Seroprevalence of Toxoplasma gondii among HIV Positive Patients under Surveillance in Greek Infectious Disease Units: A Screening Study with Comparative Evaluation of Serological Methods.

This study aims to screen for IgG antibodies against Toxoplasma gondii (T. gondii) in the sera of 155 newly diagnosed Human Immunodeficiency Virus (HIV) positive patients under surveillance in Greek Infectious Disease Units. Additionally, risk factors based on patient demographics were examined, and a comparative evaluation of commercially available serological methods was conducted. Three methods were employed to detect IgG antibodies against T. gondii: Enzyme-Linked Immunosorbent Assay (ELISA), Indirect Immunofluorescence Antibody Test (IFAT), and Western Blot (WB), which was used as a reference here. Forty-nine sera samples were true-positive for IgG antibodies against T. gondii, resulting in a 31.61% positivity rate, and the immunoassay test statistical reliability analysis resulted in higher IFAT accuracy (90.97%) compared to ELISA (76.26%). Furthermore, statistical analysis of demographic and immunological data included in the study placed female and foreign/non-Greek individuals at 2.24 (p = 0.0009) and 2.34 (p = 0.0006) times higher risk of positive T. gondii IgG testing compared to their male and Greek counterparts, respectively. Our findings on positivity rates and comparative serology underscore the importance of early and suitable screening measures for newly diagnosed HIV+ patients to mitigate the life-threatening outcomes that may arise from a potential subsequent T. gondii activation.

Transitions of blood immune endotypes and improved outcome by anakinra in COVID-19 pneumonia: an analysis of the SAVE-MORE randomized controlled trial.

BACKGROUND: Endotype classification may guide immunomodulatory management of patients with bacterial and viral sepsis. We aimed to identify immune endotypes and transitions associated with response to anakinra (human interleukin 1 receptor antagonist) in participants in the SAVE-MORE trial. METHODS: Adult patients hospitalized with radiological findings of PCR-confirmed severe pneumonia caused by SARS-CoV-2 and plasma-soluble urokinase plasminogen activator receptor levels of ≥ 6 ng/ml in the SAVE-MORE trial (NCT04680949) were characterized at baseline and days 4 and 7 of treatment using a previously defined 33-messenger RNA classifier to assign an immunological endotype in blood. Endpoints were changes in endotypes and progression to severe respiratory failure (SRF) associated with anakinra treatment. RESULTS: At baseline, 23.2% of 393 patients were designated as inflammopathic, 41.1% as adaptive, and 35.7% as coagulopathic. Only 23.9% were designated as the same endotype at days 4 and 7 compared to baseline, while all other patients transitioned between endotypes. Anakinra-treated patients were more likely to remain in the adaptive endotype during 7-day treatment (24.4% vs. 9.9%; p < 0.001). Anakinra also protected patients with coagulopathic endotype at day 7 against SRF compared to placebo (27.8% vs. 55.9%; p = 0.013). CONCLUSION: We identify an association between endotypes defined using blood transcriptome and anakinra therapy for COVID-19 pneumonia, with anakinra-treated patients shifting toward endotypes associated with a better outcome, mainly the adaptive endotype. Trial registration ClinicalTrials.gov, NCT04680949, December 23, 2020.

Efficacy and safety of early soluble urokinase plasminogen receptor plasma-guided anakinra treatment of COVID-19 pneumonia: A subgroup analysis of the SAVE-MORE randomised trial.

Background: The SAVE-MORE trial demonstrated that anakinra treatment in COVID-19 pneumonia with plasma soluble urokinase plasminogen activator (suPAR) levels of 6 ng/mL or more was associated with 0.36 odds for a worse outcome compared to placebo when expressed by the WHO-Clinical Progression Scale (CPS) at day 28. Herein, we report the results of subgroup analyses and long-term outcomes. Methods: This prospective, double-blind, randomised clinical trial, recruited patients with a confirmed SARS-CoV-2 infection, in need of hospitalisation, lower respiratory tract infection and plasma suPAR ≥6 ng/mL from 37 academic and community hospitals in Greece and Italy. Patients were 1:2 randomised to subcutaneous treatment with placebo or anakinra (100 mg) once daily for 10 days. Pre-defined subgroups of Charlson's comorbidity index (CCI), sex, age, level of suPAR, and time from symptom onset were analysed for the primary endpoint (overall comparison of distribution of frequencies of the scores from the WHO-CPS between treatments on day 28), by multivariable ordinal regression analysis in the intention to treat (ITT) population. This trial is registered with the EU Clinical Trials Register (2020-005828-11) and ClinicalTrials.gov (NCT04680949). Findings: Patients were enrolled between 23 December 2020 and 31 March 2021; 189 patients in the placebo arm and 405 patients in the anakinra arm were the ITT population. Multivariable analysis showed that anakinra treatment was accompanied by significantly lower odds for worse outcome compared to placebo at day 28 for all studied subgroups (CCI ≥ 2, OR: 0.34, 95% confidence intervals [CI] 0.22-0.50; CCI < 2, OR: 0.38, 95% CI 0.21-0.68; suPAR > 9 ng/mL, OR: 0.35, 95% CI 0.19-0.66; suPAR 6-9 ng/mL, OR: 0.35, 95% CI 0.24-0.52; patients ≥65 years, OR: 0.41, 95% CI 0.25-0.66; and patients <65 years, OR: 0.29, 95% CI 0.19-0.45). The benefit was uniform, irrespective of the time from start of symptoms until the start of the study drug. At days 60 and 90, anakinra treatment had odds of 0.40 (95% CI 0.28-0.57) and 0.46 (95% CI 0.32-0.67) respectively, for a worse outcome compared to placebo. The costs of general ward stay, ICU stay, and drugs were lower with anakinra treatment. Interpretation: Anakinra represents an important therapeutic tool in the management of COVID-19 that may be administered in all subgroups of patients; benefits are maintained until day 90. Funding: Hellenic Institute for the Study of Sepsis; Swedish Orphan Biovitrum AB.

Interferon gamma-induced protein 10 (IP-10) for the early prognosis of the risk for severe respiratory failure and death in COVID-19 pneumonia.

OBJECTIVES: Elevated concentrations of soluble urokinase plasminogen activator receptor (suPAR) predict progression to severe respiratory failure (SRF) or death among patients with COVID-19 pneumonia and guide early anakinra treatment. As suPAR testing may not be routinely available in every health-care setting, alternative biomarkers are needed. We investigated the performance of C-reactive protein (CRP), interferon gamma-induced protein-10 (IP-10) and TNF-related apoptosis-inducing ligand (TRAIL) for predicting SRF or death in COVID-19. METHODS: Two cohorts were studied; one discovery cohort with 534 patients from the SAVE-MORE clinical trial; and one validation cohort with 364 patients from the SAVE trial including also 145 comparators. CRP, IP-10 and TRAIL were measured by the MeMed Key® platform in order to select the biomarker with the best prognostic performance for the early prediction of progression into SRF or death. RESULTS: IP-10 had the best prognostic performance: baseline concentrations 2000 pg/ml or higher predicted equally well to suPAR (sensitivity 85.0 %; negative predictive value 96.6 %). Odds ratio for poor outcome among anakinra-treated participants of the SAVE-MORE trial was 0.35 compared to placebo when IP-10 was 2,000 pg/ml or more. IP-10 could divide different strata of severity for SRF/death by day 14 in the validation cohort. Anakinra treatment decreased this risk irrespective the IP-10 concentrations. CONCLUSIONS: IP-10 concentrations of 2,000 pg/ml or higher are a valid alternative to suPAR for the early prediction of progression into SRF or death the first 14 days from hospital admission for COVID-19 and they may guide anakinra treatment. CLINICALTRIALS: gov, NCT04680949 and NCT04357366.

Development and validation of SCOPE score: A clinical score to predict COVID-19 pneumonia progression to severe respiratory failure.

Most patients infected with SARS-CoV-2 (COVID-19) experience mild, non-specific symptoms, but many develop severe symptoms associated with an excessive inflammatory response. Elevated plasma concentrations of soluble urokinase plasminogen activator receptor (suPAR) provide early warning of progression to severe respiratory failure (SRF) or death, but access to suPAR testing may be limited. The Severe COvid Prediction Estimate (SCOPE) score, derived from circulating concentrations of C-reactive protein, D- dimers, interleukin-6, and ferritin among patients not receiving non-invasive or invasive mechanical ventilation during the SAVE-MORE study, offers predictive accuracy for progression to SRF or death within 14 days comparable to that of a suPAR concentration of ≥6 ng/mL (area under receiver operator characteristic curve 0.81 for both). The SCOPE score is validated in two similar independent cohorts. A SCOPE score of 6 or more is an alternative to suPAR for predicting progression to SRF or death within 14 days of hospital admission for pneumonia, and it can be used to guide treatment decisions.

Early treatment of COVID-19 with anakinra guided by soluble urokinase plasminogen receptor plasma levels: a double-blind, randomized controlled phase 3 trial.

Early increase of soluble urokinase plasminogen activator receptor (suPAR) serum levels is indicative of increased risk of progression of coronavirus disease 2019 (COVID-19) to respiratory failure. The SAVE-MORE double-blind, randomized controlled trial evaluated the efficacy and safety of anakinra, an IL-1α/ß inhibitor, in 594 patients with COVID-19 at risk of progressing to respiratory failure as identified by plasma suPAR ≥6 ng ml-1, 85.9% (n = 510) of whom were receiving dexamethasone. At day 28, the adjusted proportional odds of having a worse clinical status (assessed by the 11-point World Health Organization Clinical Progression Scale (WHO-CPS)) with anakinra, as compared to placebo, was 0.36 (95% confidence interval 0.26-0.50). The median WHO-CPS decrease on day 28 from baseline in the placebo and anakinra groups was 3 and 4 points, respectively (odds ratio (OR) = 0.40, P < 0.0001); the respective median decrease of Sequential Organ Failure Assessment (SOFA) score on day 7 from baseline was 0 and 1 points (OR = 0.63, P = 0.004). Twenty-eight-day mortality decreased (hazard ratio = 0.45, P = 0.045), and hospital stay was shorter.

Molecular Epidemiology of SARS-CoV-2 in Greece Reveals Low Rates of Onward Virus Transmission after Lifting of Travel Restrictions Based on Risk Assessment during Summer 2020.

The novel coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spread rapidly during the first months of 2020 and continues to expand in multiple areas across the globe. Molecular epidemiology has provided an added value to traditional public health tools by identifying SARS-CoV-2 clusters or providing evidence that clusters based on virus sequences and contact tracing are highly concordant. Our aim was to infer the levels of virus importation and to estimate the impact of public health measures related to travel restrictions to local transmission in Greece. Our phylogenetic and phylogeographic analyses included 389 full-genome SARS-CoV-2 sequences collected during the first 7 months of the pandemic in Greece and a random collection in five replicates of 3,000 sequences sampled globally, as well as the best hits to our data set identified by BLAST. Phylogenetic trees were reconstructed by the maximum likelihood method, and the putative source of SARS-CoV-2 infections was inferred by phylogeographic analysis. Phylogenetic analyses revealed the presence of 89 genetically distinct viruses identified as independent introductions into Greece. The proportion of imported strains was 41%, 11.5%, and 8.8% during the three periods of sampling, namely, March (no travel restrictions), April to June (strict travel restrictions), and July to September (lifting of travel restrictions based on thorough risk assessment), respectively. The results of phylogeographic analysis were confirmed by a Bayesian approach. Our findings reveal low levels of onward transmission from imported cases during summer and underscore the importance of targeted public health measures that can increase the safety of international travel during a pandemic. IMPORTANCE Our study based on current state-of-the-art molecular epidemiology methods suggests that virus screening and public health measures after the lifting of travel restrictions prevented SARS-CoV-2 onward transmission from imported cases during summer 2020 in Greece. These findings provide important data on the efficacy of targeted public health measures and have important implications regarding the safety of international travel during a pandemic. Our results can provide a roadmap about prevention policy in the future regarding the reopening of borders in the presence of differences in vaccination coverage, the circulation of the virus, and the presence of newly emergent variants across the globe.

Development of a novel risk score for the prediction of critical illness amongst COVID-19 patients.

OBJECTIVES: Coronavirus disease-19 (COVID-19) is associated with various clinical manifestations, ranging from asymptomatic infection to critical illness. The aim of this study is to evaluate the clinical and laboratory characteristics of hospitalised COVID-19 patients and construct a predictive model for the discrimination of patients at risk of disease progression. METHODS: A single-centre cohort study was conducted including consecutively patients with COVID-19. Demographic, clinical and laboratory findings were prospectively collected at admission. The primary outcome of interest was the intensive care unit admission. A risk model was constructed by applying a Cox's proportional hazard's model with elastic net penalty. Its diagnostic performance was assessed by receiver operating characteristic analysis and was compared with conventional pneumonia severity scores. RESULTS: From a total of 67 patients 15 progressed to critical illness. The risk score included patients' gender, presence of hypertension and diabetes mellitus, fever, shortness of breath, serum glucose, aspartate aminotransferase, lactate dehydrogenase, C-reactive protein and fibrinogen. Its predictive accuracy was estimated to be high (area under the curve: 97.1%), performing better than CURB-65, CRB-65 and PSI/PORT scores. Its sensitivity and specificity were estimated to be 92.3% and 93.3%, respectively, at the optimal threshold of 1.6. CONCLUSIONS: A10-variable risk score was constructed based on clinical and laboratory characteristics in order to predict critical illness amongst hospitalised COVID-19 patients, achieving better discrimination compared with traditional pneumonia severity scores. The proposed risk model should be externally validated in independent cohorts in order to ensure its prognostic efficacy.

SARS-CoV-2 Molecular Transmission Clusters and Containment Measures in Ten European Regions during the First Pandemic Wave.

BACKGROUND: The spatiotemporal profiling of molecular transmission clusters (MTCs) using viral genomic data can effectively identify transmission networks in order to inform public health actions targeting SARS-CoV-2 spread. METHODS: We used whole genome SARS-CoV-2 sequences derived from ten European regions belonging to eight countries to perform phylogenetic and phylodynamic analysis. We developed dedicated bioinformatics pipelines to identify regional MTCs and to assess demographic factors potentially associated with their formation. RESULTS: The total number and the scale of MTCs varied from small household clusters identified in all regions, to a super-spreading event found in Uusimaa-FI. Specific age groups were more likely to belong to MTCs in different regions. The clustered sequences referring to the age groups 50-100 years old (y.o.) were increased in all regions two weeks after the establishment of the lockdown, while those referring to the age group 0-19 y.o. decreased only in those regions where schools' closure was combined with a lockdown. CONCLUSIONS: The spatiotemporal profiling of the SARS-CoV-2 MTCs can be a useful tool to monitor the effectiveness of the interventions and to reveal cryptic transmissions that have not been identified through contact tracing.

Chest CT severity score and radiological patterns as predictors of disease severity, ICU admission, and viral positivity in COVID-19 patients.

BACKGROUND: Chest computed tomography (CT) is a useful tool for the diagnosis of coronavirus disease-2019 (COVID-19), although its exact value for predicting critical illness remains unclear. This study evaluated the efficacy of chest CT to predict disease progression, pulmonary complications, and viral positivity duration. METHODS: A single-center cohort study was conducted by consecutively including hospitalized patients with confirmed COVID-19. The chest CT patterns were described and a total severity score was calculated. The predictive accuracy of the severity score was evaluated using the receiver operating characteristic analysis, while a Cox proportional hazards regression model was implemented to identify the radiological features that are linked to prolonged duration of viral positivity. RESULTS: Overall, 42 patients were included with 10 of them requiring intensive care unit admission. The most common lesions were ground glass opacities (92.9%), consolidation (66.7%), and crazy-paving patterns (61.9%). The total severity score significantly correlated with inflammatory and respiratory distress markers, as well as with admission CURB-65 and PSI/PORT scores. It was estimated to predict critical illness with a sensitivity and specificity of 75% and 70%, respectively. Time-to-event analysis indicated that patients without ground-glass opacities presented significantly shorter median viral positivity (16 vs. 27 days). CONCLUSIONS: Chest CT severity score positively correlates with markers of COVID-19 severity and presents promising efficacy in predicting critical illness. It is suggested that ground-glass opacities are linked to prolonged viral positivity. Further studies should confirm the efficacy of the severity score and elucidate the long-term pulmonary effects of COVID-19.

A Patient with Cryoglobulinemic Membranoproliferative GN (MPGN) Who Survived COVID-19 Disease: Case Presentation and Current Data of COVID-19 Infection in Dialysis and Transplanted Patients in Greece.

The evolving pandemic of Coronavirus Disease 2019 has posed a substantial health risk worldwide. However, there is a paucity of data regarding the clinical course and the therapeutic management of patients with chronic kidney disease and COVID-19 infection. To date, most evidence has come from renal transplantation, with about 45 patients reported thus far, and the current data from the ERA-EDTA (ERACODA) registry for transplanted patients and patients on Renal Replacement Therapy (RRT); as for those with glomerular diseases, data are lacking. Herein, we report the case of a 62-year-old patient with severe membranoproliferative glomerulonephritis who had been receiving a high burden of immunosuppression until four months before the COVID-19 infection. He developed severe disease with acute respiratory failure requiring mechanical ventilation. After treatment with hydroxychloroquine and azithromycin, despite his low chances, he gradually recovered and survived. To the best of our knowledge, this is one of the few reported patients with glomerulonephritis who had COVID-19 Besides our single case with glomerulonephritis early during the disease outbreak, the very low prevalence of COVID-19 infection in the country's transplant recipients (0.038%) and dialysis patients (0.24%) reflects the impact of the rapid implementation of social distancing rules as well as of preventive measures for disease control in the hospitals and dialysis units in our country.

The aesthetic nature of the birthing room environment may alter the need for obstetrical interventions - an observational retrospective cohort study.

The concept of sensory delivery rooms was introduced in 2013. These rooms offer programmable calming lights, restful blurred pictures displayed on a wall-sized big screen, and sound effects. The primary aim of this observational study was to analyse the risk of obstetrical interventions among women giving birth for the first-time in a sensory delivery room vs. a standard delivery room. We included nulliparous, term pregnant women having a single baby with a cephalic presentation who were in spontaneous labour and gave birth between March 1st 2014 and July 1st 2015 in North Zealand Hospital, Hillerød. A total of 789 women were included in the study, 313 gave birth in a sensory room and 476 in a standard delivery room. The risk of a caesarean delivery was significantly decreased when giving birth in a sensory room compared with a standard delivery room (OR, multiple adjusted: 0.44; 95% CI 0.22-0.87); furthermore, the use of oxytocin infusion was also reduced (OR, multiple adjusted: 0.71; 95% CI 0.50-1.03). This observational cohort study suggests that giving birth in a sensory delivery room could lower the risk of caesarean delivery, potentially reducing the number of such deliveries by one for every 23 patients.

UV light assisted antibiotics for eradication of in vitro biofilms.

The overuse of antibiotics is accelerating the bacterial resistance, and therefore there is a need to reduce the amount of antibiotics used for treatment. Here, we demonstrate in vitro that specific wavelengths in a narrow range around 296 nm are able to eradicate bacteria in the biofilm state (grown for 24 hours) more effectively, than antibiotics and the combination of irradiation and antibiotics is even better, introducing a novel concept light assisted antibiotics. The investigated wavelength range was 249 nm to 338 nm with an approximate step of 5 nm. The novel concept that consists of a UV irradiation treatment followed by a tobramycin treatment can significantly reduce the amount of antibiotics needed for eradicating mature bacterial biofilms. The efficiency of the proposed light assisted antibiotics method was compared to combinatory antibiotic treatment and highly concentrated antibiotic monotherapy. The eradication efficacies, on mature biofilms, achieved by light assisted antibiotic and by the antibiotic monotherapy at approximately 10-fold higher concentration, were equivalent. The present achievement could motivate the development of light assisted antibiotic treatments for treating infections.

Inactivation of Pseudomonas aeruginosa biofilm after ultraviolet light-emitting diode treatment: a comparative study between ultraviolet C and ultraviolet B.

The objective of this study was to test the inactivation efficiency of two different light-based treatments, namely ultraviolet B (UVB) and ultraviolet C (UVC) irradiation, on Pseudomonas aeruginosa biofilms at different growth stages (24, 48, and 72 h grown). In our experiments, a type of AlGaN light-emitting diodes (LEDs) was used to deliver UV irradiation on the biofilms. The effectiveness of the UVB at 296 nm and UVC at 266 nm irradiations was quantified by counting colony-forming units. The survival of less mature biofilms (24 h grown) was studied as a function of UV-radiant exposure. All treatments were performed on three different biological replicates to test reproducibility. It was shown that UVB irradiation was significantly more effective than UVC irradiation in inactivating P. aeruginosa biofilms. UVC irradiation induced insignificant inactivation on mature biofilms. The fact that the UVB at 296 nm exists in daylight and has such disinfection ability on biofilms provides perspectives for the treatment of infectious diseases.

Investigation of the effect of UV-LED exposure conditions on the production of vitamin D in pig skin.

The dietary intake of vitamin D is currently below the recommended intake of 10-20µg vitamin D/day. Foods with increased content of vitamin D or new products with enhanced vitamin D are warranted. Light-emitting diodes (LEDs) are a potential new resource in food production lines. In the present study the exposure conditions with ultraviolet (UV) LEDs were systematically investigated in the wavelength range 280-340nm for achieving optimal vitamin D bio-fortification in pig skin. A wavelength of 296nm was found to be optimal for vitamin D3 production. The maximum dose of 20kJ/m(2) produced 3.5-4µg vitamin D3/cm(2) pig skin. Vitamin D3 produced was independent on the combination of time and intensity of the LED source. The increased UV exposure by UV-LEDs may be readily implemented in existing food production facilities, without major modifications to the process or processing equipment, for bio-fortifying food products containing pork skin.

Does correlated color temperature affect the ability of humans to identify veins?

In the present study we provide empirical evidence and demonstrate statistically that white illumination settings can affect the human ability to identify veins in the inner hand vasculature. A special light-emitting diode lamp with high color rendering index (CRI 84-95) was developed and the effect of correlated color temperature was evaluated, in the range between 2600 and 5700 K at an illuminance of 40±9 lx on the ability of adult humans to identify veins. It is shown that the ability to identify veins can, on average, be increased up to 24% when white illumination settings that do not resemble incandescent light are applied. The illuminance reported together with the effect of white illumination settings on direct visual perception of biosamples are relevant for clinical investigations during the night.

Can sleep quality and wellbeing be improved by changing the indoor lighting in the homes of healthy, elderly citizens?

The study investigated the effect of bright blue-enriched versus blue-suppressed indoor light on sleep and wellbeing of healthy participants over 65 years. Twenty-nine participants in 20 private houses in a uniform settlement in Copenhagen were exposed to two light epochs of 3 weeks with blue-enriched (280 lux) and 3 weeks blue-suppressed (240 lux) indoor light or vice versa from 8 to 13 pm in a randomized cross-over design. The first light epoch was in October, the second in November and the two light epochs were separated by one week. Participants were examined at baseline and at the end of each light epoch. The experimental indoor light was well tolerated by the majority of the participants. Sleep duration was 7.44 (95% CI 7.14-7.74) hours during blue-enriched conditions and 7.31 (95% CI 7.01-7.62) hours during blue-suppressed conditions (p = 0.289). Neither rest hours, chromatic pupillometry, nor saliva melatonin profile showed significant changes between blue-enriched and blue-suppressed epochs. Baseline Pittsburgh Sleep Quality Index (PSQI) was significantly worse in females; 7.62 (95% CI 5.13-10.0) versus 4.06 (95% CI 2.64-5.49) in males, p = 0.009. For females, PSQI improved significantly during blue-enriched light exposure (p = 0.007); no significant changes were found for males. The subjective grading of indoor light quality doubled from participants habitual indoor light to the bright experimental light, while it was stable between light epochs, although there were clear differences between blue-enriched and blue-suppressed electrical light conditions imposed. Even though the study was carried out in the late autumn at northern latitude, the only significant difference in Actiwatch-measured total blue light exposure was from 8 to 9 am, because contributions from blue-enriched, bright indoor light were superseded by contributions from daylight.