Nanomembrane-based Apheresis as Safe and Effective Therapy for Cytomegalovirus and Epstein-Barr Virus Reactivation

BACKGROUND: Heavy training schedules or endurance competitions in marathon are forms of extreme physical stress and lead to immunodepression in runners which could be associated with increased susceptibility to viral reactivation by ubiquitous viral infection such as cytomegalovirus (CMV) and Epstein-Barr virus (EBV). Lately, it was confirmed presence of elevated CMV and EBV loads and the lower antibody titers in competitive athletes. The most common clinical features are fatigue and adynamia accompanied with liver damage, varying from mild and transient elevation of aminotransferases to serious acute hepatitis and liver failure. CASE REPORT: Bearing in mind that a professional practice of marathon running is hazardous for the liver, therapeutic action is necessary as soon as possible to avoid serious complications and even cessation of professional competition. In our case report of professional female marathon runner, we need to treat CMV and EBV reactivation which caused liver damage, prevented regular trainings, and upcoming competitions. We opted for four sessions of nanomembrane based apheresis performed every other day for removal pathological products resulting from virus reactivation to break through the course of the disease and to prevent complications. After completing the whole procedure control laboratory tests and abdominal ultrasound were in physiological ranges. CONCLUSION: Hence, nanomembrane based apheresis can be effective and safe treatment of liver damages for elite marathon runners as well as for athletes.

Time Course of Perceived Visual Distortion and Axial Length Growth in Myopic Children Undergoing Orthokeratology

Purpose: To establish the time course of the subjective visual function changes during the first month of orthokeratology treatment in myopic children, and to investigate how the time course variations are associated with the objective optical quality changes and the axial length growth (ALG) after 1 year of treatment.Methods: A total of 58 myopic children aged from 8 to 16 years participated in this self-controlled prospective study. All subjects were fitted with designed spherical four-zone orthokeratology lenses. Subjective visual function was evaluated with orientation discrimination threshold (ODT), and objective optical quality was quantified with the high-order aberration root-mean-square (HOA-RMS) and the changing speed of HOA. The measurements were done before the lens fitting and 1 day, 1-, 2-, and 4-weeks after lens wear. Axial length was obtained at baseline and 1-year follow-up, and ALG was defined as the difference. One-way ANOVA was conducted to compare the difference for statistical analysis.Results: After lens fitting, the ODT time courses peaked on day 1 in 28 children, 1 week in 15 children, 2 weeks in 11 children, and 4 weeks in 4 children. In contrast, the HOA-RMS steadily rose during the first month, and the changing speed of HOA was only transiently elevated on day 1 after the initial lens wear. The ALG was 0.12 ± 0.20 mm in subjects whose ODT peaked at day 1, 0.08 ± 0.09 mm in subjects whose ODT peaked on 1-week, and 0.12 ± 0.15 mm in subjects whose ODT peaked on 2-week or later. There was no difference in axial growth among the subjects whose ODT peaked at different days (P = 0.734).Conclusion: While half ODT time course resembled the changing speed of HOA with a transient elevation on day 1, about a quarter of the ODT time course resemble the steadily rising of HOA-RMS, and the rest was located in the middle. The ALGs in children with different types of ODT time courses were similar.

Thrombopoietin receptor agonists for the treatment of severe persistent and chronic immune trombocytopenia in children: clinical data of Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology, Immunology

Thrombopoietin receptor agonists (TPO-RA) – romiplostim and eltrombopag – changed considerably the therapeutic options for severe persistent and chronic immune thrombocytopenia (ITP). The article presents the results of a retrospective study of TPO-RA efficacy and safety in patients under 18 years of age. The study was approved by the Independent Ethics Committee and Scientific Council of the Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology. Sixty-eight children had a total of 89 courses of TPO-RA (44 romiplostim and 45 eltrombopag). Their median age was 6.5 years. The median ITP duration was 15.8 months. All patients received previous ITP therapy (1–6 lines). Before the initiation of TPO-RA, the majority of patients had thrombocytopenia with bleeding. In most cases, the platelet response was achieved within the first 2 months of treatment. The average effective doses of romiplostim and eltrombopag were 10 mg/kg per week and 75 mg per day, respectively. Half of patients in romiplostim group and 62% of patients in eltrombopag group did not require extra therapy. The majority of patients (75.6–81.8%) achieved an overall response, but only near 50% achieved a durable (more than 24 weeks) platelet response. Six patients sustained the response after TPO-RA discontinuation. The most common adverse events (AE) of TPO-RA therapy were transient elevation in hepatic enzymes in eltrombopag group (28.9%) and thrombocytosis (18.2–22.2%) in both groups. In 6 cases the therapy was discontinued due to AEs. Two AEs were serious. Our results demonstrate that TPO-RA could safely increase platelet counts and decrease the risk of spontaneous life-threatening bleeding in nearly half of children with severe persistent and chronic ITP. TPO-RA could help to avoid long-term immunosuppressive therapy and splenectomy or delay them and the ITP remission is possible in some cases.

Crosstalk Between Dysfunctional Mitochondria and Inflammation in Glaucomatous Neurodegeneration

Mitochondrial dysfunction and excessive inflammatory responses are both sufficient to induce pathology in age-dependent neurodegenerations. However, emerging evidence indicates crosstalk between damaged mitochondrial and inflammatory signaling can exacerbate issues in chronic neurodegenerations. This review discusses evidence for the interaction between mitochondrial damage and inflammation, with a focus on glaucomatous neurodegeneration, and proposes that positive feedback resulting from this crosstalk drives pathology. Mitochondrial dysfunction exacerbates inflammatory signaling in multiple ways. Damaged mitochondrial DNA is a damage-associated molecular pattern, which activates the NLRP3 inflammasome; priming and activation of the NLRP3 inflammasome, and the resulting liberation of IL-1β and IL-18 via the gasdermin D pore, is a major pathway to enhance inflammatory responses. The rise in reactive oxygen species induced by mitochondrial damage also activates inflammatory pathways, while blockage of Complex enzymes is sufficient to increase inflammatory signaling. Impaired mitophagy contributes to inflammation as the inability to turnover mitochondria in a timely manner increases levels of ROS and damaged mtDNA, with the latter likely to stimulate the cGAS-STING pathway to increase interferon signaling. Mitochondrial associated ER membrane contacts and the mitochondria-associated adaptor molecule MAVS can activate NLRP3 inflammasome signaling. In addition to dysfunctional mitochondria increasing inflammation, the corollary also occurs, with inflammation reducing mitochondrial function and ATP production; the resulting downward spiral accelerates degeneration. Evidence from several preclinical models including the DBA/2J mouse, microbead injection and transient elevation of IOP, in addition to patient data, implicates both mitochondrial damage and inflammation in glaucomatous neurodegeneration. The pressure-dependent hypoxia and the resulting metabolic vulnerability is associated with mitochondrial damage and IL-1β release. Links between mitochondrial dysfunction and inflammation can occur in retinal ganglion cells, microglia cells and astrocytes. In summary, crosstalk between damaged mitochondria and increased inflammatory signaling enhances pathology in glaucomatous neurodegeneration, with implications for other complex age-dependent neurodegenerations like Alzheimer’s and Parkinson’s disease.

A Longitudinal, Observational Analysis of Neuronal Injury Biomarkers in a Case Report of a Patient With Paraneoplastic Anti-CRMP5 Antibody-Associated Transverse Myelitis

Biomarkers are needed to guide therapeutic decision making in autoimmune and paraneoplastic neurologic disorders. Here, we describe a case of paraneoplastic collapsing response-mediator protein-5 (CRMP5)-associated transverse myelitis (TM) where plasma neurofilament light (NfL) chain and glial fibrillary protein (GFAP) levels were observed over a 14-month clinical course, correlating with radiographical and clinical outcome measures in response to treatment. Blood and CSF samples obtained at diagnosis as well as 7 and 14 months into treatment. At the time of initial diagnosis, both plasma NfL (782.62 pg/ml) and GFAP (283.26 pg/ml) were significantly elevated. Initial treatment was with IV steroids and plasma exchange (PLEX) followed by neuroendocrine tumor removal, chemotherapy, and radiation. After initial improvement with chemotherapy, the patient experienced clinical worsening and transient elevation of plasma NfL (103.27 pg/ml and GFAP (211.58 pg/ml) levels. Whole body positron emission tomography PET scan did not demonstrate recurrence of malignancy. Repeat PLEX and rituximab induction resulted in improvements in patient function, neurologic exam, and plasma biomarker levels. To our knowledge, this is the first described longitudinal, prospective analysis of neuronal injury biomarkers and association of clinical treatment outcomes in CRMP5 myelitis. Our findings suggest that clinical improvement correlates with NfL and GFAP concentrations.

Temporal changes in blood pressure following prehospital rapid sequence intubation

BackgroundRapid Sequence intubation (RSI) is an airway procedure that uses sedative and paralytic drugs to facilitate endotracheal intubation. It is known that RSI could impact blood pressure in the peri-intubation period. However, little is known about blood pressure changes in longer time frames. Therefore, this analysis aims to describe the changes in systolic blood pressure in a large cohort of paramedic-led RSI cases over the whole prehospital timespan.MethodsIntensive Care Paramedics in Victoria, Australia, are authorised to use RSI in medical or trauma patients with a Glasgow Coma Scale <10. This retrospective cohort study analysed data from patientcare records for patients aged 12 years and above that had received RSI, from 1 January 2008 to 31 December 2019. This study quantifies the systolic blood pressure changes using regression with fractional polynomial terms. The analysis is further stratified by high versus Low Shock Index (LSI). The shock index is calculated by dividing pulse rate by systolic blood pressure.ResultsDuring the study period RSI was used in 8613 patients. The median number of blood pressure measurements was 5 (IQR 3–8). Systolic blood pressure rose significantly by 3.4 mm Hg (p<0.001) and then returned to baseline in the first 5 min after intubation for LSI cases. No initial rise in blood pressure is apparent in High Shock Index (HSI) cases. Across the whole cohort, systolic blood pressure decreased by 7.1 mm Hg (95% CI 7.9 to 6.3 mm Hg; p<0.001) from the first to the last blood pressure measured.ConclusionsOur study shows that in RSI patients a small transient elevation in systolic blood pressure in the immediate postintubation period is found in LSI, but this elevation is not apparent in HSI. Blood pressure decreased over the prehospital phase in RSI patients with LSI, but increased for HSI cases.

Incorporation of unnatural amino acid for the tagging of cannabinoid receptors 1 and 2 reveals receptor roles in regulating cAMP levels

The role of CB1/CB2 co-expression in cell signaling remains elusive. We established a simplified mammalian cell model system in which expression of CB1 or CB2 can be easily monitored under a confocal microscope. For this, we applied amber codon suppression in live cells to incorporate a single trans-cyclooctene (TCO) bearing amino acid in one of the extracellular loops of CB1 or CB2, followed by fluorescent labeling via click chemistry. We employed genetically encoded biosensors to measure the roles of CB1 and/or CB2 in regulating intracellular calcium ([Ca2+]i) and cAMP ([cAMP]i) levels. We show that the agonist-mediated activation of tagged-CB1 or -CB2 can transiently elevate [Ca2+]i levels. However, when the two receptors were co-expressed in the same cell, CB2 no longer signaled through calcium although CB1-mediated transient elevation of [Ca2+]i levels was unaffected. Because of the existence of crosstalk between calcium and cAMP signaling, we measured the effects of CB1 and/or CB2 in regulating adenylate cyclase activity. We found that the expression of CB1 increased forskolin-induced [cAMP]i levels compared to non-transfected cells. Conversely, CB2 expression decreased stimulated [cAMP]i levels under the same conditions. Finally, co-expressed CB1 and CB2 receptors showed additive yet opposing effects on stimulated [cAMP]i levels. These observations suggest that co-expressed CB1/CB2 act locally as a pair in regulating cell excitability by modulating stimulated [cAMP]i levels.

An elevated serum creatinine in a patient receiving palbociclib

Background: Serum Creatinine (SCr) is the most widely used parameter in clinical practice to estimate glomerular filtration rate (GFR). Various drugs have been reported to cause a reversible and transient elevation in SCr without a true reduction in overall kidney function Case Presentation: We describe a case of a 66-year-old-woman with right breast poorly differentiated invasive ductal carcinoma, hormone receptor positive and HER2 negative who received treatment with fulvestrant and palbociclib. The initial dose of Palbociclib was 125 mg orally a day, then reduce to 100 mg orally a day. She presented for evaluation of elevated serum creatinine with decreased eGFR. Upon evaluation an estimated glomerular filtration rate by cystatin C showed kidney function at her baseline for the past year. Conclusion: Palbocicib is a selective inhibitor of the cyclin-dependent kinases CDK4 and CDK6. Physicians should be aware that patients undergoing therapy with palbociclib require monitoring of kidney function and an increase in serum creatinine from baseline, might represent an inhibitory effect of the secretion of creatinine.

Serial Lipocalin 2 and Oncostatin M levels reflect inflammation status and treatment response in axial spondyloarthritis

Background Informative serum biomarkers for monitoring inflammatory activity and treatment responses in axial spondyloarthritis (axSpA) are lacking. We assessed whether Lipocalin 2 (LCN2) and Oncostatin M (OSM), both having roles in inflammation and bone remodeling, may accurately reflect chronic joint inflammation and treatment response in axSpA. Previous reports in animal models showed involvement of LCN2 and OSM in joint/gut inflammation. We asked whether they also play a role in human axSpA. Methods We analyzed a longitudinal observational axSpA cohort (286 patients) with yearly clinical assessments and concurrent measurements of serum LCN2 and OSM (1204 serum samples) for a mean of 4 years. Biomarker levels were correlated with MRI scoring and treatment response. Results Persistent and transient elevation of LCN2 and OSM were observed in axSpA patients. Persistent elevation of LCN2 or OSM, but not CRP, correlated with sacroiliac joint (SIJ) MRI SPARCC scores (Pearson’s correlation p = 0.0005 and 0.005 for LCN2 and OSM respectively), suggesting that LCN2/OSM outperforms CRP as reflective of SIJ inflammation. We observed both concordant and discordant patterns of LCN2 and OSM in relationship to back pain, the cardinal clinical symptom in axSpA. Twenty-six percent (73/286) of the patients remained both clinically and serologically active (CASA). Sixty percent (173/286) of the patients became clinically quiescent, with back pain resolved, but 53% (92/173) of them were serologically active (CQSA), indicating that pain control may not indicate control of joint inflammation, as reflected by positive MRI imaging of SIJ. With respect to treatment responses, transient elevation of LCN2 or OSM over time was predictive of better response to all treatments. Conclusion In axSpA, persistent LCN2 and/or OSM elevation reflects chronic SIJ inflammation and suboptimal treatment response. In our cohort, half of the currently deemed clinically quiescent patients with back pain resolved continued to demonstrate chronic joint inflammation. LCN2 and OSM profiling outperforms CRP as a predictive measure and provides an objective assessment of chronic local inflammation in axSpA patients.

3D Mapping Reveals a Complex and Transient Interstitial Matrix During Murine Kidney Development

BackgroundThe extracellular matrix (ECM) is a network of proteins and glycosaminoglycans that provides structural and biochemical cues to cells. In the kidney, the ECM is critical for nephrogenesis; however, the dynamics of ECM composition and how it relates to 3D structure during development is unknown.MethodsUsing embryonic day 14.5 (E14.5), E18.5, postnatal day 3 (P3), and adult kidneys, we fractionated proteins based on differential solubilities, performed liquid chromatography–tandem mass spectrometry, and identified changes in ECM protein content (matrisome). Decellularized kidneys were stained for ECM proteins and imaged in 3D using confocal microscopy.ResultsWe observed an increase in interstitial ECM that connects the stromal mesenchyme to the basement membrane (TNXB, COL6A1, COL6A2, COL6A3) between the embryo and adult, and a transient elevation of interstitial matrix proteins (COL5A2, COL12A1, COL26A1, ELN, EMID1, FBN1, LTBP4, THSD4) at perinatal time points. Basement membrane proteins critical for metanephric induction (FRAS1, FREM2) were highest in abundance in the embryo, whereas proteins necessary for integrity of the glomerular basement membrane (COL4A3, COL4A4, COL4A5, LAMB2) were more abundant in the adult. 3D visualization revealed a complex interstitial matrix that dramatically changed over development, including the perinatal formation of fibrillar structures that appear to support the medullary rays.ConclusionBy correlating 3D ECM spatiotemporal organization with global protein abundance, we revealed novel changes in the interstitial matrix during kidney development. This new information regarding the ECM in developing kidneys offers the potential to inform the design of regenerative scaffolds that can guide nephrogenesis in vitro.