AKR1B10, One of the Triggers of Cytokine Storm in SARS-CoV2 Severe Acute Respiratory Syndrome.

Preventing the cytokine storm observed in COVID-19 is a crucial goal for reducing the occurrence of severe acute respiratory failure and improving outcomes. Here, we identify Aldo-Keto Reductase 1B10 (AKR1B10) as a key enzyme involved in the expression of pro-inflammatory cytokines. The analysis of transcriptomic data from lung samples of patients who died from COVID-19 demonstrates an increased expression of the gene encoding AKR1B10. Measurements of the AKR1B10 protein in sera from hospitalised COVID-19 patients suggests a significant link between AKR1B10 levels and the severity of the disease. In macrophages and lung cells, the over-expression of AKR1B10 induces the expression of the pro-inflammatory cytokines Interleukin-6 (IL-6), Interleukin-1ß (IL-1ß) and Tumor Necrosis Factor a (TNFα), supporting the biological plausibility of an AKR1B10 involvement in the COVID-19-related cytokine storm. When macrophages were stressed by lipopolysaccharides (LPS) exposure and treated by Zopolrestat, an AKR1B10 inhibitor, the LPS-induced production of IL-6, IL-1ß, and TNFα is significantly reduced, reinforcing the hypothesis that the pro-inflammatory expression of cytokines is AKR1B10-dependant. Finally, we also show that AKR1B10 can be secreted and transferred via extracellular vesicles between different cell types, suggesting that this protein may also contribute to the multi-organ systemic impact of COVID-19. These experiments highlight a relationship between AKR1B10 production and severe forms of COVID-19. Our data indicate that AKR1B10 participates in the activation of cytokines production and suggest that modulation of AKR1B10 activity might be an actionable pharmacological target in COVID-19 management.

Monitoring of Unfractionated Heparin Therapy in the Intensive Care Unit Using a Point-of-Care aPTT: A Comparative, Longitudinal Observational Study with Laboratory-Based aPTT and Anti-Xa Activity Measurement.

Continuous intravenous unfractionated heparin (UFH) is administered routinely in the intensive care unit (ICU) for the anticoagulation of patients, and monitoring is performed by the activated partial thromboplastin time (APTT) or anti-Xa activity. However, these strategies are associated with potentially large time intervals before dose adjustments, which could be detrimental to the patient. The aim of the study was to compare a point-of-care (POCT) version of the APTT to (i) laboratory-based APTT and (ii) measurements of anti-Xa activity in terms of correlation, agreement and turnaround time (TAT). Thirty-five ICU patients requiring UFH therapy were prospectively included and followed longitudinally for a maximum duration of 15 days. UFH was administered according to a local adaptation of Raschke and Amanzadeh's aPTT nomograms. Simultaneous measurements of POCT-APTT (CoaguCheck® aPTT Test, Roche Diagnostics) on a drop of fresh whole blood, laboratory-based APTT (C.K. Prest®, Stago) and anti-Xa activity (STA®Liquid anti-Xa, Stago) were systematically performed two to six times a day. Antithrombin, C-reactive protein, fibrinogen, factor VIII and lupus anticoagulant were measured. The time tracking of sampling and analysis was recorded. The overall correlation between POCT-APTT and laboratory APTT (n = 795 pairs) was strongly positive (rs = 0.77, p < 0.0001), and between POCT-APTT and anti-Xa activity (n = 729 pairs) was weakly positive (rs = 0.46, p < 0.0001). Inter-method agreement (Cohen's kappa (k)) between POCT and laboratory APTT was 0.27, and between POCT and anti-Xa activity was 0.30. The median TATs from sample collection to the lab delivery of results for lab-APTT and anti-Xa were 50.9 min (interquartile range (IQR), 38.4−69.1) and 66.3 min (IQR, 49.0−91.8), respectively, while the POCT delivered results in less than 5 min (p < 0.0001). Although the use of the POCT-APTT device significantly reduced the time to results, the results obtained were poorly consistent with those obtained by lab-APTT or anti-Xa activity, and therefore it should not be used with the nomograms developed for lab-APTT.

Helicobacter canis: A Review of Microbiological and Clinical Features.

Helicobacter canis, an enterohepatic Helicobacter, has proven its role in human diseases and has been rediscussed in recent years as its zoonotic potential is increasingly described. Routine microbiological detection of this pathogen is a difficult task as its culture may fail due to fastidious growth. It is therefore supposed that many clinical laboratories under-recognize H. canis infections. A review of all clinical and microbiological literature currently available from previous relevant H. canis human clinical cases, mainly bacteremia, added with a clinical case observed at the Cliniques universitaires Saint-Luc, was performed. Clinical features of H. canis reports show the presence of underlying clinical conditions in 89% of the cases, bacteremia in 83%, associated fever in 58%, and recent close contact with pets in 83%, especially dogs. The observed microbiological trends from 10 cases of bacteremia were a median of 4 days until positive blood culture bottle detection, subcultures showing a thin layer of small colonies under microaerophilic atmosphere at 35-42°C after 3-4 days of growth, and an identification requiring 16S rRNA sequencing given the difficulties observed with MALDI-TOF MS. Low MICs were observed for penicillins, amoxicillin/clavulanic acid, carbapenems, and metronidazole in opposition to high MICs for ciprofloxacin. A frequent association of H. canis and bacteremia in immunocompromised patients with recurrent fever in contact with pets, especially dogs, was identified. Considering the fastidious growing capacities, final identification from blood cultures may not be expected before 7 days. Intravenous ceftriaxone, oral doxycycline, or metronidazole has been suggested as efficient therapeutic choices.

Pseudothrombocytopenia-A Review on Causes, Occurrence and Clinical Implications.

Pseudothrombocytopenia (PTCP), a relative common finding in clinical laboratories, can lead to diagnostic errors, overtreatment, and further (even invasive) unnecessary testing. Clinical consequences with potential life-threatening events (e.g., unnecessary platelet transfusion, inappropriate treatment including splenectomy or corticosteroids) are still observed when PTCP is not readily detected. The phenomenon is even more complex when occurring with different anticoagulants. In this review we present a case of multi-anticoagulant PTCP, where we studied different parameters including temperature, amikacin supplementation, measurement methods, and type of anticoagulant. Prevalence, clinical risk factors, pre-analytical and analytical factors, along with clinical implications, will be discussed. The detection of an anticoagulant-dependent PTCP does not necessarily imply the presence of specific disorders. Conversely, the incidence of PTCP seems higher in patients receiving low molecular weight heparin, during hospitalization, or in men aged 50 years or older. New analytical technologies, such as fluorescence or optical platelet counting, will be soon overturning traditional algorithms and represent valuable diagnostic aids. A practical laboratory approach, based on current knowledge of PTCP, is finally proposed for overcoming spuriously low platelet counts.

Leptomeningeal Carcinomatosis: A Call for Optimizing Diagnostic Sensitivity by the Hematology Laboratory.

In the cerebrospinal fluid (CSF), the demonstration of malignant cells by cytological examination is currently the gold standard for the diagnosis of leptomeningeal carcinomatosis (LC). However, a positive cytology is observed in only 50-60% of patients with LC and highly dependent on pre-analytical factors. The hematology laboratory could provide an immediate and accurate diagnosis, but diagnostic sensitivity is not always optimized once the sample is received. We hereby report a 49-year-old woman with a 3-year grade III invasive ductal carcinoma who was admitted to the emergency department due to headaches, nausea, and vomiting. The CSF revealed pleocytosis with suspicious high fluorescent cells on the hematology analyzer concomitantly with biochemical alterations. Cytomorphological examination confirmed tumor cells, thus diagnosing a leptomeningeal metastasis of her breast cancer. The patient was eventually transferred to palliative care. Cytological examination is a valuable tool for a rapid diagnosis of LC if diagnostic performance is optimized. In addition to repeated CSF collections with a sufficient volume (5-10 mL), this could be reached by processing the CSF as soon as possible, taking into account the fluorescence information from the analyzer, proceeding systematically to microscopic examination even with normal CSF white blood cell count, and providing quality improvement of the staff to identify malignant cells.

Analytical and clinical evaluation of new automated chemiluminescent immunoassays for the detection of IgG and IgM anti-Bartonella henselae antibodies.

Serological diagnosis of Bartonella henselae infection mainly rely on microscopic immunofluorescence assays (IFA), which are however time-consuming and poorly standardized. The aim of the study was to assess the use of the new fully automated VirClia® chemiluminescent immunoassays for the detection of IgG and IgM anti-B. henselae antibodies. Eighty-one patients with a well-defined B. henselae infection as well as 80 patients with an alternative disease were included. The VirClia® IgG antibody assay showed a sensitivity of 79.0% and a specificity of 93.8% for the diagnosis of B. henselae infection. For the VirClia® IgM assay, results were more conflicting with a sensitivity of 42.0% and a specificity of 98.2% to predict IFA IgM results. In 11 additional patients with uninterpretable IFA due to autoimmune antibodies, VirClia® assays were able to deliver valuable quantitative results. The VirClia® IgG assay shows good analytical and clinical performances and could be easily integrated in the diagnostic workflow of B. henselae infection.

Long-term Physicochemical Stability of Concentrated Solutions of Isosorbide Dinitrate in Polypropylene Syringes for Administration in the Intensive Care Unit.

In order to avoid fluid overload, more concentrated drug solutions in intensive care units are commonly used. This study evaluated the physicochemical stability of concentrated solution of isosorbide dinitrate in polypropylene syringes during 28 days at 5°C ± 3°C with protection from light. Five syringes of 50 mL, containing 0.60 mg/mL of isosorbide dinitrate in sodium chloride 0.9% were prepared and stored at 5°C ± 3°C with protection from light during 28 days. Immediately after preparation and periodically during the storage, isosorbide dinitrate concentration was measured by an ultra-performance liquid chromatography. Spectrophotometric absorbance at different wavelengths, pH measurements, and microscopic observations were also performed. All solutions were physicochemically stable during the whole period storage at 5°C ± 3°C. No color change, turbidity, precipitation or opacity, significant pH variations, or optic densities were observed in the solutions. Any crystals were seen by microscopic analysis. The concentration of isosorbide dinitrate remained above 90% of the initial concentration during the 28 days of storage. Solutions of isosorbide dinitrate 0.60 mg/mL in syringe of sodium chloride 0.9 % injection can be considered physically and chemically stable for 28 days when stored in syringes at 5°C ± 3°C with protection from light and may be prepared in advance by a centralized intravenous additive service.

Long-term Physicochemical Stability of Concentrated Solutions of Sodium Valproate in Polypropylene Syringes for Administration in the Intensive Care Unit.

In some situations, drug solutions in higher concentrations are used in intensive care units. The objective of this study was to evaluate the physicochemical stability of concentrated solutions of valproate sodium in polypropylene syringes during 30 days at 5°C ± 3°C. Five syringes of 40 mL containing 20 mg/mL of sodium valproate in 0.9% sodium chloride were prepared and stored at 5°C ± 3°C during 30 days. Immediately after preparation and periodically during the storage, valproate concentrations were measured by high-performance liquid chromatography. Spectrophotometric absorbance at different wavelengths, pH measurement, and microscopic observations were also performed. All solutions were physically stable during the study period storage at 5°C ± 3°C. No color change, turbidity, precipitation, or opacity at visual observation was noticed. No significant pH variations or optic densities were observed. No crystals were seen by microscopic analysis. Concentrations of valproate remained stable during the period of storage. Solutions of sodium valproate 20 mg/mL in syringes of 0.9% sodium chloride were physically and chemically stable for at least 30 days when stored in syringes at 5°C ± 3°C. These solutions may be prepared in advance by a centralized intravenous additive service.

Evaluation of the Physicochemical Stability of Amiodarone Hydrochloride in Syringes for the Intensive Care Unit.

In some emergency clinical situations, the injection of a more concentrated drug solution in the intensive care units is common. The purpose of this study was to evaluate the physicochemical stability of concentrated solutions of amiodarone hydrochloride in polypropylene syringes during 28 days of storage at 5°C ± 3°C, with protection from light. Five syringes of 50 mL, containing 25 mg/mL of amiodarone in dextrose 5%, were prepared and stored at 5°C ± 3°C with protection from light during 28 days. Immediately after preparation and periodically during the storage, amiodarone hydrochloride concentrations were measured by ultra-performance liquid chromatography. Spectrophotometric absorbance at different wavelengths, pH measurement, and microscopic observations were also performed. All solutions were physicochemically stable during the study period when stored at 5°C ± 3°C. No color change, turbidity, precipitation, opacity, significant pH variations, or optic densities were observed in the solutions. No crystals were seen by microscopic analysis. The concentration of amiodarone did not decrease during the 28 days of storage. Solutions of amiodarone 25 mg/mL in syringes of dextrose 5% are physically and chemically stable for at least 28 days when stored in syringes at 5°C ± 3°C with protection from light and may be prepared in advanced by a centralized intravenous additive service.

Long-term Physiochemical Stability of Concentrated Solutions of Salbutamol (Albuterol) in Polypropylene Syringes for Use in the Intensive Care Unit and in Obstetrics.

In order to avoid fluid overload, the use of more concentrated drug solutions in intensive care units and obstetrics is common. The objective of this study was to quantify the physicochemical stability of a concentrated solution of salbutamol (albuterol) in polypropylene syringes during 30 days of storage at 5°C ± 3°C with protection from light. Four 50-mL syringes containing 0.060mg/mL of salbutamol (albuterol) in 0.9% NaCl were prepared and stored at 5°C ± 3°C with protection from light during 30 days of storage. Immediately after preparation and periodically during the storage, salbutamol (albuterol) concentrations were measured by an ultra-performance liquid chromatography. Spectrophotometric absorbance at different wavelengths, pH measurement, and microscopic observations were also performed. All solutions were physicochemically stable during the entire period of storage at 5°C ± 3°C: no color change, turbidity, precipitation or opacity, significant pH variations, or optic densities were observed in the solutions. No crystals were seen by microscopic analysis. Concentrations of salbutamol remained stable during the storage period. Solutions of salbutamol (albuterol) 0.060 mg/mL in syringes of 0.9% NaCl are physically and chemically stable for at least 30 days when stored in syringes at 5°C ± 3°C with protection from light and may be prepared in advance by a centralized intravenous additive service.

A reminder of the place of morphology and the H-score in the diagnosis of hemophagocytic lymphohistiocytosis (HLH).

This case report reminds the reader of the place of hemophagocytosis and the H-Score in the diagnosis of secondary hemophagocytic lymphohistiocytosis.

Long-term Physicochemical Stability of Concentrated Solutions of Noradrenaline Bitartrate in Polypropylene Syringes for Administration in the Intensive Care Unit.

Intensive care units use drug solutions within higher concentrations to avoid fluid overload. The purpose of this study was to evaluate the physicochemical stability of concentrated solutions of noradrenaline bitartrate in polypropylene syringes during 30 days of storage at 5°C ± 3°C. Five 50-mL syringes containing 0.240 mg/mL of noradrenaline bitartrate in 0.9% sodium chloride were prepared and stored at 5°C ± 3°C during 30 days. Immediately after preparation and periodically during the storage, noradrenaline concentrations were measured by high-performance liquid chromatography. Spectrophotometric absorbance at different wavelengths, pH measurement, and microscopic observations were also performed. The results showed that all solutions were physicochemically stable during the entire storage period at 5°C ± 3°C, and no color change, turbidity, precipitation, opacity, significant pH variations, nor optic densities were observed. Microscopic analysis was used to determine if there was any formation of crystals. The concentration of noradrenaline was not found to decrease during the 30 days of storage. Solutions of noradrenaline bitartrate 0.240 mg/mL in syringes of 0.9% sodium chloride were physically and chemically stable for at least 30 days when stored in syringes at 5°C ± 3°C and may be prepared in advanced by a centralized intravenous additive service.

Evaluation of the DOAC-Stop® Procedure to Overcome the Effect of DOACs on Several Thrombophilia Screening Tests.

The impact of direct oral anticoagulants (DOACs) on laboratory assays used for thrombophilia testing (e.g., antithrombin, protein S, protein C, lupus anticoagulant and activated protein-C resistance) is a well-known issue and may cause false-positive and -negative results. Therefore, the correct interpretation of tests that are performed in patients taking DOACs is mandatory to prevent misclassification and the subsequent clinical consequences. We aimed at evaluating the efficiency of a new and simple procedure (DOAC-Stop®; Haematex Research, Hornsby, Australia) to overcome the effect of all DOACs in real-life settings and to assess the percentage of erroneous results due to the presence of DOACs on thrombophilia screening tests. For this purpose, 135 DOAC-treated patients (38 apixaban, 40 dabigatran, 15 edoxaban, and 42 rivaroxaban) and 20 control patients were enrolled. A significant drop in apixaban, dabigatran, edoxaban, and rivaroxaban plasma concentrations following the DOAC-Stop® treatment was observed (74.8-8.2 ng/mL [ p < 0.0001], 95.9-4.7 ng/mL [ p < 0.0001], 102.1-8.8 ng/mL [ p = 0.001], and 111.3-7.0 ng/mL [ p < 0.0001], respectively). The DOAC-Stop® treatment was mostly effective to overcome the effect of DOACs on PTT-LA, dilute Russell's viper venom time (dRVVT) screen, and dRVVT confirm tests. Using our procedures, false-positive results due to DOACs were observed only with lupus anticoagulant tests (up to 75%) and fell to zero after the DOAC-Stop® procedure, regardless of the DOAC considered. In conclusion, the DOAC-Stop® adsorbent procedure appeared to be an effective and simple way to overcome the interference of DOAC on coagulation tests and should facilitate the interpretation of thrombophilia screening tests in patients taking DOACs.