Untreated Sites Research Articles

Ecosystem recovery at hypersaline salt marsh remediation projects impacted by the Gulf War Oil Spill

ABSTRACT #1141405 The 1990 Gulf War Oil Spill was the largest marine oil spill in history, spilling an estimated 6–11 million barrels of crude oil into the Arabian Gulf and impacting approximately 800 km of shoreline in Saudi Arabia between the border with Kuwait and Abu Ali Island near Jubail. Many intertidal areas were heavily oiled, severely damaging salt marsh ecosystems and facilitating the growth of thick algal mats, which, in turn, created a physical barrier that restricted tidal exchange and prevented recolonization of marsh flora and fauna at these sites. Remediation projects have been ongoing since early 2010 at 19 project areas throughout the area of impact. Remediation activities are focused on refreshing existing or creating new tidal channels to restore hydrology and tilling the substrate to expose moderately oiled sediment and enhance the degradation of oil and break up laminate algal mats. Additional activities include removal of heavily oiled sediments, re-use of excavated unoiled and lightly oiled sediments to create additional topography in restored marshes, and vegetation planting in selected areas. Monitoring plots for long-term study were established at remediation sites, impacted but untreated sites (set-asides), and nearby reference (comparison) sites. Initial monitoring data were collected before and after remediation activities, which were primarily undertaken between 2010 and 2013. Biannual (spring and fall) monitoring was resumed in fall 2018 and will continue through spring 2022 to characterize ecosystem recovery at these sites. Based on results from fall 2018 and spring 2019, most remediation sites show some level of recovery, indicated by the return of annual vegetation and resident marsh invertebrates in lower elevation plots, and thinner algal mat types and insect burrowing activity at the upper elevations. While the lower portions of untreated sites are beginning to show recruitment of annual vegetation and invertebrates (e.g. crabs), thick, laminated algal mat cover occurs in the middle-upper elevations at untreated sites. Multimetric indices (MMIs) summarizing ecosystem health and stressors were developed to summarize recovery of these projects and evaluate progress at remediation sites relative to set-aside and comparison sites. MMI results indicate that remediation work was effective at reducing stressors in remediation sites compared to set-asides; however, the level of ecosystem recovery is variable across sites, in part depending on the time since restoration.

134 Correlation of 12r-lox activity and hsp70 with barrier function

The evaluation of the skin health is an essential step in the skincare industry in order to assess the safety and the efficacy of active ingredients. 12R-LOX (gene ALOX12B) is a lipoxygenase expressed in keratinocytes and is implicated in the oxygenation of the esterified omega-hydroxyacyl-sphingosine (EOS) ceramides, a required process to their covalent linkage to proteins of the cornified envelope. It is an important step in establishing the water barrier by preventing unnecessary evaporation through epithelial cells. HSP70 family members are among the most abundant HSPs in the skin expressed constitutively within keratinocytes. HSP70 expression is elevated in both epidermis and dermis for cytoprotection after skin samples are heat shocked or after stressors like UVB. Traditionally transepidermal water loss (TEWL) has been widely used as a way to evaluate skin barrier function and tape stripping as a way to evaluate biological markers (biomarkers). Our aim was to understand how TEWL, 12R-LOX and HSP70 could be correlated clinically. Subjects were treated with a hydrating cream for 3 weeks on their lower inner forearm. After treatment, TEWL was measured and tape strips were collected from the treatment area and untreated site. 12R-LOX enzyme activity was measured with a fluorescence-based assay and HSP70 by ELISA using the protein samples isolated from the tape stripes. The 12R-LOX enzyme activity had a negative correlation with TEWL as previously described and had a positive correlation with HSP70. Moreover, HSP70 also had a negative correlation with TEWL confirming the correlation between the two biomarkers 12R-LOX and HSP70 with barrier function. The study of these two biomarkers is a new interesting and non-invasive approach for studying skin under different conditions.

Restoration of a shrub‐encroached semi‐arid grassland: Implications for structural, hydrologic, and sediment connectivity

AbstractCross‐scale structural and functional connectivity feedbacks can amplify exogenous forces in dryland environments leading to ecosystem state change (e.g., from grassland to shrubland). Attenuation of these connectivity feedbacks would ostensibly be required to restore transitioned ecosystems to their former state. We compared structural, hydrologic, and sediment connectivity on a shrub‐encroached semi‐arid grassland in south‐eastern Arizona, USA, to that of a nearby site experiencing an increase in non‐native perennial grass (Lehmann lovegrass) abundance 5‐year following treating shrubs with tebuthiuron herbicide. Soil/vegetation attributes were quantified and paired with hydrologic experiments at fine (0.5 m2) to hillslope (50 m2) scales. Fine‐scale rainfall simulations (120 mm·h−1rainfall intensity; 45 min) showed interspaces between shrubs were hydrologically similar on the treated and control sites, whereas herbicided shrub patches were more resource conserving than those within the control (terminal infiltration rates of 105 and 71 mm·h−1, respectively). High structural connectivity of bare ground (basal gap lengths >200 cm) was correlated with increased concentrated flow run‐off and accompanied by greater sediment yields within the untreated site at a coarse scale (~9 m2). Hillslope‐scale modelling suggested a divergence between hydrologic and sediment connectivity: run‐off from high intensity rainfall was similar between sites, while predicted sediment yield was 44% less within the tebuthiuron‐treated site. Our results indicate (i) hydraulic properties of soils between shrubs are unresponsive to herbicide treatment, (ii) disruption of structural connectivity of these interspaces associated with grass cover increases subsequent to herbicide application attenuated run‐off and the energy needed for sediment transport, and (iii) sediment connectivity is reduced by conversion to a novel grassland ecological state.

Multiple lines of evidence for predator and prey responses to caribou habitat restoration

How to mitigate the impacts of anthropogenic habitat alteration and restore ecological processes has become an imperative question facing applied ecology. One high-profile example in Canada is boreal woodland caribou, which are declining across North America largely due to anthropogenic habitat alteration and associated changes to predator-prey dynamics. Habitat restoration is increasingly being implemented to recover habitat, as is mandated by federal law. But given the extent of the disturbance and the cost to conduct restoration, evaluating the effectiveness of restoration treatments is needed for effective recovery of caribou populations. We evaluated the effectiveness of silvicultural treatments to reduce predator (wolf and bear) and prey (moose and caribou) use of linear features using a multiple lines of evidence approach. All four species were less likely to be present at treated sites than untreated sites, and daily photo capture rates of moose and wolves when present at treated sites also declined; though effect sizes were typically small. Complimenting the camera-based results, individual moose, bears, and wolves monitored with GPS collars also showed a decline in the use of treated linear features, particularly those with higher intensity treatments, though the response was non-significant. The reduction in the use of treated lines did not scale-up into a significant decline in overall line-use within the treatment area. While we found more evidence than not supporting that animals reduced use of the restoration sites, our study highlights the complexity of monitoring and evaluating the success of habitat restoration. Understanding long-term responses is imperative to ensure habitat restoration is effective.

Spatio-temporal impacts of aerial adulticide applications on populations of West Nile virus vector mosquitoes

BackgroundAerial applications of insecticides that target adult mosquitoes are widely used to reduce transmission of West Nile virus to humans during periods of epidemic risk. However, estimates of the reduction in abundance following these treatments typically focus on single events, rely on pre-defined, untreated control sites and can vary widely due to stochastic variation in population dynamics and trapping success unrelated to the treatment.MethodsTo overcome these limitations, we developed generalized additive models fitted to mosquito surveillance data collected from CO2-baited traps in Sacramento and Yolo counties, California from 2006 to 2017. The models accounted for the expected spatial and temporal trends in the abundance of adult female Culex (Cx.) tarsalis and Cx. pipiens in the absence of aerial spraying. Estimates for the magnitude of deviation from baseline abundance following aerial spray events were obtained from the models.ResultsAt 1-week post-treatment with full spatial coverage of the trapping area by pyrethroid or pyrethrin products, Cx. pipiens abundance was reduced by a mean of 52.4% (95% confidence intrval [CI] − 65.6, − 36.5%) while the use of at least one organophosphate pesticide resulted in a mean reduction of 76.2% (95% CI − 82.8, − 67.9%). For Cx. tarsalis, at 1-week post-treatment with full coverage there was a reduction in abundance of 30.7% (95% CI − 54.5, 2.5%). Pesticide class was not a significant factor contributing to the reduction. In comparison, repetition of spraying over three to four consecutive weeks resulted in similar estimates for Cx. pipiens and estimates of somewhat smaller magnitude for Cx. tarsalis.ConclusionsAerial adulticides are effective for achieving a rapid short-term reduction of the abundance of the primary West Nile virus vectors, Cx. tarsalis and Cx. pipiens. A larger magnitude of reduction was estimated in Cx. pipiens, possibly due to the species’ focal distribution. Effects of aerial sprays on Cx. tarsalis populations are likely modulated by the species’ large dispersal ability, population sizes and vast productive larval habitat present in the study area. Our modeling approach provides a new way to estimate effects of public health pesticides on vector populations using routinely collected observational data and accounting for spatio-temporal trends and contextual factors like weather and habitat. This approach does not require pre-selected control sites and expands upon past studies that have focused on the effects of individual aerial treatment events.Graphical

FIELD EVALUATIONS OF ATTRACTIVE TOXIC SUGAR BAIT STATION AND VEGETATION SPRAY APPLICATIONS FOR CONTROL OF AEDES AEGYPTI IN KEY LARGO, FLORIDA

Aedes aegypti and Aedes albopictus, vectors of many arboviruses including Zika, dengue, and chikungunya, are difficult to control with traditional methods. We tested two novel approaches utilizing attractive toxic sugar baits (ATSB) against Ae. aegypti in the upper Florida Keys. Residential sites on the island of Key Largo were systematically selected using Google maps. Sites received either bait stations or vegetation spray application with ATSB. An untreated control site was selected to monitor mosquito populations. Adult and egg counts were monitored through baited BiogentsSentinel and oviposition traps. The treatment evaluation lasted 28 days following a 14-day pre-treatment evaluation. Treatment efficacy was evaluated using regression models to estimate the percent reduction of mosquitoes over time. Post-treatment, Ae. aegypti mosquito populations were reduced by 81% and 74% at days 7 and 28 (p<0.05) at the bait station site, while mosquito populations at the spray treatment site for the same period (7 to 28 days) were reduced by 66% and 82% (p<0.05), respectively. Treatment and time had no significant effect on the proportion of eggs collected after the application of the ATSB treatments. This is the first residential field trial against the Zika vector, Ae. aegypti, in South Florida that demonstrated successful reduction of female and males using both ATSB stations and vegetation spray treatments. The findings suggest that 1) ATSB stations and vegetation spray applications can reduce populations of Ae. aegypti in residential and semi-tropical areas at least up to 28 days and 2) Ae. aegypti female mosquitoes in South Florida feed on sugar, and their sugar-feeding behavior can be exploited to enhance control strategies.

Targeted dry skin treatment using a multifunctional topical moisturizer.

The development of dry skin is a complex process, with a wide variety of factors each playing different roles in its evolution. Given this, it is important when designing a formulation to tackle dry skin that these varied aspects of skin behaviour are addressed. Presented here are the results of a 3-week moisturization study carried out on dry legs. A wide range of traditional and more recently developed biophysical measurement methods have been combined with visual assessment of skin condition to enable multiple aspects of skin function to be determined. The observed changes in the skin are discussed in terms of the ingredients used in the moisturizing formulation. A range of novel and traditional skin assessment methods and techniques were used to assess the effects of an oil in water-based moisturizing product compared to an untreated site during a 3-week in vivo study on dry lower leg skin. Statistically significant improvements were observed in a range of skin parameters as a result of product usage. Skin hydration assessed using Corneometer®, Epsilon® and visual dry skin grading all increased after 3weeks of use. Skin barrier function measured using transepidermal water loss also improved. Levels of cholesterol, free fatty acids and Ceramide NH increased, as well as the average length of the stratum corneum (SC) lipid lamella bilayers, and the ratio of lipid to protein increased (measured using Lipbarvis® and in vivo Confocal Raman Spectroscopy). Increases in the levels of Ceramide EOS and NP were also observed, along with an improvement in corneocyte maturity, although these were not statistically significant. Using a variety of traditional and novel skin assessment techniques, a wide range of factors associated with the evolution of dry skin have been assessed upon treatment with a new topical moisturizer. Product usage resulted in significant improvements to skin hydration and barrier function, the levels and morphology of SC barrier lipids, and overall epidermal differentiation. As a result there was a significant reduction in the characteristics associated with the development of dry skin after use of the test product.

Formulation and Characterization of Glucosamine Sulphate Loaded Carbopol Based Hydrogels for the Management of Osteoarthritis

Osteoarthritis is emerging as the most ordinary form of arthritis, affecting 22-39% of the Indian population. A wide range of medications and therapies are available for the treatment of osteoarthritis. With a desire to develop a therapeutically effective dosage form, the present study was carried out to formulate glucosamine sulfate loaded carbopol based hydrogel. Hydrogels H1 to H6 were formulated without permeation enhancers while formulations H7 to H12 were developed with a different class of permeation enhancers such as PEG400, oleic acid, Tween 40, DMSO and PG. Based on viscosity, it was detected that formulation H4 containing polymer 1% was ideal for incorporating drug. Considering H4 as a placebo, H6 was used for further evaluation. Drug content was found to be 99.2±0.64, with in vitro drug release of 15±0.86, 22±1.59, 28±0.72, 35±0.68, 40±0.31, 47±0.83, 58±1.59, and 70±0.9 % at a duration of 1, 2, 3, 4, 5, 6, 7, 8 hours respectively. Skin irritation tests carried out on Wistar rats revealed that skin was intact with no inflammation or erythema detected, compared to untreated site. By diffusion disc method, it was evident that the levels of microbial load were relatively low, and no harmful microorganisms were identified. There were no significant changes in physicochemical properties on stability studies. Due to a simple method of preparation and effective drug delivery, glucosamine sulfate loaded hydrogels could be contemplated as a prominent formulation in the beneficiary treatment of osteoarthritis.

Elevated Skin pH Is Associated With an Increased Permeability to Synthetic Urine.

The aim of this study was to investigate the permeability of the skin following cleansing activities and its susceptibility to synthetic urine penetration. Ten healthy volunteers (aged 22-58 years) participated in the study, which was conducted in a university bioengineering laboratory. Tape stripping and sodium lauryl sulfate were used to simulate the physical and chemical irritation exacerbated by frequent cleansing activities, respectively. An untreated site also was selected to evaluate responses of intact skin. Synthetic urine was then applied for a period of 2 hours. Measurements of transepidermal water loss and skin pH were taken at baseline and after each challenge. To quantify the permeability of the skin following exposure, desorption curves of transepidermal water loss were measured and skin surface water loss was calculated. Chemically irritated skin, characterized by increased pH (7.34 ± 0.22), demonstrated an increased permeability to urine, as reflected by a significant increase in mean skin surface water loss (46,209 ± 15,596 g/m2) compared to both the intact (14,631 ± 6164 g/m2) and physically irritated (14,545 ± 4051 g/m2) skin (P = .005 in both cases). In contrast, the differences between the intact and physically irritated skin were not significant (P = .88). Permeability of the skin to irritants is influenced by the status of the skin and its acid mantle. These highlight the need to reevaluate the frequency of cleansing activities, along with the choice of product in clinical settings, favoring the use of pH-balanced cleansers.

Sickle Cell Pain: Intervention with Capsaicin Exposure (SPICE)

Sickle Cell Pain: Intervention with Capsaicin Exposure (SPICE)

Retinol Has a Skin Dehydrating Effect That Can Be Improved by a Mixture of Water-Soluble Polysaccharides

It is common that retinoids used in skincare can cause skin dryness, irritation and redness which is a complaint for the use of these molecules in skincare formulations. Objective: to investigate the influence of a mixture of polysaccharides to improve retinol-based formulations in a 12-day inner volar forearm study. Methods: in total, 22 inner volar forearms were treated over a 12-day topical application of a Placebo formulation containing 0.5% retinol verses a formulation containing 0.5% retinol and 3.0% of a complex of polysaccharides. Application occurred 2X/day in the morning and evening. Skin testing included barrier disruption, erythema, and skin hydration. After a 3-day regression of treatment, skin hydration was measured again. Results: the 0.5% retinol Placebo formulation showed a significant impact on skin dehydration compared to untreated control or polysaccharide-treated areas. The formulation containing retinol and 3.0% of the polysaccharides, maintained skin hydration levels comparable to the untreated control. Neither formulation had a statistically significant impact on skin erythema or barrier disruption. After the 3-day regression, the polysaccharide mixture continued to demonstrate significant moisturization benefits superior to the untreated and active-treated sites. Conclusions: a mixture of polysaccharides was able to mitigate the short-term skin drying effects of retinol and continued to moisturize the skin after a 3-Day regression.

SOIL BIOLOGICAL QUALITY UNDER CHEMICAL AGENTS OF PLANT PROTECTION USAGE. ASSESSMENT OF PEDOFAUNA BIODIVERSITY

This study, carried out under natural conditions, aims to understand the effect of the long-term use of phytosanitary treatments on the pedofauna biodiversity. Our results show that the influence of pesticides was clearer on the total abundance of the pedofauna in autumn than in spring. In total, we caught 620 individuals in the treated site and 868 individuals in the untreated site for the two work months October 2014 and April 2015. And the abundance of pedofauna was higher in the fall and lower in the spring for the untreated site. A total of 11 orders were identified for the site treated against 13 orders for the untreated site for the month of October. The impact of pesticides was more evident in the mesofauna where there was a total absence of springtails in the site treated, compared with only 5 individuals for the untreated site.

The effect of 0.5% topical timolol cream on regression of superficial infantile haemangioma in the involuting phase: a prospective, randomized, self-controlled study.

Infantile haemangiomas are the most common benign tumours affecting infants. Over time, the tumours may involute to some extent. However, sequelae, such as telangiectasia or fibrofatty tissue, often occur following this condition, which may cause disfigurement and influence patients' psychosocial development. This prospective, randomized, self-controlled study aimed to assess the effects of topical timolol (0.5%) on involuting infantile haemangiomas. Each involuting superficial infantile haemangioma (n = 29) was randomly divided into two regions; one region was treated with topical timolol (0.5%) cream, three times daily, and the other region was untreated. The comparative treatments continued for three months. Five independent assessors, blinded to the treatment regimen, judged the treated and untreated regions by comparing photographs before and after treatment. The topical timolol-treated tumour sections showed no difference compared with the untreated sites (p = 0.355) after three months of treatment, and by the end of the treatment, the untreated lesions showed significant differences relative to pre-treatment (p<0.001). Topical timolol was not observed to have any effect on the regression of infantile haemangiomas in the involuting phase.

Toll-Like Receptor 9 Agonists in Cancer.

Toll-like receptor 9 (TLR9) is a pattern recognition receptor that is predominantly located intracellularly in immune cells, including dendritic cells, macrophages, natural killer cells, and other antigen-presenting cells (APC). The primary ligands for TLR9 receptors are unmethylated cytidine phosphate guanosine (CpG) oligodinucleotides (ODN). TLR9 agonists induce inflammatory processes that result in the enhanced uptake and killing of microorganisms and cancer cells as well as the generation of adaptive immune responses. Preclinical studies of TLR9 agonists suggested efficacy both as monotherapy and in combination with several agents, which led to clinical trials in patients with advanced cancer. In these studies, intravenous, intratumoral, and subcutaneous routes of administration have been tested; with anti-tumor responses in both treated and untreated metastatic sites. TLR9 agonist monotherapy is safe, although efficacy is minimal in advanced cancer patients; conversely, combinations appear to be more promising. Several ongoing phase I and II clinical trials are evaluating TLR9 agonists in combination with a variety of agents including chemotherapy, radiotherapy, targeted therapy, and immunotherapy agents. In this review article, we describe the distribution, structure and signaling of TLR9; discuss the results of preclinical studies of TLR9 agonists; and review ongoing clinical trials of TLR9 agonists singly and in combination in patients with advanced solid tumors.

Comparative Evaluation of Two Hyaluronic Acid Gel Products for the Treatment of Interdental Papillary Defects.

ObjectivesThe aim was to investigate the efficacy of single injections of two different hyaluronic acid products, Flex Barrier and Revident, in reducing the size of black triangles to treat Nordland-Tarnow Class I and II recessions.Materials and MethodsForty adult patients were recruited with at least two upper and two lower interdental papilla defects in the front region between canine teeth. According to the Nordland-Tarnow classification of papillary defects, both Class I and Class II recessions were included in the investigation. Patients were randomly assigned to experimental groups to receive single injections of two different hyaluronic acid products, either Flex Barrier or Revident. The untreated sites served as controls. Photographs were taken before and immediately after the treatment, and again after one week and one month. To determine the size of the black triangles, Image J software was used. For statistical analysis, a mixed-design ANOVA was applied.ResultsBoth Flex Barrier and Revident significantly decreased the size of the treated defects immediately after the treatment and also one week later (p<0.001). The beneficial effect of Revident lasted longer than Flex Barrier as it remained significant even after one month in Revident-treated patients, however, not in the Flex Barrier-treated group. Furthermore, Nordland-Tarnow Class I lesions generally showed a greater improvement than Class II lesions.ConclusionIn this proof-of-concept, randomized clinical trial we have demonstrated the clinical applicability of both Flex Barrier and Revident, although Revident gave longer-lasting improvements than Flex Barrier. Further trials are needed to optimize multiple-application protocols for treating gingival black triangles.

Acellular dermal matrix for root coverage procedures: 9-year assessment of treated isolated gingival recessions and their adjacent untreated sites.

The long-term outcomes of acellular dermal matrix (ADM) for the treatment of isolated gingival recessions has not yet been evaluated. Thus, the aim of this study was to observe the root coverage outcomes of coronally advanced flap with ADM over time, and compare them with their adjacent untreated sites. Twelve patients (from 20) were available at the 9-year recall. Clinical parameters (recession depth, mean root coverage [mRC], keratinized tissue width [KTW], and gingival thickness) were evaluated and compared with the 1-year results, and the ADM-adjacent untreated sites (on mesial and distal) via mixed-modeling regression analyses. From 1 to 9 years, the ADM-treated isolated recessions showed a relapse from 77% to 62% mRC (P<0.05). A similar pattern toward apical shift of the gingival margin was noticed for the ADM-adjacent untreated sites without baseline recession. However, ADM-adjacent untreated sites which had presented with recession at baseline but were not treated showed a significantly more apical shift of the gingival margin (almost two-fold). A significant increase in KTW was noted for all sites. Baseline KTW ≥2mm was a significant predictor for the stability of the gingival margin at the ADM-treated, and the ADM-adjacent sites with baseline recession. ADM-treated sites displayed recession relapse from 1 to 9 years. The untreated adjacent sites with a recession at baseline, showed a higher apical displacement of the gingival margin compared with the ADM-treated sites, and ADM-adjacent sites without a recession at baseline.

Intra-hepatic Abscopal Effect Following Radioembolization of Hepatic Metastases

PurposeTo search for abscopal effects (AE) distant to the site of radiation after sequential Yittrium-90 (Y-90) radioembolization (RE) of liver malignancies.Methods and MaterialsIn this retrospective analysis, all patients treated by RE between 2007 and 2018 (n = 907) were screened for the following setting/conditions: sequential RE of left and right liver lobe in two sessions, liver-specific MRI (MRI1) acquired max. 10 days before or after first RE (RE1), liver-specific MRI (MRI2) acquired with a minimum time interval of 20 days after MRI1, but before second RE (RE2). No systemic tumor therapies between MRI1 and MRI2. No patients with liver cirrhosis. Metastases > 5 mm in untreated liver lobes were compared in MRI1 and MRI2 and rated as follows: same size or larger in MRI2 = no abscopal effect (NAE); > 30% shrinkage without Y-90 contamination in SPECT/CT = abscopal effect (AE).ResultsNinety six of 907 patients met aforementioned criteria. Median time-frame between RE1 and MRI2 was 34 (20–64) days. These 96 cases had 765 metastases which were evaluable (median 5(1–40) metastases per patient). Four patients could be identified with at least one shrinking metastasis of the untreated site: one patient with breast cancer (3 metastases: 0 NAE; 3 AE), one patient with prostate cancer (6 metastases: 3 NAE; 3 metastases > 30% shrinkage but possible Y-90 contamination) and two patients with shrinkage of one metastasis each but less than 30%.ConclusionOur retrospective study documents AE after RE of liver tumors in 1 out of 96 cases, 3 other cases remain unclear.

Quercetin postconditioning attenuates gastrocnemius muscle ischemia/reperfusion injury in rats

Quercetin, an antioxidant derived from plants, can play a beneficial role in the protection of various tissues against ischemia-reperfusion injuries (IRI). The purpose of the present research was to investigate the protective effects of quercetin on gastrocnemius muscle ischemia-reperfusion. A total of 80 adult male Wistar rats (weights: 250-300 g) were divided into ten groups (n = 8 per group). We used silk 6.0 surgical thread to create a knit to occlude the femoral artery and vein for 3 hr. The treated groups, which comprised half of each experimental group, received intraperitoneal injections of 150 mg/kg quercetin after the ischemia. Blood flow was subsequently reestablished in the reperfusion phase. The rats were kept in reperfusion for 3, 7, 14, or 28 days after which they were killed with high doses of anesthetic drugs, and the gastrocnemius muscles were removed and fixed. Tissue processing, hematoxylin and eosinand toluidine blue staining, and immunohistochemistry were used to assess tumor necrosis factor-α (TNF-α) and nuclear factor κB (NF-κB) levels. A comparison between treated and untreated ischemic sites showed that on the third day of reperfusion, the severity of edema and NF-κB level decreased significantly; on the 7th day of reperfusion, the severity of edema and the levels of TNF-α and NF-κB decreased significantly; and on the 14th day of reperfusion, all of the parameters showed significant decreases. On the 28th day of reperfusion, there were significantly decreased levels of TNF-α and NF-κB, and decreased mast cell infiltration when compared with the untreated groups. According to the results, administration of quercetin after ischemia could significantly prevent gastrocnemius muscle IRI.

Enhancement of the therapeutic effects of oncolytic vaccinia virus by using autologous and allogeneic cell-based delivery platforms.

e15271 Background: Clinical trials with oncolytic viruses for cancer treatment have shown limited efficacy due to viruses’ rapid clearance by patients’ innate and adaptive immune systems. In a recent first-in-human clinical trial, we confirmed the safety and feasibility of our approach to enhance oncolytic vaccinia virus (OVV) delivery and improve tumor targeting by utilizing an autologous cell-based cell delivery system (auto-OVV). We also developed an allogeneic cell-based platform (SuperNova1c - SNV1c) aiming to protect and potentiate OVV’s antitumor effects in large patient populations. Methods: We evaluated the immunomodulatory potential of auto-OVV by an extensive time-course analysis of cytokines in patients’ plasma (Luminex profiling) and peripheral blood immune cells (flow cytometry). We also analyzed the ability of SNV1c to protect the OVV from antibody/complement inactivation in vitro and in vivo following intratumoral injection in various mouse tumors. The immune cell infiltrations of the injected tumors were also analyzed. Results: Therapy with auto-OVV induced a coordinated activation of cytokine, T cell and NK responses in patients as early as 1 day, peaking around 1-week and lasting for up to 1-month post treatment. Effective OVV amplification in cancer patients correlated with significant changes of multiple innate and adaptive immune parameters. Patient stratification into groups with transient versus persistent viral DNA was linked to opposing and mutually exclusive patterns of robust activation of NK versus T cell responses, respectively. SNV1c showed significantly enhanced protection of OVV in vitro and led to statistically significant tumor growth inhibition as compared to control non-treated tumors or to naked OVV-treated tumors. Importantly, local administration of SNV1c induced systemic therapeutic effects. Five days after SNV1c administration, tumor infiltrating lymphocytes from both treated and untreated tumors showed increased CD4 and CD8 T-cell infiltrations, decreased Tregs, and improved effector to Treg ratios, associated with tumor growth inhibition at both treated and untreated tumor sites. Conclusions: This study establishes the timeline of treatment-related immunological changes and identifies potential immunological correlates associated with the OVV persistence in vivo. We also demonstrate the ability of our cell-based platforms to protect and potentiate OVV by circumventing innate and adaptive immune barriers, resulting in enhanced oncolytic virotherapy.

PH-Sensitive Carbon Dots for Enhancing Photomediated Antitumor Immunity.

Recent cancer immunotherapy has attracted much attention due to high specificity and recurrence prevention of tumor. Nevertheless, its therapeutic effects are still challenging in solid cancer. To establish superior antitumor immunity, chlorin e6 (Ce6)-loaded pH sensitive carbon dots were investigated (Ce6@IDCDs). At tumoral pH 6.5, Ce6 was released four times compared with the release at physiological pH 7.4 due to an imbalance between hydrophilic and hydrophobic forces via protonation of imidazole groups in Ce6@IDCDs. This result led to the superior singlet oxygen generating activity of Ce6@IDCDs without Ce6 quenching. The maturation effects of dendritic cells after co-incubation with supernatant media obtained from Ce6@IDCDs with laser-treated cells at pH 6.5 were much higher than at physiological pH. Furthermore, Ce6@IDCDs following a laser at pH 6.5 significantly promoted calreticulin exposure and high-mobility group box 1 release, as major immunogenic cell death markers. In bilateral CT-26-bearing mice model, the Ce6@IDCDs elicited significant antitumoral effects at laser treated-primary tumor regions via therapeutic reactive oxygen species. Furthermore, Ce6@IDCDs upon laser irradiation induced a large amount of activated CD8+ T cells, natural killer cells, and mature dendritic cells recruitment into tumoral tissue and hampered tumor growth even at untreated sites approximately four-fold compared with those of others. Overall, this pH-sensitive immunoinducer can accomplish primary and distant tumor ablation via photomediated cancer immunotherapy.