Phase II evaluation of VDC-1101 in canine cutaneous T-cell lymphoma.

BackgroundCanine cutaneous T-cell lymphoma (CTCL) is an uncommon disease for which efficacious therapies are lacking. The novel anticancer nucleotide prodrug VDC-1101 (formerly known as GS-9219) has shown efficacy in dogs with multicentric lymphoma. One of the observed adverse effects with this drug was a skin change characterized by hair loss, erythema, and pruritus, implying delivery of VDC-1101 to the skin.Hypothesis/objectivesThe primary study objective was to identify the objective response rate (ORR) to VDC-1101 in canine CTCL; secondary objectives included characterization of progression-free survival (PFS) and adverse events (AEs).AnimalsTwelve dogs with chemotherapy-naïve or relapsed, histologically and immunohistochemically confirmed CTCL.MethodsDogs received VDC-1101 as a 30-minute IV infusion once every 21 days. Prednisone (1 mg/kg PO q48h) was administered concurrently.ResultsIn 11 evaluable patients, responses included 1 complete response (CR), 4 partial responses (PR), 2 stable disease (SD), and 4 progressive disease for an ORR of 45% and biologic response rate (CR/PR/SD) of 64%. The median PFS was 37.5 days (26 to >399 days), which includes 1 durable and ongoing CR (>1 year). Gastrointestinal and hematologic AEs were mild; no dogs developed grade 3 or 4 AEs. Three dogs developed dermatopathies and 1 of these dogs was removed from the study as a result of this AE.Conclusions and clinical importanceVDC-1101 has activity against canine CTCL and could provide another treatment option in a disease process with a poor prognosis

The role of cardiac troponin T quantity and function in cardiac development and dilated cardiomyopathy

Background: Hypertrophic (HCM) and dilated (DCM) cardiomyopathies results from sarcomeric protein mutations, including cardiac troponin T (cTnT, TNNT2). We determined whether TNNT2 mutations cause cardiomyopathies by altering cTnT function or quantity; whether the severity of DCM is related to the ratio of mutant to wildtype cTnT; whether Ca2+ desensitization occurs in DCM; and whether absence of cTnT impairs early embryonic cardiogenesis. Methods and Findings: We ablated Tnnt2 to produce heterozygous Tnnt2+/ mice, and crossbreeding produced homozygous null Tnnt2-/-embryos. We also generated transgenic mice overexpressing wildtype (TGWT) or DCM mutant (TGK210Δ) Tnnt2. Crossbreeding produced mice lacking one allele of Tnnt2, but carrying wildtype (Tnnt2+/-/TGWT) or mutant (Tnnt2+/-/TGK210Δ) transgenes. Tnnt2+/-mice relative to wildtype had significantly reduced transcript (0.82 ± 0.06 [SD] vs. 1.00 ± 0.12 arbitrary units; p = 0.025), but not protein (1.01 ± 0.20 vs. 1.00 ± 0.13 arbitrary units; p = 0.44). Tnnt2+/-mice had normal hearts (histology, mass, left ventricular end diastolic diameter [LVEDD], fractional shortening [FS]). Moreover, whereas Tnnt2+/-/ TGK210Δ mice had severe DCM, TGK210Δ mice had only mild DCM (FS 18 ± 4 vs. 29 ± 7%; p < 0.01). The difference in severity of DCM may be attributable to a greater ratio of mutant to wildtype Tnnt2 transcript in Tnnt2+/-/TGK210Δ relative to TGK210Δ mice (2.42±0.08, p = 0.03). Tnnt2+/-/TGK210Δ muscle showed Ca2+ desensitization (pCa50 = 5.34 ± 0.08 vs. 5.58 ± 0.03 at sarcomere length 1.9 μm. p<0.01), but no difference in maximum force generation. Day 9.5 Tnnt2-/-embryos had normally looped hearts, but thin ventricular walls, large pericardial effusions, noncontractile hearts, and severely disorganized sarcomeres. Conclusions: Absence of one Tnnt2 allele leads to a mild deficit in transcript but not protein, leading to a normal cardiac phenotype. DCM results from abnormal function of a mutant protein, which is associated with myocyte Ca2+ desensitization. The severity of DCM depends on the ratio of mutant to wildtype Tnnt2 transcript. cTnT is essential for sarcomere formation, but normal embryonic heart looping occurs without contractile activity. © 2008 Ahmad et al

Enhancing fragility of zero-based text watermarking utilizing effective characters list

Text is an important medium used for sharing information worldwide. For a text document, digital watermarking is an efficient way for copyright protection, authentication, tamper proofing, to name but a few. In this paper, a zero-based watermarking approach is proposed for document authentication and tamper detection. To enhance the fragility of watermark, the proposed text watermarking approach can be comfortably utilized – based on the Effective Characters List (ECL) for watermark generation. The ECL method is generated for English text zero-watermarking by maintaining the contents of the original document and constructing the watermark by formulating the smooth transition between the selected characters in the documents. The evaluation of the proposed watermarking approach is based on three famous watermarking attacks including deletion, insertion, and reordering with an accuracy of 80.76%, 80.36%, and 88.1%, respectively. For a fair evaluation, a comparison is put forth with a recent zero-based watermarking method - clearly showing that the proposed method outperforms existing with greater accuracy. © 2019, Springer Science+Business Media, LLC, part of Springer Nature

Alteration of renal respiratory Complex-III during experimental type-1 diabetes

<p>Abstract</p> <p>Background</p> <p>Diabetes has become the single most common cause for end-stage renal disease in the United States. It has been established that mitochondrial damage occurs during diabetes; however, little is known about what initiates mitochondrial injury and oxidant production during the early stages of diabetes. Inactivation of mitochondrial respiratory complexes or alteration of their critical subunits can lead to generation of mitochondrial oxidants, mitochondrial damage, and organ injury. Thus, one goal of this study was to determine the status of mitochondrial respiratory complexes in the rat kidney during the early stages of diabetes (5-weeks post streptozotocin injection).</p> <p>Methods</p> <p>Mitochondrial complex activity assays, blue native gel electrophoresis (BN-PAGE), Complex III immunoprecipitation, and an ATP assay were performed to examine the effects of diabetes on the status of respiratory complexes and energy levels in renal mitochondria. Creatinine clearance and urine albumin excretion were measured to assess the status of renal function in our model.</p> <p>Results</p> <p>Interestingly, of all four respiratory complexes only cytochrome c reductase (Complex-III) activity was significantly decreased, whereas two Complex III subunits, Core 2 protein and Rieske protein, were up regulated in the diabetic renal mitochondria. The BN-PAGE data suggested that Complex III failed to assemble correctly, which could also explain the compensatory upregulation of specific Complex III subunits. In addition, the renal F₀F₁-ATPase activity and ATP levels were increased during diabetes.</p> <p>Conclusion</p> <p>In summary, these findings show for the first time that early (and selective) inactivation of Complex-III may contribute to the mitochondrial oxidant production which occurs in the early stages of diabetes.</p

RNA based approaches to profile oncogenic pathways from low quantity samples to drive precision oncology strategies

Precision treatment of cancer requires knowledge on active tumor driving signal transduction pathways to select the optimal effective targeted treatment. Currently only a subset of patients derive clinical benefit from mutation based targeted treatment, due to intrinsic and acquired drug resistance mechanisms. Phenotypic assays to identify the tumor driving pathway based on protein analysis are difficult to multiplex on routine pathology samples. In contrast, the transcriptome contains information on signaling pathway activity and can complement genomic analyses. Here we present the validation and clinical application of a new knowledge-based mRNA-based diagnostic assay platform (OncoSignal) for measuring activity of relevant signaling pathways simultaneously and quantitatively with high resolution in tissue samples and circulating tumor cells, specifically with very small specimen quantities. The approach uses mRNA levels of a pathway\u27s direct target genes, selected based on literature for multiple proof points, and used as evidence that a pathway is functionally activated. Using these validated target genes, a Bayesian network model has been built and calibrated on mRNA measurements of samples with known pathway status, which is used next to calculate a pathway activity score on individual test samples. Translation to RT-qPCR assays enables broad clinical diagnostic applications, including small analytes. A large number of cancer samples have been analyzed across a variety of cancer histologies and benchmarked across normal controls. Assays have been used to characterize cell types in the cancer cell microenvironment, including immune cells in which activated and immunotolerant states can be distinguished. Results support the expectation that the assays provide information on cancer driving signaling pathways which is difficult to derive from next generation DNA sequencing analysis. Current clinical oncology applications have been complementary to genomic mutation analysis to improve precision medicine: (1) prediction of response and resistance to various therapies, especially targeted therapy and immunotherapy; (2) assessment and monitoring of therapy efficacy; (3) prediction of invasive cancer cell behavior and prognosis; (4) measurement of circulating tumor cells. Preclinical oncology applications lie in a better understanding of cancer behavior across cancer types, and in development of a pathophysiology-based cancer classification for development of novel therapies and precision medicine

THE EFFECTS OF SILICA FERTILIZER AS AN ANTI-HERBIVORE DEFENSE IN CUCUMBER

This study aims to improve our understanding of silicon’s role in deterring herbivores from Cucumis sativa. We hypothesized that silicon’s role in plant defense is due to the presence of silica augmenting other physical and/or chemical defenses used by the plant. Using C. sativa plants treated with either a silica fertilizer treatment (Si+) or a control solution (Si-), we monitored feeding preferences of two types of her- bivores, a chewing herbivore (Diabrotica balteata) and a piercing/sucking herbivore (Bemisia tabaci). Leaves from treatment plants were visited less and eaten less than leaves from control plants. We then assessed the differences in physical defenses by comparing the leaf structural components, nutrient and water content, and trichome density between treatment and control plants. For chemical plant defenses, we measured leaf carbon and nitrogen levels in, and volatile organic compounds (VOCs) from treatment and control plants. We found no significant difference between treatment and control plants in: lignin content, most elemental plant nutrients, water content, trichome density, and quantity of carbon and nitrogen. We did see an increase in the VOC Indole, known for plant defense priming, an increase in phosphorous levels and a decrease in cellulose levels in silica treated plants

Experiences and lessons learned from 29 HPV vaccination programs implemented in 19 low and middle-income countries, 2009-2014

BACKGROUND: Cervical cancer is the greatest cause of age-weighted years of life lost in the developing world. Human papillomavirus (HPV) infection is associated with a high proportion of cervical cancers, and HPV vaccination may help to reduce the incidence of cancer. The aim of the study was to identify barriers, obstacles, and strategies and to analyze key concerns and lessons learned with respect to the implementation of HPV vaccination program in low- and middle-income countries. METHODS: The Gardasil Access Program (GAP) is a donation program established to enable organizations and institutions in eligible low-resource countries to gain operational experience designing and implementing HPV vaccination programs. This study used an online survey to capture the experiences and insights of program managers participating in the GAP. Different factors related to HPV vaccination program management were collected. A mixed-method approach enabled the presentation of both quantitative measurements and qualitative insights. RESULTS: Twenty-nine programs implemented by 23 institutions in 19 low- and middle-income countries were included. Twenty programs managers (97.7 %) reported that their institution implemented sensitization strategies about vaccination prior to the launch of vaccination campaign. The most frequently reported obstacles to HPV vaccination by the program managers were erroneous perceptions of population related to the vaccine’s safety and efficacy. Reaching and maintaining follow-up with target populations were identified as challenges. Insufficient infrastructure and human resources financing and the vaccine delivery method were identified as significant health system barriers. Coupling HPV vaccination with other health interventions for mothers of targeted girls helped to increase vaccination and cervical cancer screening. The majority of program managers reported that their programs had a positive impact on national HPV vaccination policy. The majority of institutions had national and international partners that provided support for human resources, technical assistance, and training and financial support for health professionals. CONCLUSION: Local organizations and institutions can implement successful HPV vaccination campaigns. Adequate and adapted planning and resources that support information sharing, sensitization, and mobilization are essential for such success. These results can inform the development of programs and policies related to HPV vaccination in low- and middle-income countries

Interaction between NANOS2 and the CCR4-NOT Deadenylation Complex Is Essential for Male Germ Cell Development in Mouse

Nanos is one of the evolutionarily conserved proteins implicated in germ cell development and we have previously shown that it interacts with the CCR4-NOT deadenylation complex leading to the suppression of specific RNAs. However, the molecular mechanism and physiological significance of this interaction have remained elusive. In our present study, we identify CNOT1, a component of the CCR4-NOT deadenylation complex, as a direct factor mediating the interaction with NANOS2. We find that the first 10 amino acids (AAs) of NANOS2 are required for this binding. We further observe that a NANOS2 mutant lacking these first 10 AAs (NANOS2-ΔN10) fails to rescue defects in the Nanos2-null mouse. Our current data thus indicate that the interaction with the CCR4-NOT deadenylation complex is essential for NANOS2 function. In addition, we further demonstrate that NANOS2-ΔN10 can associate with specific mRNAs as well as wild-type NANOS2, suggesting the existence of other NANOS2-associated factor(s) that determine the specificity of RNA-binding independently of the CCR4-NOT deadenylation complex

Howard County Farmers Association (HCFA) Business Concept

Final project for AREC489N: Economics of Local Agriculture: Food Hubs (Spring 2016). University of Maryland, College Park.The Howard County Office of Community Sustainability is seeking food hub-based solutions that connect small and mid-sized farmers in Howard County with restaurants and retailers to expand the market for locally produced agricultural products. Under the supervision of instructor Philip Gottwals, the University of Maryland’s PALS-affiliated AREC 489N team studied successes and failures of organizations that have undertaken similar projects to determine the most appropriate and feasible solutions to the issues raised. A review of public data quickly revealed that Howard County's agricultural sector is both small and highly diversified. As such it offers both opportunities for, and challenges to, the creation of a food hub. Vegetable production, for example, is limited to 110 acres of production with the largest crop acreage devoted to an ornamental crop, pumpkins. The next largest vegetable crops are sweet corn and tomatoes, with all others representing negligible commercial, fresh market acreage. For a food hub to be viable, an increase in production and diversity would be required. Otherwise, it would be impractical to gather and distribute wholesale quantities of vegetables to restaurants, retailers, or wholesalers. Interviews with County farmers confirmed that market opportunity is not limited by facility-based services, such as aggregation, but instead were limited by programmatic and policy restrictions. Chief among their concerns are the impending food safety certification requirements imposed by the federal Food Safety Modernization Act audited by Good Agricultural Practices (GAP) certification. These regulations, which were previously enforced only on the largest farms, now apply to all fruit and vegetable producers, regardless of farm size or program cost. Unless these standards are adopted at the farm level, local producers would be barred from many, if not all, commercial sales. Further evidence that a facility-based food hub is unlikely to be successful in Howard County can be found in the depth and breadth of the existing food supply chain. Howard County is the epicenter of a 100-mile radius supply chain that includes approximately 4,000 firms that are involved in all aspects of the food industry, including logistics, manufacturing, and distribution. After interviewing businesses operating in these sectors, it was determined that sufficient options to aggregate, ship, or value add were available in the market, but that critical services, such as quality assurance, food safety certification, and marketing support were lacking, putting local farmers at a marked disadvantage over farms from outside the area. Given the above, the project team concluded that it would be a greater benefit to our client if further research and project development efforts were directed toward designing a multi-pronged Quality Assurance Program for the County combined with an associated marketing and brand management program to raise awareness of Howard County farm products. This multifaceted approach strives to implement the newly emerging food safety requirements as a method of expansion into new markets. This allows growers to participate in the supply-chain where well-documented demand for local, GAP-certified produce and quality assured beef cattle exists. The second facet of the approach involves creating a suite of strategic marketing initiatives designed to bolster consumer demand for local food.Howard Count