bluuebunny/crossref_metadata_2025 huggingface dataset:
Relationship between neuromelanin and dopamine terminals within the Parkinson’s nigrostriatal system
Authors: Martín-Bastida Antonio , Lao-Kaim Nicholas P , Roussakis Andreas Antonios , Searle Graham E , Xing Yue , Gunn Roger N , Schwarz Stefan T , Barker Roger A , Auer Dorothee P , Piccini Paola
Published: April 2019

Parkinson’s disease is characterized by the progressive loss of pigmented dopaminergic neurons in the substantia nigra and associated striatal deafferentation. Neuromelanin content is thought to reflect the loss of pigmented neurons, but available data characterizing its relationship with striatal dopaminergic integrity are not comprehensive or consistent, and predominantly involve heterogeneous samples. In this cross-sectional study, we used neuromelanin-sensitive MRI and the highly specific dopamine transporter PET radioligand, 11C-PE2I, to assess the association between neuromelanin-containing cell levels in the substantia nigra pars compacta and nigrostriatal terminal density in vivo, in 30 patients with bilateral Parkinson’s disease. Fifteen healthy control subjects also underwent neuromelanin-sensitive imaging. We used a novel approach taking into account the anatomical and functional subdivision of substantia nigra into dorsal and ventral tiers and striatal nuclei into pre- and post-commissural subregions, in accordance with previous animal and post-mortem studies, and consider the clinically asymmetric disease presentation. In vivo, Parkinson’s disease subjects displayed reduced neuromelanin levels in the ventral (−30 ± 28%) and dorsal tiers (−21 ± 24%) as compared to the control group [F(1,43) = 11.95, P = 0.001]. Within the Parkinson’s disease group, nigral pigmentation was lower in the ventral tier as compared to the dorsal tier [F(1,29) = 36.19, P < 0.001] and lower in the clinically-defined most affected side [F(1,29) = 4.85, P = 0.036]. Similarly, lower dopamine transporter density was observed in the ventral tier [F(1,29) = 76.39, P < 0.001] and clinically-defined most affected side [F(1,29) = 4.21, P = 0.049]. Despite similar patterns, regression analysis showed no significant association between nigral pigmentation and nigral dopamine transporter density. However, for the clinically-defined most affected side, significant relationships were observed between pigmentation of the ventral nigral tier with striatal dopamine transporter binding in pre-commissural and post-commissural striatal subregions known to receive nigrostriatal projections from this tier, while the dorsal tier correlated with striatal projection sites in the pre-commissural striatum (P < 0.05, Benjamini-Hochberg corrected). In contrast, there were no statistically significant relationships between these two measures in the clinically-defined least affected side. These findings provide important insights into the topography of nigrostriatal neurodegeneration in Parkinson’s disease, indicating that the characteristics of disease progression may fundamentally differ across hemispheres and support post-mortem data showing asynchrony in the loss of neuromelanin-containing versus tyrosine hydroxylase positive nigral cells.
Financialisation, institutions and financing constraints in developing countries
Authors: Mertzanis Charilaos
Published: April 2019

The paper uses a consistent firm-level data from the World Banks Enterprise Surveys to explore the impact of financialisation in the economy on firms’ access to finance in 138 developing countries. Access to finance reflects survey-based firms’ perceptions of external financing constraints. Financialisation is proxied by consistent cross-country measures of financial depth. These proxies capture separately the role of bank-based versus market-based financing. Firm-, sector- and country-level information is jointly used for the analysis. Firm-specific characteristics and economic and non-economic national factors are included as controls. The results show that the proxies of financialisation are broadly robust predictors of financing constraints of firms in developing countries. However, the magnitude of the financialisation effect varies between bank-based and market-based channels of financing as well as between low- and high-income countries, and it is influenced by social, institutional and religious factors.
Parallel Progress in Perceived Age and Life Expectancy
Authors: Steiner Ulrich K , Larsen Lisbeth A , Christensen Kaare
Published: April 2019

Background

Human life expectancy continues to rise in most populations. This rise not only leads to longer lives but also is accompanied by improved health at a given age, that is, recent cohorts show a reduction of biological age for a given chronological age. Despite or even because of the diversity of biomarkers of aging, an accurate quantification of a general shift in biological age across time has been challenging.

Methods

Here, we compared age perception of facial images taken in 2001 over a decade and related these changes in age perception to changes in life expectancy.

Results

We show that age perception changes substantially across time and parallels the progress in life expectancy. In 2012, people aged more than 70 years needed to look 2.3 years younger to be rated the same age as in 2002.

Conclusions

Our results suggest that age perception reflects the past life events better than predicts future length of life, that is, it is written in your face how much you have aged so far. We draw this conclusion as age perception among elderly individuals paralleled changes in life expectancy at birth but not changes in remaining life expectancies. We suggest that changes in age perception should be explored for younger age classes to inform on aging processes, including whether aging is delayed or slowed with increasing life expectancy.
Spread through air spaces is an independent predictor of recurrence in stage III (N2) lung adenocarcinoma
Authors: Terada Yuriko , Takahashi Tsuyoshi , Morita Shigeki , Kashiwabara Kosuke , Nagayama Kazuhiro , Nitadori Jun-Ichi , Anraku Masaki , Sato Masaaki , Shinozaki-Ushiku Aya , Nakajima Jun
Published: April 2019

OBJECTIVES

Spread through air space (STAS) is recognized as a pattern of invasion in lung adenocarcinoma and has been reported to be a predictor of recurrence and survival in patients with early-stage lung adenocarcinoma. However, this parameter has not been studied well in stage III (N2) lung adenocarcinoma. In this study, we evaluated the association between STAS invasion patterns and recurrence and survival in stage III (N2) lung adenocarcinoma.

METHODS

We retrospectively reviewed data from 76 patients at University of Tokyo with stage III (N2) lung adenocarcinoma who underwent surgery from August 1998 to December 2013. Statistical analysis was performed to determine the impact of STAS invasion clinicopathological features and clarify the relationship between this pattern of invasion and survival.

RESULTS

Tumour STAS was observed in 46 of 76 patients (60.5%) and was significantly associated with the presence of lymphatic invasion (P < 0.001), papillary components (P < 0.001) and micropapillary components (P < 0.001). STAS was also significantly associated with recurrence (5-year recurrence-free probability: 19.0% vs 46.1%, P < 0.05). Univariate analyses showed that STAS was a significant risk factor for recurrence (hazard ratio 1.94, 95% confidence interval 1.07–3.51; P = 0.029).

CONCLUSIONS

The presence of STAS invasion pattern is a significant risk factor for recurrence in stage III (N2) lung adenocarcinoma.
Plasma and tissue pharmacokinetics of fosfomycin in morbidly obese and non-obese surgical patients: a controlled clinical trial
Authors: Dorn Christoph , Petroff David , Neumann Nancy , Kratzer Alexander , El-Najjar Nahed , Dietrich Arne , Kloft Charlotte , Zeitlinger Markus , Kees Martin G , Kees Frieder , Wrigge Hermann , Simon Philipp
Published: April 2019

Objectives

To assess the pharmacokinetics and tissue penetration of fosfomycin in obese and non-obese surgical patients.

Methods

Fifteen obese patients undergoing bariatric surgery and 15 non-obese patients undergoing major intra-abdominal surgery received an intravenous single short infusion of 8 g of fosfomycin. Fosfomycin concentrations were determined by LC-MS/MS in plasma and microdialysate from subcutaneous tissue up to 8 h after dosing. The pharmacokinetic analysis was performed in plasma and interstitial fluid (ISF) by non-compartmental methods.

Results

Thirteen obese patients (BMI 38–50 kg/m2) and 14 non-obese patients (BMI 0–29 kg/m2) were evaluable. The pharmacokinetics of fosfomycin in obese versus non-obese patients were characterized by lower peak plasma concentrations (468 ± 139 versus 594 ± 149 mg/L, P = 0.040) and higher V (24.4 ± 6.4 versus 19.0 ± 3.1 L, P = 0.010). The differences in AUC∞ were not significant (1275 ± 477 versus 1515 ± 352 mg·h/L, P = 0.16). The peak concentrations in subcutaneous tissue were reached rapidly and declined in parallel with the plasma concentrations. The drug exposure in tissue was nearly halved in obese compared with non-obese patients (AUC∞ 1052 ± 394 versus 1929 ± 725 mg·h/L, P = 0.0010). The tissue/plasma ratio (AUCISF/AUCplasma) was 0.86 ± 0.32 versus 1.27 ± 0.34 (P = 0.0047).

Conclusions

Whereas the pharmacokinetics of fosfomycin in plasma of surgical patients were only marginally different between obese and non-obese patients, the drug exposure in subcutaneous tissue was significantly lower in the obese patients.
Rethinking antimicrobial stewardship paradigms in the context of the gut microbiome
Authors: Shahi Farah , Redeker Kelly , Chong James
Published: April 2019

Ongoing concerns over the presence and persistence of antimicrobial resistance (AMR), particularly in Gram-negative bacteria, continue to have significant global health impacts. The gastrointestinal tract, or ‘gut’, environment amplifies AMR in the human gut microbiome, even in the absence of antibiotics. It constitutes a complex and diverse community of organisms, and patterns and alterations within it are increasingly being found to be associated with states of health and disease. Our understanding of the effects of routes of administration of antimicrobials on the gut microbiome is still lacking despite recent advances in metagenomics. In this article we review current evidence for antibiotic effects on gut microbiota and explore possible prescribing and stewardship approaches that would seek to minimize these effects. If we are to preserve existing and new antimicrobials, we need to consider their use in the context of their effect on gut ecology, and the human microbiome in general.
The effects of incomplete milking and increased milking frequency on milk production rate and milk composition1
Authors: Kuehnl Jordan M , Connelly Meghan K , Dzidic Alen , Lauber Megan , Fricke Hannah P , Klister Marisa , Olstad Emma , Balbach Maria , Timlin Emily , Pszczolkowski Virginia , Crump Peter M , Reinemann Doug J , Hernandez Laura L
Published: April 2019

Increased milking frequency and incomplete milking have differential effects on milk yield and mammary gland physiology that are important for optimization of milking practices in dairy herds. The objectives of this experiment were to determine the effects of increased milking frequency and incomplete milking on milk production rate (MPR) and milk composition and to determine if milking 3 times daily (3×) could rescue the negative production effects of incomplete milking. Twenty-two multiparous cows were enrolled onto this experiment beginning at 5 days in milk (DIM) and continuing through 47 DIM. A split-plot design was used to randomize the 2 treatments, which were milking frequency and incomplete milking. Eleven cows were randomly assigned to be milked 2 times (2×) daily and 11 cows were randomly assigned to be milked 3×. Within each cow, a contralateral half-udder was randomly assigned to be incompletely milked (30% milk remaining in the gland; IM), and the other half-udder was randomly assigned to be milked completely (CM). Quarter-level milk yields were recorded at each milking session. Milk samples from all quarters were collected twice weekly at the beginning of the morning milking for analysis. Cows milked 2× tended to have reduced MPR compared with 3× milked cows (1.81 ± 0.06 vs. 1.97 ± 0.06 kg milk/h; P = 0.06). Half-udders that were CM and IM produced 1.09 ± 0.03 and 0.80 ± 0.03 kg milk/h, respectively. There was an interaction between incomplete milking treatment and week of lactation (P = 0.04). No interaction was detected between milking frequency and incomplete milking for MPR or milk components. Cows milked 3× had increased milk fat percent (1.93 ± 0.09% vs. 1.65 ± 0.09%, P = 0.047), decreased milk lactose percent (4.80 ± 0.04% vs. 4.93 ± 0.04%, P = 0.04), and exhibited no differences in milk protein percent or milk somatic cell count (SCC) compared with cows milked 2×. Half-udders that were IM had increased milk fat percent (2.15 ± 0.07% vs. 1.43 ± 0.07%, P < 0.0001), decreased lactose percent (4.75 ± 0.03% vs. 4.99 ± 0.03%, P < 0.0001), increased milk log10SCC (4.22 ± 0.05 vs. 4.41 ± 0.05, P = 0.0004), and no differences in milk protein percent compared with CM half-udders. These results indicate that a 3× milking frequency in IM half-udders was not able to improve milk production compared with IM half-udders milked 2×. Our results indicate that 30% milk remaining in the gland had an irreversible impact on milk yield as increased milking frequency was not able to reverse the milk yield lost.
A hyperintensional logical framework for deontic reasons
Authors: Faroldi Federico L G , Protopopescu Tudor
Published: April 2019

In this paper we argue that normative reasons are hyperintensional and put forward a formal account of this thesis. That reasons are hyperintensional means that a reason for a proposition does not imply that it is also a reason for a logically equivalent proposition. In the first part we consider three arguments for the hyperintensionality of reasons: (i) an argument from the nature of reasons, (ii) an argument from substitutivity and (iii) an argument from explanatory power. In the second part we describe a hyperintensional logic of reasons based on justification logics. Eventually we discuss the philosophical import of this proposal and highlight some limitations and possible developments.
‘Why genes in pieces?’—revisited
Authors: Smithers Ben , Oates Matt , Gough Julian
Published: April 2019

The alignment between the boundaries of protein domains and the boundaries of exons could provide evidence for the evolution of proteins via domain shuffling, but literature in the field has so far struggled to conclusively show this. Here, on larger data sets than previously possible, we do finally show that this phenomenon is indisputably found widely across the eukaryotic tree. In contrast, the alignment between exons and the boundaries of intrinsically disordered regions of proteins is not a general property of eukaryotes. Most interesting of all is the discovery that domain–exon alignment is much more common in recently evolved protein sequences than older ones.
A centrally concentrated sub-solar-mass starless core in the Taurus L1495 filamentary complex
Authors: Tokuda Kazuki , Tachihara Kengo , Saigo Kazuya , André Phillipe , Miyamoto Yosuke , Zahorecz Sarolta , Inutsuka Shu-ichiro , Matsumoto Tomoaki , Takashima Tatsuyuki , Machida Masahiro N , Tomida Kengo , Taniguchi Kotomi , Fukui Yasuo , Kawamura Akiko , Tatematsu Ken’ichi , Kandori Ryo , Onishi Toshikazu
Published: April 2019

The formation scenario of brown dwarfs is still unclear because observational studies to investigate its initial condition are quite limited. Our systematic survey of nearby low-mass star-forming regions using the Atacama Compact Array (aka the Morita array) and the IRAM 30-m telescope in 1.2 mm continuum has identified a centrally concentrated starless condensation with a central H2 volume density of ∼106 cm−3, MC5-N, connected to a narrow (width ∼0.03 pc) filamentary cloud in the Taurus L1495 region. The mass of the core is $\sim {0.2\!-\!0.4}\, M_{\odot }$, which is an order of magnitude smaller than typical low-mass pre-stellar cores. Taking into account a typical core to star formation efficiency for pre-stellar cores (∼20%–40%) in nearby molecular clouds, brown dwarf(s) or very low-mass star(s) may be going to be formed in this core. We have found possible substructures at the high-density portion of the core, although much higher angular resolution observation is needed to clearly confirm them. The subsequent N2H+ and N2D+ observations using the Nobeyama 45-m telescope have confirmed the high-deuterium fractionation (∼30%). These dynamically and chemically evolved features indicate that this core is on the verge of proto-brown dwarf or very low-mass star formation and is an ideal source to investigate the initial conditions of such low-mass objects via gravitational collapse and/or fragmentation of the filamentary cloud complex.
A Biosensor-Based Assay (GlnLux-Agar) Shows Defoliation Triggers Rapid Release of Glutamine from Nodules and Young Roots of Forage Legumes
Authors: Thilakarathna Malinda S. , Raizada Manish N.
Published: April 2019

Forage legumes experience defoliation from grazing and injury in both natural and agricultural ecosystems. Defoliation induces rhizodeposition of nitrogen (N) compounds from root systems that can feed microbes and plants that depend on the rhizosphere. The literature suggests that N exudates are primarily released from root tips, and those from legume nodules are released via slow nodule decomposition. However, the early timing and precise locations of N release postdefoliation are poorly characterized. The objectives of this study were to directly image tissue-specific N exudation sites in forage legumes, specifically for glutamine, and to do so at early time points postdefoliation. Glutamine is the primary assimilate of symbiotic nitrogen fixation in nodules and a key transport form of fixed N in amide-exporting legumes. Three amide-exporting forages, alfalfa (Medicago sativa), red clover (Trifolium pretense), and white clover (Trifolium repens), were defoliated or not, and placed on agar embedded with whole cell biosensor cells (GlnLux) that detect glutamine. There were two unexpected findings. First, Gln release occurred rapidly, starting within 2 h postdefoliation, depleting rapidly. Second, the sources of early Gln release were primarily nodules in addition to the expected young lateral roots/root tips. Lux quantification statistically confirmed the key findings. These observations suggest that N exudate release should be added to the list of defoliation stress early responses in nodules, and may have implications for our understanding of how defoliation impacts the rhizosphere microbiome. Furthermore, GlnLux-agar imaging represents a new assay to explore the proposed but yet unknown mechanisms underlying organic N exudation in plants.
Stimulatory Effects of Sublethal Doses of Carbendazim on the Virulence and Sclerotial Production of Botrytis cinerea
Authors: Cong Menglong , Zhang Bao , Zhang Kunyu , Li Guoqing , Zhu Fuxing
Published: April 2019

Stimulatory effects of low doses of fungicides on the virulence of phytopathogens have profound implications for applications of fungicides. The present study demonstrated that carbendazim sprayed at 0.001 to 0.03 μg/ml had stimulatory effects on the virulence of mycelia of Botrytis cinerea, and the maximum percent stimulations were 15.5 and 21.4% for isolates HB459 and HB536, respectively. Potato dextrose agar (PDA) amended with carbendazim at 0.01, 0.02, and 0.05 μg/ml inhibited mycelial growth of isolate HB536 by 0.8, 10.0, and 30.6%, respectively. However, after the inhibited mycelia were inoculated on cucumber leaves, virulence increased by 10.1, 12.9, and 10.8%, respectively. With respect to sclerotial production, carbendazim at 0.005 and 0.02 μg/ml in PDA significantly (P < 0.05) increased, while at 0.1 μg/ml significantly (P < 0.05) reduced the sclerotial number and weight of both isolates compared with nontreated controls. Conidia germination percentages slightly yet statistically significantly (P < 0.05) increased after being inoculated on PDA amended with carbendazim at 0.001 and 0.005 μg/ml. Carbendazim at 0.001∼0.02 μg/ml, either sprayed on cucumber leaves or cosuspended with conidia, exerted significantly (P < 0.05) stimulatory effects on the virulence of B. cinerea conidia. Mechanism studies showed that sublethal doses of carbendazim did not increase the expression levels of pathogenicity-related pectin methylesterase gene Bcpme1, endopolygalacturonase gene Bcpg2, cutinase gene CutA, xylanase gene Xyn11A, or NADPH oxidase gene BcnoxA.
A-102 Do biological differences exist between HTLV-1A and HTLV-1C?
Authors: Fujikawa Dai , Moles Ramona , Omsland Maria , Purcell Damian , Galli Veronica , Franchini Genoveffa
Published: April 2019

HTLV-1A, the most common HTLV-1 subtype, and HTLV-1C, the most divergent type, differ substantially. The apparent differences in susceptibility to infection and disease manifestation suggest that the 2 viral subtypes may affect their host's immune system differently. HTLV-1A and C have similar genetic organization: the structural Gag and Envelope proteins, viral enzymes, Reverse Transcriptase, Integrase and Protease, and the transcriptional and post-transcriptional regulators Tax and Rex proteins are highly conserved between the 2 viral subtypes. Strikingly, however, the RNA sequence of orf-I and orf-II, encoding the p12, p8, and p30 proteins, are highly divergent in HTLV-1C. The proteins encoded by HTLV-1A orf-I and orf-II are essential for HTLV-1A to persist in the host and escape immune recognition. The p8/p12 proteins target MHC-I and ICAM-1 expression and inibit the killing of infected cells by Cytotoxic T cells, Natural Killer cells. The p30 protein, via the PU.1 transcription machinery, inhibits interferon responses in monocytes. It is thus conceivable that the expression of these proteins in T cells in individuals with a high number of infected cells may affect the overall host response to pathogens. We therefore focused on orf-I to investigate potential biological differences between HTLV-1A and C. In HTLV-1C, we found that a putative doubly-spliced mRNA produces a 16KD protein (p16) by juxtaposing the first exon of Rex in frame with orf-I. We found that p16 inhibits inflammasome activation and cytokine production, an essential step in monocytes to respond to pathogens. We constructed chimeric HTLV-1A/C viruses whereby either the entire 3′ end or the Orf-I of HTLV-1C was swapped into the HTLV-1A backbone. The overall aim of our studies is to investigate the effect of the orf-I proteins of both viral subtypes on immune cells in vitro and in vivo using isogenic HTLV-1A/C chimeric viruses orf-I mutants.
B-102 Exosomal protein signatures: mechanistic insights and biomarker potential
Authors: Hoshino Ayuko , Lyden David
Published: April 2019

In 1889, Stephen Paget first proposed that organ distribution of metastases is a non-random event, yet metastatic organotropism remains one of the greatest mysteries in cancer biology. A growing number of studies demonstrate that tumor-derived microvesicles, referred to as exosomes, may alter the tumor microenvironment at future sites of metastasis promoting pre-metastatic niche formation, and thus creating a favorable “soil” for incoming metastatic “seeds.” However, by what mechanism, and whether their role is significant in tumor progression remains unknown. We have recently demonstrated that exosomes released by lung-, liver- and brain-tropic tumor cells preferentially fuse with resident cells at their future metastatic sites, such as fibroblasts and epithelial cells in the lung, Kupffer cells in the liver, and endothelial cells in the brain. We found that tumor-derived exosome homing to organ-specific cell types prepares the pre-metastatic niche. Moreover, treatment with exosomes derived from lung-tropic models can redirect to the lung the metastatic distribution of cells that normally lack the capacity to form lung metastasis. Proteomic profiling of exosomes revealed distinct integrin expression patterns associated with each organ-specific metastatic site. Whereas exosomal α6β4 integrins were associated with lung metastasis, exosomal integrin αvβ5 was linked to liver metastasis and αvβ3 to brain metastasis. Targeting α6β4 and αvβ5 integrins decreased exosome uptake and metastasis in the lung and liver, respectively. Moreover, our clinical data indicate that integrin expression profiles in circulating plasma exosomes from cancer patients could be used to predict organ-specific metastasis. To gain a more comprehensive understanding of the relationship between exosomal protein cargo and tumor progression, we have built one of the largest exosome proteomics databases, including human and murine normal, pre-cancer and cancer exosomes from a variety sources, from cell lines to plasma and other bodily fluids. Out of 490 human samples, 206 were isolated from normal subjects and 284 were from cancer patients. This dataset has allowed us to extend the list of markers that can be used to identify and isolate exosomes from human samples. When comparing the protein cargo of blood plasma exosomes isolated from normal subjects versus cancer patients with early stage pancreatic, colorectal, breast and lung cancer, we were able to define a cancer-specific proteomic signature that distinguishes cancer exosomes. Using paired samples from tumor tissue and plasma of the same cancer patients with breast, lung or pancreatic cancer, we were able to derive cancer type-specific exosomal signatures, which can be used as biomarkers to identify and classify tumors of unknown origin. Therefore, proteomic profiling of exosomes has great clinical potential as a non-invasive, rapid liquid biopsy that could aid in tumor detection classification.
C-101 Sialoside-based pattern recognition and its application in immunotherapy
Authors: Liu Yang , Zheng Pan
Published: April 2019

It is well established that the innate immune system recognizes both pathogen-associated and danger-associated molecular patterns (PAMPs and DAMPs respectfully). We have demonstrated that CD24-Siglec interactions selective represses innate immune response to DAMPs but not PAMPs. Since this pathway is based on sialic acid on the ligands that interact with Siglecs, we termed the it sialoside-based pattern recognition. Given the intrinsic connection between innate and adaptive immune responses, we have explored the possibility to suppress T-cell-mediated immune damages to host tissues. Our data from both preclinical and clinical investigation support the utility of bolstering the sialoside-based pattern recognition to treat autoimmune diseases, sequela of viral infection and graft vs host diseases.
C-103 Multimodal approach to cancer immunotherapy
Authors: Schlom Jeff
Published: April 2019

The use of checkpoint inhibitor monoclonal antibodies (MAbs) has transformed the landscape of cancer immunotherapy. Consequently, human tumors can be characterized as either “hot” or “cold.” Hot tumors are defined by the presence of tumor infiltrating T-cell lymphocytes (TILs); melanoma is the prototype of a hot tumor. Studies have shown that the presence of TILs results in tumor cells expressing molecules, termed “checkpoints,” with the ability to anergize T cells. The clinical use of these checkpoint inhibitor MAbs has resulted in durable anti-tumor responses in approximately 50% of patients with melanoma and in 10%–15% of patients with some solid tumors such as lung cancer. The vast majority of solid tumors, that is, carcinomas of the breast, prostate, colon, and lung, as well as others, however, can be characterized as cold in that they do not respond to checkpoint inhibitor therapy. Preclinical studies and ongoing clinical studies are now revealing that a multi-modal approach to cancer immunotherapy can result in increased anti-tumor activity. This includes the use of agents designed to (a) activate the immune response with the use of vaccines, (b) potentiate the immune response with cytokines such as IL-15 and IL-12; these cytokines also have the potential to activate the innate immune system, for example, natural killer (NK) cells, (c) eliminate or reduce immunosuppressive entities such as TGF-beta, IL-8, and checkpoint molecules, and (d) modify the tumor phenotype to render tumor cells more susceptible to T-cell‒mediated attack. Recent preclinical and clinical studies have also shown that the use of bifunctional agents capable of bringing both immunostimulatory molecules as well as molecules to inhibit immunosuppressive entities to the tumor microenvironment results in enhanced anti-tumor activity.
C-105 Clinical trial of HER2 cancer vaccine shows benefit in 45% of evaluable patients in phase I study
Authors: Berzofsky Jay , Wood Lauren , Maeng Hoyoung , Trepel Jane , Stroncek David , Morris John
Published: April 2019

We have developed and tested therapeutic cancer vaccine targeting HER2-expressing cancers based on autologous dendritic cells (DCs) transduced with an adenovirus expressing the non-signaling extracellular and transmembrane domains of HER2, a driver oncogene in many cancers including breast, ovarian, lung, colorectal, gastroesophageal, and bladder, among others. In mice, the homologous vaccine cured virtually all mice with large established tumors up to 2 cm and with established macroscopic lung metastases. In mice, the protection was dependent on antibodies to HER2 that inhibited HER2 phosphorylation, but was FcR (ADCC) independent, and thus different from the main recognized mechanism of the clinically approved anti-HER2 monoclonal antibody, trastuzumab. A phase I trial is ongoing in patients with advanced metastatic cancers who have failed at least one line of standard therapy, whose tumors scored HER2 1+, 2+ or 3+ by immunohistochemistry or have a HER2 FISH score ≥1.8. Part 1 of the trial was carried out in patients naïve to trastuzumab or other HER2-directed therapies, as it is important to demonstrate safety in the absence of complicating effects of prior HER2-targeted therapies. In Part 1, although no responses were seen in 6 patients at the lowest dose of 5 million DCs, at the second and third dose escalations (10 and 20 million DCs), of 11 evaluable patients, 5 (45%) had clinical benefit, including one CR lasting 89 weeks, one PR lasting 24 weeks, and 3 cases of stable disease, seen in patients with metastatic ovarian, gastroesophageal or colorectal cancers. Adverse reactions were mainly injection site reactions that did not require treatment in most patients. Serial echocardiograms did not show any evidence of cardiotoxicity. The number of circulating tumor cells also decreased in 40% (8/20), 83% (5/6) and 100% (2/2) at 12, 28 and 48 weeks on study, respectively. Based on these results and safety data, the FDA and IRB have approved increasing the maximum dose to 40 million DCs and opening Part 2 of the trial in patients who have failed prior HER2-directed therapies like trastuzumab, mostly metastatic breast and gastric cancer patients. Early results in Part II starting at 20 million DCs already showed 2/6 patients with stable disease. Thus, we have translated a cancer vaccine from mice to human clinical trials with very promising early results, and intend to combine this vaccine with checkpoint inhibitors, as vaccines can induce T cell responses that turn “cold” tumors into “hot” ones amenable to checkpoint blockade immunotherapy.
C-107 How checkpoint blockade alters the repertoire and function of tumor specific T-cells
Authors: Pardoll Drew
Published: April 2019

The past 10 years have seen a transformation in cancer treatment, as immunotherapy has come to fruition after many decades of research. Interestingly, this inflexion point has not been based on stimulation of anti-tumor immunity by agents like vaccines or systemically delivered cytokines, but rather by blockade of inhibitory receptors or their ligands—generally referred to as checkpoints. These advances were highlighted in Science's 2013 “Breakthrough of the Year” and by this year's Nobel Prizes. The success of checkpoint-blocking antibodies demonstrates that many, arguably most, cancer patients contain T cells capable of recognizing and killing their cancer. The most significant checkpoint to be blocked is PD-1, whose ligands are PD-L1 and PD-L2. Three anti-PD-1 and 3 anti-PD-1 antibodies have been approved for treatment of 13 different cancer types. The activity of these antibodies depends on activation of T cells. There are 2 FDA approved biomarkers to identify patients more likely to respond to anti-PD-1. One is expression of PD-L1 and the other is mismatch repair deficiency, which is the cause of 4% of cancers. Mismatch repair deficiency (MMRd also called micro-satellite instability-MSI) results from homozygous inactivation of one of 4 mismatch repair genes. These cancers typically have over 1000 non-synonymous mutations/exome. Because a major target of anti-tumor T cells is mutation-associated “neoantigenic” peptides, patients with MMRd cancers have huge numbers of neoantigens that can be recognized but they are not killed because they are restrained by engaging PD-1. Anti-PD-1 antibodies induce major cancer regressions in over 50% of MMRd cancers, regardless of tissue of origin. Three major endeavors are advancing cancer immunotherapy for the 75%–80% or cancer patients NOT benefitted by monotherapy with anti-PD-1/PD-L1 antibodies: (1) Therapeutic combinations with anti-PD-1 or anti-PD-L1 being foundational, (2) Reversal of T cell dysfunction and (3) Refined development of biomarkers to identify patients most likely to respond to various immunotherapies and their combinations. Recent technologies, including TCRseq, single cell analyses and multispectral imaging are accelerating the identification of new drug targets and biomarkers to promote this endeavor.
E-104 Intravital microscopy of the tumor microenvironment: Escape and relevance for immunotherapy
Authors: Friedl Peter
Published: April 2019

Cancer cell invasion results from coordination of cell shape, deformability and actin dynamics relative to the tumor microenvironment. When monitored in vivo, using intravital multiphoton second and third harmonic generation and fluorescence microscopy, tissue microniches provide invasion-promoting tracks that enable collective migration along tracks of least resistance. As main routes, non-destructive contact-guidance is mediated by preformed multi-interface perimuscular, vascular and –neural tracks of 1D, 2D and 3D topography with form constitutive tissue conduit-like tracks with defined geometry, nanotropography and molecular composition as predominant routes of invasion (“highways”). During adoptive T cell therapy, interstitial conduits further represented a preferential route for cytotoxic T cell (CTL) distribution, which allowed CTL accumulation around invading tumor nests and effective killing, by CTL swarming and cooperation. In conclusion, both cancer invasion and CTL trafficking occurs in predefined tissue niches of the tumor microenvironment indicating common themes of cell guidance by encountered physical space and molecular cues which can be exploited for adoptive T cell therapy.
E-105 Mesenchymal stromal cells: Mediators of inflammation in cancer
Authors: DeClerck Yves
Published: April 2019

Cancer-associated fibroblasts (CAF) have been identified in the tumor microenvironment (TME) of many cancers. CAF represent a heterogeneous population of cells that can have pro-tumorigenic and anti-tumorigenic functions. We have isolated from primary human neuroblastoma tumors, a population of CAF that share phenotypic and functional characteristics of bone marrow-derived mesenchymal stromal cells (MSC). These cells are CD73+, CD90+, CD105+, CD34−, CD45−, CD14−, CD19−, CD31−, and VEGFR2−, and retain the ability to differentiate into osteoblasts, chondrocytes and adipocytes, a feature of MSC. However, we show that these cells also express fibroblast activation protein alpha (FAP/DPPIV), fibroblast specific protein 1 (FSP1/S1004A) and alpha smooth muscle actin (SMA), markers of cancer-associated fibroblasts. When in the presence of tumor cells, these MSC/CAF stimulate the production of multiple pro-tumorigenic cytokines and chemokines including interleukin (IL)-6, IL-8, CCL2/monocyte chemoattractant protein 1, CXCL-12/stromal-derived factor 1 (SDF1), CXCL1, vascular endothelial growth factor (VEGF-A), plasminogen activator inhibitor 1 (PAI-1) and transforming growth factor (TGF) β. The production of these cytokines/chemokines by MSC/CAF is stimulated by the production of soluble factors by tumor cells including galectin-3 binding protein and extracellular vesicles. In addition to stimulating angiogenesis, the recruitment and M2 polarization of macrophages, and to suppress immune cells, MSC/CAF have a direct effect on tumor cells by stimulating tumor cell growth, survival and resistance to chemotherapy in an ERK1/2 and STAT3-dependent manner. We then show that targeting ERK1/2 and STAT3 or targeting MSC in combination with chemotherapy or immunotherapy in mice orthotopically implanted with human neuroblastoma tumors, inhibits tumor growth and extends survival. In contrast, targeting a single cytokine like IL-6 has no significant effect. These data thus identify a new type of CAF sharing characteristics of bone marrow-derived MSCs that contribute to a pro-tumorigenic inflammatory reaction in the TME and provide preclinical evidence that targeting these cells or the pathways they activated in tumor cells is a valuable therapeutic approach.
F-101 A hemagglutinin stalk-based universal influenza virus vaccine
Authors: Krammer Florian
Published: April 2019

Influenza virus infections are a significant cause of morbidity and mortality worldwide. Current vaccines show acceptable efficacy against antigenically matched viruses by inducing strain specific antibodies against the membrane-distal globular head domain of the viral hemagglutinin, but fail to protect against drifted and pandemic strains. The membrane-proximal stalk domain of the viral hemagglutinin exhibits a high degree of conservation across influenza virus subtypes, and monoclonal antibodies directed against this region typically show broad neutralizing activity. However, these antibodies are rare and usually not induced/boosted by regular seasonal vaccines. Here, we developed a universal influenza virus vaccination strategy based on the conserved stalk domain of group 1 and group 2 hemagglutinins. By sequential vaccination of mice and ferrets with chimeric hemagglutinin constructs that share the same stalk domain but have divergent head domains we were able to specifically boost broadly neutralizing antibody titers against conserved epitopes in the hemagglutinin stalk. Animals vaccinated with these constructs were protected from morbidity and mortality induced by infection with a panel of heterologous and heterosubtypic influenza A viruses, including avian influenza virus subtypes of concern like H5N1 and H7N9. The induced antibody response was long-lived and also interfered with virus transmission between animals. The present data suggest that this vaccine strategy has the potential to provide broad influenza virus protection in humans and clinical trials are currently ongoing. A universal influenza virus vaccine would represent a major advance towards the control of influenza worldwide and would significantly enhance our pandemic preparedness.
H-102 The potential role of “defective” proviruses in the pathogenesis of HIV-1 infection
Authors: Lane H. Clifford , Imamichi Hiromi
Published: April 2019

A pool of CD4+T cells harboring HIV-1 proviruses is established early during HIV-1 infection and persists despite years of suppressive combination antiretroviral therapy (cART). Greater than 95% of the proviruses in these pools are “defective” due to large internal deletions and lethal genetic mutations. Although unable to encode intact viruses, these “defective” proviruses are biologically active, transcribe HIV-RNA and produce HIV-1 proteins in vivo. We hypothesize that ongoing production of foreign proteins from these translationally-competent yet defective HIV-1 proviruses contribute to the persistent immune activation and systemic inflammation during suppressive ART.
I-102 Viral fitness costs and benefits of disrupting the host cell actin cytoskeleton in mucosal HIV-infection
Authors: Mempel Thorsten , Usmani Shariq , Murooka Thomas
Published: April 2019

HIV-1 disrupts multiple processes that support the motility of infected host cells, including expression and proper localization of chemokine receptors, their downstream signal transduction machinery, and proteins regulating actin cytoskeletal turnover. While models have been developed to explain how these conserved lentiviral activities support HIV replication, their role and relative importance for HIV infection in vivo are largely unexplored. Recently, intravital microscopy studies in advanced humanized mouse models of HIV infection have enabled the visualization of the dynamic behavior of HIV-infected cells in the environmental context of lymphoid tissues, such as spleen and lymph node. Our lab is studying how HIV alters the migratory behavior of HIV-infected T cells in lymph nodes of BLT humanized mice, and how these alterations affect viral replication and persistence. We will present and discuss our recent findings on the roles of the HIV envelope, and the accessory Vpu, and Nef proteins in this context.
I-107 Long-range allosteric effects induced by Tax–HLA-A2 binding to an anti-HTLV-1 TCR: Implications for early T cell signaling
Authors: Rangaranjan Sneha , He Yanan , Kerzic Melissa , Ma Buyong , Pierce Brian , Nussinov Ruth , Orban John , Mariuzza Roy
Published: April 2019

The mechanism whereby αβ T cell receptor (TCR) engagement by peptide–MHC (pMHC) is first communicated to the CD3 signaling apparatus of the TCR–CD3 complex, a process termed early T cell activation, is not well understood. To address the possibility that pMHC binding induces allosteric changes in TCR conformation and/or dynamics that are relayed to CD3, we carried out NMR analysis and molecular dynamics (MD) simulations of both the α and β chains of a human antiviral TCR (A6) that recognizes the Tax antigen from HTLV-1 bound to HLA-A2. We observed pMHC-induced NMR signal perturbations in the TCR V domains that propagate to 3 distinct sites in the C domains: (1) the Cβ FG loop that projects from the Vβ/Cβ interface, (2) a cluster of Cβ residues near the Cβ αA helix, a region involved in interactions with CD3, and (3) the Cα AB loop located at the membrane-proximal base of the TCR. A biological role for each of these allosteric sites is supported by previous mutational and functional studies of TCR signaling. In addition, the pattern of long-range ligand-induced changes in TCR A6 seen by NMR is broadly similar to that predicted by MD simulations. We propose that the unique structure of the TCR β chain enables allosteric communication between the TCR binding sites for pMHC and CD3.
J-102 Approaches to eliminate persistent viruses
Authors: Wilson Eleanor
Published: April 2019

With new directly acting antiviral treatments for Hepatitis C, we are learning how to directly target and cure a persistent viral infection for the first time. Dr. Wilson will discuss approaches to eliminating persistent viral infections and promising future research directions.
J-103 The International Coalition to Eliminate Hepatitis B (ICE-HBV)
Authors: Revill Peter , Penicaud Capucine
Published: April 2019

Background:

Over 257 million people worldwide are chronically infected with hepatitis B virus (HBV), resulting in over 880,000 deaths per year from cirrhosis and liver cancer. There is no known cure for chronic HBV, due in part to the continued presence of transcriptionally active DNA in the nucleus which is not directly targeted by current antiviral therapies. Our aim is to inspire and support the discovery of a safe, scalable and effective cure for the benefit of all people living with CHB.

Methods:

To achieve this, we have created an international research driven forum, the International Coalition to Eliminate Hepatitis B (ICE-HBV), which is coordinating collaborative partnerships among researchers and stakeholders to accelerate the search of an HBV cure.

Results:

ICE-HBV working groups have developed a joint global scientific strategy for HBV cure, with input from key stakeholders including the HBV affected community. Through engagement with key stakeholders, ICE-HBV aims to drive changes in governmental policy to ensure more funds are channelled to HBV cure research and drug development. ICE-HBV fosters new collaborations among HBV researchers and industry worldwide and initiate new projects to fast-track the discovery of an HBV cure.

Conclusions:

The push for a cure for chronic HBV infection is particularly timely thanks to the recent development of cell culture infection models that, for the first time, empower truly curative research. Through its global network, ICE-HBV is striving to promote effective collaboration and facilitate opportunities that can produce a cure for chronic HBV infection.
PB-1 7, 8 Dihydroxyflavone reduces mitochondrial dysregulation in the HIV-1 transgenic mouse model as a treatment for HIV associated Neurocognitive disorder (HAND)
Authors: Bryant Joseph , Andhavarapu Sanketh , Arvas Muhammed Ikbal , Gupta Udit , Katuri Akhil , Asemu Girma , Makar Tapas , Davis Harry
Published: April 2019

The introduction of combined antiretroviral therapy (cART) significantly increased the lifespan of HIV patients, turning a fatal disease to a chronic one. However, as a lower but persistent level of HIV infection remains, the susceptibility of HIV-associated neurocognitive disorder (HAND) is increased. Therefore, research is currently seeking improved treatment for this complication of HIV. 7, 8 Dihydroxyflavone (DHF) is a TrkB agonist that can cross the blood brain barrier and mimic the effect of Brain Derived Neurotrophic Factor (BDNF). Previous studies have described its potential as a treatment in a myriad of neurological diseases including Alzheimer's disease, peripheral sensory neuropathies, amyotrophic lateral sclerosis, Parkinson's disease, and Multiple Sclerosis. This study investigates mitochondrial dysregulation in the brain of HIV-1 Tg26 mice, a murine model of HIV, and the potential therapeutic effects of DHF treatment on this dysregulation in HAND. The mice were divided into 3 groups: wild type, untreated-Tg26, and DHF-treated Tg26 (Tg+DHF). Mice were sacrificed after 28 days and 7 µm thick paraffin sections of the brain of each mice were prepared. Tissues were immunohistochemically stained to detect the expression of mitochondria regulating proteins—SIRT3, PGC1-a, FIS-1, MFN-2, Citrate Synthase, SUR1, TRPM4—in the hippocampal and cortex regions. Our results show, that in these regions of Tg26 mice, SIRT3, PGC1-a, FIS-1, MFN-2, and Citrate Synthase were present in decreased levels, implicating the extensive role of mitochondrial dysfunction in HAND pathology. However, following DHF treatment, Tg26 mice expressed these cells at greater levels, suggesting a possible mechanism of DHF. Furthermore, both SUR1 and TRPM4 were upregulated in Tg26 mice. Previous studies have shown that SUR1 and TRPM4 often co-assemble to form an ion channel that plays a significant role in central nervous system injury and calcium homeostasis. Calcium is a regulator of mitochondrial dynamics, suggesting that the upregulation of SUR1 and TRPM4 may contribute to the mitochondrial changes that are characteristic to HAND. Interestingly, administration of DHF resulted in decreased expression of SUR1 and TRPM4. Therefore, the present study suggests that DHF treatment in the hippocampal and cortex regions of the TG26 model plays a role in reducing mitochondrial dysfunction by regulating calcium balance via the SUR1-TRPM4 ion channels, indicating its therapeutic potential as a treatment for HIV-associated neurocognitive disorder.