition receptor of the innate immune system that is currently under clinical investigation as a therapeutic target in inflammatory diseases. Here we investigated the role of TLR9 in atherosclerosis. Methods and Results: Newly generated double-knockout ApoE−/−:TLR9−/− mice and control ApoE−/− mice were fed a high-fat diet from 8 weeks of age and effects on lesion size, cellular composition, inflammatory status, and plasma lipids were assessed after 8, 12, 15 and 20 weeks. All four time points demonstrated exacerbated atherosclerotic lesion severity in ApoE−/−:TLR9−/− mice, with a corresponding increase in lipid deposition and accumulation of macrophages, dendritic cells and CD4+ T cells. Although ApoE−/−:TLR9−/− mice exhibited an increase in plasma VLDL/LDL cholesterol, the VLDL/LDL:HDL ratio was unaltered because of a parallel increase in plasma HDL cholesterol. As a potential mechanism accounting for plaque progression in ApoE−/−:TLR9−/− mice, CD4+ T cell accumulation was further investigated and depletion of these cells in ApoE−/−:TLR9−/− mice significantly reduced lesion severity. As a final translational approach, administration of a TLR9 agonist (type B CpG ODN) to ApoE−/− mice resulted in a reduction of lesion severity. Conclusions: Genetic deletion of the innate immune receptor TLR9 exacerbated atherosclerosis in ApoE−/− mice. CD4+ T-cells were identified as potential mediators of this effect. A type B CpG ODN TLR9 agonist reduced lesion severity, thus identifying a novel therapeutic approach in atherosclerosis.

Enhanced Efficacy of Cidofovir Combined with Vaccinia Immune Globulin in Treating Progressive Cutaneous Vaccinia Virus Infections in Immunosuppressed Hairless Mice

The treatment of progressive vaccinia in individuals has involved antiviral drugs, such as cidofovir (CDV), brincidofovir, and/or tecovirimat, combined with vaccinia immune globulin (VIG). VIG is costly, and its supply is limited, so sparing the use of VIG during treatment is an important objective. VIG sparing was modeled in immunosuppressed mice by maximizing the treatment benefits of CDV combined with VIG to determine the effective treatments that delayed the time to death, reduced cutaneous lesion severity, and/or decreased tissue viral titers. SKH-1 hairless mice immunosuppressed with cyclophosphamide and hairless SCID mice (SHO strain) were infected cutaneously with vaccinia virus. Monotherapy, dual combinations (CDV plus VIG), or triple therapy (topical CDV, parenteral CDV, and VIG) were initiated 2 days postinfection and were given every 3 to 4 days through day 11. The efficacy assessment included survival rate, cutaneous lesion severity, and viral titers. Delays in the time to death and the reduction in lesion severity occurred in the following order of efficacy: triple therapy had greater efficacy than double combinations (CDV plus VIG or topical plus parenteral CDV), which had greater efficacy than VIG alone. Parenteral administration of CDV or VIG was necessary to suppress virus titers in internal organs (liver, lung, and spleen). The skin viral titers were significantly reduced by triple therapy only. The greatest efficacy was achieved by triple therapy. In humans, this regimen should translate to a faster cure rate, thus sparing the amount of VIG used for treatment.

Co-infection with HPV types from the same species provides natural cross-protection from progression to cervical cancer.

BackgroundThe worldwide administration of bivalent and quadrivalent HPV vaccines has resulted in cross-protection against non-vaccine HPV types. Infection with multiple HPV types may offer similar cross-protection in the natural setting. We hypothesized that infections with two or more HPV types from the same species, and independently, infections with two or more HPV types from different species, associate with protection from high-grade lesions.FindingsWe recruited a cohort of 94 HIV, HPV-positive women from Botswana, with Grade 2 or higher cervical intraepithelial neoplasia. Infections with 2 or more HPV types from a single species associated with reduced lesion severity in univariate analysis (OR = 0.41, 95% CI 0.18-0.97, p = 0.042), when adjusted for the presence of HPV 16 or 18 types (OR = 0.41, 95% CI 0.17-1.00, p = 0.049), or all high-risk HPV type infections (OR = 0.38, 95% CI 0.16-0.90, p = 0.028). Infections with 2 or more HPV types from different species did not associate (OR = 0.68, 95% CI 0.25-1.81, p = 0.435).ConclusionsOur findings show that co-infections with genetically similar HPV types reduce the likelihood of progression to high-grade lesions in HIV positive women, an effect not observed in co-infections with taxonomically different HPV types. This observation is possibly caused by an immune cross-protection through a similar mechanism to that observed after HPV vaccination.

<b>Effects of wounding, humidity, temperature, and inoculum concentrations on the severity of corky dry rot caused by <i>Fusarium semitectum</i> in melon fruits

Corky dry rot, caused by Fusarium semitectum, is the main postharvest disease of melons in Brazil. This study investigated the effects of wounding, humidity, temperature, and inoculum concentration on the severity of corky dry rot under controlled conditions. Cantaloupe and honeydew melon types were inoculated by spraying conidial suspensions of three F. semitectum isolates. In all experiments, the tested F. semitectum isolates did not differ in relation to disease severity, but, the cantaloupe melon showed higher levels of severity. No lesions appeared on fruits that lacked wounds, and increasing wound age reduced lesion severity. Melons that were inoculated with F. semitectum developed symptoms regardless of the presence or absence of a moist chamber at the post-inoculation stage, but the lesions were larger under moist chamber conditions. There were no symptoms at 10°C, but a temperature increase from 15 to 25°C resulted in a disease severity increase. The largest lesions were observed when both melon types were inoculated with a concentration of 106 conidia mL-1, but even the lowest concentration (101 conidia mL-1) was sufficient for causing lesions. Injury reduction and/or the acceleration of melon healing, as well as environmental variable control and a reduction of inoculum sources, are essential to reducing corky dry rot severity.

Protective Role for Toll-Like Receptor-9 in the Development of Atherosclerosis in Apolipoprotein E–Deficient Mice

Atherosclerosis is driven by inflammatory reactions that are shared with the innate immune system. Toll-like receptor-9 (TLR9) is an intracellular pattern recognition receptor of the innate immune system that is currently under clinical investigation as a therapeutic target in inflammatory diseases. Here, we investigated whether TLR9 has a role in the development of atherosclerosis in apolipoprotein E-deficient (ApoE(-/-)) mice. Newly generated double-knockout ApoE(-/-):TLR9(-/-) mice and control ApoE(-/-) mice were fed a high-fat diet from 8 weeks and effects on lesion size, cellular composition, inflammatory status, and plasma lipids were assessed after 8, 12, 15, and 20 weeks. All 4 time points demonstrated exacerbated atherosclerotic lesion severity in ApoE(-/-):TLR9(-/-) mice, with a corresponding increase in lipid deposition and accumulation of macrophages, dendritic cells, and CD4(+) T cells. Although ApoE(-/-):TLR9(-/-) mice exhibited an increase in plasma very low-density lipoprotein/low-density-lipoprotein cholesterol, the very low-density lipoprotein/low-density lipoprotein:high-density lipoprotein ratio was unaltered because of a parallel increase in plasma high-density lipoprotein cholesterol. As a potential mechanism accounting for plaque progression in ApoE(-/-):TLR9(-/-) mice, CD4(+) T-cell accumulation was further investigated and depletion of these cells in ApoE(-/-):TLR9(-/-) mice significantly reduced lesion severity. As a final translational approach, administration of a TLR9 agonist (type B CpG oligodeoxynucleotide 1668) to ApoE(-/-) mice resulted in a reduction of lesion severity. Genetic deletion of the innate immune receptor TLR9 exacerbated atherosclerosis in ApoE(-/-) mice fed a high-fat diet. CD4(+) T cells were identified as potential mediators of this effect. A type B CpG oligodeoxynucleotide TLR9 agonist reduced lesion severity, thus identifying a novel therapeutic approach in atherosclerosis.

Platelet-Leukocyte Conjugation Mediated By GPIbα-αXβ2 (CD11c/CD18) Interaction

BackgroundThe formation of platelet-leukocyte conjugates has been implicated in a variety of pathologies, including thrombosis, atherosclerosis, glomerulonephritis, multiple sclerosis and other inflammatory diseases. In prior studies (Simon et al., J. Exp Med, 2000, 193-204), platelet GPIbα and leukocyte αMβ2 (CD11b/CD18) had been implicated as receptor:counter-receptor pairing in mediating formation of platelet:leukocyte complexes. A specific segment (αM201-217) in the I domain of αM (CD11b) subunit of αMβ2 engaged GPIbα, and a peptide (M2) corresponding to this segment blocked neutrophil recognition of platelets and anti-M2 blocked conjugate formation (Ehlers et al., J. Exp. Med, 2003, 1077-88). The I-domains of αM and αX are highly conserved (74% homology), but αX has different amino acids at two positions shown to be critical for recognition of GPIbα by the αM I-domain. This study considers the role of a particular receptor:counter-receptor pairing, GPIb on platelets and αXβ2 (CD11c/CD18) on leukocytes, in mediating this heterotypic cell interaction. Methods and ResultsTo directly address whether αXβ2 recognizes GPIbα, αXβ2 was expressed in HEK cells and its corresponding I-domain was expressed and purified. αXβ2 cells adhered specifically to GPIbα, and purified αX I-domain bound to platelets by flow cytometry and to GPIbα by surface plasmon resonance. However, this interaction was readily distinguished from that of αM I-domain binding to GPIbα. αX binding to GPIbα was not inhibited by: 1) M2 or anti-M2; 2) a peptide from a region of GPIbα implicated in vWF interaction and that blocked adhesion of αMβ2 cells to GPIbα; 3) small molecules that blocked αM I-domain binding; and 4) mutations in GPIbα that blocked αM I-domain binding. Furthermore, by mutational analyses, a region of αX I domain was implicated binding GPIbα that is structurally distant and unrelated to the αM I domain binding site.Next, we examined the role of αXβ2 in vivo in a cytokine- and neutrophil-dependent inflammatory response. The local Shwartzman-like reaction (LSR), induced by successive LPS and cytokine injections into the skin, produces a thrombo-hemorrhagic vasculitis, which depends on leukocytes and their interaction with activated platelets and endothelial cells. In prior work, we have shown that the LSR is dependent upon leukocyte αM and platelet GPIba. Leukocyte-derived microparticles promote the accumulation of fibrin, generation of occlusive thrombi, and eventual hemorrhage from inflamed blood vessels. The LSR produces hemorrhage in the intact skin 24 hours after TNF-a injection. CD11c-/- mice subjected to LSR exhibited a significant reduction in the development of hemorrhagic lesions. In analysis blinded to genotype, CD11c-/- mice had a 76% reduction in lesion severity (lesion hemorrhage score: WT, 3.4 ± 1.0, n=12 vs. CD11c-/-, 0.8 ± 1.0, n=11; P<0.001). Histopathology of vasculitic lesions demonstrated substantial reductions in erythrocyte extravasation, edema, leukocyte accumulation, and occlusive thrombi in CD11c-/- mice. ConclusionsLeukocyte αXβ2 (CD11c/CD18) recognizes GPIbα on platelets in a way distinct from αMβ2 (CD11bCD18). This interaction appears to be functionally significant in vivo and could be targeted for suppressing adverse inflammatory responses. Disclosures:Simon:Cordis/J&J: Consultancy; Janssen/J&J: Consultancy; Medtronic Vascular: Consultancy; Merck: Consultancy; Medtronic Foundation: Research Funding.

MBX-500 Is Effective for Treatment of Clostridium difficile Infection in Gnotobiotic Piglets

The novel antibiotic MBX-500, dosed at 100, 200, or 400 mg/kg twice daily for 7 days, was evaluated for the treatment of Clostridium difficile infection (CDI) in the gnotobiotic pig model. MBX-500 increased survival at all doses and at high doses improved clinical signs and reduced lesion severity, similar to vancomycin. Our results show that MBX-500 is an effective antibiotic for the treatment of diarrhea associated with CDI and prevents severe systemic disease.

Intranasal delivery of pGDNF nanoparticles provides neuroprotection in the rat 6‐hydroxydopamine model of Parkinson's disease

Glial cell line‐derived neurotrophic factor (GDNF) has been shown to be neuroprotective toward substantia nigra (SN) dopamine neurons in animal models of Parkinson's disease (PD) and in some clinical trials. However, its delivery to brain has required invasive surgical routes that are untenable for many patients with PD. The goal of these experiments was to test whether intranasal delivery of a GDNF expression plasmid could protect dopamine cells in the rat 6‐hydroxydopamine (6‐OHDA) model of PD. If successful, intranasal delivery of pGDNF could provide a renewable source of GDNF within the brain without need for surgical injections or frequent re‐dosing.Rats were given intranasal saline, naked pGDNF or pGDNF in a PEGylated polylysine nanoparticle (NP) preparation 7 days prior to receiving a unilateral 6‐OHDA lesion. Three weeks after 6‐ OHDA, rotational behavior to 5 mg/kg d‐amphetamine was assessed, followed by sacrifice. Tyrosine hydroxylase (TH) immunohistochemistry revealed a significant reduction in lesion severity in the SN of rats given intranasal pGDNF NPs versus those given naked pGDNF or saline. In addition, rotational behavior was reduced in rats treated with pGDNF NPs. These results demonstrate the utility of intranasal delivery of pGDNF NPs as a non‐invasive means of gene therapy for PD and possibly other diseases of the brain. Support provided by: 2011–12 Northeastern University Tier 1 Seed Grant and the M.J. Fox Foundation.

Evaluation of homologous and heterologous protection induced by a virulent field strain of orf virus and an orf vaccine in goats

To evaluate cross protection provided by administration of contagious ecthyma vaccines against strains of orf virus in goats. 126 Boer-Spanish crossbred goats (3 to 20 days old). 85 goats were vaccinated with a goat-derived contagious ecthyma vaccine. Of these, 41 were challenge exposed with the virus strain for the contagious ecthyma vaccine, 40 were challenge exposed with a more virulent field strain of orf virus, and 4 were lost to predation or died. Another 41 goats were vaccinated with a vaccine produced from a more virulent field strain of orf virus; of these, 18 were challenge exposed with the virus strain of the goat-derived contagious ecthyma vaccine, 18 were challenge exposed with the more virulent field strain of orf virus, and 5 were lost to predation or died. Vaccination with the goat-derived contagious ecthyma vaccine did not significantly reduce the number of goats with lesions or lesion severity caused by challenge exposure with the more virulent field strain of orf virus. Vaccination with the vaccine produced from the more virulent field strain of orf virus significantly reduced the number of goats with lesions attributable to challenge exposure with the virus strain of the goat-derived contagious ecthyma vaccine, but it failed to significantly reduce lesion severity. Vaccination did not result in cross protection for the 2 strains of orf virus. This may have been attributable to antigenic differences and may be a factor in outbreaks of contagious ecthyma in vaccinated goats.

The value to patients of reducing lesion severity in plaque psoriasis

Plaque psoriasis is associated with significant psychosocial, quality-of-life, and economic burden. The objective of this study was to quantify the value to patients of reducing the severity and size of plaque psoriasis lesions. Subjects included individuals with a self-reported diagnosis of plaque psoriasis from a nationally representative US household panel. Subjects completed a web-based conjoint analysis survey and chose between hypothetical treatments in a series of questions. Each alternative was defined by lesion severity, percentage of body surface area (BSA) covered by lesions, type of treatment, injection discomfort or pain (if treatment included injections), risk of serious lung infection, and monthly out-of-pocket cost. 28,200 panelists were invited to participate. 18,330 responded, 503 qualified, and 419 completed the survey. Mean age was 54.5 years and 52% were female. 64% (35%) of patients reported psoriasis severity as mild or mild to moderate (moderate to very severe). Patients were willing to pay $486.73 out-of-pocket per month to eliminate severe lesions covering 25% BSA on the arms and legs and $444.80 out-of-pocket per month to eliminate moderate lesions covering 4% BSA on the face. Individuals with plaque psoriasis are willing to pay substantial amounts to reduce lesion severity and percentage of BSA covered by lesions.

Topical Treatment of Cutaneous Vaccinia Virus Infections in Immunosuppressed Hairless Mice with Selected Antiviral Substances

Certain nucleoside, nucleotide and pyrophosphate analogues may be useful for treating severe complications arising as a result of virus dissemination following smallpox (live vaccinia virus) vaccinations, especially in immunocompromised individuals. We used an immunosuppressed hairless mouse model to study the effects of 10 antiviral agents on progressive vaccinia infections. Hairless mice were immunosuppressed by treatment with cyclophosphamide (100 mg/kg) every 4 days starting 1 day prior to vaccinia virus (WR strain) infection of wounded skin. Topical treatments with antiviral agents were applied twice a day for 7 days starting 5 days after virus exposure. Topical 1% cidofovir cream treatment was effective in significantly reducing primary lesion severity and decreasing the number of satellite lesions. Topical 1% cyclic HPMPC and 1% phosphonoacetic acid were not quite as active as cidofovir. Ribavirin (5%) treatment reduced lesion severity and diminished the numbers of satellite lesions, but the mice died significantly sooner than placebos. 2-Amino-7-[(1,3,-dihydroxy-2-propoxy)methyl]purine (compound S2242; 1%) moderately reduced primary lesion sizes. Ineffective treatments included 5% arabinosyladenine, 1% arabinosylcytosine, 1% 5-chloro-arabinosylcytosine, 5% arabinosylhypoxanthine 5-monophosphate and 5% viramidine. Of the compounds tested, topically applied cidofovir was the most effective treatment of cutaneous vaccinia virus infections in immunosuppressed mice. Topical treatment with cidofovir could be considered as an adjunct to intravenous drug therapy for serious infections.

Water, water, everywhere: sodium and water balance and the injured brain

This review focuses on water shift and oedema in acute brain injury, with particular aspects on pathophysiology of water movements, the role of aquaporins and the potential of new therapies. This review reports on update of both significant experimental and clinical findings on factors implicated in oedema formation. The main inputs came from the demonstrated role of aquaporins (especially AQP4) in brain oedema control. The absence of aquaporin agonist or antagonist does not help to clarify the net effect of aquaporins on brain oedema. The clinical practice of osmotherapy, especially with hypertonic saline failed to improve neurological outcome in a large randomized clinical trial. Colloid treatment was not proven efficient and potentially dangerous. Some hopes might come from targeting inflammatory cascade and neurogenic mediators to reduce lesion severity and to limit the blood-brain barrier dysrupture. Water content control and partition can be better assessed in clinic with NMR helping to make decisions, but with limited proven therapies. The timing for such interventions might be crucial and future biomarkers might be very helpful.

Conditioned Pharmacotherapeutic Effects: A Preliminary Study

To test the hypothesize that psoriasis patients treated under a partial schedule of pharmacologic (corticosteroid) reinforcement would show less severe symptoms and relapse than those given the same amount of drug under standard conditions. Behavioral conditioning as an inherent component of many pharmacotherapeutic protocols has never been examined. A double-blind, simple randomization intervention was conducted with 46 patients from California and New York. Initially, lesions were treated with 0.1% acetonide triamcinolone under standard treatment conditions. Thereafter, a Standard Therapy group continued on continuous reinforcement (active drug every treatment) with 100% of the initial dose; Partial Reinforcement patients received a full dose 25% to 50% of the time and placebo medication other times; Dose Control patients received continuous reinforcement with 25% to 50% of the initial dose. Severity of disease scores in California neither supported nor refuted the hypothesis. In New York, where there was no difference between Partial Reinforcement and Dose Control groups at baseline, partial reinforcement effected a greater reduction in lesion severity than Dose Control conditions and did not differ from Standard Therapy patients receiving two to four times more drug. For the entire population, the frequency of relapse under partial reinforcement (26.7%) was lower than in Dose Control patients (61.5%) and did not differ from full-dose treatment (22.2%). A partial schedule of pharmacotherapeutic reinforcement could maintain psoriasis patients with a cumulative amount of corticosteroid that was relatively ineffective when administered under standard treatment conditions. Conceivably, corticosteroid administration only one quarter or half as frequently as currently prescribed is sufficient to treat psoriasis. We posit, however, that these preliminary observations implicate conditioning processes in-and for the design of-regimens of pharmacotherapy.

Abstract 3373: MRP-8/14 is Critical for the Biological Response to Vascular Injury

Background : MRP-8 (S100A8) and MRP-14 (S100A9) are members of the S100-family of calcium-modulated proteins that regulate myeloid cell function and control inflammation, in part, through activation of toll-like receptor-4 and RAGE. Using a transcriptional profiling approach in patients with acute coronary syndromes, we made the novel discovery that MRP-14 is associated with atherothrombosis, and demonstrated that elevated plasma levels of MRP-8/14 heterodimer predict increased risk of first and recurrent cardiovascular events. Despite functioning as a pro-inflammatory mediator, the role of MRP-8/14 in vascular inflammation and disease are unknown. Methods and Results : We evaluated vascular inflammation in wild-type (WT) and MRP-14-deficient (MRP-14 −/− ) mice that lack MRP-8/14 complexes with experimental arterial injury, vasculitis, or atherosclerosis. After femoral artery wire injury, MRP-14 −/− mice had a 33% reduction in leukocyte accumulation ( P =.0004) and a 72% reduction in cellular proliferation ( P =0.001) in the arterial wall followed by a 45% reduction in neointima formation ( P =0.004) compared to WT mice. In a cytokine-induced local Schwartzman-like reaction that produces thrombohemorrhagic vasculitis, MRP-14 −/− mice had a 59% reduction in lesion severity ( P &lt;0.0001), an 82% smaller hemorrhagic area ( P =0.015), and a 45% decline in neutrophil accumulation ( P =0.013). Finally, in response to high-fat feeding, mice doubly deficient in ApoE and MRP-8/14 complexes had attenuation in atherosclerotic lesion area by 34% ( P =0.023) and a 60% decrease in macrophage accumulation in plaques ( P =0.021) compared to mice deficient in ApoE alone. Conclusions: This study demonstrates that MRP-8/14 broadly regulates vascular inflammation and contributes to the biological response to vascular injury. Targeting MRP-8/14 may provide a novel therapeutic approach to modulate diverse forms of vascular injury, including restenosis, vasculitis, and atherosclerosis.

Mannitol biosynthesis is required for plant pathogenicity byAlternaria alternata

Mannitol has been hypothesized to play a role in antioxidant defense. In previous work, we confirmed the presence of the two mannitol biosynthetic enzymes, mannitol dehydrogenase (MtDH) and mannitol 1-phosphate 5-dehydrogenase (MPDH), in the fungus Alternaria alternata and created disruption mutants for both enzymes. These mutants were used to investigate the role of mannitol in pathogenicity of A. alternata on its host, tobacco. Conidia of all mutants were viable and germinated normally. GC-MS analysis demonstrated elevated levels of trehalose in the mutants, suggesting that trehalose may substitute for mannitol as a storage compound for germination. Tobacco inoculation showed no reduction in lesion severity caused by the MtDH mutant as compared with wild type; however, the MPDH mutant and a mutant in both enzymes caused significantly less disease. Microscopy analysis indicated that the double mutant was unaffected in the ability to germinate and produce appressoria on tobacco leaves and elicited a defense response from the host, indicating that it was able to penetrate and infect the host. We conclude that mannitol biosynthesis is required for pathogenesis of A. alternata on tobacco, but is not required for spore germination either in vitro or in planta or for initial infection.

Effects of high hydrostatic pressure on Eimeria acervulina pathogenicity, immunogenicity and structural integrity

Eimeria acervulina is a protozoan parasite that can cause intestinal lesions and reduced weight gain in chickens. E. acervulina oocysts were treated by high hydrostatic pressure and evaluated for pathogenicity, immunogenicity , and structural integrity. Pressure treatment of E. acervulina oocysts at 550 MPa for 2 min at 4, 20 or 40 °C rendered the parasites nonpathogenic to chickens. Pressure treatment at 40 °C also prevented fecal shedding of oocysts. Upon challenge with non-pressurized E. acervulina oocysts, partial immunity was observed with a reduction in lesion severity in chickens that had been inoculated with pressure-treated oocysts. No changes to the fragility and permeability of the oocyst wall or excystation of sporocysts were observed as a result of pressure treatment. Light and scanning electron microscopy revealed no changes to the whole oocyst or sporocysts. Recovery and the morphology of excysted sporozoites were altered by pressure treatment. These results suggest that pressure affects sporozoite integrity. High-hydrostatic pressure processing has been shown to inactivate various microorganisms and is utilized commercially for enhanced food safety and quality. Some pathogenic microorganisms have been inactivated by HPP yet retain immunogenic properties suggesting potential application for vaccine development. Eimeria acervulina is a poultry pathogen for which new vaccines are sought. E. acervulina is also closely related to Cyclospora cayetanensis , a foodborne human pathogen. HPP was explored for effect on E. acervulina for potential vaccine development for chickens and for insight on HPP effects on parasites for enhanced safety of human foods.

IL-7 Is a Critical Factor in Modulating Lesion Development in Skn-Directed Autoimmunity

In a murine model of autoimmunity targeted against the epidermal cell Ags, Skn, adoptive transfer of Skn-immune T cells to immunosuppressed recipients elicits skin lesions in areas of mild epidermal trauma. In this study, we examined peripheral regulation of Skn-induced autoreactivity disrupted by rendering the mice immunoincompetent. We found that regulation of Skn-directed autoimmunity was restored by cotransfer of normal syngeneic spleen cells at twice the concentration of Skn-immune cells and was evidenced by significantly reduced lesion severity by days 5-7 post-cotransfer compared with animals given injections of Skn-immune cells alone. Enrichment and depletion of normal CD4(+) or CD8(+) spleen cells and RT-PCR analysis of selected cytokines identified CD4(+) cells as the regulatory cells in the cotransfer inoculum; however, significant reduction in lesion severity was observed only when there was a concomitant increase in levels of IL-7. The role of IL-7 was further supported in that mice cotransferred with Skn-immune cells plus normal spleen cells, but also treated with anti-IL-7 Ab, no longer exhibited reduced lesion severity. To determine whether IL-7 expression without normal spleen cell cotransfer could modulate lesion development, an IL-7-encoding plasmid (pCMV-Tag1-IL-7) was topically delivered to sites flanking the stressed skin site in Skn-induced autoimmune mice. Daily application of 15 mug of pCMV-Tag1-IL-7 significantly suppressed lesion severity. Our results support a mechanism for CD4(+) T cells and IL-7 in contributing to the control of autoreactivity.

DNA vaccines against chronic lung infections by Pseudomonas aeruginosa

Vaccines containing outer membrane protein F (OprF) of Pseudomonas aeruginosa are effective in reducing lesion severity in a mouse pulmonary chronic infection model. One OprF-based vaccine, called F/I, contains carboxy oprF sequences fused to oprI in an expression vector. When delivered three times biolistically by gene gun, the F/I vaccine induces protection that is antibody-mediated in outbred mice. To demonstrate the role of F/I-induced antibody-mediated immunity, B-cell-deficient [B(−)] and B-cell-intact [B(+)] mice were immunized with F/I, challenged with Pseudomonas, and examined for lesion severity. As expected, F/I-immunized B(+) mice had fewer and less severe lesions than vector-immunized B(+) mice. However, surprisingly, F/I- and vector-immunized B(−) mice were equally protected to levels similar to F/I-immunized B(+) mice. Examination of immune cell populations and cytokine levels indicated a relative increase in the quantity of CD3+ T-lymphocytes in vector- or F/I-immunized and challenged B(−) mice compared to B(+) mice. These data indicate the protective role played by cell-mediated immunity in B(−) mice, which supports our hypothesis that cell-mediated immunity can play an important role in protection against P. aeruginosa.