Alter Disease Progression Research Articles

Do Frogs Infected with Batrachochytrium dendrobatidis Avoid Water While Sloughing?

Chytridiomycosis is a skin infection caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd), that has been responsible for amphibian declines and extinctions worldwide. Chytridiomycosis increases the permeability of amphibian skin leading to low plasma ion levels and loss of physiological homeostasis. Because temporary increases in cutaneous salt loss and water uptake occur during sloughing, potentially exacerbating the effects of Bd infection, avoiding water while sloughing might help to alleviate these effects, and could contribute to intra- and interspecific differences in pathophysiology. Furthermore, because the amount of time an infected individual spends in contact with water could affect the severity of ion loss, we questioned whether amphibian behavior is altered in response to infection with Bd. This project utilized continuous infrared video footage of captive Australian Green Tree Frogs (Litoria caerulea) as a method of behavioral analysis, to determine whether infection with Bd altered the duration of water contact, activity levels, and the location of sloughing events. We found that although frogs became less active once infected, there was no change in the duration of time spent in contact with water. Infected frogs showed a decrease in the proportion of sloughing events occurring in contact with water as the experiment progressed. Our study is novel in that it provides insight into the behavioral effects of chytridiomycosis, demonstrating that Bd infection can induce behaviors that might alter disease progression.

Preventing Rheumatoid Arthritis: A Global Challenge

Significant advancements have been made in the discovery of gene signatures, biomarkers, novel therapeutic targets, diagnostic tools, and risk factors that predict the development of rheumatoid arthritis (RA). There is also overwhelming evidence that treatment of early RA can prevent or alter disease progression and potentially lead to drug-free remission. Despite these advancements, there are significant challenges to identifying patients at risk of developing RA on a global scale. This commentary provides an overview of challenges related to the primary, secondary, and tertiary prevention of RA in the context of health care systems. Patient-level, provider-level, and health care system–level barriers to implementing prevention strategies are discussed. Strategies and opportunities to address these challenges, on both a local and global scale, are reported. Benefits as well as potential negative consequences that may be associated with implementation of prevention strategies for RA are discussed in the context of individuals and public health.

A dual amylin and calcitonin receptor agonist inhibits pain behavior and reduces cartilage pathology in an osteoarthritis rat model

Pain and disability are the main clinical manifestations of osteoarthritis, for which only symptomatic therapies are available. Hence, there is a need for therapies that can simultaneously alter disease progression and provide pain relief. KBP is a dual amylin- and calcitonin-receptor agonist with antiresorptive and chondroprotective properties. In this study we investigated the effect of KBP in a rat model of osteoarthritis. Medial meniscectomy (MNX) was performed in 39 rats, while 10 underwent sham surgery. Rats were treated with KBP and/or naproxen. Nociception was assessed by mechanical and cold allodynia, weight bearing asymmetry, and burrowing behavior. Blood samples were collected for biomarker measurements, and knees for histology. Cartilage histopathology was evaluated according to the advanced Osteoarthritis Research International (OARSI) score and KBPs invitro antiresorptive effects were assessed using human osteoclasts cultured on bone. The MNX animals displayed an increased nociceptive behavior. Treatment with KBP attenuated the MNX-induced osteoarthritis-associated joint pain. The cartilage histopathology was significantly lower in rats treated with KBP than in MNX animals. Bone and cartilage degradation, assessed by CTX-I and CTX-II plasma levels, were decreased in all KBP-treated groups and KBP potently inhibited bone resorption invitro. Our study demonstrates the effectiveness of KBP in ameliorating osteoarthritis-associated joint pain and in protecting the articular cartilage, suggesting KBP as a potential drug candidate for osteoarthritis.

Platelet function in HIV plus dengue coinfection associates with reduced inflammation and milder dengue illness

HIV-infected subjects under virological control still exhibit a persistent proinflammatory state. Thus, chronic HIV infection changes the host homeostasis towards an adapted immune response that may affect the outcome of coinfections. However, little is known about the impact of HIV infection on inflammatory amplification and clinical presentation in dengue. Platelets have been shown to participate in immune response in dengue and HIV. We hypothesized that altered platelet responses in HIV-infected subjects may contribute to altered inflammatory milieu and disease progression in dengue. We prospectively followed a cohort of 84 DENV-infected patients of whom 29 were coinfected with HIV under virological control. We report that dengue and HIV coinfection progress with reduced inflammation and milder disease progression with lower risk of vascular instability. Even though the degree of thrombocytopenia and platelet activation were similar between dengue-infected and HIV plus dengue-coinfected patients, plasma levels of the platelet-derived chemokines RANTES/CCL5 and PF4/CXCL4 were lower in coinfection. Consistently, platelets from coinfected patients presented defective secretion of the stored-chemokines PF4 and RANTES, but not newly synthesized IL-1β, when cultured ex vivo. These data indicate that platelets from HIV-infected subjects release lower levels of chemokines during dengue illness, which may contribute to milder clinical presentation during coinfection.

Abstract P2-09-02: Modulation of the immune microenvironment in high risk DCIS by intralesional injection of anti-PD-1 (pembrolizumab)

Abstract Background. Ductal carcinoma in situ (DCIS) is a risk factor for the subsequent development of invasive breast cancer. Features of DCIS that are associated with a high risk of a subsequent event include large size (> 5 cm), high grade, comedo necrosis, palpable mass, hormone receptor negativity, and HER2 positivity. We have previously shown that immune infiltrates are positively associated with these high-risk features, suggesting that manipulating the immune microenvironment in high-risk DCIS, for example via checkpoint blockade, could potentially alter disease progression. Methods. In this phase 1 pilot study we investigated changes in the immune microenvironment of high risk DCIS after intralesional injection of anti-PD-1 (pembrolizumab). Study participants received 2 intralesional injections of pembrolizumab, 3 weeks apart, with surgery approximately 3 weeks after the 2nd dose. The study started with a dose of 2 mg/injection (1/100 of the standard 200 mg iv dose), then escalated to 4 mg and 8 mg, with 3 patients at each dose. Tissue samples from pre-treatment biopsies and post-treatment surgical resections were stained with two 6-plex immune panels using Opal immunofluorescence reagents (Perkin Elmer) on a fully automated Ventana Discovery platform, imaged with a Vectra 3 system and analyzed with inForm software (Perkin Elmer). An algorithm for tumor/stroma segmentation developed in inForm was used to randomly select 10 high power fields (hpfs) for imaging. Cell phenotype maps were generated for each of these hpfs for each sample. Cell densities were determined per area of stroma, DCIS, or total tissue and averaged across all hpfs for a given case. Spatial analyses were performed to quantitate co-localization of immune cells with DCIS cells. Results. The intralesional injections were easily administered and well tolerated. No systemic toxicities were observed at any dose. MRI imaging demonstrated no change in the size of lesions after treatment. Multiplex immunofluorescence analyses demonstrated heterogeneous responses ranging from dramatic increases in T cells, in particular CD8+ T cells, in cases which had a T cell infiltrate prior to therapy, to no post-therapy T cell infiltrate in cases with a pre-therapy immune desert. We also observed increases in B cells and macrophages and a decrease in the ratio of FoxP3+ T cells to CD8+ T cells, the latter mainly due to a significant increase in CD8+ cells, as opposed to a decrease in FoxP3+ cells. Spatial analyses indicated that in some cases, despite a marked increase in T cells post therapy, these cells did not co-localize with DCIS cells, indicating a state of immune exclusion. Conclusions. We have demonstrated the safety and feasibility of intralesional injection of an immune checkpoint inhibitor (pembrolizumab) in high risk DCIS. In some patients we observed a dramatic change in the immune microenvironment, with an increase in T cells, B cells, and macrophages, and a decrease in the FoxP3:CD8 ratio, even at a dose that is 1/100 of the standard intravenous clinical dose. An expansion study is underway in which patients will receive 4 injections of pembrolizumab at 3 week intervals prior to going to surgery to determine if more injections/time will increase response rate. Citation Format: Campbell MJ, McCune E, Rothschild H, Bolen J, VandenBerg S, Chien J, Wong J, Esserman L. Modulation of the immune microenvironment in high risk DCIS by intralesional injection of anti-PD-1 (pembrolizumab) [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-09-02.

Rasagiline for amyotrophic lateral sclerosis: A randomized, controlled trial.

Rasagiline is a monoamine oxidase B (MAO-B) inhibitor with possible neuroprotective effects in patients with amyotrophic lateral sclerosis (ALS). We performed a randomized, double-blind, placebo-controlled trial of 80 ALS participants with enrichment of the placebo group with historical controls (n = 177) at 10 centers in the United States. Participants were randomized in a 3:1 ratio to 2 mg/day rasagiline or placebo. The primary outcome was average slope of decline on the ALS Functional Rating Scale-Revised (ALSFRS-R). Secondary measures included slow vital capacity, survival, mitochondrial and molecular biomarkers, and adverse-event reporting. There was no difference in the average 12-month ALSFRS-R slope between rasagiline and the mixed placebo and historical control cohorts. Rasagiline did not show signs of drug-target engagement in urine and blood biomarkers. Rasagiline was well tolerated with no serious adverse events. Rasagiline did not alter disease progression compared with controls over 12 months of treatment. Muscle Nerve 59:201-207, 2019.

242 - Role of thiocyanate in the repair of myeloperoxidase-derived thiol oxidation during chronic inflammation

Myeloperoxidase is a vital component of the human immune system, producing the potent oxidant hypochlorous acid (HOCl), to kill bacteria and other invading pathogens. However, an overproduction of HOCl causes oxidative stress and damage to host cells, which promotes the development of numerous inflammatory diseases, particularly atherosclerosis. Thiocyanate (SCN-) is the favoured substrate for MPO and a potent competitive inhibitor of HOCl formation. This promotes the formation of hypothiocyanous acid (HOSCN), a selective oxidant that targets thiols, resulting in the formation of reversible oxidation products. This has led to the hypothesis that supplementation with SCN- may have therapeutic value to reduce oxidation during chronic inflammation. In this study, we compare the reactivity of HOCl and HOSCN with macrophages, and examine whether the addition of SCN- can alter the extent and nature of cellular damage. Exposure of the J774A.1 murine macrophage cell line to HOCl resulted in loss of cellular thiols, inactivation of enzymes, activation of pro-inflammatory and stress-related signaling pathways and cell death. Treatment of macrophages with HOSCN also resulted in extensive thiol oxidation and enzyme inactivation, though in this case, reversible thiol oxidation products, including sulfenic acids, were seen on a range of cellular proteins. By targeting glycolytic enzymes within cells, HOSCN could also re-route glycolytic flux through the pentose phosphate pathway to elevate the production of NADPH. SCN- decreased the extent of HOCl-induced cell death, and altered the nature of thiol oxidation from non-reversible to reversible. These data support a potential role for SCN- to reduce the extent of oxidative damage during chronic inflammation, and help to rationalize in vivo observations, whereby supplementation with SCN- prevents tissue damage and alters disease progression in animal models of inflammation, including cystic fibrosis and atherosclerosis.

FAN1 modifies Huntington's disease progression by stabilizing the expanded HTT CAG repeat.

Huntington’s disease (HD) is an inherited neurodegenerative disease caused by an expanded CAG repeat in the huntingtin (HTT) gene. CAG repeat length explains around half of the variation in age at onset (AAO) but genetic variation elsewhere in the genome accounts for a significant proportion of the remainder. Genome-wide association studies have identified a bidirectional signal on chromosome 15, likely underlain by FANCD2- and FANCI-associated nuclease 1 (FAN1), a nuclease involved in DNA interstrand cross link repair. Here we show that increased FAN1 expression is significantly associated with delayed AAO and slower progression of HD, suggesting FAN1 is protective in the context of an expanded HTT CAG repeat. FAN1 overexpression in human cells reduces CAG repeat expansion in exogenously expressed mutant HTT exon 1, and in patient-derived stem cells and differentiated medium spiny neurons, FAN1 knockdown increases CAG repeat expansion. The stabilizing effects are FAN1 concentration and CAG repeat length-dependent. We show that FAN1 binds to the expanded HTT CAG repeat DNA and its nuclease activity is not required for protection against CAG repeat expansion. These data shed new mechanistic insights into how the genetic modifiers of HD act to alter disease progression and show that FAN1 affects somatic expansion of the CAG repeat through a nuclease-independent mechanism. This provides new avenues for therapeutic interventions in HD and potentially other triplet repeat disorders.

Medical Management of Rheumatic Heart Disease: A Systematic Review of the Evidence.

Rheumatic heart disease (RHD) is an important cause of heart disease globally. Its management can encompass medical and procedural (catheter and surgical) interventions. Literature pertaining to the medical management of RHD from PubMed 1990-2016 and via selected article reference lists was reviewed. Areas included symptom management, left ventricular dysfunction, rate control in mitral stenosis, atrial fibrillation, anticoagulation, infective endocarditis prophylaxis, and management in pregnancy. Diuretics, angiotensin blockade and beta-blockers for left ventricular dysfunction, and beta-blockers and If inhibitors for rate control in mitral stenosis reduced symptoms and improved left ventricular function, but did not alter disease progression. Rhythm control for atrial fibrillation was preferred, and where this was not possible, rate control with beta-blockers was recommended. Anticoagulation was indicated where there was a history of cardioembolism, atrial fibrillation, spontaneous left atrial contrast, and mechanical prosthetic valves. While warfarin remained the agent of choice for mechanical valve implantation, non-vitamin K antagonist oral anticoagulants may have a role in RHD-related AF, particularly with valvular regurgitation. Evidence for anticoagulation after bioprosthetic valve implantation or mitral valve repair was limited. RHD patients are at increased risk of endocarditis, but the evidence supporting antibiotic prophylaxis before procedures that may induce bacteremia is limited and recommendations vary. The management of RHD in pregnancy presents particular challenges, especially regarding decompensation of previously stable disease, the choice of anticoagulation, and the safety of medications in both pregnancy and breast feeding.

Abstract 4058: Race-associated breast cancer disparities: The role of CRYBB2 and its pseudogene

Abstract Research demonstrates young African American (AA) women have higher mortality rates for triple-negative breast cancer (TNBC), despite lower incidence rates, when compared to Caucasian American (CA) women. These disparate poor patient outcomes may be attributed to access to quality care, socioeconomic status, education, or geographic location. However, numerous other studies have demonstrated that gene expression differences may also contribute to the higher mortality rates experienced by AA women, suggestive of biological differences among these two patient populations. Consequently, CRYBB2 is one gene that has consistently been associated with AA race in normal and malignant tissues. Although, its basic function in these tissues is currently unknown, higher expression of CRYBB2 is significantly correlated with poor outcome in breast, colorectal and prostate cancers in AAs. We analyzed all publically available gene expression datasets from studies focused on breast cancer health disparities and discovered that all probes detected CRYBB2 as well as its pseudogene, CRYBB2P1. It is becoming increasingly clear that pseudogenes have the ability to modify the mechanisms of their functional parental genes. However, it remains unclear whether the high expression of CRYBB2, CRYBB2P1, or both, affect cell behaviors to alter disease progression. Herein, we investigated the biological consequences of CRYBB2 and CRYBB2P1 in three basal-like TNBC cell lines (SUM159, MDA-MB-231, Hs578t) using CRISPR/Cas9 and overexpression technologies. The functional consequence of CRYBB2 and CRYBB2P1 expression were evaluated using in vitro and biochemical assays, as well as in vivo tumorigenesis studies. Our findings demonstrate overexpression of CRYBB2 and CRYBB2P1 alter cell proliferation, cytokine and growth factor production, and tumorigenesis. This data highlights the potential molecular mechanisms by which CRYBB2 and CRYBB2P1 contribute to poor patient outcome. Citation Format: Maya A. Barrow, Megan E. Martin, Denise K. Reaves, Breanna Jeffcoat, Ginger R. Smith, Michael Tarpley, Kevin P. Williams, Jodie M. Fleming. Race-associated breast cancer disparities: The role of CRYBB2 and its pseudogene [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4058.

Abstract 961: Intralesional injection of anti-PD-1 (pembrolizumab) results in increased T cell infiltrate in high risk DCIS

Abstract Background. Accumulating evidence shows a correlation between tumor-infiltrating lymphocytes (TILs), in particular the presence of CD8+ T cells, in cancer tissue and favorable prognosis in various malignancies. Less is know about the role of the immune microenvironment in the context of pre-malignant lesions such as ductal carcinoma in situ (DCIS). In a previous study of women with DCIS, we observed that high numbers of activated CD8+ T cells within the DCIS lesions were associated with good outcomes while low numbers associated with high risk features and poor outcomes. These results suggest that manipulating the immune microenviroment in DCIS, for example via checkpoint blockade, could potentially alter disease progression. Methods. We recently initiated a phase 1 pilot study to investigate changes in the immune microenvironment of high risk DCIS after intralesional injection of anti-PD-1 (pembrolizumab). Study participants were given 2 intralesional injections of pembrolizumab 3 weeks apart with surgery approximately 3 weeks after the 2nd dose. The study started with a dose of 2 mg/ml (1/100 of the standard 200 mg/ml iv dose), then escalated to 4mg/ml and 8 mg/ml, with 3 patients at each dose. FFPE tissue samples from pre-treatment biopsies and post-treatment surgical resections were immunostained using Opal reagent kits (Perkin Elmer) on a fully automated Ventana Discovery platform, imaged with a Vectra automated imaging system, and analyzed with inForm software (Perkin Elmer). A 7-plex panel, consisting of mAbs to CD3, CD8, CD68, PD-1, PD-L1, Ki67, and cytokeratins, was used. An algorithm for tumor/stroma segmentation developed in inForm was used to randomly select 7-10 high power fields (hpfs) for imaging. Cell phenotype maps were generated for each of these hpfs for each sample. Cell densities were determined per area of stroma, DCIS, or total tissue and averaged across all hpfs for a given case. Results. We are completing our 2nd cohort of patients, and there have been no adverse clinical events. The injections are easily administered and well tolerated. By MRI imaging, there was no change in the size of lesions after the treatment. All patients have proceeded to surgical resection. Following intralesional pembrolizumab injection, multiplex fIHC analyses indicated an increase in CD8+ T cells, which in some areas was striking. This increase was mainly observed within the epithelial DCIS regions, as opposed to a lesser increase in stromal CD8+ T cells. We also observed an increase in CD68+ macrophages. Conclusions. We have demonstrated the safety and feasibility of intralesional injection of an immune checkpoint inhibitor (pembrolizumab) in high risk DCIS. Further, we have shown that this immunotherapeutic intervention results in a change in the immune microenvironment, with an increase in both T cells and macrophages, even at a dose that is 1/100 of the standard intravenous clinical dose. Citation Format: Michael J. Campbell, Emma McCune, Jennifer Bolen, Scott VandenBerg, Jo Chien, Jasmine Wong, Laura Esserman. Intralesional injection of anti-PD-1 (pembrolizumab) results in increased T cell infiltrate in high risk DCIS [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 961.

Bdnf variant is associated with milder motor symptom severity in early-stage Parkinson's disease

IntroductionParkinson's disease (PD) progression is heterogeneous. Variants in PD-related genes may alter disease progression or severity. We examined if the single nucleotide variant rs6265 in the gene Bdnf alters clinical phenotype in early-stage, unmedicated PD. MethodsA retrospective analysis was conducted using data collected in the Deprenyl And Tocopherol Antioxidative Therapy Of Parkinsonism (DATATOP) study. DNA samples (n = 217) were genotyped for the Bdnf rs6265 variant, and the primary endpoint was time to initiate levodopa. The Parkinson's Progression Markers Initiative (PPMI) was used for validation (n = 383). ResultsThe primary endpoint of time to initiate levodopa was associated with a delay in subjects with two copies of the rs6265 minor (Met66) allele (HR: 4.9; 95% CI: 1.3–18.8). Secondary endpoints were not different among genotypes. PPMI subjects with two Met66 alleles demonstrated significantly lower total and part III Movement Disorder Society – United Parkinson's Disease Rating Scale (MDS-UPDRS) scores at baseline, as well as more tremor-related symptoms, but not a delay in initiation of maintenance pharmacotherapy. ConclusionsData from two distinct, unmedicated, early-stage PD cohorts suggest that carrying two copies of the rs6265 Met66 allele (∼4% of the population) is associated with less severity in motor symptoms and potentially a slower rate of progression.

VEN-α, an engineered mammalian milk protein, demonstrates potent proinflammatory cytokine neutralization implicated in Inflammatory Bowel Disease

Abstract Inflammatory Bowel Disease, which is comprised of Crohn’s disease (CD) and ulcerative colitis (UC), are chronic and debilitating diseases. Traditional therapies, which consist of anti-inflammatories and immunomodulators, are palliative at best and do not alter the progressive course of the disease. Advances in the understanding IBD pathology have led to new therapeutic strategies that are centered around targeted biologics to significantly alter disease progression. Although these targeted biologics demonstrate significant improvements in the quality of life for patients, systemic administration can have significant negative side effects such as risk of infection and malignancies. Ventria Bioscience utilizes a nonanimal-based expression system known as ExpressTec to express at commercially viable levels a recombinant human milk protein that has potent anti-inflammatory activity. This protein, known as VEN-α, exhibits a protective effect when T84 monolayers are exposed to proinflammatory cytokines TNF-alpha and Interferon-gamma in the presence of VEN-α in vitro. Further, when orally delivered once daily to mice subjected to DSS treatment ad libitum, VEN-α improves clinical and histopathological outcomes that is comparable to cyclosporin. Oral delivery of VEN-α may reduce unwanted effects associated with systemic administration and at the same time concentrate the active biologic at the site of pathology. Future studies planned will focus on further detailing efficacy in other models of IBD and understanding the pharmacokinetic profile of this orally delivered biologic. However, collectively the data to date suggests the orally delivered VEN-α has potential as a novel targeted biologic in the treatment of IBD.

Decreased medial compartment loading and increased medial femorotibial articular cartilage thickness 12 months after limb realignment surgery

Purpose: High tibial osteotomy (HTO) is a limb realignment surgery for patients with medial compartment knee osteoarthritis (OA) and varus alignment. The surgery is proposed to improve aberrant knee joint loading and thereby alter disease progression. The purposes of the present study were to explore changes after HTO in: [1] MRI-based measures of articular cartilage thickness and [2] 3D gait analysis-based measures of medial-to-lateral knee load distribution (the external knee adduction moment (KAM) and angular impulse (KAAI)). Methods: Patients with clinical and radiographic evidence of medial compartment knee OA were recruited from an orthopaedic clinic specializing in HTO. Participants were included if they had varus malalignment, early to moderate disease severity (defined using Kellgren and Lawrence Grades 1–3) and knee pain localized to the medial compartment. All patients received an HTO plate and screws made of polyetheretherketone (PEEK) that does not create artifact on MRI. MR images were assessed preoperatively, 6 weeks postoperatively to determine any immediate changes due to surgery, then again at 12 months postoperatively to assess the effects of long-term altered loads. Gait analysis was performed preoperatively and 12 months postoperatively. MR images were acquired on a 3T scanner using sagittal double-echo steady state (DESS) imaging. The medial and lateral femorotibial cartilage compartments were segmented manually and the reader was blinded to disease severity (including masking all but the articular surfaces on the MRIs), alignment, and the order of image acquisition (baseline, 6 weeks and 12 months). The sums of the mean thickness in the medial tibia (MT) and central weight-bearing region of the medial femoral condyle (cMF), and the lateral tibia (LT) and central weight-bearing region of the lateral femoral condyle (cLF), were used to calculate the cartilage thickness in the medial (MFTC) and lateral femorotibial compartments (LFTC) respectively. 3D gait analysis was performed and the external knee moment in the frontal plane was calculated using inverse dynamics to represent medial-to-lateral knee load distribution. The mean changes with 95% confidence intervals (95%CI) were calculated for each follow-up time point, minus baseline. The number of individual patients achieving previously suggested smallest detectable changes (SDC) in medial articular cartilage thickness and peak knee adduction moments were also determined. Results: The demographic and clinical characteristics for eight participants are presented in Table 1. Changes in articular cartilage thickness at 6 weeks and 12 months after surgery are shown in Fig. 1. At 12 months, the KAM decreased 1.3%BWxHt (95%CI: −1.8 to −0.7) and had a moderate-to-high correlation (r = −0.64) with the observed increases in articular cartilage thickness. Similar results were found when the KAM was replaced with the KAAI (−0.7%BWxHts; 95%CI: −1.1 to −0.4; r = −0.76). At the 12-month follow-up, 75% of participants (6 of 8) had an increase in medial articular cartilage thickness greater than the smallest detectable change (SDC) threshold in MFTC (111 μm) previously reported for cartilage thickness loss. Similarly, 63% of participants (5 of 8) had a decrease in KAM greater than the previously reported SDC threshold (1%BWxHt). Conclusions: This is the first study to quantify increases in medial femorotibial articular cartilage thickness that are correlated to decreases in medial knee loading after limb realignment surgery.View Large Image Figure ViewerDownload Hi-res image Download (PPT)

Spectroscopic Studies of Buffer and Metal Ion Effects on Amyloid-Beta Peptide Structure and Aggregation

Amyloid-β peptides are found in brains obtained from Alzheimer's patients. These peptides aggregate forming insoluble Amyloid-β plaques. It is uncertain how the presence of these peptide plaques correlate with the onset of the disease, however environmental conditions are known to alter disease progression. Previous studies have proven that metal ions such as zinc and copper are co-localized within Aβ plaques. Recent studies suggest that these metal ions may serve a role in inducing Aβ peptide misfolding and aggregation. The Cu2+ and Zn2+ ions are believed to bind to the Aβ peptide structure and increase aggregation. The metal ion concentrations play a factor as higher metal ion concentrations correlate with a higher aggregation rate. Understanding the environmental conditions that affect Aβ peptide structure and aggregation will provide a greater understanding about the role of metals in disease. Current studies are aimed at understanding how metal ions influence peptide structure, solvation, and aggregation. Infrared spectroscopy was used to monitor aggregation and structural changes of control Aβ peptide and Aβ peptide in the presence of Cu2+ and Zn2+ over time. Infrared spectra show that peptide length, buffer and metal concentration influence Aβ peptide structure, solvation, and aggregation. AcknowledgementsNSF-REU #1461175JMU Department of Chemistry and Biochemistry

Disruptor of telomeric silencing 1-like (DOT1L): disclosing a new class of non-nucleoside inhibitors by means of ligand-based and structure-based approaches.

Chemical inhibition of chromatin-mediated signaling involved proteins is an established strategy to drive expression networks and alter disease progression. Protein methyltransferases are among the most studied proteins in epigenetics and, in particular, disruptor of telomeric silencing 1-like (DOT1L) lysine methyltransferase plays a key role in MLL-rearranged acute leukemia Selective inhibition of DOT1L is an established attractive strategy to breakdown aberrant H3K79 methylation and thus overexpression of leukemia genes, and leukemogenesis. Although numerous DOT1L inhibitors have been several structural data published no pronounced computational efforts have been yet reported. In these studies a first tentative of multi-stage and LB/SB combined approach is reported in order to maximize the use of available data. Using co-crystallized ligand/DOT1L complexes, predictive 3-D QSAR and COMBINE models were built through a python implementation of previously reported methodologies. The models, validated by either modeled or experimental external test sets, proved to have good predictive abilities. The application of these models to an internal library led to the selection of two unreported compounds that were found able to inhibit DOT1L at micromolar level. To the best of our knowledge this is the first report of quantitative LB and SB DOT1L inhibitors models and their application to disclose new potential epigenetic modulators.

Leishmania-Derived Trimannose Modulates the Inflammatory Response To Significantly Reduce Leishmania major-Induced Lesions

Leishmania lipophosphoglycan (LPG) is a key virulence factor, initiating inflammation resulting in cutaneous lesions. LPG is capped by various oligosaccharides. How these glycans are recognized and how they alter the course of Leishmania infection are poorly understood. Previous studies synthesized α-1,2-trimannose cap sugars on latex beads and demonstrated that C57BL/6 mice coinoculated with Leishmania major and trimannose-coated beads produced significantly higher levels of interleukin-12p40 (IL-12p40) and other proinflammatory, type 1 cytokines than mice inoculated with L. major alone within the first 48 h of infection. However, as L. major infection typically progress over weeks to months, the role of trimannose in altering disease progression over the course of infection was unknown. Wild-type mice were inoculated with either trimannose-coated or carrier (uncoated) beads, infected with L. major alone, coinoculated with carrier beads and L. major, or coinoculated with trimannose-coated beads and L. major Trimannose treatment of L. major-infected mice decreased the parasite load and significantly decreased the lesion size at 14 days postinfection (p.i.) compared to results for nontreated, infected mice. Infected, trimannose-treated mice had decreased IL-12p40 and IL-10 secretion and increased interferon gamma secretion at 14 days p.i. Mannose receptor knockout (MR-/-) mice lack the ability to detect trimannose. When MR-/- mice were infected with L. major and treated with trimannose beads, they did not have decreased lesion size. Leishmania-derived trimannose represents a novel immunomodulator that provides early type 1-skewed cytokine production to control the parasite load and alter the course of cutaneous leishmaniasis.

Modification of the natural progression of epileptogenesis by means of biperiden in the pilocarpine model of epilepsy

Brain injuries are often associated with the later development of epilepsy. Evidence suggests that morphological and functional changes occur in the remaining neural tissue during a silent (or latent) period in which no seizures are expressed. It is believed that this silent (reorganization) period may provide a therapeutic window for modifying the natural history of disease progression. Here we provide evidence that biperiden, a muscarinic anticholinergic agent, is able to alter disease progression in an animal model of epilepsy. We observed that biperiden was capable of slowing the manifestation of the first spontaneous epileptic seizure and effectively reduced the severity and number of recurrent, spontaneous epileptic seizures during the animals’ lifespan. Biomolecular (microdialysis) and electrophysiological (extracellular field recordings) studies determined that biperiden was capable of elevating the threshold of hippocampal excitability, thereby making the hippocampal glutamatergic pathways less responsive to stimuli when high concentrations of potassium were used in vivo or in vitro. Notably, there was no hindrance of long-term memory or learning (a potential problem given the amnestic nature of biperiden). We conclude that biperiden has antiepileptogenic potential and may represent an opportunity for the prevention of post-traumatic epilepsy.

Epidemiological consequences of immune sensitisation by pre-exposure to vector saliva.

Blood-feeding arthropods—like mosquitoes, sand flies, and ticks—transmit many diseases that impose serious public health and economic burdens. When a blood-feeding arthropod bites a mammal, it injects saliva containing immunogenic compounds that facilitate feeding. Evidence from Leishmania, Plasmodium and arboviral infections suggests that the immune responses elicited by pre-exposure to arthropod saliva can alter disease progression if the host later becomes infected. Such pre-sensitisation of host immunity has been reported to both exacerbate and limit infection symptoms, depending on the system in question, with potential implications for recovery. To explore if and how immune pre-sensitisation alters the effects of vector control, we develop a general model of vector-borne disease. We show that the abundance of pre-sensitised infected hosts should increase when control efforts moderately increase vector mortality rates. If immune pre-sensitisation leads to more rapid clearance of infection, increasing vector mortality rates may achieve greater than expected disease control. However, when immune pre-sensitisation prolongs the duration of infection, e.g., through mildly symptomatic cases for which treatment is unlikely to be sought, vector control can actually increase the total number of infected hosts. The rising infections may go unnoticed unless active surveillance methods are used to detect such sub-clinical individuals, who could provide long-lasting reservoirs for transmission and suffer long-term health consequences of those sub-clinical infections. Sensitivity analysis suggests that these negative consequences could be mitigated through integrated vector management. While the effect of saliva pre-exposure on acute symptoms is well-studied for leishmaniasis, the immunological and clinical consequences are largely uncharted for other vector-parasite-host combinations. We find a large range of plausible epidemiological outcomes, positive and negative for public health, underscoring the need to quantify how immune pre-sensitisation modulates recovery and transmission rates in vector-borne diseases.

Disease Modifying Therapies Not Beneficial in Secondary Progressive Multiple Sclerosis

To examine whether medications for multiple sclerosis (MS) alter disease progression in secondary progressive MS (SPMS), researchers assessed outcomes