Use of exogenic phosphocreatine in ICU rehabilitation of patients with COVID-19 (pilot study)

The objective: to establish the possible effectiveness of exogenous phosphocreatine as a component of pharmacological support during the resuscitation stage of rehabilitation measures in critically ill patients with COVID-19.Subjects and Methods. Within a randomized study, 21 patients diagnosed with COVID-19 were divided into two groups: Group 1 (patients received the infusion of exogenous phosphocreatine as part of intensive care) and Group 2 (patients received standard intensive care) against the background of rehabilitation measures. Patients were assessed for muscle strength using the MRC scale, exercise tolerance by Borg rating, oxygenation parameters, routine clinical laboratory blood tests, dependence on respiratory support, outcome on day 10 of therapy, and hospital outcome.Results. The effectiveness of the use of the exogenous phosphocreatine as a component of pharmacological support during the resuscitation stage of rehabilitation measures in critical patients has been confirmed by positive dynamics: an increase in muscle strength (the MRC score in the group receiving exogenous phosphocreatine on day 10 was 0.5 points higher) and an increase in exercise tolerance (Borg rating in the group receiving exogenous phosphocreatine on day 10 was 1.5 points higher), significant increase in oxygenation based on arterial blood saturation data, and significant increase in lymphocyte count by 25% in the group receiving exogenous phosphocreatine.Conclusion. Exogenous phosphocreatine is a candidate drug for pharmacological support during resuscitation stage of rehabilitation of critical patients with COVID-19.

Empyema caused by Streptococcus constellatus: a case report and literature review

Background Streptococcus constellatus is a member of Streptococcus anginosus group (SAG) that tends to cause pyogenic infections in various sites. However, Streptococcus constellatus is easily ignored by routine clinical laboratory tests for its prolonged anaerobic culture environment. Case presentation A 71-year-old man was admitted to our hospital due to productive cough, fever, chest pain and shortness of breath for 3 weeks. Chest computed tomography showed patchy opacities and right-sided pleural effusion, so a chest tube was inserted and purulent and hemorrhagic fluid was aspirated. The routine etiological examinations of the pleural effusion were all negative, and next-generation sequencing (NGS) detected Streptococcus constellatus. Intravenous piperacillin-tazobactam 4.5 g every 8 h was used accordingly. The patient recovered and subsequent chest computed tomography confirmed the improvement. Conclusions We reported a case of empyema secondary to Streptococcus constellatus infection, which was identified by NGS, instead of bacterial culture. This case highlights the utility of NGS in detecting pathogens negative in traditional bacterial tests.

Development of a rapid SARS-CoV-2 neutralisation test by detecting antibodies that block interaction of spike protein receptor-binding domain (RBD) with angiotensin-converting enzyme 2 (ACE2)

The urgent need for a rapid and reliable Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) neutralising antibody detection test compatible with routine clinical laboratory testing currently exists. This is necessary to provide accurate estimates of immunity and to monitor vaccine effectiveness. Utilising Biochip Array Technology (BAT), the Randox SARS-CoV-2 Biochip proxy virus neutralisation test (pVNT) has been developed. Immobilising SARS-CoV-2 Spike RBD on the Biochip surface, innovative assay design enables direct sample addition to the Biochip well without the need for off-board sample pre-incubation step. Results are reported within 1.5 hours and testing is carried out without the prerequisite of live virus or biosafety level 3 (BSL3) laboratory facilities. In this study, assay validation is performed using recombinant antibodies and clinical samples and an excellent correlation against conventional virus neutralisation methods is established (100% clinical specificity and 98% clinical sensitivity). Serial dilution of samples with high neutralising antibody levels demonstrate end-point sample dilutions comparable with those obtained using the SARS-CoV-2 microneutralisation test. Species independent neutralising antibody detection capacity of the SARS-CoV-2 Biochip pVNT is also demonstrated. The findings of this study exemplifying the utility of the SARS-CoV-2 Biochip pVNT as a robust and reliable method for the accurate measurement of neutralising antibodies against SARS-CoV-2 and the availability of this test can now positively impact current testing deficiencies in this area.

Metabolomics strategy for diagnosing urinary tract infections

Metabolomics has emerged as a mainstream approach for investigating complex metabolic phenotypes. Despite its obvious suitability to diagnostics, it has yet to be integrated into routine clinical laboratory applications. Metabolomics-based diagnosis of infectious diseases is a logical application of this technology since microbial metabolic waste products are concentrated at the site of infection. In the case of urinary tract infections (UTIs), one of the most common bacterial infections, microbial catabolites are trapped in the bladder and thus could enable rapid diagnosis of UTIs. We conducted an untargeted metabolomics screen of 110 clinical specimens and identified two metabolites, agmatine and N6-methyladenine, that are predictive of a wide transect of pathogen species that collectively account for over 90% of UTI infections. We showed that these metabolites were consistently observed in multiple independent cohorts, including a blinded trial of over 587 clinical specimens (95% sensitivity, 86% specificity; PPV, 0.68; NPV, 0.98). These findings demonstrate the potential utility of metabolomics for diagnosing infectious diseases. Moreover, the rapid analysis times enabled through our metabolomics diagnostic approach - six minutes per sample - could curtail the inappropriate UTI clinical prescribing practices that are currently contributing to the selection of antimicrobial resistance.

Assessing the Need for Multiplex and Multifunctional Tick-Borne Disease Test in Routine Clinical Laboratory Samples from Lyme Disease and Febrile Patients with a History of a Tick Bite

Human polymicrobial infections in tick-borne disease (TBD) patients is an emerging public health theme. However, the requirement for holistic TBD tests in routine clinical laboratories is ambiguous. TICKPLEX® PLUS is a holistic TBD test utilized herein to assess the need for multiplex and multifunctional diagnostic tools in a routine clinical laboratory. The study involved 150 specimens categorized into Lyme disease (LD)-positive (n = 48), LD-negative (n = 30), and febrile patients from whom borrelia serology was requested (n = 72, later “febrile patients”) based on reference test results from United Medix, Finland. Reference tests from DiaSorin, Immunetics, and Mikrogen Diagnostik followed the two-tier LD testing system. A comparison between the reference tests and TICKPLEX® PLUS produced 86%, 88%, and 87% positive, negative, and overall agreement, respectively. Additionally, up to 15% of LD and 11% of febrile patients responded to TBD related coinfections and opportunistic microbes. The results demonstrated that one (TICKPLEX® PLUS) test can aid in a LD diagnosis instead of four tests. Moreover, TBD is not limited to just LD, as the specimens produced immune responses to several TBD microbes. Lastly, the study indicated that the screening of febrile patients for TBDs could be a missed opportunity at reducing unreported patient cases.

A Multicenter Study to Evaluate Harmonization of Assays for C-Terminal Telopeptides of Type I Collagen (ß-CTX): A Report from the IFCC-IOF Committee for Bone Metabolism (C-BM)

Background Biochemical bone turnover markers are useful tools to assess bone remodeling. C-terminal telopeptide of type I collagen (ß-CTX) has been recommended as a reference marker for bone resorption in research studies. Methods We describe the results of a multicenter study for routine clinical laboratory assays for ß-CTX in serum and plasma. Four centers (Athens GR, Copenhagen DK, Liege BE and Sheffield UK) collected serum and plasma (EDTA) samples from 796 patients presenting to osteoporosis clinics. Specimens were analyzed in duplicate with each of the available routine clinical laboratory methods according to the manufacturers’ instructions. Passing-Bablok regressions, Bland–Altman plots, V-shape evaluation method, and Concordance correlation coefficient for ß-CTX values between serum and plasma specimens and between methods were used to determine the agreement between results. A generalized linear model was employed to identify possible variables that affected the relationship between the methods. Two pools of serum were finally prepared and sent to the four centers to be measured in 5-plicates on 5 consecutive days with the different methods. Results We identified significant variations between methods and between centers although comparison results were generally more consistent in plasma compared to serum. We developed univariate linear regression equations to predict Roche Elecsys®, IDS-iSYS, or IDS ELISA ß-CTX results from any other assay and a multivariable model including the site of analysis, the age, and weight of the patient. The coefficients of determination (R2) increased from approximately 0.80 in the univariate model to approximately 0.90 in the multivariable one, with the site of analysis being the major contributing factor. Results observed on the pools also suggest that long-term storage could explain the difference observed with the different methods on serum. Conclusion Our results show large within- and between-assay variation for ß-CTX measurement, particularly in serum. Stability of the analyte could be one of the explanations. More studies should be undertaken to overcome this problem. Until harmonization is achieved, we recommend measuring ß-CTX by the same assay on EDTA plasma, especially for research purposes in large pharmacological trials where samples can be stored for long periods before they are assayed.

Hematologic indices in individuals with pathogenic germline DICER1 variants

Pathogenic germline variants in DICER1 underlie an autosomal dominant, pleiotropic tumor-predisposition disorder. Murine models with the loss of DICER1 in hematopoietic stem cell progenitors demonstrate hematologic aberrations that include reductions in red and white blood cell counts, hemoglobin volume, and impaired maturation resulting in dysplasia. We investigated whether hematologic abnormalities such as those observed in DICER1-deficient mice were observed in humans with a pathogenic germline variant in DICER1. A natural history study of individuals with germline pathogenic DICER1 variants and family controls conducted through the National Cancer Institute (NCI) evaluated enrollees at the National Institutes of Health Clinical Center during a comprehensive clinical outpatient visit that included collecting routine clinical laboratory studies. These were compared against normative laboratory values and compared between the DICER1 carriers and controls. There were no statistical differences in routine clinical hematology laboratory studies observed in DICER1 carriers and family controls. A review of the medical history of DICER1 carriers showed that none of the individuals in the NCI cohort developed myelodysplastic syndrome or leukemia. Query of the International Pleuropulmonary Blastoma/DICER1 Registry revealed 1 DICER1 carrier who developed a secondary leukemia after treatment of pleuropulmonary blastoma. We found limited evidence that the hematologic abnormalities observed in murine DICER1 models developed in our cohort of DICER1 carriers. In addition, no cases of myelodysplastic syndrome were observed in either the NCI cohort or the International Pleuropulmonary Blastoma/DICER1 Registry; 1 case of presumed secondary leukemia was reported. Abnormalities in hematologic indices should not be solely attributed to DICER1. This trial was registered at www.clinicaltrials.gov as #NCT01247597.

Comparison of Three Skin Sampling Methods and Two Media for Culturing Malassezia Yeast

Malassezia is a lipid-dependent commensal yeast of the human skin. The different culture media and skin sampling methods used to grow these fastidious yeasts are a source of heterogeneity in culture-based epidemiological study results. This study aimed to compare the performances of three methods of skin sampling, and two culture media for the detection of Malassezia yeasts by culture from the human skin. Three skin sampling methods, namely sterile gauze, dry swab, and TranswabTM with transport medium, were applied on 10 healthy volunteers at 5 distinct body sites. Each sample was further inoculated onto either the novel FastFung medium or the reference Dixon agar for the detection of Malassezia spp. by culture. At least one colony of Malassezia spp. grew on 93/300 (31%) of the cultures, corresponding to 150 samplings. The positive culture rate was 67%, 18%, and 15% (P < 10−3), for samples collected with sterile gauze, TranswabTM, and dry swab, respectively. The positive culture rate was 62% and 38% (P < 0.003) by using the FastFung and the Dixon media, respectively. Our results showed that sterile gauze rubbing skin sampling followed by inoculation on FastFung medium should be implemented in the routine clinical laboratory procedure for Malassezia spp. cultivation.

Comparison of Three Skin Sampling Methods and Two Media for Culturing Malassezia Yeast

Malassezia is lipid-dependent commensal yeast of the human skin. The different culture media and skin sampling methods used to grow these fastidious yeasts are a source of heterogeneity in culture-based epidemiological study results. This study aimed to compare the performances of three methods of skin sampling, and two culture media for the detection of Malassezia yeasts by culture from the human skin. Three skin sampling methods, namely sterile gauze, dry swab and TranswabTM with transport medium, were applied on 10 healthy volunteers. Each sample was further inoculated onto either the novel FastFung medium or the reference Dixon agar for the detection of Malassezia spp. by culture. At least one colony of Malassezia spp. grew on 93/300 (31%) of the cultures, corresponding to 150 samplings. The positive culture rate was 67%, 18%, and 15% (P &amp;lt; 10-3), for samples collected with sterile gauze, TranswabTM, and dry swab, respectively. The positive culture rate was 62% and 38% (P &amp;lt; 0.003) by using the FastFung and the Dixon media, respectively. Our results showed that sterile gauze rubbing skin sampling followed by inoculation on FastFung medium should be implemented in the routine clinical laboratory procedure for Malassezia spp. cultivation.