Surveillance Functions Research Articles

Enhanced Immune Functions of In Vitro Human Natural Killer Cells and Splenocytes in Immunosuppressed Mice Supplemented with Mature Silkworm Products

Objectives: The immune-enhancing properties of steamed mature silkworm, known as HongJam (HJ), were investigated using human interleukin-2-independent Natural Killer 92 (NK92-MI) cells and a cyclophosphamide intraperitoneal injection-induced immunosuppressed mice model (CPA-IP). White Jade variety mature silkworm HJ (WJ-HJ) was used to prepare WJ-HJ supercritical fluid extracts (WJ-SCE) and WJ-HJ-supplemented feeds. Results: Treatment with WJ-SCE significantly enhanced proliferation, migration, and cytotoxicity of NK92-MI cells against various cancer cells while improving mitochondrial function and ATP production (p < 0.05). In CPA-IP mice, consumption of WJ-HJ-supplemented feeds restored immune function by improving body weight, immune organ indices, immunoglobulin levels, and blood cytokines. Splenocyte proliferation and cytotoxicity were significantly elevated in both saline intraperitoneal injection (Sal-IP) and CPA-IP groups with WJ-HJ supplementation, independent of mitogen activation (p < 0.05). Conclusions: These results suggest that WJ-HJ enhances immune modulation and immune surveillance functions of NK cells by improving mitochondrial and cytotoxic functions. WJ-HJ holds promise as a functional food for immune enhancement, pending clinical validation.

SLAM/SAP signaling regulates discrete γδ T cell developmental checkpoints and shapes the innate-like γδ TCR repertoire.

During thymic development, most γδ T cells acquire innate-like characteristics that are critical for their function in tumor surveillance, infectious disease, and tissue repair. The mechanisms, however, that regulate γδ T cell developmental programming remain unclear. Recently, we demonstrated that the SLAM/SAP signaling pathway regulates the development and function of multiple innate-like γδ T cell subsets. Here, we used a single-cell proteogenomics approach to identify SAP-dependent developmental checkpoints and to define the SAP-dependent γδ TCR repertoire in mice. SAP deficiency resulted in both a significant loss of an immature Gzma+Blk+Etv5+Tox2+ γδT17 precursor population and a significant increase in Cd4+Cd8+Rorc+Ptcra+Rag1+ thymic γδ T cells. SAP-dependent diversion of embryonic day 17 thymic γδ T cell clonotypes into the αβ T cell developmental pathway was associated with a decreased frequency of mature clonotypes in neonatal thymus, and an altered γδ TCR repertoire in the periphery. Finally, we identify TRGV4/TRAV13-4(DV7)-expressing T cells as a novel, SAP-dependent Vγ4 γδT1 subset. Together, the data support a model in which SAP-dependent γδ/αβ T cell lineage commitment regulates γδ T cell developmental programming and shapes the γδ TCR repertoire.

SLAM/SAP signaling regulates discrete γδ T cell developmental checkpoints and shapes the innate-like γδ TCR repertoire

During thymic development, most γδ T cells acquire innate-like characteristics that are critical for their function in tumor surveillance, infectious disease, and tissue repair. The mechanisms, however, that regulate γδ T cell developmental programming remain unclear. Recently, we demonstrated that the SLAM/SAP signaling pathway regulates the development and function of multiple innate-like γδ T cell subsets. Here, we used a single-cell proteogenomics approach to identify SAP-dependent developmental checkpoints and to define the SAP-dependent γδ TCR repertoire in mice. SAP deficiency resulted in both a significant loss of an immature Gzma+Blk+Etv5+Tox2+ γδT17 precursor population and a significant increase in Cd4+Cd8+Rorc+Ptcra+Rag1+ thymic γδ T cells. SAP-dependent diversion of embryonic day 17 thymic γδ T cell clonotypes into the αβ T cell developmental pathway was associated with a decreased frequency of mature clonotypes in neonatal thymus, and an altered γδ TCR repertoire in the periphery. Finally, we identify TRGV4/TRAV13-4(DV7)-expressing T cells as a novel, SAP-dependent Vγ4 γδT1 subset. Together, the data support a model in which SAP-dependent γδ/αβ T cell lineage commitment regulates γδ T cell developmental programming and shapes the γδ TCR repertoire.

Identification and Phenotypic Characterization of Neoantigen-Specific Cytotoxic CD4+ T Cells in Endometrial Cancer.

Tumor-reactive CD4+ T cells often accumulate in the tumor microenvironment (TME) in human cancer, but their functions and roles in antitumor responses remain elusive. Here, we investigated the immunopeptidome of HLA class II-positive (HLA-II+) endometrial cancer with an inflamed TME using a proteogenomic approach. We identified HLA-II neoantigens, one of which induced polyclonal CD4+ tumor-infiltrating lymphocyte responses. We then experimentally demonstrated that neoantigen-specific CD4+ tumor-infiltrating lymphocytes lyse target cells in an HLA-II-dependent manner. Single-cell transcriptomic analysis of the TME coupled with T-cell receptor sequencing revealed the presence of CD4+ T-cell clusters characterized by CXCL13 expression. The CXCL13+ clusters contained two subclusters with distinct cytotoxic gene expression patterns. The identified neoantigen-specific CD4+ T cells were found exclusively in one of the CXCL13+ subclusters characterized by granzyme B and CCL5 expression. These results demonstrate the involvement of tumor-reactive CD4+ T cells with cytotoxic function in immune surveillance of endometrial cancer and reveal their transcriptomic signature.

Family Policing and the Carceral State: How Carceral Violence Persists Through the Surveillance, Punishment, and Regulation of Families

ABSTRACT Although conceptualizations of the carceral state have become more expansive in recent years to include systems beyond prisons and policing, the child welfare system has often eluded this conceptualization due to a societal perception that it protects vulnerable children from harm. Despite this, the system has continued to function as a system of social control informed by carceral logics. Families become trapped within this extensive system of surveillance, many who receive little reprieve from the issues that brought them to the system’s attention. Once involved in the system, families are forced to comply with services focused on regulation of the behaviors deemed to be harmful or risky, while the system employs an escalating series of punishments to ensure compliance including the forcible separation of children from their families. In this paper, we analyze these functions of surveillance, regulation, and punishment, adapting and extending penal techniques described by Kohler-Hausmann (2019) to position the child welfare system as an arm of the carceral continuum used to maintain the oppression of Black communities. As such, we use the term family policing to more accurately capture the carceral functions of the child welfare system and the role it plays in the lives of families,

Towards a More Democratic Forum: Source Analysis of News Reporting on Older People's Problems in Delo during the Slovenian Parliamentary Elections from 2004 to 2018

While an increasing number of studies provide important insights into media construction of old age, the structure of news sources is seldom examined. The latter is especially important in democratic societies, during election years, when people elect representatives who tackle older people's social problems. Using content analysis, we examined the source structure of news articles reporting on older people's problems in the Slovenian newspaper Delo in five sequential parliamentary election years from 2004 to 2018. The findings showed that news coverage was predominantly constructed by elite sources rarely including representatives of older people. This characteristic strengthens the surveillance function of Delo but undermines its role as a democratic forum. However, during the last three observed election years, Delo's news reporting showed signs of transitioning towards a more democratic forum, which holds potential for fostering older people's civic inclusion.

Circular RNA circWBSCR22 facilitates colorectal cancer metastasis by enhancing CHD4's protein stability

Widespread metastases continue to be a massive challenge for colorectal cancer (CRC) therapy and contribute to the high mortality rate in patients with CRC. Circular RNAs (circRNAs) are a novel group of endogenous RNAs identified as agents modulating tumorigenesis and aggressiveness with tissue heterogeneity. However, the biological functions of circRNAs in CRC metastasis remain largely unknown. Here, we identified that circWBSCR22, a novel circRNA back-spliced from the WBSCR22 pre-mRNA, was significantly elevated in CRC tissue compared with adjacent normal tissue. Further gain- and loss-of-function assays manifested that circWBSCR22 promotes epithelial-mesenchymal transition (EMT) and metastasis in vitro and in vivo, which are mediated by the chromodomain helicase-DNA-binding protein 4 (CHD4) protein. Mechanically, circWBSCR22 binds directly to up-frameshift protein 1 (UPF1), an RNA binding protein recognized for its function in RNA surveillance, and hinders its role in directing CHD4 protein ubiquitination for degradation. Consequently, by stabilizing the CHD4 protein, circWBSCR22 hastens the development and metastasis of CRC. Therefore, our findings first delineate the oncogenic role and mechanism of circWBSCR22 in CRC growth and metastasis and its potential to serve as a therapeutic target for CRC.

6.F. Scientific session: Understanding the impact of climate change on health: the latest reports from EEA and NPHIs

Abstract Climate change poses a significant threat to human life and wellbeing worldwide. Extreme weather events like heatwaves, storms or floods can have adverse impacts on health, as can many other factors influenced by climate change, such as allergens, UV radiation and vector-borne diseases. In order to support the action needed to protect and promote health and wellbeing, many National Public Health Institutes (NPHIs) aim to keep relevant stakeholders informed on the scientific evidence of the impacts of climate change on health. Reports that synthesise the evidence can help in the development of suitable and health-relevant climate change adaptation and mitigation measures. In this workshop, European Environment Agency assessments on climate and health are presented, including the European Climate Risk Assessment Report and recent outputs of the European Climate and Health Observatory. Next, three NPHIs introduce their recent national reports on the impact of climate change on health, sharing lessons learned and valuable insights into their content, process and reception, highlighting similarities and differences. The workshop will feature presentations by NPHIs from the United Kingdom (UK Health Security Agency), Germany (Robert Koch Institute), and the Netherlands (National Institute for Public Health and the Environment) as well as the European Climate and Health Observatory. The perspective of the European Climate and Health Observatory offers Europe-wide insights as a provider of tools and knowledge that support actions from NPHIs and other health actors. Our aim is to highlight the value of such reports, and inform and inspire NPHIs and other stakeholders that may be considering the publication of similar reports. The format of the workshop will be informative and interactive, with opportunity for Q&A, discussion, and engagement with the audience. By showcasing the recent reports, the workshop aims to increase awareness of climate change as a public health issue and establish NPHIs as key climate actors. Furthermore, it aims to highlight the importance of having a national, up-to-date evidence base to drive climate measures and enhance resilience. Key messages • National evidence-based reports on climate change and health are a useful tool for informing health-relevant climate change adaptation and mitigation measures. • Through the public health functions of surveillance, monitoring, responding and evaluating, NPHIs play a crucial role in collating and assessing the evidence on the health effects of climate change.

Designing an Intervention to Improve Medication Safety for Nursing Home Residents Based on Experiential Knowledge Related to Patient Safety Culture at the Nursing Home Front Line: Cocreative Process Study.

Despite years of attention, avoiding medication-related harm remains a global challenge. Nursing homes provide essential health care for frail older individuals, who often experience multiple chronic diseases and polypharmacy, increasing their risk of medication errors. Evidence of effective interventions to improve medication safety in these settings is inconclusive. Focusing on patient safety culture is a potential key to intervention development as it forms the foundation for overall patient safety and is associated with medication errors. This study aims to develop an intervention to improve medication safety for nursing home residents through a cocreative process guided by integrated knowledge translation and experience-based codesign. This study used a cocreative process guided by integrated knowledge translation and experience-based co-design principles. Evidence on patient safety culture was used as an inspirational source for exploration of medication safety. Data collection involved semistructured focus groups to generate experiential knowledge (stage 1) to inform intervention design in a multidisciplinary workshop (stage 2). Research validation engaging different types of research expertise and municipal managerial representatives in finalizing the intervention design was essential. Acceptance of the final intervention for evaluation was aimed for through contextualization focused on partnership with a municipal advisory board. An abductive, rapid qualitative analytical approach to data analysis was chosen using elements from analyzing in the present, addressing the time-dependent, context-bound aspects of the cocreative process. Experiential knowledge was represented by three main themes: (1) closed systems and gaps between functions, (2) resource interpretation and untapped potential, and (3) community of medication safety and surveillance. The main themes informed the design of preliminary intervention components in a multidisciplinary workshop. An intervention design process focused on research validation in addition to contextualization resulted in the Safe Medication in Nursing Home Residents (SAME) intervention covering (1) campaign material visualizing key roles and responsibilities regarding medication for nursing home residents and (2) "Medication safety reflexive spaces" focused on social and health care assistants. The cocreative process successfully resulted in the multifaceted SAME intervention, grounded in lived experiences shared by some of the most important (but often underrepresented in research) stakeholders: frontline health care professionals and representatives of nursing home residents. This study brought attention toward closed systems related to functions in medication management and surveillance, not only informing the SAME intervention design but as opportunities for further exploration in future research. Evaluation of the intervention is an important next step. Overall, this study represents an important contribution to the complex field of medication safety. RR2-10.2196/43538.

Defenders of the Transcriptome: Guard Protein-Mediated mRNA Quality Control in Saccharomyces cerevisiae.

Cell survival depends on precise gene expression, which is controlled sequentially. The guard proteins surveil mRNAs from their synthesis in the nucleus to their translation in the cytoplasm. Although the proteins within this group share many similarities, they play distinct roles in controlling nuclear mRNA maturation and cytoplasmic translation by supporting the degradation of faulty transcripts. Notably, this group is continuously expanding, currently including the RNA-binding proteins Npl3, Gbp2, Hrb1, Hrp1, and Nab2 in Saccharomyces cerevisiae. Some of the human serine-arginine (SR) splicing factors (SRSFs) show remarkable similarities to the yeast guard proteins and may be considered as functional homologues. Here, we provide a comprehensive summary of their crucial mRNA surveillance functions and their implications for cellular health.

Iron overload impact on peripheral blood cells, lymphocytes and karyotypes in newly diagnosed myelodysplastic syndromes (MDS) patients: a single-center retrospective analysis

We conducted a retrospective analysis of 68 newly diagnosed myelodysplastic syndrome (MDS) patients who were treated in the Department of Hematology at Yuncheng Central Hospital affiliated to Shanxi Medical University from October 2017 to March 2022. Based on the serum ferritin (SF) level, the newly diagnosed MDS patients were divided into two groups: iron overload (IOL) group (SF > 1000 ng/mL) and non-iron overload (NIOL) group(SF ≤ 1000 ng/mL). General clinical data, peripheral blood cells, lymphocyte subsets and marrow karyotypes were compared. We found that MDS patients with IOL had lower hemoglobin (Hb), lower reticulocyte (RET) counts, and lower absolute natural killer (NK) cell counts compared with NIOL group (p < 0.05). The correlation analysis also showed a negative correlation between the NK cell counts and SF level (r = −0.270, P = 0.026). Furthermore, we found IOL group had higher incidence of complex and poor karyotypes. In conclusion, we found that IOL is correlated with anemia in MDS patients, which may lead to a decrease in T lymphocyte cells, especially in NK cells, that play a role in the immune pathogenesis of MDS, thereby suppressing immune surveillance function. Additionally, IOL may contribute to the pathogenesis of MDS by affecting karyotypes.

Family communal space as a natural surveillance function to create resilient coastal settlements

ABSTRACT Family communual space (FCS) can become a natural surveillance site for residents to monitor their environment. This study, of a fishing village in Indonesia, aims to determine the extent to which FCS plays a role as natural surveillance for its environment. The method uses a combination of quantitative (space syntax) with connectivity, visibility, and isovist approaches, as well as qualitative, such as support for data on kinship level, time of FCS use, and FCS location. The spatial analysis results show that visibility is low because the housing is dense and irregular, axial connectivity is low because there are many bends in roads, and the isovist has a wide observation range. The isovist was able to demonstrate the FCS’s role in natural surveillance. Findings: FCS and kinship ties can optimize environmental security even though visibility and connectivity are low to achieve resilience and sustainability and these are new prospects for researching the same thing but with different spatial methods or different places.

Mouse brain contains age-dependent extraparenchymal granular structures and astrocytes, both reactive to natural IgM antibodies, linked to the fissura magna

BackgroundMouse brains can contain specific polyglucosan aggregates known as Periodic Acid-Schiff (PAS)-granules. Generated in astrocytes, these granules increase with age and exhibit neo-epitopes of carbohydrate nature that are recognized by natural IgM antibodies (IgMs). The existence of neoepitopes on PAS granules suggests the presence of neoepitopes in other brain structures, and this is investigated here. To this end, brain sections from SAMP8 and ICR-CD1 mice were examined at different ages.ResultsWe have identified two novel structures that, apart from PAS granules, are recognized by natural IgMs. On one side, IgM reactive (IgM+) granular structures which are placed in the longitudinal fissure, the quadrigeminal cistern, and a region that extends from the quadrigeminal cistern to the interpeduncular cistern. This last region, located between the telencephalon and both the mesencephalon and diencephalon, is designated henceforth as the fissura magna, as it is indeed a fissure and the largest in the brain. As all these regions are extraparenchymal (EP), the IgM+ granules found in these zones have been named EP granules. These EP granules are mainly associated with fibroblasts and are not stained with PAS. On the other side, some IgM+ astrocytes have been found in the glia limitans, near the above-mentioned fissures. Remarkably, EP granules are more prevalent at younger ages, while the number of IgM+ astrocytes increases with age, similarly to the already described evolution of PAS granules.ConclusionsThe present work reports the presence of two brain-related structures that, apart from PAS granules, contain neo-epitopes of carbohydrate nature, namely EP granules and IgM+ astrocytes. We suggest that EP granules, associated to fibroblasts, may be part of a physiological function in brain clearance or brain-CSF immune surveillance, while both PAS granules and IgM+ astrocytes may be related to the increasing accumulation of harmful materials that occurs with age and linked to brain protective mechanisms. Moreover, the specific localisation of these EP granules and IgM+ astrocytes suggest the importance of the fissura magna in these brain-related cleaning and immune functions. The overall results reinforce the possible link between the fissura magna and the functioning of the glymphatic system.

Underreporting of Cases in the COVID-19 Outbreak of Borriana (Spain) during Mass Gathering Events in March 2020: A Cross-Sectional Study.

Determining the number of cases of an epidemic is the first function of epidemiological surveillance. An important underreporting of cases was observed in many locations during the first wave of the COVID-19 pandemic. To estimate this underreporting in the COVID-19 outbreak of Borriana (Valencia Community, Spain) in March 2020, a cross-sectional study was performed in June 2020 querying the public health register. Logistic regression models were used. Of a total of 468 symptomatic COVID-19 cases diagnosed in the outbreak through anti-SARS-CoV-2 serology, 36 cases were reported (7.7%), resulting in an underreporting proportion of 92.3% (95% confidence interval [CI], 89.5-94.6%), with 13 unreported cases for every reported case. Only positive SARS-CoV-2 polymerase chain reaction cases were predominantly reported due to a limited testing capacity and following a national protocol. Significant factors associated with underreporting included no medical assistance for COVID-19 disease, with an adjusted odds ratio [aOR] of 10.83 (95% CI 2.49-47.11); no chronic illness, aOR = 2.81 (95% CI 1.28-6.17); middle and lower social classes, aOR = 3.12 (95% CI 1.42-6.85); younger age, aOR = 0.97 (95% CI 0.94-0.99); and a shorter duration of illness, aOR = 0.98 (95% CI 0.97-0.99). To improve the surveillance of future epidemics, new approaches are recommended.

SLAM/SAP signaling regulates discrete γδ T cell developmental checkpoints and shapes the innate-like γδ TCR repertoire.

During thymic development, most γδ T cells acquire innate-like characteristics that are critical for their function in tumor surveillance, infectious disease, and tissue repair. The mechanisms, however, that regulate γδ T cell developmental programming remain unclear. Recently, we demonstrated that the SLAM-SAP signaling pathway regulates the development and function of multiple innate-like γδ T cell subsets. Here, we used a single-cell proteogenomics approach to identify SAP-dependent developmental checkpoints and to define the SAP-dependent γδ TCR repertoire. SAP deficiency resulted in both a significant loss of an immature Gzma + Blk + Etv5 + Tox2 + γδT17 precursor population, and a significant increase in Cd4 + Cd8+ Rorc + Ptcra + Rag1 + thymic γδ T cells. SAP-dependent diversion of embryonic day 17 thymic γδ T cell clonotypes into the αβ T cell developmental pathway was associated with a decreased frequency of mature clonotypes in neonatal thymus, and an altered γδ TCR repertoire in the periphery. Finally, we identify TRGV4/TRAV13-4(DV7)-expressing T cells as a novel, SAP-dependent Vγ4 γδT1 subset. Together, the data suggest that SAP-dependent γδ/αβ T cell lineage commitment regulates γδ T cell developmental programming and shapes the γδ TCR repertoire.

Re-thinking the sustainability of sovereign debt

Abstract This article explores the contributions that law can make to the development of a holistic approach to sovereign debt sustainability. We focus on debt sustainability assessments (DSAs) conducted by the International Monetary Fund (IMF), which are linked to the IMF’s surveillance and lending functions, and determine whether it is necessary to restructure the debt of a country in debt distress and the timing, process, and terms of such a debt restructuring. While the precise causes of each country’s debt situation are unique, all countries are grappling with the rising costs and growing risks posed by climate change and other environmental and social factors. We suggest that the IMF’s current treatment of these environmental and social factors is opaque, unpredictable, and hard for the citizens of affected countries and other outsiders to understand. It also obscures the true burden that debt obligations impose on a sovereign and the country’s residents, and, thus, the amount of debt relief that it may need in order to achieve a sustainable debt position. To address these shortcomings, we identify financial, economic, environmental, and social (FEES) factors that we contend should be incorporated in the design of the frameworks governing DSAs and the operating principles and practices of the IMF through which DSAs are conducted. We argue that the IMF should draw on various hard and soft sources of international and transnational law to develop a FEES-based approach to sovereign debt sustainability that is more consistent, predictable, and legitimate than the current approach.

Органы политической полиции Саратовской губернии в конце XIX – начале XX века: институциональные изменения, направления работы и специфика межгубернского взаимодействия

Introduction. The relevance of the chosen research topic is of lasting importance, since the place, role and importance of the political police is always of paramount importance in order to ensure the security of the state and its power structures. Theoretical analysis. During the second half of the 19th and the beginning of the 20th centuries, the gendarmerie bodies underwent gradual reform and serious intradepartmental and interregional reorganization. As a result, by the beginning of the last century, two main, interconnected and interacting structures of the political police operated in the Russian Empire – a separate corps of gendarmes (with a network of provincial gendarmerie departments and gendarmerie police departments of railways) and Security (search) departments (points). Empirical analysis. It was revealed that the political police bodies operating in the early twentieth century on the territory of the Saratov province (Saratov Provincial Gendarmerie Department, Tambov-Ural Gendarmerie Police Department of Railways, Saratov Security Department and Volga Region Security Department), which underwent organizational and structural changes in accordance with the directive acts of the authorities, purposefully performed their functions of search, covert supervision and surveillance of unreliable elements, inquiry and political investigation, as well as the execution of court sentences in political cases. Results. The process of reorganization and restructuring of political police bodies at the local level, in particular, in the Saratov province, testifies to the contradictory and ambiguous nature of the ongoing transformations. The empirical material studied shows that the authorities of late imperial Russia failed, to the necessary extent, to create an effective system of political investigation, which was supposed to resist the rapidly developing revolutionary movement at the beginning of the 20th century.

Synergistic antitumor immune response mediated by paclitaxel-conjugated nanohybrid oncolytic adenovirus with dendritic cell therapy.

Dendritic cell (DC)-based vaccines have emerged as a promising strategy in cancer immunotherapy due to low toxicity. However, the therapeutic efficacy of DC as a monotherapy is insufficient due to highly immunosuppressive tumor environment. To address these limitations of DC as immunotherapeutic agent, we have developed a polymeric nanocomplex incorporating (1) oncolytic adenovirus (oAd) co-expressing interleukin (IL)-12 and granulocyte-macrophage colony-stimulating factor (GM-CSF) and (2) arginine-grafted bioreducible polymer with PEGylated paclitaxel (APP) to restore antitumor immune surveillance function in tumor milieu and potentiate immunostimulatory attributes of DC vaccine. Nanohybrid complex (oAd/APP) in combination with DC (oAd/APP+DC) induced superior expression level of antitumor cytokines (IL-12, GM-CSF, and interferon gamma) than either oAd/APP or DC monotherapy in tumor tissues, thus resulting in superior intratumoral infiltration of both endogenous and exogenous DCs. Furthermore, oAd/APP+DC treatment led superior migration of DC to secondary lymphoid organs, such as draining lymph nodes and spleen, in comparison with either monotherapy. Superior migration profile of DCs in oAd/APP+DC treatment group resulted in more prolific activation of tumor-specific T cells in these lymphoid organs and greater intratumoral infiltration of T cells. Additionally, oAd/APP+DC treatment led to lower subset of tumor infiltrating lymphocytes and splenocytes being immunosuppressive regulatory T cells than any other treatment groups. Collectively, oAd/APP+DC led to superior induction of antitumor immune response and amelioration of immunosuppressive tumor microenvironment to elicit potent tumor growth inhibition than either monotherapy.

Transgenerational increases in DNA methylation in Arabidopsis plants defective in active DNA demethylation

Spontaneous gain or loss of DNA methylation occurs in plant and animal genomes, and DNA methylation changes can lead to meiotically stable epialleles that generate heritable phenotypic diversity. However, it is unclear whether transgenerational epigenetic stability may be regulated by any cellular factors. Here, we examined spontaneously occurring variations in DNA methylation in wild-type and ros1 mutant Arabidopsis plants that were propagated for ten generations from single-seed descent. We found that the ros1 mutant, which is defective in active DNA demethylation, showed an increased transgenerational epimutation rate. The ros1 mutation led to more spontaneously gained methylation than lost methylation at individual cytosines, compared to the wild type which had similar numbers of spontaneously gained and lost methylation cytosines. Consistently, transgenerational differentially methylated regions were also biased toward hypermethylation in the ros1 mutant. Our results reveal a genetic contribution of the ROS1 DNA demethylase to transgenerational epigenetic stability and suggest that ROS1 may have an unexpected surveillance function in preventing transgenerational DNA methylation increases.

Machine learning based DNA melt curve profiling enables automated novel genotype detection

Surveillance for genetic variation of microbial pathogens, both within and among species, plays an important role in informing research, diagnostic, prevention, and treatment activities for disease control. However, large-scale systematic screening for novel genotypes remains challenging in part due to technological limitations. Towards addressing this challenge, we present an advancement in universal microbial high resolution melting (HRM) analysis that is capable of accomplishing both known genotype identification and novel genotype detection. Specifically, this novel surveillance functionality is achieved through time-series modeling of sequence-defined HRM curves, which is uniquely enabled by the large-scale melt curve datasets generated using our high-throughput digital HRM platform. Taking the detection of bacterial genotypes as a model application, we demonstrate that our algorithms accomplish an overall classification accuracy over 99.7% and perform novelty detection with a sensitivity of 0.96, specificity of 0.96 and Youden index of 0.92. Since HRM-based DNA profiling is an inexpensive and rapid technique, our results add support for the feasibility of its use in surveillance applications.