tbuckley/cabot-search huggingface dataset:
Longitudinal analysis of the gut microbiota during anti-PD-1 therapy reveals stable microbial features of response in melanoma patients
Authors: Angeli D G Macandog, Carlotta Catozzi, Mariaelena Capone, Amir Nabinejad, Padma P Nanaware, Shujing Liu, Smita Vinjamuri, Johanna A Stunnenberg, Serena Galiè, Maria Giovanna Jodice, Francesca Montani, Federica Armanini, Ester Cassano, Gabriele Madonna, Domenico Mallardo, Benedetta Mazzi, Salvatore Pece, Maria Tagliamonte, Vito Vanella, Massimo Barberis, Pier F Ferrucci, Christian U Blank, Marlene Bouvier, Miles C Andrews, Xiaowei Xu, Laura Santambrogio, Nicola Segata, Luigi Buonaguro, Emilia Cocorocchio, Paolo A Ascierto, Teresa Manzo, Luigi Nezi
Journal: Cell Host & Microbe (2024)

Immune checkpoint inhibitors (ICIs) improve outcomes in advanced melanoma, but many patients are refractory or experience relapse. The gut microbiota modulates antitumor responses. However, inconsistent baseline predictors point to heterogeneity in responses and inadequacy of cross-sectional data. We followed patients with unresectable melanoma from baseline and during anti-PD-1 therapy, collecting fecal and blood samples that were surveyed for changes in the gut microbiota and immune markers. Varying patient responses were linked to different gut microbiota dynamics during ICI treatment. We select complete responders by their stable microbiota functions and validate them using multiple external cohorts and experimentally. We identify major histocompatibility complex class I (MHC class I)-restricted peptides derived from flagellin-related genes of Lachnospiraceae (FLach) as structural homologs of tumor-associated antigens, detect FLach-reactive CD8
Quantitative flow ratio versus fractional flow reserve for coronary revascularisation guidance (FAVOR III Europe): a multicentre, randomised, non-inferiority trial
Authors: Birgitte Krogsgaard Andersen, Martin Sejr-Hansen, Luc Maillard, Gianluca Campo, Truls Råmunddal, Barbara E Stähli, Vincenzo Guiducci, Luigi Di Serafino, Javier Escaned, Ignacio Amat Santos, Ramón López-Palop, Ulf Landmesser, Ruthe Storgaard Dieu, Hernán Mejía-Rentería, Lukasz Koltowski, Greta Žiubrytė, Laura Cetran, Julien Adjedj, Youssef S Abdelwahed, Tommy Liu, Lone Juul Hune Mogensen, Ashkan Eftekhari, Jelmer Westra, Karsten Lenk, Gianni Casella, Eric Van Belle, Simone Biscaglia, Niels Thue Olsen, Paul Knaapen, Janusz Kochman, Ramón Calviño Santos, Roberto Scarsini, Evald Høj Christiansen, Niels Ramsing Holm
Journal: The Lancet (2024)
The evolutionary ecology of ethanol
Authors: Anna C Bowland, Amanda D Melin, David J Hosken, Kimberley J Hockings, Matthew A Carrigan
Journal: Trends in Ecology & Evolution (2024)

The consumption of ethanol has frequently been seen as largely restricted to humans. Here, we take a broad eco-evolutionary approach to understanding ethanol's potential impact on the natural world. There is growing evidence that ethanol is present in many wild fruits, saps, and nectars and that ethanol ingestion offers benefits that favour adaptations for its use in multiple taxa. Explanations for ethanol consumption span both the nutritional and non-nutritional, with potential medicinal value or cognitive effects (with social-behavioural benefits) explored. We conclude that ethanol is ecologically relevant and that it has shaped the evolution of many species and structured symbiotic relationships among organisms, including plants, yeast, bacteria, insects, and mammals.
Lnc-H19-derived protein shapes the immunosuppressive microenvironment of glioblastoma
Authors: Junju Chen, Yixin Gao, Jian Zhong, Xujia Wu, Zhaojie Leng, Ming Liu, Yesheng Wang, Yuan Wang, Xuesong Yang, Nunu Huang, Feizhe Xiao, Maolei Zhang, Xuesong Liu, Nu Zhang
Journal: Cell Reports Medicine (2024)
Quality of Life After Transcatheter Tricuspid Valve Replacement
Authors: Suzanne V Arnold, Rebecca T Hahn, Vinod H Thourani, Raj Makkar, Moody Makar, Rahul P Sharma, Christiane Haeffele, Charles J Davidson, Akhil Narang, Brian O'Neill, James Lee, Pradeep Yadav, Firas Zahr, Scott Chadderdon, Mackram Eleid, Sorin Pislaru, Robert Smith, Molly Szerlip, Brian Whisenant, Nishant Sekaran, Santiago Garcia, Terri Stewart-Dehner, Paul A Grayburn, Anna Sannino, Clayton Snyder, Yiran Zhang, Michael J Mack, Martin B Leon, Philipp Lurz, Susheel Kodali, David J Cohen
Journal: Journal of the American College of Cardiology (2024)
Coronary Plaque Radiomic Phenotypes Predict Fatal or Nonfatal Myocardial Infarction
Authors: Márton Kolossváry, Andrew Lin, Jacek Kwiecinski, Sebastien Cadet, Piotr J Slomka, David E Newby, Marc R Dweck, Michelle C Williams, Damini Dey
Journal: JACC. Cardiovascular imaging (2024)
FLZ attenuates Parkinson’s disease pathological damage by increasing glycoursodeoxycholic acid production via down-regulating Clostridium innocuum
Authors: Meiyu Shang, Jingwen Ning, Caixia Zang, Jingwei Ma, Yang Yang, Yueqi Jiang, Qiuzhu Chen, Yirong Dong, Jinrong Wang, Fangfang Li, Xiuqi Bao, Dan Zhang
Journal: Acta Pharmaceutica Sinica B (2024)

Increasing evidence shows that the early lesions of Parkinson's disease (PD) originate from gut, and correction of microbiota dysbiosis is a promising therapy for PD. FLZ is a neuroprotective agent on PD, which has been validated capable of alleviating microbiota dysbiosis in PD mice. However, the detailed mechanisms still need elucidated. Through metabolomics and 16S rRNA analysis, we identified glycoursodeoxycholic acid (GUDCA) was the most affected differential microbial metabolite by FLZ treatment, which was specially and negatively regulated by Clostridium innocuum, a differential microbiota with the strongest correlation to GUDCA production, through inhibiting bile salt hydrolase (BSH) enzyme. The protection of GUDCA on colon and brain were also clarified in PD models, showing that it could activate Nrf2 pathway, further validating that FLZ protected dopaminergic neurons through promoting GUDCA production. Our study uncovered that FLZ improved PD through microbiota-gut-brain axis, and also gave insights into modulation of microbial metabolites may serve as an important strategy for treating PD.
Reverse Magnetic Resonance Tuning Nanoplatform with Heightened Sensitivity for Non-invasively Multiscale Visualizing Ferroptosis-based Tumor Sensitization Therapy
Authors: Yi Zhu, Jiali Deng, Hongwei Lu, Zhu Mei, Ziwei Lu, Jiajing Guo, An Chen, Rong Cao, Xinyi Ding, Jingyi Wang, Helen Forgham, Ruirui Qiao, Zhongling Wang
Journal: Biomaterials (2024)
Nanoparticles and bone microenvironment: a comprehensive review for malignant bone tumor diagnosis and treatment
Authors: Yujing Guan, Wei Zhang, Yuling Mao, Shenglong Li
Journal: Molecular Cancer (2024)

Malignant bone tumors, which are difficult to treat with current clinical strategies, originate from bone tissues and can be classified into primary and secondary types. Due to the specificity of the bone microenvironment, the results of traditional means of treating bone tumors are often unsatisfactory, so there is an urgent need to develop new treatments for malignant bone tumors. Recently, nanoparticle-based approaches have shown great potential in diagnosis and treatment. Nanoparticles (NPs) have gained significant attention due to their versatility, making them highly suitable for applications in bone tissue engineering, advanced imaging techniques, and targeted drug delivery. For diagnosis, NPs enhance imaging contrast and sensitivity by integrating targeting ligands, which significantly improve the specific recognition and localization of tumor cells for early detection. For treatment, NPs enable targeted drug delivery, increasing drug accumulation at tumor sites while reducing systemic toxicity. In conclusion, understanding bone microenvironment and using the unique properties of NPs holds great promise in improving disease management, enhancing treatment outcomes, and ultimately improving the quality of life for patients with malignant bone tumors. Further research and development will undoubtedly contribute to the advancement of personalized medicine in the field of bone oncology.
1,25-Dihydroxyvitamin D3 protects against placental inflammation by suppressing NLRP3-mediated IL-1β production via Nrf2 signaling pathway in preeclampsia
Authors: Xueqing Liu, Xinyu Zhang, Linlin Ma, Na Qiang, Jiao Wang, Yujia Huang, Xiaolei Yuan, Chunmei Lu, Yang Cao, Jie Xu
Journal: Metabolism (2024)
Challenges and Opportunities in the Clinical Translation of High-Resolution Spatial Transcriptomics
Authors: Tancredi Massimo Pentimalli, Nikos Karaiskos, Nikolaus Rajewsky
Journal: Annual Review of Pathology Mechanisms of Disease (2024)

Pathology has always been fueled by technological advances. Histology powered the study of tissue architecture at single-cell resolution and remains a cornerstone of clinical pathology today. In the last decade, next-generation sequencing has become informative for the targeted treatment of many diseases, demonstrating the importance of genome-scale molecular information for personalized medicine. Today, revolutionary developments in spatial transcriptomics technologies digitalize gene expression at subcellular resolution in intact tissue sections, enabling the computational analysis of cell types, cellular phenotypes, and cell–cell communication in routinely collected and archival clinical samples. Here we review how such molecular microscopes work, highlight their potential to identify disease mechanisms and guide personalized therapies, and provide guidance for clinical study design. Finally, we discuss remaining challenges to the swift translation of high-resolution spatial transcriptomics technologies and how integration of multimodal readouts and deep learning approaches is bringing us closer to a holistic understanding of tissue biology and pathology.
Surface-engineered mesenchymal stem cell for refractory asthma therapy: Reversing airway remodeling
Authors: Xing-Yue He, Meng-Meng Han, Yu-Chen Zhao, Ling Tang, Yi Wang, Lei Xing, Ning Wei, Jing Wang, Guang-Ji Wang, Fang Zhou, Jee-Heon Jeong, Hu-Lin Jiang
Journal: Journal of Controlled Release (2024)
Extracellular vesicles powered cancer immunotherapy: Targeted delivery of adenovirus-based cancer vaccine in humanized melanoma model
Authors: Sara Mathlouthi, Lukasz Kuryk, Marta Prygiel, Maria Giovanna Lupo, Aleksandra Anna Zasada, Cristiano Pesce, Nicola Ferri, Beate Rinner, Stefano Salmaso, Mariangela Garofalo
Journal: Journal of Controlled Release (2024)

Malignant melanoma, a rapidly spreading form of skin cancer, is becoming more prevalent worldwide. While surgery is successful in treating early-stage melanoma, patients with advanced disease have only a 20 % chance of surviving beyond five years. Melanomas with mutations in the NRAS gene are characterized for a more aggressive tumor biology, poorer prognosis and shorter survival. Hence, new therapeutic strategies are needed, especially for this specific group of patients. Novel approaches, such as cancer vaccines, offer promising solutions by stimulating the anti-tumor immune response. Nevertheless, their clinical efficacy is still modest and more effective approaches are required. Herein, we propose the systemic administration of the adenovirus-based cancer vaccine complexed in extracellular vesicles (EVs) with the aim of achieving a targeted therapeutic effect. The vaccine was based on previously tested oncolytic adenovirus Ad5/3-D24-ICOSL-CD40L in combination with melanoma-specific antigens targeting NRAS mutations to enhance the anticancer effect. The antineoplastic properties of the oncolytic vaccine were evaluated in xenograft MUG Mel-2 melanoma BALB/c nude mice. Moreover, to mimic the tumor microenvironment, while investigating at the same time immune cell infiltration and drug penetration, we established a 3D co-culture model based on human NRAS mutated MUG Mel-2 spheroids and PBMCs (HLA matched), which displayed a synergistic effect when treated with the cancer vaccine compared to relative controls. Subsequently, we investigated the systemic delivery of the vaccine in EV formulations in a humanized NSG MUG Mel-2 melanoma mouse model. Our study provides a promising strategy for a tumor-targeted vaccine delivery by EVs, resulting in improved anticancer efficacy and increased infiltration of tumor-infiltrating lymphocytes. This study explores the potential of EVs for the selective delivery of cancer vaccines against malignancies, such as NRAS melanoma. Overall, this research could pave the way for applying autologous EVs as a safe and efficacious tool for targeted cancer therapy.
Auto-Abs neutralizing type I IFNs in patients with severe Powassan, Usutu, or Ross River virus disease
Authors: Adrian Gervais, Paul Bastard, Lucy Bizien, Céline Delifer, Pierre Tiberghien, Chaturaka Rodrigo, Francesca Trespidi, Micol Angelini, Giada Rossini, Tiziana Lazzarotto, Francesca Conti, Irene Cassaniti, Fausto Baldanti, Francesca Rovida, Alessandro Ferrari, Davide Mileto, Alessandro Mancon, Laurent Abel, Anne Puel, Aurélie Cobat, Charles M. Rice, Dániel Cadar, Jonas Schmidt-Chanasit, Johannes F. Scheid, Jacob E. Lemieux, Eric S. Rosenberg, Marianna Agudelo, Stuart G. Tangye, Alessandro Borghesi, Guillaume André Durand, Emilie Duburcq-Gury, Braulio M. Valencia, Andrew R. Lloyd, Anna Nagy, Margaret M. MacDonald, Yannick Simonin, Shen-Ying Zhang, Jean-Laurent Casanova
Journal: The Journal of Experimental Medicine (2024)

Arboviral diseases are a growing global health concern. Pre-existing autoantibodies (auto-Abs) neutralizing type I interferons (IFNs) can underlie encephalitis due to West Nile virus (WNV) (∼40% of patients) and tick-borne encephalitis (TBE, due to TBE virus [TBEV]) (∼10%). We report here that these auto-Abs can also underlie severe forms of rarer arboviral infections. Auto-Abs neutralizing high concentrations of IFN-α2, IFN-β, and/or IFN-ω are present in the single case of severe Powassan virus (POWV) encephalitis studied, two of three cases of severe Usutu virus (USUV) infection studied, and the most severe of 24 cases of Ross River virus (RRV) disease studied. These auto-Abs are not found in any of the 137 individuals with silent or mild infections with these three viruses. Thus, auto-Abs neutralizing type I IFNs underlie an increasing list of severe arboviral diseases due to Flaviviridae (WNV, TBEV, POWV, USUV) or Togaviridae (RRV) viruses transmitted to humans by mosquitos (WNV, USUV, RRV) or ticks (TBEV, POWV).
AI-empowered perturbation proteomics for complex biological systems
Authors: Liujia Qian, Rui Sun, Ruedi Aebersold, Peter Bühlmann, Chris Sander, Tiannan Guo
Journal: Cell Genomics (2024)

SummaryThe insufficient availability of comprehensive protein-level perturbation data is impeding the widespread adoption of systems biology. In this perspective, we introduce the rationale, essentiality, and practicality of perturbation proteomics. Biological systems are perturbed with diverse biological, chemical, and/or physical factors, followed by proteomic measurements at various levels, including changes in protein expression and turnover, post-translational modifications, protein interactions, transport, and localization, along with phenotypic data. Computational models, employing traditional machine learning or deep learning, identify or predict perturbation responses, mechanisms of action, and protein functions, aiding in therapy selection, compound design, and efficient experiment design. We propose to outline a generic PMMP (perturbation, measurement, modeling to prediction) pipeline and build foundation models or other suitable mathematical models based on large-scale perturbation proteomic data. Finally, we contrast modeling between artificially and naturally perturbed systems and highlight the importance of perturbation proteomics for advancing our understanding and predictive modeling of biological systems.
Isolation and escape mapping of broadly neutralizing antibodies against emerging delta-coronaviruses
Authors: Megi Rexhepaj, Daniel Asarnow, Lisa Perruzza, Young-Jun Park, Barbara Guarino, Mathew Mccallum, Katja Culap, Christian Saliba, Giada Leoni, Alessio Balmelli, Courtney N. Yoshiyama, Miles S. Dickinson, Joel Quispe, Jack T. Brown, M. Alejandra Tortorici, Kaitlin R. Sprouse, Ashley L. Taylor, Davide Corti, Tyler N. Starr, Fabio Benigni, David Veesler
Journal: Immunity (2024)

Porcine delta-coronavirus (PDCoV) spillovers were recently detected in febrile children, underscoring the recurrent zoonoses of divergent CoVs. To date, no vaccines or specific therapeutics are approved for use in humans against PDCoV. To prepare for possible future PDCoV epidemics, we isolated PDCoV spike (S)-directed monoclonal antibodies (mAbs) from humanized mice and found that two, designated PD33 and PD41, broadly neutralized a panel of PDCoV variants. Cryoelectron microscopy (cryo-EM) structures of PD33 and PD41 in complex with the S receptor-binding domain (RBD) and ectodomain trimer revealed the epitopes recognized by these mAbs, rationalizing their broad inhibitory activity. We show that both mAbs competitively interfere with host aminopeptidase N binding to neutralize PDCoV and used deep-mutational scanning epitope mapping to associate RBD antigenic sites with mAb-mediated neutralization potency. Our results indicate a PD33-PD41 mAb cocktail may heighten the barrier to escape. PD33 and PD41 are candidates for clinical advancement against future PDCoV outbreaks.
Hyperglycemia-triggered lipid peroxidation destabilizes STAT4 and impairs anti-viral Th1 responses in type 2 diabetes
Authors: Victor Gray, Weixin Chen, Rachael Julia Yuenyinn Tan, Jia Ming Nickolas Teo, Zhihao Huang, Carol Ho-Yi Fong, Tommy Wing Hang Law, Zi-Wei Ye, Shuofeng Yuan, Xiucong Bao, Ivan Fan-Ngai Hung, Kathryn Choon-Beng Tan, Chi-Ho Lee, Guang Sheng Ling
Journal: Cell Metabolism (2024)
Oral administration of <i>Lactobacillus casei</i> DG® after ileostomy closure in restorative proctocolectomy: a randomized placebo-controlled trial (microbiota and immune microenvironment in pouchitis -MEP1)
Authors: Imerio Angriman, Melania Scarpa, Edoardo Savarino, Ilaria Patuzzi, Alessandra Rigo, Andromachi Kotsafti, Astghik Stepanyan, Elisa Sciuto, Francesco Celotto, Silvia Negro, Antonino Caruso, Cesare Ruffolo, Romeo Bardini, Salvatore Pucciarelli, Brigida Barberio, Gaya Spolverato, Fabiana Zingone, Renata D’Incà, Ignazio Castagliuolo, Marco Scarpa
Journal: Gut Microbes (2024)

Pouchitis is an idiopathic inflammatory disease that may occur in ileal pouches, and it can lead to ileal pouch failure. This was a single-center, randomized, double-blinded, placebo-controlled trial that assessed the effect of Lactobacillus casei (L. casei) DG®, a probiotic strain, on the ileal pouch mucosa to determine the crosstalk between microbiota and mucosal immune system. Fifty-two patients undergoing restorative proctocolectomy were recruited and randomly assigned to receive a daily oral supplementation of L. casei DG® (n = 26) or placebo (n = 26) for 8 weeks from the ileostomy closure (T0) to a pouch endoscopy after 8 weeks (T1) and 1 year (T2). Ileal pouch mucosa samples were collected at T0, T1, and T2. At T1, the L. casei DG®-supplemented group showed a significant reduction of inflammatory cytokines levels compared to T0 baseline levels in the pouch mucosa, whereas in the placebo group cytokines levels resulted stable. In conclusion, probiotic manipulation of mucosal microbiota by L. casei DG®-supplementation after stoma closure in patients who underwent restorative proctocolectomy has a beneficial impact on the ileal pouch microenvironment. Registration number: NCT03136419 (http://www.clinicaltrials.gov).
Transfer of microbiota from lean donors in combination with prebiotics prevents excessive weight gain and improves gut-brain vagal signaling in obese rats
Authors: Dulce M. Minaya, Jiyoung S Kim, Rebecca Kirkland, Jillian Allen, Sitara Cullinan, Neil Maclang, Guillaume de Lartigue, Claire de La Serre
Journal: Gut Microbes (2024)

Gastrointestinal (GI) microbiota plays an active role in regulating the host's immune system and metabolism, as well as certain pathophysiological processes. Diet is the main factor modulating GI microbiota composition and studies have shown that high fat (HF) diets induce detrimental changes (dysbiosis) in the GI bacterial makeup. HF diet induced dysbiosis has been associated with structural and functional changes in gut-brain vagally mediated signaling system, associated with overeating and obesity. Although HF-driven changes in microbiota composition are sufficient to alter vagal signaling, it is unknown if improving microbiota composition after diet-induced obesity has been established can ameliorate gut-brain signaling and metabolic outcomes. In this study, we evaluated the effect of lean gut microbiota transfer in obese, vagally compromised, rats on gut-brain communication, food intake, and body weight. Male rats were maintained on regular chow or 45% HF diet for nine weeks followed by three weeks of microbiota depletion using antibiotics. The animals were then divided into four groups (n = 10 each): LF – control fed regular chow, LF-LF – chow fed animals that received microbiota from chow fed donors, HF-LF – HF fed animals that received microbiota from chow fed donors, and HF-HF – HF fed animals that received microbiota from HF fed donors. HF-LF animals received inulin as a prebiotic to aid the establishment of the lean microbiome. We found that transferring a LF microbiota to HF fed animals (HF-LF) reduced caloric intake during the light phase when compared with HF-HF rats and prevented additional excessive weight gain. HF-LF animals displayed an increase in postprandial activation of both primary sensory neurons innervating the GI tract and brainstem secondary neurons. We concluded from these data that improving microbiota composition in obese rats is sufficient to ameliorate gut-brain communication and restore normal feeding patterns which was associated with a reduction in weight gain.
Meal Timing Interventions for Weight Loss and Metabolic Health—What Does the Evidence Tell Us So Far?
Authors: Collin J. Popp, Emily N. C. Manoogian, Blandine Laferrère
Journal: JAMA Network Open (2024)

Hiu Yee Liu, MNDP; Ashley A. Eso, MNDP; Nathan Cook, MNDP; Hayley M. O'Neill, PhD; Loai Albarqouni, MD, MSc, PhD
Saving Children’s Lives Through Universal Pediatric Readiness Is a Wise Investment
Authors: Laura G. Burke, Jeanette I. Beaute, Kenneth A. Michelson
Journal: JAMA Network Open (2024)

Craig D. Newgard, MD, MPH; Amber Lin, MS; Jeremy D. Goldhaber-Fiebert, PhD; Katherine E. Remick, MD; Marianne Gausche-Hill, MD; Randall S. Burd, MD, PhD; Susan Malveau, MS; Jennifer N. B. Cook, GCPH; Peter C. Jenkins, MD, MSc; Stefanie G. Ames, MD, MS; N. Clay Mann, PhD, MS; Nina E. Glass, MD; Hilary A. Hewes, MD; Mary Fallat, MD; Apoorva Salvi, MS; Brendan G. Carr, MD, MS; K. John McConnell, PhD; Caroline Q. Stephens, MD, MPH; Rachel Ford, MPH; Marc A. Auerbach, MD; Sean Babcock, MS; Nathan Kuppermann, MD, MPH
Smoking Cessation and Incident Cardiovascular Disease
Authors: Jun Hwan Cho, Seung Yong Shin, Hoseob Kim, Mina Kim, Kyeongmin Byeon, Moonki Jung, Ki-Woon Kang, Wang-Soo Lee, Sang-Wook Kim, Gregory Y. H. Lip
Journal: JAMA Network Open (2024)

The association between smoking cessation and cardiovascular disease (CVD) risk in relation to cumulative smoking exposure remains poorly understood.
State and National Estimates of the Cost of Emergency Department Pediatric Readiness and Lives Saved
Authors: Craig D. Newgard, Amber Lin, Jeremy D. Goldhaber-Fiebert, Katherine E. Remick, Marianne Gausche-Hill, Randall S. Burd, Susan Malveau, Jennifer N. B. Cook, Peter C. Jenkins, Stefanie G. Ames, N. Clay Mann, Nina E. Glass, Hilary A. Hewes, Mary Fallat, Apoorva Salvi, Brendan G. Carr, K. John McConnell, Caroline Q. Stephens, Rachel Ford, Marc A. Auerbach, Sean Babcock, Nathan Kuppermann
Journal: JAMA Network Open (2024)

High emergency department (ED) pediatric readiness is associated with improved survival among children receiving emergency care, but state and national costs to reach high ED readiness and the resulting number of lives that may be saved are unknown.