Non-selective Primary Human Tumor Cell Line Generation from Surgical Resections to be Paired With Flash Frozen and Paraffin Embedded Tissue: Advancements in Democratizing Translational Research Materials to Rural Institutions

Translational cancer research relies on the availability of human patient tissue demonstrating the specific disease process under investigation. Biobanks of human tissue have historically been and remain to date the primary access point for cancer research samples. Biorepositories routinely supply researchers with varying sample types for use in biomedical studies; most commonly formalin-fixed and paraffin-embedded (FFPE) tissue, or fresh snap-frozen tissue. In conjunction with preserved tissue samples, viable tumor cell lines derived from patient tissue have emerged to be a new gold standard in cancer research particularly in drug discovery and functional prognostic assays. Tissue banks providing these samples are being termed “next-generation” and are adapting to directly assist researchers by performing high throughput technical studies such as routine histology and immunostaining of donor tissue. These high quality, next-generation biorepositories are a relatively scarce resource in the broader research community in the United States and have traditionally been associated with large centralized and very well established university medical centers. This article describes the next-generation resources now available at the Edwards Comprehensive Cancer Center with its association with the Marshall University Joan C. Edwards School of Medicine in Huntington, West Virginia. This manuscript details the procedures, protocols, and success rates of the Tissue Procurement Program (TPP) to generate a growing cohort of viable primary human tumor cell lines representing varying malignancies to be used in conjunction with matched formalin-fixed and paraffin-embedded (FFPE) and snap-frozen tissue samples for comprehensive translational research

Minimally manipulative method for the expansion of human bone marrow mesenchymal stem cells to treat osseous defects

Copyright © 2019 Hamerly, Tweedell, Hritzo, Nyasembe, Tekwani, Nanayakkara, Walker and Dinglasan. Malaria is a major global health threat, with nearly half the world\u27s population at risk of infection. Given the recently described delayed clearance of parasites by artemisinincombined therapies, new antimalarials are needed to facilitate the global effort toward elimination and eradication. NPC1161 is an 8-aminoquinoline that is derived from primaquine with an improved therapeutic profile compared to the parent compound. The (R)-(-) enantiomer (NPC1161B) has a lower effective dose that results in decreased toxic side effects such as hemolysis compared to the (S)-(+)-enantiomer, making it a promising compound for consideration for clinical development. We explored the effect of NPC1161B on Plasmodium falciparum oocyst and sporozoite development to evaluate its potential transmission-blocking activity viz. its ability to cure mosquitoes of an ongoing infection. When mosquitoes were fed NPC1161B 4 days after P. falciparum infection, we observed that total oocyst numbers were not affected by NPC1161B treatment. However, the sporozoite production capacity of the oocysts was impaired, and salivary gland sporozoite infections were completely blocked, rendering the mosquitoes non-infectious. Importantly, NPC1161B did not require prior liver metabolism for its efficacy as is required in mammalian systems, suggesting that an alternative metabolite is produced in the mosquito that is active against the parasite. We performed liquid chromatography-mass spectrometry (LC-MS)/MS analysis of methanol extracts from the midguts of mosquitoes fed on an NPC1161B (434.15 m/z)-treated blood meal and identified a compound with a mass of 520.2 m/z, likely a conjugate of NPC1161B or an oxidized metabolite. These findings establish NPC1161B, and potentially its metabolites, as transmission-blocking candidates for the treatment of P. falciparum

Expression of poly-ADP-ribose polymerase (PARP) in endometrial adenocarcinoma: Prognostic potential

© 2020 Background: In the United States endometrial carcinoma is the most common female gynecologic malignancy. An average of more than 60,000 new cases of endometrial carcinomas have been diagnosed yearly over the past 5 years, with a higher incidence occurring in the central Appalachian states of Ohio and West Virginia. In the U.S., the national average of newly diagnosed endometrial carcinomas is 26.8 in every 100,000 women, while in the states of Ohio and West Virginia the average is 30.5 and 31.1 in every 100,000 women, respectively. This notable increase in the incidence of endometrial carcinomas may be due a variety of elevated risk factors including but not limited to: tobacco use, obesity, and genetic predisposition of the predominant demographic. The American Cancer Society estimates that approximately 55,000 new cases of endometrial carcinoma will be diagnosed in 2020 yet, this disease is widely considered understudied and under-represented in mainstream cancer research circles. Methods: The aim of this study was to quantitate the co-expression of two DNA repair proteins poly-ADP-ribose polymerase 1 and 2 (Parp-1 and Parp-2) by enzyme- linked immuno-sorbent assay (ELISA) in 60 endometrioid endometrial tumor samples and compare their expression to matched non-malignant endometrial tissue from the same corresponding donors from central Appalachia. Results: We found that Parp-1 was significantly overexpressed in endometrial carcinoma relative to corresponding normal tissue. This overexpression implicates Parp inhibition therapy as a possible treatment for the disease. Our results also found a protective effect of native Parp-2 expression in non-malignant endometrial tissue with each 1 ng/mL increase in PARP-2 concentration in normal tissue was associated with a 10 % reduction in the hazard of tumor progression (HR = 0.90; p = 0.039) and a 21 % reduction in the hazard of death (HR = 0.79; p = 0.044). Conclusions: This study demonstrated the over-expression of the druggable target Parp-1 in endometrial adenocarcinoma and observed a strong negative correlation of native Parp-2 expression and disease progression via the quantification of the Parp proteins using enzyme- linked immuno-sorbent assay (ELISA) assays

Assessing the Influence of Strategic Tillage on Crop Yields and Soil Properties in Dryland No-Tillage Systems

Implementing strategic tillage (ST) in otherwise long-term no-till (NT) systems could control herbicide resistant weeds and increase profitability of crop production in semi-arid dryland cropping systems. For the purpose of this study, ST is defined as a single tillage event (once every 6-10 years) in an otherwise NT system to reduce density of herbicide tolerant grass weeds. However, there is little information on the long-term (\u3e5 years) effects of ST on soil health parameters and crop yields. This study used long-term tillage and crop rotation plots established in 1976 at the Kansas State University Agriculture Research Center in Hays, KS. Treatments include three rotations: continuous wheat (WW), wheat-sorghum-fallow (WSF), and wheat-fallow (WF); and two tillage regimes: no-till (NT) and reduced tillage (RT). In 2016, a new tillage treatment, ST, was added to control herbicide resistant (HR) grass weeds and to mix soil to reduce nutrient and pH stratification. Soil samples were collected following wheat harvest in 2022 to investigate soil properties after 5 years of ST. Results of the 2022 sampling showed rotation and tillage had no significant effect (P \u3e 0.05) on bulk density. However, bulk density was least in the 0- to-2-inch soil depth compared to the 2- to 6-inch and 6- to 12-inch depths, with values of 1.16, 1.44, and 1.39 g/cm3, respectively. Soil organic carbon (SOC) was greatest in the 0- to 2-inch soil depth. The SOC concentration in soils under NT was not different compared to ST, whereas soils under RT had 8% less SOC than NT. Wind-erodible fraction (WEF) was not different among tillage treatments. Tillage treatments had a significant effect on mean weight diameter (MWD), with NT having the highest MWD followed by ST. Winter wheat yield was greatest in RT across the crop rotations. Strategic tillage increased wheat yields in WW compared to NT. Crop rotation, tillage intensity, and depth were all important with pH because pH was highest in WW rotation, RT tillage, and the 6- to 12-inch soil depth. Phosphorus concentrations were highest in WW and the 0- to 2-inch soil depth. Potassium had the greatest concentrations in RT and the 0- to 2-inch soil depth. Grain sorghum yield was not different between NT and ST, but yields for both were greater than RT. Overall, ST had no negative effect on soil properties or crop yield and can be a mitigation option to control herbicide resistant weeds and increase profitability of dryland crop production

Selective expansion of viral variants following experimental transmission of a reconstituted feline immunodeficiency virus quasispecies

Following long-term infection with virus derived from the pathogenic GL8 molecular clone of feline immunodeficiency virus (FIV), a range of viral variants emerged with distinct modes of interaction with the viral receptors CD134 and CXCR4, and sensitivities to neutralizing antibodies. In order to assess whether this viral diversity would be maintained following subsequent transmission, a synthetic quasispecies was reconstituted comprising molecular clones bearing envs from six viral variants and its replicative capacity compared in vivo with a clonal preparation of the parent virus. Infection with either clonal (Group 1) or diverse (Group 2) challenge viruses, resulted in a reduction in CD4+ lymphocytes and an increase in CD8+ lymphocytes. Proviral loads were similar in both study groups, peaking by 10 weeks post-infection, a higher plateau (set-point) being achieved and maintained in study Group 1. Marked differences in the ability of individual viral variants to replicate were noted in Group 2; those most similar to GL8 achieved higher viral loads while variants such as the chimaeras bearing the B14 and B28 Envs grew less well. The defective replication of these variants was not due to suppression by the humoral immune response as virus neutralising antibodies were not elicited within the study period. Similarly, although potent cellular immune responses were detected against determinants in Env, no qualitative differences were revealed between animals infected with either the clonal or the diverse inocula. However, in vitro studies indicated that the reduced replicative capacity of variants B14 and B28 in vivo was associated with altered interactions between the viruses and the viral receptor and co-receptor. The data suggest that viral variants with GL8-like characteristics have an early, replicative advantage and should provide the focus for future vaccine development

Preliminary Vehicle Impact Simulation Technology Advancement (Pre-VISTA)

DTFH61-90-C-00000The Lawrence Livermore National Laboratory (LLNL) has completed a preliminary evaluation of current finite element technology capability applied to roadside hardware design and vehicle crashworthiness research using the nonlinear structural code, DYNA3D. Independently developed vehicle models were evaluated, modified, and then used in several vehicle crash simulations. Criteria were specified for improving the development of vehicle models. Simulation capabilities were increased, and future direction identified for the roadside safety community

Impaired decisional impulsivity in pathological videogamers

Abstract Background Pathological gaming is an emerging and poorly understood problem. Impulsivity is commonly impaired in disorders of behavioural and substance addiction, hence we sought to systematically investigate the different subtypes of decisional and motor impulsivity in a well-defined pathological gaming cohort. Methods Fifty-two pathological gaming subjects and age-, gender- and IQ-matched healthy volunteers were tested on decisional impulsivity (Information Sampling Task testing reflection impulsivity and delay discounting questionnaire testing impulsive choice), and motor impulsivity (Stop Signal Task testing motor response inhibition, and the premature responding task). We used stringent diagnostic criteria highlighting functional impairment. Results In the Information Sampling Task, pathological gaming participants sampled less evidence prior to making a decision and scored fewer points compared with healthy volunteers. Gaming severity was also negatively correlated with evidence gathered and positively correlated with sampling error and points acquired. In the delay discounting task, pathological gamers made more impulsive choices, preferring smaller immediate over larger delayed rewards. Pathological gamers made more premature responses related to comorbid nicotine use. Greater number of hours played also correlated with a Motivational Index. Greater frequency of role playing games was associated with impaired motor response inhibition and strategy games with faster Go reaction time. Conclusions We show that pathological gaming is associated with impaired decisional impulsivity with negative consequences in task performance. Decisional impulsivity may be a potential target in therapeutic management

Outcomes in patients with gunshot wounds to the brain.

Introduction:Gunshot wounds to the brain (GSWB) confer high lethality and uncertain recovery. It is unclear which patients benefit from aggressive resuscitation, and furthermore whether patients with GSWB undergoing cardiopulmonary resuscitation (CPR) have potential for survival or organ donation. Therefore, we sought to determine the rates of survival and organ donation, as well as identify factors associated with both outcomes in patients with GSWB undergoing CPR. Methods:We performed a retrospective, multicenter study at 25 US trauma centers including dates between June 1, 2011 and December 31, 2017. Patients were included if they suffered isolated GSWB and required CPR at a referring hospital, in the field, or in the trauma resuscitation room. Patients were excluded for significant torso or extremity injuries, or if pregnant. Binomial regression models were used to determine predictors of survival/organ donation. Results:825 patients met study criteria; the majority were male (87.6%) with a mean age of 36.5 years. Most (67%) underwent CPR in the field and 2.1% (n=17) survived to discharge. Of the non-survivors, 17.5% (n=141) were considered eligible donors, with a donation rate of 58.9% (n=83) in this group. Regression models found several predictors of survival. Hormone replacement was predictive of both survival and organ donation. Conclusion:We found that GSWB requiring CPR during trauma resuscitation was associated with a 2.1% survival rate and overall organ donation rate of 10.3%. Several factors appear to be favorably associated with survival, although predictions are uncertain due to the low number of survivors in this patient population. Hormone replacement was predictive of both survival and organ donation. These results are a starting point for determining appropriate treatment algorithms for this devastating clinical condition. Level of evidence:Level II

Towards a belief revision based adaptive and context sensitive information retrieval system

In an adaptive information retrieval (IR) setting, the information seekers' beliefs about which terms are relevant or nonrelevant will naturally fluctuate. This article investigates how the theory of belief revision can be used to model adaptive IR. More specifically, belief revision logic provides a rich representation scheme to formalize retrieval contexts so as to disambiguate vague user queries. In addition, belief revision theory underpins the development of an effective mechanism to revise user profiles in accordance with information seekers' changing information needs. It is argued that information retrieval contexts can be extracted by means of the information-flow text mining method so as to realize a highly autonomous adaptive IR system. The extra bonus of a belief-based IR model is that its retrieval behavior is more predictable and explanatory. Our initial experiments show that the belief-based adaptive IR system is as effective as a classical adaptive IR system. To our best knowledge, this is the first successful implementation and evaluation of a logic-based adaptive IR model which can efficiently process large IR collections