Journal articles on the topic 'Thomas Keller'

Written in the run-up to the COP26 summit held in Glasgow, this review essay reflects on theological tools for the climate justice movement in conversation with five recent books. Reviewed works: Catherine Keller, Facing Apocalypse: Climate, Democracy, and Other Last Chances (Maryknoll, N.Y.: Orbis Books, 2021) Thomas Lynch, Apocalyptic Political Theology: Hegel, Taubes and Malabou. Political Theologies (London: Bloomsbury, 2019) Alastair McIntosh, Riders on the Storm: The Climate Crisis and the Survival of Being (Edinburgh: Birlinn, 2020) Hannah Malcolm, ed., Words for a Dying World: Stories of Grief and Courage from the Global Church (London: SCM Press, 2020) Frances Ward, Like There’s No Tomorrow: Climate Crisis, Eco-Anxiety and God. (Durham: Sacristy Press, 2020)

O trabalho que agora apresentamos corresponde a algumas inquietações decorrentes de um dos aspectos fundamentais da nossa dissertação de doutoramento - a possibilidade de modelar a performance desportivomotora, a partir da integração das perspectivas de Fleishman e Quaintance (1984) e da metodologia da Modelação da Estrutura da Covariância. Trata-se sobretudo de uma exposição sem outras pretensões que não sejam a da estimulação da discussão e da crítica às idéias que avançaremos. A possibilidade de modelar a performance desportivo motora tem exercido um enorme fascínio em investigadores de diferentes áreas do conhecimento. A prová-lo estão as suas variadas e incessantes propostas. Thomas, Eclache e Keller (1 989) oferecem uma visão lata deste movimento no contexto do desporto de rendimento. Convém referir, no entanto e de acordo com a nossa perspectiva, que as propostas pictográficas dos vários autores nem sempre correm em paralelo com o vigor que deveria ser exigido aos enunciados teóricos que lhes servem de base, excepção feita para o modelo de Bouchard e Godbout (1 973).

Abstract Cardiovascular health is related to cognition in later life (Samieri, 2018). Psychological factors, such as depressive symptoms, have been linked with cardiovascular health (Thomas, Kalaria, & O’Brien, 2004). Marital quality, an important indicator of social connection, has been linked with cardiovascular response (Seider et al., 2009), and both depression and marital satisfaction are linked with a quicker recovery from heart attacks (Keller, 1998). Depressive symptoms and marital quality may buffer links between cardiovascular health and cognitive functioning. The purpose of this study was to examine cardiovascular links with cognition, in connection with depressive symptoms and marital quality. Using data from 864 participants of the Life and Family Legacy study (Mean age = 61.78), we examined predictors from 2010 in relation to cognition measured in 2017/2018. Word recall and computation subscales of the Minnesota Cognitive Acuity Screen (MCAS) were used to assess cognitive functioning. Results from multiple regression models indicated that after controlling for age, gender, education, income, and marital status, having hypertension and higher depressive symptoms were predictive of word recall. Lower depressive symptoms were also predictive of higher computation scores. Depression did not moderate links between cardiovascular health and cognitive functioning. Among married participants (n=632), positive marital quality had no main effect nor moderating association with cardiovascular health predicting cognitive functioning. Further research is needed to better understand how biological, psychological, and social factors interact to affect cognition in later life. Longitudinal work should track these associations in context of cognitive changes with aging.

The effects of thermal conductivity which depend on temperature are conversely proportional with the linear function of temperature on free convective flow where the fluid is viscous and incompressible along a heated uniform and the vertical wavy surface has been examined in this study. The boundary layer equations with the associated boundary conditions that govern the flow are converted into a nondimensional form by using an appropriate transformation. In the domain of a vertical plate that is flat, the resulting method of nonlinear PDEs is mapped and then worked out numerically by applying the implicit central finite difference technique with Newton’s quasilinearization method, and the block Thomas algorithm is well known as the Keller-box method. The outputs are obtained in the terms of the heat transferring rate, the frictional coefficient of skin, the isotherms, and streamlines. The outcomes showed that the local heat transferring rate, the local skin friction coefficient, the temperature, and the velocity all are decreasing, and both the thermal layer of boundary and velocity become narrower with the rising values of reciprocal variation of temperature-dependent thermal conductivity. On the other hand, the friction coefficient of skin, the velocity, and the temperature decrease where the friction coefficient of skin and velocity decrease by 43% and 64%, respectively, but the heat transfer rate increases by 61% approximately, and both the boundary layer thermal and velocity become thinner when the Prandtl number increases.

Indonesian Archery Association (PERPANI) is the national organization of the sport of archery. Archery is an Olympic sport that should be competed in every Olympic Games. Indonesia has the opportunity to win a medal at the Olympics because it has won a silver medal which was the first medal for Indonesia at the 1988 Olympics. The main challenge for the Indonesian Archery Association (PERPANI) is the slow regeneration of outstanding athletes. The phenomenon that entering the world of sports as a profession for teenagers is still considered a field that cannot promise a good future. This study aims to identify and analyze the marketing public relations Strategy of the Indonesian Archery Association (PERPANI) in developing archery among teenagers. The theory used in this research is Thomas L Harris' Three Ways Strategy, namely Push, Pull and Pass and Major Tools of Marketing Public Relations Phillip Kotler & Kevin Lane Keller. The Indonesian Archery Association (PERPANI) applies Push, Pull and Push strategies, including by: (a). Produce a movie (Film) entitled 3 Srikandi, about the struggle of the trio of Three Indonesian womens archer to get Indonesia's first medal in the Olympics; (b). Invite important figures, one of which is the President as one of the participants in the archery competition; (c). Incorporating archery scenes in the opening ceremony of the Asean Para Games (Opening Ceremony of the Asian Para Games); (d). Organizing the Olympic Movement In Actions. This study uses a qualitative approach with a qualitative descriptive method using a paradigm. The research data were obtained through in-depth interview techniques. Sources of data or informants are structural officials at the Indonesian Archery Association, technical data analysis using analysis from Mile and Huberman. The Indonesian Archery Association also implements Major Tools of Marketing Public Relations which consists of Publications, events, sponsorship, news, speeches, public services, identity media, event activities are activities that greatly affect the target.

-Richard Price, C.G.A. Oldendorp, C.G.A. Oldendorp's history of the Mission of the Evangelical Brethren on the Caribbean Islands of St. Thomas, St. Croix, and St. John. Edited by Johann Jakob Bossard. English edition and translation by Arnold R. Highfield and Vladimir Barac. Ann Arbor MI: Karoma, 1987. xxxv + 737 pp.-Peter J. Wilson, Lawrence E. Fisher, Colonial madness: mental health in the Barbadian social order. New Brunswick, N.J.: Rutgers University Press, 1985. xvi + 215 pp.-George N. Cave, R.B. le Page ,Acts of identity: Creloe-based approaches to language and ethnicity. Cambridge: Cambridge University Press, 1985. x + 275 pp., Andree Tabouret-Keller (eds)-H. Hoetink, Julia G. Crane, Saba silhouettes: life stories from a Caribbean island. Julia G. Crane (ed), New York: Vantage Press, 1987. x + 515 pp.-Sue N. Greene, Anne Walmsley ,Facing the sea: a new anthology from the Caribbean region. London and Kingston: Heinemann, 1986. ix + 151 pp., Nick Caistor, 190 (eds)-Melvin B. Rahming, Mark McWatt, West Indian literature and its social context. Cave Hill, Barbados, Department of English, 1985.-David Barry Gaspar, Rebecca J. Scott, Slave emancipation in Cuba: the transition to free labor, 1860-1899. Princeton, New Jersey: Princeton University Press, 1985. xviii + 319 pp.-Mary Butler, Louis A. Perez Jr., Cuba under the Platt agreement, 1902-1934. Pittsburgh: University of Pittsburgh Press, 1986. xvii + 410 pp.-Ana M. Rodríguez-Ward, Idsa E. Alegria Ortega, La comisión del status de Puerto Rico: su historia y significación. Río Piedras, Puerto Rico: Editorial Universitaria. 1982. ix + 214 pp.-Alain Buffon, Jean Crusol, Changer la Martinique: initiation a l'économie des Antilles. Paris: Editions Caribeennes, 1986. 96 pp.-Klaus de Albuquerque, Bonham C. Richardson, Panama money in Barbados, 1900-1920. Knoxville: University of Tennesse Press, 1985. xiv + 283 pp.-Steven R. Nachman, Marcel Fredericks ,Society and health in Guyana: the sociology of health care in a developing nation. Authors include Janet Fredericks. Durham: Carolina Academic Press, 1986. xv + 173 pp., John Lennon, Paul Mundy (eds)

Abstract Multiple myeloma (MM) is the second most common hematological malignancy and remains incurable, thus demanding for new therapeutic targets. While the pathophysiology of MM is poorly understood, the substantial responsiveness of MM patients to proteasomal inhibitors (PIs) like bortezomib or carfilzomib hints towards a central role of the ubiquitin proteasome system (UPS). Deubiquitylases (DUBs) are therapeutically targetable components of the UPS, whose inhibition can destabilize oncoproteins. However, the identities of oncoprotein-regulating DUBs remain largely elusive. To identify new vulnerabilities in MM, a CRISPR/Cas9 screen targeting all human DUBs was performed. For validated candidates, phenotypical analysis regarding proliferation and cell cycle progression was performed, as well affinity and non-affinity mass spectrometry-based screens to identify substrates. We thereby identified OTUD6B as a novel oncogene that drives G1/S-transition. LIN28B, a suppressor of microRNA biogenesis, was delineated as both a cell cycle-specific deubiquitylation substrate and activator of OTUD6B. RNA-Seq and qPCR analyses of OTUD6B and LIN28B depleted MM cells revealed that the stabilization of LIN28B drives MYC expression and activity at the G1/S transition, which in turn allows for rapid S-phase entry. Thus, silencing of OTUD6B as well as LIN28B inhibited MM outgrowth in xenograft experiments. Analyses of large MM patient cohorts revealed a progressive increase of OTUD6B expression along the transition from normal plasma cells to MGUS to MM and that high expression of OTUD6B was associated with a significantly adverse overall survival. Furthermore, OTUD6B expression was found to strongly correlate with MYC expression and significantly reduced progression-free survival in patients treated with the PI bortezomib. Knockout of OTUD6B in MM cells significantly enhance the anti-myeloma activity of the drug when using sub-lethal doses. Together, these results validate OTUD6B as a new therapeutically targetable oncogene, dependency, and prognostic factor in MM, that eventually serves as a master regulator of MYC activity to drive cell cycle progression. Citation Format: Ria Spallek, Carmen Paulmann, Oleksandra Karpiuk, Jana Zecha, Susan Klaeger, Isabell Schaeffer, Rupert Öllinger, Thomas Engleitner, Jan Krönke, Matthias Wirth, Ullrich Keller, Roland Rad, Bernahrd Kuster, Florian Bassermann. OTUD6B is a dependency in multiple myeloma that drives S-phase entry via MYC activation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2298.

The Midyear Council Meeting of The Paleontological Society (PS) was called to order by President Patricia H. “Tricia” Kelley on 10 March 2001 at 8:10 hrs in Room 291 of Mendenhall Laboratory on the campus of The Ohio State University. Attending were Council members William I. Ausich, Thomas W. Kammer, Carl W. Stock, Christopher G. Maples, Nigel C. Hughes, Seth Finnegan, Ann F. “Nancy” Budd, Mark A. Wilson, Dale A. Springer, Russell H. “Tim” White, Jed Day, and Elizabeth “Liz” Nesbitt. Steven J. Hageman, and Peter J. Harries were also present.

Carp edema virus disease is killing wild and cultured varieties of carp and worrying koi enthusiasts and carp aquaculturists in the United States and around the world. The disease causes skin lesions and swelling and is sometimes called “koi sleepy disease” because infected fish become lethargic and unresponsive. This 6-page fact sheet describes symptoms, diagnosis, prevention, and what fish producers, wholesalers, or retailers can do if they suspect carp may have contracted the disease. Written by Shohreh Hesami, Pedro Viadanna, Natalie Steckler, Staci Spears, Patrick Thompson, Karen Kelley, Roy Yanong, Ruth Francis-Floyd, Johnny Shelley, Joseph Groff, Andy Goodwin, Olga Haenen, and Thomas Waltzek, and published by the School of Forest Resources and Conservation Program in Fisheries and Aquatic Sciences, October 2015. FA189/FA189: Carp Edema Virus Disease (CEVD) / Koi Sleepy Disease (KSD) (ufl.edu)

buy this issueOn April 8, 2016, in what has already become a historic case on the climate, Magistrate Judge Thomas Coffin of the United States District Court of Oregon ruled against a motion to dismiss, in favor of the youthful plaintiffs in the Children's Trust lawsuit (Kelley Cascade Rose Juliana, et al. v. United States of America, et al.) and against the defendants, consisting of the federal government and the fossil-fuel industry. The twenty-one young people constituting the principal plaintiffs, ranging in age from 8 to 21, insist that the federal government has an obligation to protect the climate for the future on their behalf under the public trust doctrine, based on the fifth and ninth amendments to the U.S. Constitution. They claim, as stated in Coffin's ruling, that "government action and inaction…threatens catastrophic consequences".… The plaintiffs in the suit also include climatologist James Hansen, as a guardian for future generations.… The defendants' argument to dismiss was directed principally at what they contended were limits on the federal government's public trust responsibility. It thus turned on whether the United States was obligated simply to follow capitalist precepts with respect to the natural-physical environment, or whether the government had a public trust to maintain the environment for the population and for future generations, going beyond the rules of the market.Click here to purchase a PDF version of this article at the Monthly Review website.

By February of 1858, the American abolitionist community had at least twice been exposed to a poem — attributed to Robert Browning — entitled “The King is Cold.” It appeared in January in the National Anti-Slavery Standard, a weekly newspaper published in New York City, and, one month later, it was reprinted in William Garrison's Boston paper, the Liberator. Yet aside from this brief record of publication, the poem has left no discernible traces, either before or since. The oddly one-sided (i.e., American) appearances of “The King is Cold” surely contributed to its being overlooked by generations of Browning scholars and editors, including such modern fugitive-hunters as Broughton, Honan, and Kelley. In fact, with a few notable exceptions, Browning scholarship has been reluctant to extend its efforts across the Atlantic. We still await an analysis of the poet's American transactions that would update the important research done by Louise Greer in the 1950s. For most of his life, Browning was much more popular in the United States than in England, and, as Greer puts it, “Browning must have known more Americans than any other English man of letters” (39). And, although their author never visited the United States, Browning's poems arrived by the 1840s, finding enthusiastic audiences that included such luminaries as Hawthorne, Lowell, Emerson, and Thomas Higginson. This Boston intellectual clique — transcendentalist, Unitarian, and abolitionist — recognized in Robert (and, more rapidly, in Elizabeth Barrett) the “brave translunary things that our first poets had” (Lowell qtd. in Greer 14). As the uncatalogued existence of “The King is Cold” suggests, the fruits of this special relationship remain incompletely gathered.

Abstract The non-receptor protein tyrosine phosphatase SHP2 (PTPN11) plays an important role in the regulation of RAS/MAPK signal transduction downstream of growth factor receptor activation. Loss of SHP2 activity suppresses tumor cell growth, making SHP2 a potential target for cancer therapy. Here we report the discovery of GDC-1971 (formerly RLY-1971), a highly potent, selective, and orally bioavailable small-molecule SHP2 inhibitor that stabilizes SHP2 in a closed, auto-inhibited conformation. GDC-1971 potently inhibits both wild-type SHP2 (IC50 <1nM) and the E76K activating mutant (IC50 <250nM) in biochemical assays. In standard 2-dimensional and anchorage-independent growth conditions, GDC-1971 inhibits cellular proliferation in models harboring receptor tyrosine kinases (RTKs), SHP2, NF1, KRAS, or BRAF mutations in a dose-dependent manner. GDC-1971 potently inhibits the proliferation of cellular models harboring KRAS G12C or G12A mutations (median IC50 <80 nM) compared to models harboring other KRAS G12, G13 or Q61 mutations (median IC50 >1 uM), indicating a link between KRAS GTP hydrolysis and SHP2 dependency. Despite this trend, some non-KRAS G12C or G12A cell lines harboring other KRAS mutations responded to GDC-1971 in vitro, suggesting some heterogeneity in RTK/SHP2 signaling dependence in subsets of other KRAS mutants. In vivo, GDC-1971 demonstrates dose-dependent RAS/MAPK pathway inhibition and induces significant tumor-growth inhibition in human xenograft models harboring EGFR and KRAS alterations at continuous daily doses that are well tolerated. Given the reported role of SHP2 as a critical mediator of resistance to targeted therapies, we assessed the activity of GDC-1971 combinations in multiple contexts. We observed increased suppression of the MAPK signaling cascade and anti-proliferation synergy when combining GDC-1971 with EGFR, ALK, and KRAS G12C inhibitors in vitro. The observed in vitro synergy translated to dramatic anti-tumor growth effects in vivo. GDC-1971 in combination with the KRAS G12C covalent inhibitor GDC-6036 resulted in significant regressions at doses well below those required for single agent activity in a KRAS G12C-mutant NSCLC xenograft model. In rodent and dog toxicology studies, GDC-1971 is well tolerated at exposures above those required to induce regression in xenograft models. The biochemical and cellular potency and favorable pharmaceutical properties of GDC-1971 support the further clinical development in RTK/MAPK pathway altered tumors using continuous daily dosing alone and in combination with other targeted agents, including the KRAS-G12C inhibitor GDC-6036 (clinical trial NCT04449874). Citation Format: Bret Williams, Alexander Taylor, Olivia Orozco, Christopher Owen, Elizabeth Kelley, Andre Lescarbeau, Kelley Shortsleeves, Randy Kipp, Vy Nguyen, Erin Brophy, Jeremy Wilbur, Yong Tang, David Lanzetta, Nigel Waters, Sherri Smith, Fabrizio Giordanetto, Paul Maragakis, Jack Greismann, Lindsay Willmore, Eric Therrien, Yang Xiao, Marie Evangelista, Luca Gerosa, Eva Lin, Mark Merchant, Alfonso Arrazate, Emily Chan, Pablo Sáenz-López Larrocha, Stefan Chun, Thomas Hunsaker, Gauri Deshmukh, Christine M. Bowman, David E. Shaw, Mark Murcko, Mahesh Padval, W Patrick Walters, James Watters, Donald A. Bergstrom. Discovery and characterization of the potent, allosteric SHP2 inhibitor GDC-1971 for the treatment of RTK/RAS driven tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3327.

Background:COMPACT is a non-interventional study to collect real-world evidence in European countries and Canada on effectiveness, safety and quality of life in rheumatoid arthritis (RA), ankylosing spondylitis or psoriatic arthritis patients (pts) treated with SDZ-ETN (GP2015), an approved etanercept biosimilar. The first effectiveness and safety data from the study have been reported earlier1.Objectives:This interim analysis assessed patient usage behaviour and feelings of self-administered injection in general and with the auto-injector device using the Self-Injection Assessment Questionnaire (SIAQ) at Week 12 in pts with RA.Methods:Pts aged ≥18 years for whom treatment with SDZ ETN were initiated are being enrolled. The SIAQ, a patient questionnaire validated for pts with RA, was developed to assess overall pt experience with subcutaneous self-injection2. It assesses the perceived self-confidence on self-injection, potential barriers, as well as satisfaction with self-injection via device before the first self-injection (PRE module) and after dosing (POST module). The POST module used in COMPACT includes 21 items grouped into six hypothetical domains: “feelings about injection”,“self-image”, “self-confidence”, “injection-site reactions”, “ease of use of self-injection device (SD), and “satisfaction with self-injection”. Descriptive statistics were used to summarise SIAQ POST module data. The results for “ease of use of SD” domain are reported here. The “ease of use of SD” was rated by pts on a 6-point scale: 1 (very difficult) to 6 (very easy).Results:Of the 430 pts recruited, pts with RA represented the largest group (59.5%, n=256). Majority of pts with RA (77.7%) had comorbidities. Of the 256 pts with RA, 102 (40%) pts who used SD responded to the questionnaire. Majority of the pts found usage of the SD easy or very easy, for each of the domains assessed (Table). 49 % and 14% of the patients were “comfortable” and “very comfortable”, respectively using the SD. A majority of patients reported to be bothered by pain at the injection site “not at all” or only “a little” (69.6%), and to be bothered by redness “not at all” or only “a little” (89.2%), respectively.Table.Overall patient experience with usability of self-injection device at Week 12 (RA population)Domain: Ease of use of self-injection deviceQuestionsCategory, %Very easyEasySome what easySomewhat difficultDifficultVery difficultN/ARemoval of Cap36.334.316.74.93.92.02.0To depress the device34.342.213.72.92.02.92.0To administer without any help42.235.310.82.02.05.92.0Use of self-injection device38.237.311.83.92.93.92.0Conclusion:The interim analysis results, although descriptive, show a clear trend for ease of use and good satisfaction with SDZ-ETN SD in pts with RA.References:[1]Schmalzing M, et al.Arthritis Rheumatol. 2019;71 (suppl 10).[2]Keininger D, et al.Health Qual Life Outcomes. 2011,13;9:2.Disclosure of Interests:Herbert Kellner: None declared, Ayman Askari Speakers bureau: Eli Lilly, Pfizer, Thomas Kupka: None declared, Hilke Friccius-Quecke Employee of: Sandoz Hexal AG, Fabricio Furlan Employee of: Sandoz Hexal AG, Sohaib HACHAICHI Employee of: Sandoz Hexal AG, Marc Schmalzing Consultant of: Paid consultant for Hexal AG

Abstract In the phase 3 CheckMate 459 study (NCT02576509), first-line nivolumab (NIVO) treatment demonstrated higher overall response rate (ORR) and a favorable safety profile, but no significant improvement in overall survival (OS) compared with sorafenib (SORA) in patients with advanced hepatocellular carcinoma (HCC). Here, we report exploratory biomarker analyses from the trial with a potential to identify patients who may benefit from NIVO. Archival or fresh tumor samples were collected before treatment and subjected to whole exome sequencing (WES) (400/730: 55%) for genetic alterations including high microsatellite instability (MSI-H) and WNT/beta-catenin pathway components, as well as whole transcriptome RNA sequencing (469/730: 64%) for analysis of individual genes, inflammation signatures and Gene Set Enrichment Analysis (GSEA). Associations between biomarkers and best overall response (BOR), progression free survival (PFS) and OS were evaluated at a minimum follow-up of 33·6 months. RNA sequencing analysis demonstrated that high Gajewski inflammation gene signature score of ≥0.47 (30% of tumors) was associated with an improvement in BOR (p=0.0043), PFS (p=0.00065) and OS (p=0.0073) within the NIVO arm only. Upon comparison of NIVO versus SORA, only patients whose tumors were inflammation high demonstrated NIVO benefit for BOR (p=0.017), PFS (p=0.0063) and OS (p=0.037). Responses to NIVO were associated with higher expression of CD8A, CD8B, PDCD1 (PD-1) and other inflammation-associated genes within the tumor (false discovery rate, FDR<0.01). GSEA of 50 hallmark gene sets for ORR showed that responders to NIVO and SORA were enriched in similar gene sets such as inflammatory response, interferon alpha/gamma response, E2F targets and G2M checkpoint (FDR<0.01). In contrast, GSEA for OS showed inflammatory response and IL6-JAK-STAT3 signaling gene sets were associated with benefit from NIVO, but shorter OS for SORA (FDR<0.01). WES analysis demonstrated that the prevalence of MSI-H was low (12/400: 3%) and insufficient to confirm relationship to clinical outcomes (no radiographic responses observed in 9 NIVO- and 3 SORA-treated patients). Alterations within beta-catenin gene were not associated with survival outcomes for NIVO. Overall, patients with advanced HCC whose tumors had high Gajewski inflammation gene signature demonstrated improved outcomes with NIVO versus SORA. NIVO responders had higher expression of individual inflammation-associated genes. Expression of inflammatory response and IL6-JAK-STAT3 signaling gene sets showed association with longer OS for NIVO, but not SORA. These exploratory analyses support further studies of tumor inflammation-related biomarkers to identify patients most likely to benefit from PD-1 inhibition in advanced HCC. Citation Format: Jaclyn Neely, Jin Yao, Masatoshi Kudo, Richard S. Finn, Bruno Sangro, Ignacio Melero, Anthony El-Khoueiry, Marina Tschaika, Damir Begic, Ashwin Sama, Parul Doshi, Thomas Yau, Robin Kate Kelley. Genomic and transcriptomic analyses related to the clinical efficacy of first-line nivolumab in advanced hepatocellular carcinoma from the phase 3 CheckMate 459 trial [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2145.

BackgroundAdoptive cell therapy (ACT) utilizing tumor infiltrating lymphocytes (TIL) has demonstrated durable responses in patients with metastatic melanoma and offers potential for other solid tumors. Preclinical experience with expanded TIL from soft tissue sarcoma (STS) demonstrates less frequent tumor-specific reactivity compared to melanoma samples, limiting the potential for efficacy.1 We hypothesized that CD69+ TIL have increased tumor-specific reactivity, which can be manipulated in culture, thereby offering an opportunity to enhance the antitumor effect of this cellular immunotherapy product.MethodsPatients were enrolled on an IRB-approved protocol and TIL were expanded from fresh surgical specimens. After enzymatic digestion, tumor single cell suspensions were cultured in media containing 10% human serum and IL-2 (6000IU/mL). Expanded TIL were then enriched for CD8+ using magnetic bead isolation and CD69+ by flow cytometry cell sorting (FACS). After co-culture with autologous tumor digest, functional capacity was compared between bulk TIL and enriched TIL by evaluation of IFN-gamma (IFNg) and Granzyme B (GzB) secretion. Capacity for direct tumor cytotoxicity was assessed by Cr51 assay after co-culture of autologous immortalized cell lines with expanded TIL subpopulations after enrichment.ResultsFollowing co-culture with autologous tumor digest, CD69+ TIL demonstrated increased IFNg secretion compared to CD69- TIL in 6 samples (1.4–4.2x, p<0.05). CD8+ enriched TIL (75% compared to bulk) had higher relative IFNg secretion in both CD69+ and CD69- subsets (4.2 and 5.8x, respectively, p<0.001). Maximal IFNg secretion was seen from TIL that were both CD69+ sorted and CD8+ enriched, demonstrating an synergistic effect (16.3x vs Bulk CD69-, 4.2x vs Bulk CD69+, 2.8x vs CD8 enriched CD69- ; p<0.001). Functional capacity was also assessed by GzB secretion with similar results. CD69+ TIL had increased relative secretion (1.8–2.2x) compared to CD69- TIL (p< 0.01). CD8+ enriched TIL had increased relative GzB secretion in both CD69- and CD69+ sorted fractions (1.4x, 1.2x, respectively, p<.05). CD69+ sorted and CD8+ enriched TIL demonstrated an additive effect (2.6x vs Bulk CD69-, p<0.01; 1.2x vs Bulk CD69+, p<0.05; 1.8x vs CD8 enriched CD69-, p<0.01). CD8+ enriched CD69+ sorted TIL had greater relative cytotoxicity (3x, p<0.05) at 40:1 E:T ratio against autologous tumor cell lines compared to bulk expanded TIL (figure 1).Abstract 179 Figure 1Functional capacity of CD69+ TIL is demonstrated by increased secretion of GzB (A) and IFNg (B) after co-culture with autologous tumor digest. CD69+ TIL have greater cytotoxicity against autologous immortalized cell lines compared to bulk TIL at 40:1 E:T ratio (C).ConclusionsTIL expanded from STS demonstrate greater tumor-specific functional capacity and cytotoxicity after CD8 enrichment and CD69+ FACS compared to bulk expanded TIL. These data validate the strategy to enhance CD8+CD69+ TIL during culture to yield a more efficacious cellular immunotherapy product.AcknowledgementsThis work was funded by NIH K08CA252642Trial Registration n/aReference1. Mullinax JE, Hall M, Beatty M, Weber AM, Sannasardo Z, Svrdlin T, Hensel J, Bui M, Richards A, Gonzalez RJ, Cox CA, Kelley L, Mulé JJ, Sarnaik AA, Pilon-Thomas S. Expanded Tumor-infiltrating Lymphocytes From Soft Tissue Sarcoma Have Tumor-specific Function. J Immunother 2021 Feb-Mar 01;44(2):63–70.Ethics ApprovalAbstract cites IRB-approved protocol in methods section.Consent n/a

We present a fully second order IMplicit/EXplicit (IMEX) time integration technique for solving incompressible multi-phase flow problems. A typical incompressible multi-phase flow model consists of the Navier–Stokes equations plus an interface dynamics equation (e.g., the level set equation). Our IMEX strategy is applied to such a model in the following manner. The hyperbolic terms of the Navier–Stokes equations together with the interface dynamics equation are solved explicitly (Explicit Block) making use of the well-understood explicit numerical schemes [Leveque, R. J. [1998] Finite Volume Methods for Hyperbolic Problems, “Texts in Applied Mathematics”, (Cambridge University Press); Thomas, J. W. [1999] Numerical Partial Differential Equations II (Conservation Laws and Elliptic Equations), “Texts in Applied Mathematics” (Springer-Verlag, New York)]. On the other hand, the nonhyperbolic (stiff) parts of the flow equations are solved implicitly (Implicit Block) within the framework of the Jacobian-Free Newton Krylov (JFNK) method [Knoll, D. A. and Keyes, D. E. [2004] Jacobian-free Newton Krylov methods: A survey of approaches and applications. J. Comput. Phys. 193, 357–397; Saad, Y. [2003] Iterative Methods for Sparse Linear Systems (Siam); Kelley, C. T. [2003] Solving Nonlinear Equations with Newton’s Method (Siam)]. In our algorithm implementation, the explicit block is embedded in the implicit block in a way that it is always part of the nonlinear function evaluation. In this way, there exists a continuous interaction between the implicit and explicit algorithm blocks meaning that the improved solutions (in terms of time accuracy) at each nonlinear iteration are immediately felt by the explicit block and the improved explicit solutions are readily available to form the next set of nonlinear residuals. This continuous interaction between the two algorithm blocks results in an implicitly balanced algorithm in that all nonlinearities due to coupling of different time terms are converged with the desired numerical time accuracy. In other words, we obtain a self-consistent IMEX method that eliminates the possible order reductions in time convergence that is quite common in certain types of nonlinearly coupled systems. We remark that an incompressible multi-phase flow model can be a highly nonlinearly coupled system with the involvement of very stiff surface tension source terms. These kinds of flow problems are difficult to tackle numerically. In other words, highly nonlinear surface terms may remain unconverged leading to time inaccuracies or time order reductions to the first order even though the overall numerical scheme is designed as high order (second-order or higher) [Sussman, M. and Ohta, M. [2009] A stable and efficient method for treating surface tension in incompressible two-phase flow, SIAM J. Sci. Comput. 31(4), 2447–2471; Zheng, W., Zhu, B., Kim, B. and Fedkiw, R. [2015] A new incompressibility discretization for a hybrid particle MAC grid representation with surface tension, J. Comput. Phys. 280, 96–142]. These and few more issues are addressed in this paper. We have numerically tested our newly proposed scheme by solving several multi-phase flow settings such as an air bubble rising in water, a Rayleigh–Taylor instability problem that is initiated by placing a heavy fluid on top of a lighter one, and a droplet problem in which a water droplet hits the pool of water. Our numerical results show that we have achieved the second-order time accuracy without any order reductions. Moreover, the interfaces between the fluids are captured reasonably well.

Michael B. Katz. Reconstructing American Education. Cambridge and London: Harvard University Press, 1987. Pp. viii, 212. Cloth, $22.50; E. D. Hirsch, Jr. Cultural Literacy: What Every American Needs to Know. Boston: Houghton Mifflin Co., 1987. Pp. xvii, 251. Cloth, $16.45; Diana Ravitch and Chester E. Finn, Jr. What Do Our 17-Year-Olds Know? A Report on the First National Assessment of History and Literature. New York: Harper & Row, 1987. Pp. ix, 293. Cloth, $15.95. Review by Richard A. Diem of The University of Texas at San Antonio. Henry J. Steffens and Mary Jane Dickerson. Writer's Guide: History. Lexington, Massachusetts, and Toronto: D. C. Heath and Company, 1987. Pp. x, 211. Paper, $6.95. Review by William G. Wraga of Bernards Township Public Schools, Basking Ridge, New Jersey. J. Kelley Sowards, ed. Makers of the Western Tradition: Portraits from History. New York: St. Martin's Press, 1987. Fourth edition. Vol: 1: Pp. ix, 306. Paper, $12.70. Vol. 2: Pp. ix, 325. Paper, $12.70. Review by Robert B. Luehrs of Fort Hays State University. John L. Beatty and Oliver A. Johnson, eds. Heritage of Western Civilization. Englewood Cliffs, New Jersey: Prentice-Hall, Inc., 1987. Sixth Edition. Volume I: Pp. xi, 465. Paper, $16.00; Volume II: pp. xi, 404. Paper, $16.00. Review by Dav Levinson of Thayer Academy, Braintree, Massachusetts. Lynn H. Nelson, ed. The Human Perspective: Readings in World Civilization. New York: Harcourt Brace Jovanovich, 1987. Vol. I: The Ancient World to the Early Modern Era. Pp. viii, 328. Paper, $10.50. Vol. II: The Modern World Through the Twentieth Century. Pp, x, 386. Paper, 10.50. Review by Gerald H. Davis of Georgia State University. Gerald N. Grob and George Attan Billias, eds. Interpretations of American History: Patterns and Perspectives. New York: The Free Press, 1987. Fifth Edition. Volume I: Pp. xi, 499. Paper, $20.00: Volume II: Pp. ix, 502. Paper, $20.00. Review by Larry Madaras of Howard Community College. Eugene Kuzirian and Larry Madaras, eds. Taking Sides: Clashing Views on Controversial Issues in American History. -- Volume II: Reconstruction to the Present. Guilford, Connecticut: The Dushkin Publishing Groups, Inc., 1987. Pp. xii, 384. Paper, $9.50. Review by James F. Adomanis of Anne Arundel County Public Schools, Annapolis, Maryland. Joann P. Krieg, ed. To Know the Place: Teaching Local History. Hempstead, New York: Hofstra University Long Island Studies Institute, 1986. Pp. 30. Paper, $4.95. Review by Marilyn E. Weigold of Pace University. Roger Lane. Roots of Violence in Black Philadelphia, 1860-1900. Cambridge, Massachusetts, and London: Harvard University Press, 1986. Pp. 213. Cloth, $25.00. Review by Ronald E. Butchart of SUNY College at Cortland. Pete Daniel. Breaking the Land: The Transformation of Cotton, Tobacco, and Rice Cultures since 1880. Urbana and Chicago: University of Illinois Press, 1985. Pp. xvi, 352. Paper, $22.50. Review by Thomas S. Isern of Emporia State University. Norman L. Rosenberg and Emily S. Rosenberg. In Our Times: America Since World War II. Englewood Cliffs, New Jersey: Prentice-Hall, 1987. Third edition. Pp. xi, 316. Paper, $20.00; William H. Chafe and Harvard Sitkoff, eds. A History of Our Time: Readings on Postwar America. New York: Oxford University Press, 1987. Second edition. Pp. xiii, 453. Paper, $12.95. Review by Monroe Billington of New Mexico State University. Frank W. Porter III, ed. Strategies for Survival: American Indians in the Eastern United States. New York, Westport, Connecticut, and London: Greenwood Press, 1986. Pp. xvi, 232. Cloth, $35.00. Review by Richard Robertson of St. Charles County Community College. Kevin Sharpe, ed. Faction & Parliament: Essays on Early Stuart History. London and New York: Methuen, 1985. Pp. xvii, 292. Paper, $13.95; Derek Hirst. Authority and Conflict: England, 1603-1658. Cambridge: Harvard University Press, 1986. Pp. viii, 390. Cloth, $35.00. Review by K. Gird Romer of Kennesaw College. N. F. R. Crafts. British Economic Growth During the Industrial Revolution. New York: Oxford University Press, 1985. Pp. 193. Paper, $11.95; Maxine Berg. The Age of Manufactures, 1700-1820. New York: Oxford University Press, 1985. Pp. 378. Paper, $10.95. Review by C. Ashley Ellefson of SUNY College at Cortland. J. M. Thompson. The French Revolution. New York: Basil Blackwell, 1985 reissue. Pp. xvi, 544. Cloth, $45.00; Paper, $12.95. Review by W. Benjamin Kennedy of West Georgia College. J. P. T. Bury. France, 1814-1940. London and New York: Methuen, 1985. Fifth edition. Pp. viii, 288. Paper, $13.95; Roger Magraw. France, 1815-1914: The Bourgeois Century. New York and Oxford: Oxford University Press, 1985. Pp. 375. Cloth, $24.95; Paper, $9.95; D. M.G. Sutherland. France, 1789-1815: Revolution and Counterrevolution. New York and Oxford: Oxford University Press, 1986. Pp. 242. Cloth, $32.50; Paper, $12.95. Review by Fred R. van Hartesveldt of Fort Valley State College. Woodford McClellan. Russia: A History of the Soviet Period. Englewood Cliffs, New Jersey: Prentice-Hall, 1986. Pp. xi, 387. Paper, $23.95. Review by Pasquale E. Micciche of Fitchburg State College. Ranbir Vohra. China's Path to Modernization: A Historical Review from 1800 to the Present. Englewood Cliffs, New Jersey: Prentice-Hall, 1987. Pp. xiii, 302. Paper, $22.95. Reivew by Steven A. Leibo of Russell Sage College. John King Fairbank. China Watch. Cambridge and London: Harvard University Press, 1987. Pp. viii, Cloth, $20.00. Review by Darlene E. Fisher of New Trier Township High School, Winnetka, Illinois. Ronald Takaki, ed. From Different Shores: Perspectives on Race and Ethnicity in America. New York and Oxford: Oxford University Press, 1987. Pp. 253. Paper, $13.95. Review by Robert C. Sims of Boise State University.

Hamsters were exposed to sodium arsenite (173 mg As/L) in drinking water for 6 days. Equal amounts of proteins from urinary bladder or liver extracts of control and arsenic-treated hamsters were labeled with Cy3 and Cy5 dyes, respectively. After differential in gel electrophoresis and analysis by the DeCyder software, several protein spots were found to be down-regulated and several were up regulated. Our experiments indicated that in the bladder tissues of hamsters exposed to arsenite, transgelin was down-regulated and GST-pi was up-regulated. The loss of transgelin expression has been reported to be an important early event in tumor progression and a diagnostic marker for cancer development [29-32]. Down-regulation of transgelin expression may be associated with the carcinogenicity of inorganic arsenic in the urinary bladder. In the liver of arsenite-treated hamsters, ornithine aminotransferase was up-regulated, and senescence marker protein 30 and fatty acid binding protein were down-regulated. The volume ratio changes of these proteins in the bladder and liver of hamsters exposed to arsenite were significantly different than that of control hamsters. Introduction Chronic exposure to inorganic arsenic can cause cancer of the skin, lungs, urinary bladder, kidneys, and liver [1-6]. The molecular mechanisms of the carcinogenicity and toxicity of inorganic arsenic are not well understood [7-9). Humans chronically exposed to inorganic arsenic excrete MMA(V), DMA(V) and the more toxic +3 oxidation state arsenic biotransformants MMA(III) and DMA (III) in their urine [10, 11], which are carcinogen [12]· After injection of mice with sodium arsenate, the highest concentrations of the very toxic MMA(III) and DMA(III) were in the kidneys and urinary bladder tissue, respectively, as shown by experiments of Chowdhury et al [13]. Many mechanisms of arsenic toxicity and carcinogenicity have been suggested [1, 7, 14] including chromosome abnormalities [15], oxidative stress [16, 17], altered growth factors [18], cell proliferation [19], altered DNA repair [20], altered DNA methylation patterns [21], inhibition of several key enzymes [22], gene amplification [23] etc. Some of these mechanisms result in alterations in protein expression. Methods for analyzing multiple proteins have advanced greatly in the last several years. In particularly, mass spectrometry (MS) and tandem MS (MS/MS) are used to analyze peptides following protein isolation using two-dimensional (2-D) gel electrophoresis and proteolytic digestion [24]. In the present study, Differential In Gel Electrophoresis (DIGE) coupled with Mass Spectrometry (MS) has been used to study some of the proteomic changes in the urinary bladder and liver of hamsters exposed to sodium arsenite in their drinking water. Our results indicated that transgelin was down-regulated and GST-pi was up-regulated in the bladder tissues. In the liver tissues ornithine aminotransferase was up-regulated, and senescence marker protein 30, and fatty acid binding protein were down-regulated. Materials and Methods Chemicals Tris, Urea, IPG strips, IPG buffer, CHAPS, Dry Strip Cover Fluid, Bind Silane, lodoacetamide, Cy3 and Cy5 were from GE Healthcare (formally known as Amersham Biosciences, Uppsala, Sweden). Thiourea, glycerol, SDS, DTT, and APS were from Sigma-Aldrich (St. Louis, MO, USA). Glycine was from USB (Cleveland, OH, USA). Acrylamide Bis 40% was from Bio-Rad (Hercules, CA, USA). All other chemicals and biochemicals used were of analytical grade. All solutions were made with Milli-Q water. Animals Male hamsters (Golden Syrian), 4 weeks of age, were purchased from Harlan Sprague Dawley, USA. Upon arrival, hamsters were acclimated in the University of Arizona animal care facility for at least 1 week and maintained in an environmentally controlled animal facility operating on a 12-h dark/12-h light cycle and at 22-24°C. They were provided with Teklad (Indianapolis, IN) 4% Mouse/Rat Diet # 7001 and water, ad libitum, throughout the acclimation and experimentation periods. Sample preparation and labelling Hamsters were exposed to sodium arsenite (173 mg) in drinking water for 6 days and the control hamsters were given tap water. On the 6th day hamsters were decapitated rapidly by guillotine. Urinary bladder tissues and liver were removed, blotted on tissue papers (Kimtech Science, Precision Wipes), and weighed. Hamster urinary bladder or liver tissues were homogenized in lysis buffer (30mMTris, 2M thiourea, 7M urea, and 4% w/w CHAPS adjusted to pH 8.5 with dilute HCI), at 4°C using a glass homogenizer and a Teflon coated steel pestle; transferred to a 5 ml acid-washed polypropylene tube, placed on ice and sonicated 3 times for 15 seconds. The sonicate was centrifuged at 12,000 rpm for 10 minutes at 4°C. Small aliquots of the supernatants were stored at -80°C until use (generally within one week). Protein concentration was determined by the method of Bradford [25] using bovine serum albumin as a standard. Fifty micrograms of lysate protein was labeled with 400 pmol of Cy3 Dye (for control homogenate sample) and Cy5 Dye (for arsenic-treated urinary bladder or liver homogenate sample). The samples containing proteins and dyes were incubated for 30 min on ice in the dark. To stop the labeling reaction, 1uL of 10 mM lysine was added followed by incubation for 10 min on ice in the dark. To each of the appropriate dye-labeled protein samples, an additional 200 ug of urinary bladderor liver unlabeled protein from control hamster sample or arsenic-treated hamster sample was added to the appropriate sample. Differentially labeled samples were combined into a single Microfuge tube (total protein 500 ug); protein was mixed with an equal volume of 2x sample buffer [2M thiourea, 7M urea, pH 3-10 pharmalyte for isoelectric focusing 2% (v/v), DTT 2% (w/v), CHAPS 4% (w/v)]; and was incubated on ice in the dark for 10 min. The combined samples containing 500 ug of total protein were mixed with rehydration buffer [CHAPS 4% (w/v), 8M urea, 13mM DTT, IPG buffer (3-10) 1% (v/v) and trace amount of bromophenol blue]. The 450 ul sample containing rehydration buffer was slowly pipetted into the slot of the ImmobilinedryStripReswelling Tray and any large bubbles were removed. The IPG strip (linear pH 3-10, 24 cm) was placed (gel side down) into the slot, covered with drystrip cover fluid (Fig. 1), and the lid of the Reswelling Tray was closed. The ImmobillineDryStrip was allowed to rehydrate at room temperature for 24 hours. First dimension Isoelectric focusing (IEF) The labeled sample was loaded using the cup loading method on universal strip holder. IEF was then carried out on EttanIPGphor II using multistep protocol (6 hr @ 500 V, 6 hr @ 1000 V, 8 hr @ 8000 V). The focused IPG strip was equilibrated in two steps (reduction and alkylation) by equilibrating the strip for 10 min first in 10 ml of 50mM Tris (pH 8.8), 6M urea, 30% (v/v) glycerol, 2% (w/v) SDS, and 0.5% (w/v) DTT, followed by another 10 min in 10 ml of 50mM Tris (pH 8.8), 6M urea, 30% (v/v) glycerol, 2% (w/v) SDS, and 4.5% (w/v) iodoacetamide to prepare it for the second dimension electrophoresis. Second dimension SDS-PAGE The equilibrated IPG strip was used for protein separation by 2D-gel electrophoresis (DIGE). The strip was sealed at the top of the acrylamide gel for the second dimension (vertical) (12.5% polyacrylamide gel, 20x25 cm x 1.5 mm) with 0.5% (w/v) agarose in SDS running buffer [25 mMTris, 192 mM Glycine, and 0.1% (w/v) SDS]. Electrophoresis was performed in an Ettan DALT six electrophoresis unit (Amersham Biosciences) at 1.5 watts per gel, until the tracking dye reached the anodic end of the gel. Image analysis and post-staining The gel then was imaged directly between glass plates on the Typhoon 9410 variable mode imager (Sunnyvale, CA, USA) using optimal excitation/emission wavelength for each DIGE fluor: Cy3 (532/580 nm) and Cy5 (633/670 nm). The DIGE images were previewed and checked with Image Quant software (GE Healthcare) where all the two separate gel images could be viewed as a single gel image. DeCyde v.5.02 was used to analyze the DIGE images as described in the Ettan DIGE User Manual (GE Healthcare). The appropriate up-/down regulated spots were filtered based on an average volume ratio of ± over 1.2 fold. After image acquisition, the gel was fixed overnight in a solution containing 40% ethanol and 10% acetic acid. The fixed gel was stained with SyproRuby (BioRad) according to the manufacturer protocol (Bio-Rad Labs., 2000 Alfred Nobel Drive, Hercules, CA 94547). Identification of proteins by MS Protein spot picking and digestion Sypro Ruby stained gels were imaged using an Investigator ProPic and HT Analyzer software, both from Genomic Solutions (Ann Arbor, MI). Protein spots of interest that matched those imaged using the DIGE Cy3/Cy5 labels were picked robotically, digested using trypsin as described previously [24] and saved for mass spectrometry identification. Liquid chromatography (LC)- MS/MS analysis LC-MS/MS analyses were carried out using a 3D quadrupole ion trap massspectrometer (ThermoFinnigan LCQ DECA XP PLUS; ThermoFinnigan, San Jose, CA) equipped with a Michrom Paradigm MS4 HPLC (MichromBiosources, Auburn, CA) and a nanospray source, or with a linear quadrupole ion trap mass spectrometer (ThermoFinnigan LTQ), also equipped with a Michrom MS4 HPLC and a nanospray source. Peptides were eluted from a 15 cm pulled tip capillary column (100 um I.D. x 360 um O.D.; 3-5 um tip opening) packed with 7 cm Vydac C18 (Vydac, Hesperia, CA) material (5 µm, 300 Å pore size), using a gradient of 0-65% solvent B (98% methanol/2% water/0.5% formic acid/0.01% triflouroacetic acid) over a 60 min period at a flow rate of 350 nL/min. The ESI positive mode spray voltage was set at 1.6 kV, and the capillary temperature was set at 200°C. Dependent data scanning was performed by the Xcalibur v 1.3 software on the LCQ DECA XP+ or v 1.4 on the LTQ [27], with a default charge of 2, an isolation width of 1.5 amu, an activation amplitude of 35%, activation time of 50 msec, and a minimal signal of 10,000 ion counts (100 ion counts on the LTQ). Global dependent data settings were as follows: reject mass width of 1.5 amu, dynamic exclusion enabled, exclusion mass width of 1.5 amu, repeat count of 1, repeat duration of a min, and exclusion duration of 5 min. Scan event series were included one full scan with mass range of 350-2000 Da, followed by 3 dependent MS/MS scans of the most intense ion. Database searching Tandem MS spectra of peptides were analyzed with Turbo SEQUEST, version 3.1 (ThermoFinnigan), a program that allows the correlation of experimental tandem MS data with theoretical spectra generated from known protein sequences. All spectra were searched against the latest version of the non redundant protein database from the National Center for Biotechnology Information (NCBI 2006; at that time, the database contained 3,783,042 entries). Statistical analysis The means and standard error were calculated. The Student's t-test was used to analyze the significance of the difference between the control and arsenite exposed hamsters. P values less than 0.05 were considered significant. The reproducibility was confirmed in separate experiments. Results Analysis of proteins expression After DIGE (Fig. 1), the gel was scanned by a Typhoon Scanner and the relative amount of protein from sample 1 (treated hamster) as compared to sample 2 (control hamster) was determined (Figs. 2, 3). A green spot indicates that the amount of protein from sodium arsenite-treated hamster sample was less than that of the control sample. A red spot indicates that the amount of protein from the sodium arsenite-treated hamster sample was greater than that of the control sample. A yellow spot indicates sodium arsenite-treated hamster and control hamster each had the same amount of that protein. Several protein spots were up-regulated (red) or down-regulated (green) in the urinary bladder samples of hamsters exposed to sodium arsenite (173 mg As/L) for 6 days as compared with the urinary bladder of controls (Fig. 2). In the case of liver, several protein spots were also over-expressed (red) or under-expressed (green) for hamsters exposed to sodium arsenite (173 mg As/L) in drinking water for 6 days (Fig. 3). The urinary bladder samples were collected from the first and second experiments in which hamsters were exposed to sodium arsenite (173 mg As/L) in drinking water for 6 days and the controls were given tap water. The urinary bladder samples from the 1st and 2nd experiments were run 5 times in DIGE gels on different days. The protein expression is shown in Figure 2 and Table 1. The liver samples from the 1st and 2nd experiments were also run 3 times in DIGE gels on different days. The proteins expression were shown in Figure 3 and Table 2. The volume ratio changed of the protein spots in the urinary bladder and liver of hamsters exposed to arsenite were significantly differences than that of the control hamsters (Table 1 and 2). Protein spots identified by LC-MS/MS Bladder The spots of interest were removed from the gel, digested, and their identities were determined by LC-MS/MS (Fig. 2 and Table 1). The spots 1, 2, & 3 from the gel were analyzed and were repeated for the confirmation of the results (experiments; 173 mg As/L). The proteins for the spots 1, 2, and 3 were identified as transgelin, transgelin, and glutathione S-transferase Pi, respectively (Fig. 2). Liver We also identified some of the proteins in the liver samples of hamsters exposed to sodium arsenite (173 mg As/L) in drinking water for 6 days (Fig. 3). The spots 4, 5, & 6 from the gels were analyzed and were repeated for the confirmation of the results. The proteins for the spots 4, 5, and 6 were identified as ornithine aminotransferase, senescence marker protein 30, and fatty acid binding protein, respectively (Fig. 3) Discussion The identification and functional assignment of proteins is helpful for understanding the molecular events involved in disease. Weexposed hamsters to sodium arsenite in drinking water. Controls were given tap water. DIGE coupled with LC-MS/MS was then used to study the proteomic change in arsenite-exposed hamsters. After electrophoresis DeCyder software indicated that several protein spots were down-regulated (green) and several were up-regulated (red). Our overall results as to changes and functions of the proteins we have studied are summarized in Table 3. Bladder In the case of the urinary bladder tissue of hamsters exposed to sodium arsenite (173 mg As/L) in drinking water for 6 days, transgelin was down-regulated and GST-pi was up-regulated. This is the first evidence that transgelin is down-regulated in the bladders of animals exposed to sodium arsenite. Transgelin, which is identical to SM22 or WS3-10, is an actin cross linking/gelling protein found in fibroblasts and smooth muscle [28, 29]. It has been suggested that the loss of transgelin expression may be an important early event in tumor progression and a diagnostic marker for cancer development [30-33]. It may function as a tumor suppressor via inhibition of ARA54 (co-regulator of androgen receptor)-enhanced AR (androgen receptor) function. Loss of transgelin and its suppressor function in prostate cancer might contribute to the progression of prostate cancer [30]. Down-regulation of transgelin occurs in the urinary bladders of rats having bladder outlet obstruction [32]. Ras-dependent and Ras-independent mechanisms can cause the down regulation of transgelin in human breast and colon carcinoma cell lines and patient-derived tumorsamples [33]. Transgelin plays a role in contractility, possibly by affecting the actin content of filaments [34]. In our experiments loss of transgelin expression may be associated or preliminary to bladder cancer due to arsenic exposure. Arsenite is a carcinogen [1]. In our experiments, LC-MS/MS analysis showed that two spots (1 and 2) represent transgelin (Fig. 2 and Table 1). In human colonic neoplasms there is a loss of transgelin expression and the appearance of transgelin isoforms (31). GST-pi protein was up-regulated in the bladders of the hamsters exposed to sodium arsenite. GSTs are a large family of multifunctional enzymes involved in the phase II detoxification of foreign compounds [35]. The most abundant GSTS are the classes alpha, mu, and pi classes [36]. They participate in protection against oxidative stress [37]. GST-omega has arsenic reductase activity [38]. Over-expression of GST-pi has been found in colon cancer tissues [39]. Strong expression of GST-pi also has been found in gastric cancer [40], malignant melanoma [41], lung cancer [42], breast cancer [43] and a range of other human tumors [44]. GST-pi has been up-regulated in transitional cell carcinoma of human urinary bladder [45]. Up-regulation of glutathione – related genes and enzyme activities has been found in cultured human cells by sub lethal concentration of inorganic arsenic [46]. There is evidence that arsenic induces DNA damage via the production of ROS (reactive oxygen species) [47]. GST-pi may be over-expressed in the urinary bladder to protect cells against arsenic-induced oxidative stress. Liver In the livers of hamsters exposed to sodium arsenite, ornithine amino transferase was over-expressed, senescence marker protein 30 was under-expressed, and fatty acid binding protein was under-expressed. Ornithine amino transferase has been found in the mitochondria of many different mammalian tissues, especially liver, kidney, and small intestine [48]. Ornithine amino transferase knockdown inhuman cervical carcinoma and osteosarcoma cells by RNA interference blocks cell division and causes cell death [49]. It has been suggested that ornithine amino transferase has a role in regulating mitotic cell division and it is required for proper spindle assembly in human cancer cells [49]. Senescence marker protein-30 (SMP30) is a unique enzyme that hydrolyzes diisopropylphosphorofluoridate. SMP30, which is expressed mostly in the liver, protects cells against various injuries by stimulating membrane calcium-pump activity [50]. SMP30 acts to protect cells from apoptosis [51]. In addition it protects the liver from toxic agents [52]. The livers of SMP30 knockout mice accumulate phosphatidylethanolamine, cardiolipin, phosphatidyl-choline, phosphatidylserine, and sphingomyelin [53]. Liver fatty acid binding protein (L-FABP) also was down- regulated. Decreased liver fatty acid-binding capacity and altered liver lipid distribution hasbeen reported in mice lacking the L-FABP gene [54]. High levels of saturated, branched-chain fatty acids are deleterious to cells and animals, resulting in lipid accumulation and cytotoxicity. The expression of fatty acid binding proteins (including L-FABP) protected cells against branched-chain saturated fatty acid toxicity [55]. Limitations: we preferred to study the pronounced spots seen in DIGE gels. Other spots were visible but not as pronounced. Because of limited funds, we did not identify these others protein spots. In conclusion, urinary bladders of hamsters exposed to sodium arsenite had a decrease in the expression of transgelin and an increase in the expression of GST-pi protein. Under-expression of transgelin has been found in various cancer systems and may be associated with arsenic carcinogenicity [30-33). Inorganic arsenic exposure has resulted in bladder cancer as has been reported in the past [1]. Over-expression of GST-pi may protect cells against oxidative stress caused by arsenite. In the liver OAT was up regulated and SMP-30 and FABP were down regulated. These proteomic results may be of help to investigators studying arsenic carcinogenicity. The Superfund Basic Research Program NIEHS Grant Number ES 04940 from the National Institute of Environmental Health Sciences supported this work. Additional support for the mass spectrometry analyses was provided by grants from NIWHS ES06694, NCI CA023074 and the BIOS Institute of the University of Arizona. Acknowledgement The Author wants to dedicate this paper to the memory of his former supervisor Dr. H. VaskenAposhian who passed away in September 6, 2019. He was an emeritus professor of the Department of Molecular and Cellular Biology at the University of Arizona. 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SL-279, a 6-page fact sheet by Rao Mylavarapu, Kelley Hines, and Thomas Obreza, presents the details of several types of diagnostic tests for nutrient status and management in commercial citrus groves in Florida, as well as sample submission procedures. Includes references and a copy of the sample submission form (in the pdf version). Published by the UF Soil and Water Science Department, December 2008. SL 279/SS492: Diagnostic Nutrient Testing for Commercial Citrus in Florida (ufl.edu)