Gliomas are the most common brain tumor type in children and adolescents. To date, diagnosis and therapy monitoring for these tumors rely on magnetic resonance imaging (MRI) and histopathological as well as molecular analyses of tumor tissue. Recently, liquid biopsies (LB) have emerged as promising tool for diagnosis and longitudinal tumor assessment potentially allowing for a more precise therapeutic management. However, the optimal strategy for monitoring gliomas by LB remains to be determined. In this study, we analyzed circulating tumor DNA (ctDNA) from 78 liquid biopsies (plasma n = 44, cerebrospinal fluid n = 34 (CSF)) of 35 glioma patients, determining H3F3A K28M (K27M) and BRAF V600E mutation allele frequency using droplet digital PCR (ddPCR). All results were correlated to clinically relevant parameters including diagnostic imaging and CSF aspiration site (ventricular vs lumbar) with respect to tumor localization. Regarding diagnostic accuracy, the calculated sensitivity score in the H3F3A K27M cohort was 84.61% for CSF and 73.68% for plasma. In the BRAF V600E cohort, we determined a sensitivity of 83.3% in plasma and 80% in CSF. The overall specificity was 100%. With respect to the CSF aspiration, the intra-operatively obtained CSF demonstrated 100% detection rate, followed by ventricular CSF obtained via Ommaya Reservoir/shunt puncture (93%) and CSF obtained via lumbar puncture (66%). Notably, this further correlated with the proximity of the CSF site to tumor localization. Longitudinal CSF monitoring demonstrated a good correlation to clinical and radiological disease evolution. Importantly, we show for the first time that monitoring BRAF V600E by ddPCR could serve as treatment response assessment in gliomas. In summary, our observation may inform recommendations with regard to location of CSF aspiration when incorporating LB into future treatment protocols.

• Keywords
• CSF sampling site, Droplet digital PCR, Glioma, Liquid biopsy, Longitudinal monitoring, Targeted therapy,
• MeSH
• Circulating Tumor DNA cerebrospinal fluid   genetics   MeSH
• Child MeSH
• Adult MeSH
• Glioma * genetics   pathology   diagnosis   MeSH
• Histones * genetics   MeSH
• Middle Aged MeSH
• Humans MeSH
• Adolescent MeSH
• Young Adult MeSH
• Mutation MeSH
• Biomarkers, Tumor * genetics   cerebrospinal fluid   MeSH
• Brain Neoplasms * genetics   diagnosis   pathology   MeSH
• Child, Preschool MeSH
• Proto-Oncogene Proteins B-raf * genetics   MeSH
• Liquid Biopsy methods   MeSH
• Check Tag
• Child MeSH
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Adolescent MeSH
• Young Adult MeSH
• Male MeSH
• Child, Preschool MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• BRAF protein, human MeSH Browser
• Circulating Tumor DNA MeSH
• H3-3A protein, human MeSH Browser
• Histones * MeSH
• Biomarkers, Tumor * MeSH
• Proto-Oncogene Proteins B-raf * MeSH

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is an RNA virus responsible for coronavirus disease 2019 (COVID-19). While SARS-CoV-2 primarily targets the lungs and airways, it can also infect other organs, including the central nervous system (CNS). The aim of this study was to investigate whether the choroid plexus could serve as a potential entry site for SARS-CoV-2 into the brain. Tissue samples from 24 deceased COVID-19-positive individuals were analyzed. Reverse transcription real-time PCR (RT-qPCR) was performed on selected brain regions, including the choroid plexus, to detect SARS-CoV-2 viral RNA. Additionally, immunofluorescence staining and confocal microscopy were used to detect and localize two characteristic proteins of SARS-CoV-2: the spike protein S1 and the nucleocapsid protein. RT-qPCR analysis confirmed the presence of SARS-CoV-2 viral RNA in the choroid plexus. Immunohistochemical staining revealed viral particles localized in the epithelial cells of the choroid plexus, with the spike protein S1 detected in the late endosomes. Our findings suggest that the blood-cerebrospinal fluid (B-CSF) barrier in the choroid plexus serves as a route of entry for SARS-CoV-2 into the CNS. This study contributes to the understanding of the mechanisms underlying CNS involvement in COVID-19 and highlights the importance of further research to explore potential therapeutic strategies targeting this entry pathway.

• Keywords
• COVID‐19, SARS‐CoV‐2, blood‐cerebrospinal fluid barrier, choroid plexus, neuroinvasion,
• MeSH
• COVID-19 * virology   pathology   MeSH
• Adult MeSH
• Phosphoproteins MeSH
• Spike Glycoprotein, Coronavirus metabolism   genetics   MeSH
• Blood-Brain Barrier * virology   MeSH
• Virus Internalization * MeSH
• Coronavirus Nucleocapsid Proteins MeSH
• Middle Aged MeSH
• Humans MeSH
• Brain virology   MeSH
• Choroid Plexus * virology   pathology   MeSH
• RNA, Viral genetics   MeSH
• SARS-CoV-2 * physiology   genetics   MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Phosphoproteins MeSH
• Spike Glycoprotein, Coronavirus MeSH
• Coronavirus Nucleocapsid Proteins MeSH
• nucleocapsid phosphoprotein, SARS-CoV-2 MeSH Browser
• RNA, Viral MeSH
• spike protein, SARS-CoV-2 MeSH Browser

Development of hematopoietic populations through the process of differentiation is critical for proper hematopoiesis. The transcription factor CCAAT/enhancer binding protein alpha (C/EBPα) is a master regulator of myeloid differentiation, and the identification of C/EBPα target genes is key to understand this process. Here we identified the Ecotropic Viral Integration Site 2B (EVI2B) gene as a direct target of C/EBPα. We showed that the product of the gene, the transmembrane glycoprotein EVI2B (CD361), is abundantly expressed on the surface of primary hematopoietic cells, the highest levels of expression being reached in mature granulocytes. Using shRNA-mediated downregulation of EVI2B in human and murine cell lines and in primary hematopoietic stem and progenitor cells, we demonstrated impaired myeloid lineage development and altered progenitor functions in EVI2B-silenced cells. We showed that the compromised progenitor functionality in Evi2b-depleted cells can be in part explained by deregulation of cell proliferation and apoptosis. In addition, we generated an Evi2b knockout murine model and demonstrated altered properties of hematopoietic progenitors, as well as impaired G-CSF dependent myeloid colony formation in the knockout cells. Remarkably, we found that EVI2B is significantly downregulated in human acute myeloid leukemia samples characterized by defects in CEBPA. Altogether, our data demonstrate that EVI2B is a downstream target of C/EBPα, which regulates myeloid differentiation and functionality of hematopoietic progenitors.

• MeSH
• Leukemia, Myeloid, Acute metabolism   pathology   MeSH
• Apoptosis MeSH
• Cell Differentiation drug effects   MeSH
• Bone Marrow Cells cytology   MeSH
• Down-Regulation drug effects   MeSH
• Estradiol pharmacology   MeSH
• Granulocyte Colony-Stimulating Factor pharmacology   MeSH
• Granulocytes cytology   metabolism   MeSH
• Hematopoietic Stem Cells cytology   metabolism   MeSH
• Cells, Cultured MeSH
• Humans MeSH
• RNA, Small Interfering metabolism   MeSH
• Membrane Glycoproteins antagonists & inhibitors   genetics   metabolism   MeSH
• Membrane Proteins antagonists & inhibitors   genetics   metabolism   MeSH
• Mice, Inbred C57BL MeSH
• Mice, Knockout MeSH
• Mice MeSH
• Cell Proliferation drug effects   MeSH
• Promoter Regions, Genetic MeSH
• CCAAT-Enhancer-Binding Protein-alpha genetics   metabolism   MeSH
• RNA Interference MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Estradiol MeSH
• EVI2B protein, human MeSH Browser
• Granulocyte Colony-Stimulating Factor MeSH
• RNA, Small Interfering MeSH
• Membrane Glycoproteins MeSH
• Membrane Proteins MeSH
• CCAAT-Enhancer-Binding Protein-alpha MeSH

We developed a mobile laboratory allowing field exposure of lung tissue models to ambient air at localities with various pollution sources (Background, Industrial, Traffic, Urban) in different seasons (summer/fall/winter). In samples originating from healthy and asthmatic individuals, we assessed the parameters of toxicity, lipid peroxidation and immune response; we further performed comprehensive monitoring of air pollutants at sampling sites. We measured lactate dehydrogenase (LDH) and adenylate kinase (AK) production and transepithelial electrical resistance (TEER), analyzed 15-F2t-isopostane (IsoP) and a panel of 20 cytokines/chemokines/growth factors. In the ambient air, we detected particulate matter (PM), and other relevant chemicals (benzene, benzo[a]pyrene (BaP), NOx). In the Traffic locality, we found very high concentrations of ultrafine particles and NOx and observed low TEER values in the exposed samples, indicating significant traffic-related toxicity of the ambient air. In the Urban locality, sampled in winter, we observed high PM and BaP levels. We found lower AK levels in samples from healthy individuals exposed in this locality than in the asthmatic samples. In the samples from the Industrial locality, sampled in summer, we detected higher concentrations of TNFα, MIP-1α, Eotaxin, GROα, GM-CSF, IL-6 and IL-7 than in the Urban locality samples. We hypothesize that pollen or other plant-related components of the ambient air were responsible for this response. In conclusion, our data proved the feasibility of our mobile laboratory for field measurements of the biological response of lung tissue models exposed to ambient air, reflecting not only the levels of toxic compounds, but also season-specific parameters.

• Keywords
• air-liquid interface, asthma, lung tissue model, outdoor air pollution, real-world exposure,
• MeSH
• Asthma * metabolism   MeSH
• Cytokines metabolism   MeSH
• Adult MeSH
• Air Pollutants * toxicity   analysis   MeSH
• Humans MeSH
• Environmental Monitoring * methods   MeSH
• Particulate Matter analysis   toxicity   MeSH
• Respiratory Mucosa * drug effects   metabolism   MeSH
• Seasons MeSH
• Environmental Exposure * MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Male MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Comparative Study MeSH
• Names of Substances
• Cytokines MeSH
• Air Pollutants * MeSH
• Particulate Matter MeSH

PURPOSE OF THE STUDY: Although great advances have been made in both radiological diagnosis and antibiotic therapy of microbial infections, the treatment of spinal infections remains a major clinical challenge. Many of the patients affected are referred to spinal units with long delays. The general population is ageing and the number of immunocompromised patients, as well as the number of operative procedures for spinal disorders are increasing. The aim of our study was to evaluate the clinical presentations of spinal infections, options for their diagnosis, indications for treatment and their risk factors and the results of surgery. MATERIAL AND METHODS: The group of 112 patients evaluated after the treatment of spinal infection comprised 63 men and 49 women at an average age of 59.4 years (range, 17 to 84). The average follow-up was 3.2 years (range, 6 months to 8 years). Of these, 82 patients had primary hematogenous infection, 29 had post-operative infection,and one had an infected gun shot wound. Thirty-six patients showed neurological deficit and six were paraplegic. The diagnostic methods included FBC, CRP and EST tests, examination of blood cultures, aspirates and biopsy samples from the infected site, bone scintigraphy, MRI and CT scanning. Indications for surgery included an infection not responding to conservative treatment,with existing or impending spinal instability, and with or without neurological deficit. The surgical management involved transpedicular drainage of the abscess, wound debridement from the posterior approach and instrumented spondylodesis. Surgery which included spinal decompression with radical excision of infected tissue was augmented with posterolateral instrumented fusion and/or anterior stabilization, as indicated. RESULTS: Of the 112 patients treated, seven died of uncontrollable sepsis after surgery; the remaining 105 were followed up. Another four patients died of causes unrelated to the spinal problem treated within 12 months. All patients recovered except for two in whom the infection persisted, but 13 required more than one surgical procedure. One patient with CSF leakage failed to heal after five interventions. The most frequently isolated infectious agents were Staphylococus aureus, Staphylococus epidermidis and E. coli. Of the 33 patients with neurological deficit, 24 improved by one or two Frankel grades. The neurological status of six paraplegic patients did not improve, but their functional findings did after stabilization of the spine. Clinical evaluation showed 47 (44.7 %) very good, 40 (38 %) good, eight (7.6 %) unchanged and 10 (9.5 %) poor outcomes. CONCLUSION: Early diagnosis is a prerequisite for good treatment outcomes. Clinical examination, results of laboratory tests, and scintigraphy and MRI findings play the key role. When progressing osteolysis is suspected, a CT scan is necessary. Debridement should be as radical as possible, but always in compliance with the patient's health state. At an advanced stage of disease, spinal stabilization is important because it allows us to remove infected tissue. Intravenous and then oral antibiotic therapy at 2 to 4 and 6 to 12 weeks of follow-up is mandatory. The management of spinal infections is a complex process requiring good multidisciplinary cooperation.

• MeSH
• Bacterial Infections surgery   MeSH
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Adolescent MeSH
• Spinal Diseases surgery   MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Adolescent MeSH
• Male MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Female MeSH
• Publication type
• English Abstract MeSH
• Journal Article MeSH