Bacteria are abundantly present through the body, and the proteins they secrete into the cellular microenvironment both interact with surface receptors and are internalized and interact with internal proteins responsible for important pathways and functions. One of these proteins is DnaK, a moonlighting bacterial chaperone interacting with many human proteins. Intrinsically disordered regions (IDRs) enable structural flexibility essential for multifunctionality, yet their role in DnaK and its interactors remains unclear. Analyzing data from our previous studies of Mycoplasma DnaK and human protein interactions, this study is the first to employ computational analysis to assess disorder in bacterial DnaK, bacterial proteomes, and human interactors, aiming to elucidate novel disorder-mediated mechanisms of bacterial pathogenesis. Results indicate that bacterial chaperones, including DnaK, are expected to be significantly more disordered than the proteome averages (∼27-42% chaperones are highly disordered vs. ∼7-10% proteome-wide highly disordered proteins). Human proteins interacting with bacterial DnaK cluster in chromatin organization and RNA processing networks. DnaK interactors are significantly more disordered than human HSP70/HSC70 interactors, as well as average human proteins, nuclear proteins, and proteins involved in RNA processing and DNA repair. Confirmed interactors, such as XRCC1 and USP10, feature long IDRs with many PTMs, have multiple disorder-based binding regions, and show high LLPS propensity, suggesting that disorder facilitates cellular pathways subversion. These findings are consistent with a model, in which DnaK, by virtue of its structural flexibility, may preferentially engage disordered host proteins, potentially influencing cellular pathways, paving the way for a hypothesis that warrants direct experimental validation.

Bacterial proteins released into the cellular microenvironment are increasingly recognized as pivotal modulators of host key signaling pathways, with significant implications for cellular functions. This review explores the multifaceted roles of such bacterial proteins, often functioning as virulence factors, in modulating the host cell cycle. By focusing on the interactions between selected bacterial proteins and essential components of the cell cycle machinery, we describe the mechanisms through which these interactions disrupt relevant cellular functions and contribute to disease development, with a particular focus on cancer.

Background: The death of a child due to complex congenital heart disease (CCHD) in pediatric intensive care units profoundly affects families, often resulting in lasting grief and emotional distress. Despite advancements in pediatric palliative care (PPC), significant gaps persist in communication and end-of-life (EoL) planning. This study explores the experiences and perceptions of bereaved parents to identify areas for improvement in PPC delivery. Methods: A qualitative phenomenological design was used to analyze the lived experiences of 18 bereaved parents whose children died from CCHD at a tertiary cardiac center. Semi-structured telephone interviews were conducted, incorporating five open-ended questions. Data were analyzed inductively using Colaizzi’s method to identify recurring themes and subthemes. Results: Four key thematic areas emerged: communication issues, the parental role, child care, and bereavement support. Parents highlighted inconsistent communication, lack of preparedness for EoL decisions, and emotional isolation as major challenges. Positive experiences often involved compassionate healthcare providers and structured psychological support. A significant proportion of parents identified family support and faith as key coping mechanisms, while others expressed dissatisfaction with post-mortem follow-up and the absence of long-term bereavement care. Conclusions: Bereaved parents’ experiences underscore the need for improved communication strategies, greater parental involvement in care, and enhanced bereavement support. Integrating structured decision-making pathways early in the care trajectory may help mitigate parental distress and improve the quality of EoL experiences for children with CCHD.

Aims Hypogeal environments with cultural heritage interest pose a real challenge for their preservation and conservation. The ancient Etruscan Necropolis of Tarquinia, Italy, consists of 200 tombs decorated with extraordinary mural paintings, of great artistic and historical value. Since the beginning of the restoration campaign in 2016, a regular microbiological survey has been performed in the Tomba degli Scudi. The aim of this study was to investigate the nature of an expansion of black spots on the pictorial layers recently observed.Methods and results To determine the origin of the black spots in the atrium chamber of the Tomba degli Scudi, the fungal community was sampled using various techniques: cellulose discs, swabs, and nylon membranes and investigated by a multi-analytical approach. The obtained results suggest that the identified fungal strains (e.g. Gliomastix murorum and Pseudogymnoascus pannorum) are common to many subterranean environments around the world, such as Lascaux cave.Conclusions The continuous and long-term monitoring made it possible to detect alterations at an early stage and assess the harmfulness of different fungal strains. This work is a demonstration of the effectiveness of prevention and monitoring actions within these fragile and valuable environments.

Background Chemotherapy is a primary treatment for cancer, but its efficacy is often limited by cancer-associated bacteria (CAB) that impair tumor suppressor functions. Our previous research found that Mycoplasma fermentans DnaK, a chaperone protein, impairs p53 activities, which are essential for most anti-cancer chemotherapeutic responses. Methods To investigate the role of DnaK in chemotherapy, we treated cancer cell lines with M. fermentans DnaK and then with commonly used p53-dependent anti-cancer drugs (cisplatin and 5FU). We evaluated the cells’ survival in the presence or absence of a DnaK-binding peptide (ARV-1502). We also validated our findings using primary tumor cells from a novel DnaK knock-in mouse model. To provide a broader context for the clinical significance of these findings, we investigated human primary cancer sequencing datasets from The Cancer Genome Atlas (TCGA). We identified F. nucleatum as a CAB carrying DnaK with an amino acid composition highly similar to M. fermentans DnaK. Therefore, we investigated the effect of F. nucleatum DnaK on the anti-cancer activity of cisplatin and 5FU. Results Our results show that both M. fermentans and F. nucleatum DnaKs reduce the effectiveness of cisplatin and 5FU. However, the use of ARV-1502 effectively restored the drugs' anti-cancer efficacy. Conclusions Our findings offer a practical framework for designing and implementing novel personalized anti-cancer strategies by targeting specific bacterial DnaKs in patients with poor response to chemotherapy, underscoring the potential for microbiome-based personalized cancer therapies.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has quickly become a global health pandemic. Among the viral proteins, RNA-dependent RNA polymerase (RdRp) is responsible for viral genome replication and has emerged as a promising target against SARS-CoV-2 infection. Dietary bioactive compounds represent an important source of evolutionarily optimized molecules with antiviral properties against SARS-CoV-2 RdRp. We investigated the inhibitory potential effects of different phytochemicals against SARS-CoV-2 RdRp, including andrographolide, kaempferol, resveratrol, and silibinin. Unlike the other investigated compounds, kaempferol exhibited a significant dose-dependent in vitro inhibition of SARS-CoV-2 RdRp activity. To assess the binding interactions and stability of the SARS-CoV-2 RdRp-kaempferol complex, we performed in silico techniques, including molecular docking, quantum chemical calculation, and molecular dynamics simulations. We found strong binding affinities and stability between kaempferol and SARS-CoV-2 RdRp variants (Wuhan and Omicron). These findings provide valuable insights into the antiviral properties of kaempferol as a stable inhibitor of SARS-CoV-2 RdRp.

Well-controlled repair mechanisms are involved in the maintenance of genomic stability, and their failure can precipitate DNA abnormalities and elevate tumor risk. In addition, the tumor microenvironment, enriched with factors inducing oxidative stress and affecting cell cycle checkpoints, intensifies DNA damage when repair pathways falter. Recent research has unveiled associations between certain bacteria, including Mycoplasmas, and various cancers, and the causative mechanism(s) are under active investigation. We previously showed that Mycoplasma fermentans DnaK, an HSP70 family chaperone protein, hampers the activity of proteins like PARP1 and p53, crucial for genomic integrity. Moreover, our analysis of its interactome in human cancer cell lines revealed DnaK's engagement with several components of DNA-repair machinery. Finally, in vivo experiments performed in our laboratory using a DnaK knock-in mouse model generated by our group demonstrated that DnaK exposure led to increased DNA copy number variants, indicative of genomic instability. We present here evidence that expression of DnaK is linked to increased i) incidence of tumors in vivo upon exposure to urethane, a DNA damaging agent; ii) spontaneous DNA damage ex vivo; and iii) expression of proinflammatory cytokines ex vivo, variations in reactive oxygen species levels, and increased β-galactosidase activity across tissues. Moreover, DnaK was associated with increased centromeric instability. Overall, these findings highlight the significance of Mycoplasma DnaK in the etiology of cancer and other genetic disorders providing a promising target for prevention, diagnostics, and therapeutics.

Journal Article A STEM Training Approach Focused on Microscopy at Morgan State University Get access Frank J Denaro, Frank J Denaro Department of Biology Morgan State University, Baltimore, MD, USA Corresponding author: Frank.Denaro@morgan.edu Search for other works by this author on: Oxford Academic Google Scholar Simon Nyaga, Simon Nyaga Department of Biology Morgan State University, Baltimore, MD, USA Search for other works by this author on: Oxford Academic Google Scholar Davide Zella, Davide Zella Institute of Human Virology, Baltimore, MD, USA Search for other works by this author on: Oxford Academic Google Scholar J Bryant, J Bryant Institute of Human Virology, Baltimore, MD, USA Search for other works by this author on: Oxford Academic Google Scholar Francesca Benedetti Francesca Benedetti Institute of Human Virology, Baltimore, MD, USA Search for other works by this author on: Oxford Academic Google Scholar Microscopy and Microanalysis, Volume 29, Issue Supplement_1, 1 August 2023, Pages 2111–2112, https://doi.org/10.1093/micmic/ozad067.1095 Published: 22 July 2023

Biomineralization of calcium carbonate by living organisms, including bacteria, has been studied for its potential use in conservation and restoration applications in Cultural Heritage. This study reports the carbonatogenic properties of three bacterial strains (Lysinibacillus fusiformis 3.20, Psychrobacillus psychrodurans 7Mo and Lederbergia lenta Vetro1) isolated from Il Giovane di Mozia sculpture (Mozia), the Etruscan mural paintings of the Tomba degli Scudi (Tarquinia), and a microbial community isolated from the Sant'Eustachio statue (Matera). The crystals precipitated have been investigated through a multi-analytical approach: Field Emission Scanning Electron Microscopy (FESEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). The analyses showed the formation of stable vaterite by the bacterial cultures, while the microbial community induced calcite precipitation. This approach aims to support the conservation-restoration efforts, taking into the account the requests of the conservation-restorers and the unique characteristics of each Cultural Heritage artwork.(c) 2023 Consiglio Nazionale delle Ricerche (CNR). Published by Elsevier Masson SAS. All rights reserved.

The history of the Earth is a story of the co-evolution of minerals and microbes: not only have numerous rocks arisen from life but also life itself may have formed from rocks. To understand the strong association between microbes and inorganic substrates, we investigated the moonmilk, a calcium carbonate deposit of possible microbial origin, occurring in the Iron Age Etruscan necropolis of Tarquinia, in Italy. These tombs provide a unique environment where the hypogeal walls of the tombs are covered by this speleothem. To study moonmilk formation, we investigated the bacterial community in the rock in which the tombs were carved: calcarenite and hybrid sandstone. We present the first evidence that moonmilk precipitation is driven by microbes within the rocks and not only on the rock surfaces. We also describe how the moonmilk produced within the rocks contributes to rock formation and evolution. The microbial communities of the calcarenite and hybrid sandstone displayed, at the phylum level, the same microbial pattern of the moonmilk sampled from the walls of the hypogeal tombs, suggesting that the moonmilk originates from the metabolism of an endolytic bacterial community. The calcite moonmilk is the only known carbonate speleothem on Earth with undoubted biogenic origin, thus representing a robust and credible biosignature of life. Its presence in the inner parts of rocks adds to its characteristics as a biosignature.

The COVID-19 pandemic has not only strained healthcare systems in Africa but has also intensified the impact of emerging and re-emerging diseases. Specifically in Equatorial Guinea, mirroring the situation in other African countries, unique zoonotic outbreaks have occurred during this challenging period. One notable resurgence is Marburg virus disease (MVD), which has further burdened the already fragile healthcare system. The re-emergence of the Marburg virus amid the COVID-19 pandemic is believed to stem from a probable zoonotic spill-over, although the precise transmission routes remain uncertain. Given the gravity of the situation, addressing the existing challenges is paramount. Though the genome sequences from the current outbreak were not available for this study, we analyzed all the available whole genome sequences of this re-emerging pathogen to advocate for a shift towards active surveillance. This is essential to ensure the successful containment of any potential Marburg virus outbreak in Equatorial Guinea and the wider African context. This study, which presents an update on the phylodynamics and the genetic variability of MARV, further confirmed the existence of at least two distinct patterns of viral spread. One pattern demonstrates a slower but continuous and recurring virus circulation, while the other exhibits a faster yet limited and episodic spread. These results highlight the critical need to strengthen genomic surveillance in the region to effectively curb the pathogen’s dissemination. Moreover, the study emphasizes the importance of prompt alert management, comprehensive case investigation and analysis, contact tracing, and active case searching. These steps are vital to support the healthcare system’s response to this emerging health crisis. By implementing these strategies, we can better arm ourselves against the challenges posed by the resurgence of the Marburg virus and other infectious diseases.

Antibiotic resistance is a significant global health concern that affects both human and animal populations. The One Health approach acknowledges the interconnectedness of human health, animal health, and the environment. It emphasizes the importance of collaboration and coordination across these sectors to tackle complex health challenges such as antibiotic resistance. In the context of One Health, antibiotic resistance refers to the ability of bacteria to withstand the efficacy of antibiotics, rendering them less effective or completely ineffective in treating infections. The emergence and spread of antibiotic-resistant bacteria pose a threat to human and animal health, as well as to the effectiveness of medical treatments and veterinary interventions. In particular, One Health recognizes that antibiotic use in human medicine, animal agriculture, and the environment are interconnected factors contributing to the development and spread of antibiotic resistance. For example, the misuse and overuse of antibiotics in human healthcare, including inappropriate prescribing and patient non-compliance, can contribute to the selection and spread of resistant bacteria. Similarly, the use of antibiotics in livestock production for growth promotion and disease prevention can contribute to the development of antibiotic resistance in animals and subsequent transmission to humans through the food chain. Addressing antibiotic resistance requires a collaborative One Health approach that involves multiple participants, including healthcare professionals, veterinarians, researchers, and policymakers.

The human microbiota affects critical cellular functions, although the responsible mechanism(s) is still poorly understood. In this regard, we previously showed that Mycoplasma fermentans DnaK, an HSP70 chaperone protein, hampers the activity of important cellular proteins responsible for DNA integrity. Here, we describe a novel DnaK knock-in mouse model generated in our laboratory to study the effect of M. fermentans DnaK expression in vivo. By using an array-based comparative genomic hybridization assay, we demonstrate that exposure to DnaK was associated with a higher number of DNA copy number variants (CNVs) indicative of unbalanced chromosomal alterations, together with reduced fertility and a high rate of fetal abnormalities. Consistent with their implication in genetic disorders, one of these CNVs caused a homozygous Grid2 deletion, resulting in an aberrant ataxic phenotype that recapitulates the extensive biallelic deletion in the Grid2 gene classified in humans as autosomal recessive spinocerebellar ataxia 18. Our data highlight a connection between components of the human urogenital tract microbiota, namely Mycoplasmas, and genetic abnormalities in the form of DNA CNVs, with obvious relevant medical, diagnostic, and therapeutic implications.

Monkeypox, a viral zoonotic disease, has emerged as a significant global threat in recent years. This review focuses on the importance of global monitoring and rapid response to monkeypox outbreaks. The unpredictable nature of monkeypox transmissions, its potential for human-to-human spread, and its high morbidity rate underscore the necessity for proactive surveillance systems. By analyzing the existing literature, including recent outbreaks, this review highlights the critical role of global surveillance in detecting, containing, and preventing the further spread of monkeypox. It also emphasizes the need for enhanced international collaboration, data sharing, and real-time information exchange to effectively respond to monkeypox outbreaks as a global health concern. Furthermore, this review discusses the challenges and opportunities of implementing robust surveillance strategies, including the use of advanced diagnostic tools and technologies. Ultimately, these findings underscore the urgency of establishing a comprehensive global monitoring framework for monkeypox, enabling early detection, prompt response, and effective control measures to protect public health worldwide.

Journal Article Identification of Increased Blood Brain Barrier Permeability in the Substantia Nigra of the HIV-1 Transgenic Rat Get access Frank Denaro, Frank Denaro Biology, Morgan State University, Baltimore MD frank.denaro@morgan.edu Search for other works by this author on: Oxford Academic Google Scholar Myla Worthington, Myla Worthington Biology, Morgan State University, Baltimore MD Search for other works by this author on: Oxford Academic Google Scholar Sumiko Williams, Sumiko Williams Biology, Morgan State University, Baltimore MD Search for other works by this author on: Oxford Academic Google Scholar Francesca Benedetti, Francesca Benedetti Institute of Human Virology University of Maryland Baltimore MD Search for other works by this author on: Oxford Academic Google Scholar Davide Zella, Davide Zella Institute of Human Virology University of Maryland Baltimore MD Search for other works by this author on: Oxford Academic Google Scholar Harry Davis, Harry Davis Institute of Human Virology University of Maryland Baltimore MD Search for other works by this author on: Oxford Academic Google Scholar Joseph Bryant Joseph Bryant Institute of Human Virology University of Maryland Baltimore MD Search for other works by this author on: Oxford Academic Google Scholar Microscopy and Microanalysis, Volume 28, Issue S1, 1 August 2022, Pages 3214–3215, https://doi.org/10.1017/S1431927622011953 Published: 01 August 2022