Laboratorinės medicinos klinika (K100000)

Background: May-Hegglin anomaly (MHA) is a rare autosomal dominant MYH9-related disorder characterized by thrombocytopenia, giant platelets, and Dohle-like cytoplasmic inclusions in leukocytes. Fewer than 200 cases have been reported worldwide, largely due to underrecognition and misdiagnosis. Clinical manifestations are usually mild and nonspecific, such as minor bleeding or easy bruising. Because clinical signs may be subtle, diagnosis relies heavily on laboratory evaluation. Detailed morphological analysis of peripheral blood smear remains a key tool for distinguishing MHA from other causes of thrombocytopenia. Objective: To demonstrate the diagnostic value of peripheral blood smear evaiuation in identifying suspected May-Hegglin anomaly in two related patients with idiopathic inherited thrombocytopenia. Method: Two related patients repeatedly presenting to the emergency department for unrelated complaints. They underwent complete blood count analysis and peripheral blood smear examination following automated analyzer flags indicating platelet abnormalities. Digital cell morphology was performed using a CellaVision analyzer. Morphologic findings, hematołogic parameters, and relevant clinical data were retrospectively reviewed and compared with established diagnostic features of MayHegglin anomaly. Result: Both patients had persistent mild-to-moderate thrombocytopenia (40-56x10^9/L) with elevated mean platelet volume (12-13.4 fL), and the automated analyzer repeatedly flagged,,giant platelet/ platelet clumps". Peripheral blood smear examination revealed enlarged platelets and well-defined spindle-shaped pale blue Dohle-like cytoplasmic inclusions in neutrophils, eosinophils, basophils, and monocytes. These inclusions were consistentiy observed across multiple visits over several months and were not associated with acute infection, trauma, or inflammatory changes in leukocytes. One patient underwent surgery without hemorrhagic complications. The familial occurrence, along with consistent morphological findings, strongly supports a diagnosis of MYH9-related thrombocytopenia. Conclusion: Persistent macrothrombocytopenia with clearly visible Dohle-like inclusions in leukocytes provides strong morphological evidence of MYH9-related thrombocytopenia. Systematic evaluation of peripheral blood smear allows the laboratory to identify these characteristic features. Early recognition supports accurate diagnosis, informs clinicians, and helps prevent misdiagnosis and unnecessary interventions. This report highlights the critical role of the laboratory in identifying rare inherited thrombocytopenias.

Background The incidence of bloodstream infections (BSI) caused by third-generation cephalosporin-resistant (3GCR) Enterobacterales has been increasing across Europe (1). However, it remains uncertain whether resistance is associated with worse clinical outcomes. To address this gap, we aimed to estimate the mortality associated with BSI due to 3GCR Enterobacterales compared to infections caused by susceptible strains in hospitalized patients. Methods Individual patient level data from four studies identified through a systematic search (PROSPERO CRD42022322029) were included in the analysis (2–5). A one-step individual participant data meta-analysis was performed using a multilevel (study) logistic regression model adjusted for sex, age, McCabe score, surgical procedure in the previous month, type of acquisition, BSI source and appropriateness of empiric therapy. Results A total of 7111 patients were included in the analysis, with individual study sample sizes ranging from 282 to 3452. The dataset comprised three cohort studies and one case-control study, all conducted in Spain between 1997 and 2017 (Table 1). Compared to patients with BSI caused by susceptible strains (n=6356), those with 3GCR Enterobacterales (n=755) were more frequently male (56.3% vs. 48.7%), had a higher proportion of fatal or ultimately fatal disease according to the McCabe score (43.3% vs. 34.6%), were more likely to have undergone a surgical procedure in the previous month (13.6% vs. 9.2%), and more often presented with healthcare-associated or hospital-acquired infections (68.3% vs. 39.9%). They were also less frequently prescribed appropriate empiric therapy (64.1% vs. 93.5%) and had higher mortality (14.8% vs. 9.6%) (Table 2). In the crude analysis, 3GCR was associated with increased mortality (OR 1.57; 95% CI 1.25 to 1.96; n=7111). However, after adjusting for confounders, the association was no longer statistically significant (aOR 1.13; 95% CI 0.86 to 1.48; n=6144). These findings were consistent in the two-step meta-analysis (Figure 1). Conclusions In this dataset, 3CGR was not associated with increased mortality in hospitalized patients with Enterobacterales BSI compared to susceptible strains. This study demonstrates the value of IPD meta-analysis in providing robust evidence to inform public health decisions.

Shewanella algae is an emerging marine bacterium capable of causing serious infections in immunocompromised individuals. We report the first documented case in Lithuania of bullous cellulitis caused by S. algae, complicated by necrotizing soft-tissue infection and ultimately requiring leg amputation. To determine whether similar cases had been described earlier in Lithuania, a structured literature review was conducted. We searched PubMed/MEDLINE, Embase, Scopus, Web of Science, and Google Scholar by combining the terms "Shewanella algae," "Shewanella," "Baltic Sea," "wound infection," "necrotizing infection," "sepsis," "case report," and "Lithuania." No date limits were set, and all titles and abstracts were screened. A 74-year-old woman with poorly controlled type 2 diabetes and recent seawater exposure presented with acute right leg pain and skin lesions. Blood and wound cultures confirmed S. algae infection. Despite broad-spectrum antibiotics and intensive care, the patient's condition deteriorated, requiring below-knee and later above-knee amputation. This case emphasizes the aggressive clinical course S. algae can take in vulnerable patients and highlights the importance of early recognition and multidisciplinary management of severe skin and soft tissue infections.

Antimicrobial-resistant bacteria are a major global health threat. Mobile genetic elements (MGEs) have been crucial for spreading resistance to new bacterial species, including human pathogens. Understanding how MGEs promote resistance could be essential for prevention. Here we present an investigation of MGEs and their association with resistance genes in pathogenic bacteria collected from 59 diagnostic units during 2020, representing a snapshot of clinical infections from 35 counties worldwide.

Background-Aim Accuracy of laboratory test results depends on many factors. For clinicians incomprehensible test results are thought to be ‘a problem’ of laboratory work. However, importance of preanalytical phase to final results is not taken into account seriously. Factors like poor adherence to procedures of patient preparation and sample collection can dramatically affect the result of any test. Unexplained results require step by step investigation of conditions that could have affected the sample. Our case was linked to fluorescein angiography, which is an important and frequently used diagnostic tool in ophthalmology. Intravenous fluorescein interference led to false elevated FT3 level. Methods Test results were obtained by fluorometric enzyme immunoassay (AIA-2000, Tosoh Bioscience). Results Case report. A 70-year-old man was hospitalized in Hospital of Lithuanian University of Health Sciences Kauno klinikos for examination of unspecified bacterial infection. Multidisciplinal examinations were made. Additionally, in suspicion of subacute thyroiditis, a workup for thyroid function was initiated. Serum tests were measured: TSH, FT4, Anti-TPO levels were in reference range, FT3 - higher than the upper limit of analytical range (>38.4 pmol/L). Investigation of incomprehensible results that brought more questions to clinicians and laboratory staff revealed that the serum of the patient looked fluorescent yellow-colored, and blood for the tests was drawn right after fluorescein angiography. In Instructions for Use of FT3 reagent, it is pointed out that injection of fluorescein may cause falsely elevated results. 5 days later FT3 was measured in a new serum sample and its level (4,11 pmol/L) was in reference range. Conclusions Fluorescein angiography has been used for decades, and several authors have demonstrated fluorescein interference with several laboratory tests. We must understand the value of inaccurate results to patient and hospital because it can lead to unnecessary or even harmful diagnostic procedures and/or therapy. We perpetually regain our attention to it but similar cases appearing show the problem of sampling errors remains. Solution to this requires cooperation between laboratory staff and clinicians to improve the quality of specimen collection and laboratory test results.

Background-Aim Daratumumab is a medication for multiple myeloma treatment. It is a CD38-target monoclonal antibody. CD38 protein is expressed in malignant plasma cells and red blood cells (RBC). Daratumumab causes interference with serological testing, including RBC antibody screening, antibody identification, and crossmatching based on indirect antiglobulin testing. Therefore, treatment with daratumumab can lead to pan-reactivity and difficulty in patients’ pretransfusion testing. Dithiothreitol (DTT) can interrupt the bond between anti-CD38 antibodies and CD38 receptors, so RBC treatment with DTT can help eliminate daratumumab interference in pretransfusion testing. Methods A 63-year-old woman was hospitalized in The Hospital of Lithuanian University of Health Sciences Kauno Klinikos for severe anemia (hemoglobin 61 g/L) after treatment with daratumumab. The patient required a blood transfusion, and a matching of RBC was ordered. Patient venous blood was collected with EDTA. The samples were studied for routine pretransfusion tests through the conventional methods used in the laboratory: ID-DiaCell I-II-III screening cells; and ID-Cards LISS/Coombs (BioRad, Swiss). Donors’ RBCs were treated with DTT. K+ and E+ erythrocytes were used as quality control to verify that DTT denatured only the K antigen and preserved the E antigen. Results First, the crossmatch testing was performed with the patient’s plasma and four different donors’ RBCs (without DTT); all tests were positive - RBC incompatible. Meanwhile, crossmatch tests with the patient’s plasma and the donors’ RBC previously treated with DTT presented compatibility with two donors’ erythrocytes (the other two remained incompatible). Conclusions Treatment with CD38-target monoclonal antibody-based medication, such as daratumumab, can challenge patient pretransfusion testing. However, the DTT method can help eliminate daratumumab interference in blood compatibility testing and could also help provide safe blood transfusions to patients treated with anti-CD38 monoclonal antibodies.

Background: Peripheral blood stem cells (PBSCs) are widely used for autologous blood stem cell transplantation (auto-SCT) for patients with various conditions. CD34+ cell viability evaluation after cryopreservation and thawing is an important parameter used for quality assurance of the cryopreserved cells. CD34+ cell post-thaw viability results and their correlation with clinical parameters were evaluated. Methods: The study included 209 patients, 461 aphereses, 640 products and 239 auto-SCT in the Hospital of Lithuanian University of Health Sciences Kaunas Clinics from 2015 to 2022 December. The patient’s plasma and DMSO (final concentration 10%) were used for PBSC cryopreservation. PBSC products were cryopreserved using a controlled freezer and then transferred to vapor phase nitrogen. Cell viability was evaluated after thawing PBSC in a water bath at 37 °C. Immediately after thawing viability was assessed using a BD FACS Canto flow cytometer and BD Stem Cell Enumeration Kit. A linear regression model and Pearson correlation were used for statistical analysis. The study included 209 patients, 461 aphereses, 640 products and 239 auto-SCT in the Hospital of Lithuanian University of Health Sciences Kaunas Clinics from 2015 to 2022 December. The patient’s plasma and DMSO (final concentration 10%) were used for PBSC cryopreservation. PBSC products were cryopreserved using a controlled freezer and then transferred to vapor phase nitrogen. Cell viability was evaluated after thawing PBSC in a water bath at 37 °C. Immediately after thawing viability was assessed using a BD FACS Canto flow cytometer and BD Stem Cell Enumeration Kit. A linear regression model and Pearson correlation were used for statistical analysis. Results: In total 209 patients (106 females and 103 males) with a median age of 59 (range 18–73) years were harvested after G-CSF (10g/kg) or Cyclo+G-CSF or additional mobilizing agents such as plerixafor (0.24 mg/kg) for auto-SCT. The median post-thaw CD34+ viability was 75% (range 1–96). Median leukocytes (>1.0 × 109 /l) engraftment on two consecutive days was 11 (range 8–13) days, and 15 days for platelets (>50 × 109 /l), ranging 9–23 days after SCT. Statistically significant negative correlations were observed between CD34+ cell post-thaw viability and CD45+ count in leukapheresis product (p = 0.001, R2 = 0,076); CD34+ count (p = 0.002, R2 = 0.02); a volume of leukapheresis product (p = 0.001, R2 = 0.076). Conclusions: The results suggest that cryopreservation of cells using 10% DMSO final concentration is acceptable with successful cell engraftment in all patients. Lower CD45+ and CD34+ cell counts and lower leukapheresis volume had a better impact on post-thaw cell viability. Further studies are needed to confirm these findings and improve post-thaw cell viability.

Background and Objectives: Growing antibiotic resistance among bacteria is a global issue that is becoming harder and more expensive to solve. Traditional treatment options are becoming less effective, causing more fatal outcomes of nosocomial infections. Since the development of new antibiotics has stagnated in the last decade, a novel approach is needed. Materials and Methods: Graphene-based materials are being developed and tested for various applications, and the medical field is no exception. We tested 98 clinical A. baumannii strains for antibiotic resistance, AMP-C production and the effectiveness of a graphene oxide and silver nanoparticle hybrid nanocomposite. The disc diffusion method was used to determine antibiotic susceptibility results. Antibiotic discs containing cefotaxime, cloxacillin and clavulanate were used to detect AMP-C production. The effectiveness of the GO–Ag hybrid nanocomposite was determined by counting colony forming units (CFUs) after a suspension of A. baumannii and the GO–Ag hybrid nanocomposite was plated on MH agar and incubated overnight to grow colonies. Results: In our research, we found that A. baumannii strains are resistant to the majority of commonly used antibiotics. Antibiotic resistance levels and AMP-C production can be factors, indicating the better effectiveness of the graphene oxide and silver nanoparticle hybrid nanocomposite. Conclusions: In this study, a GO–Ag hybrid nanocomposite was shown to have the potential to fight even the most problematic bacteria like A. baumannii.

The Two Weeks in the World research project has resulted in a dataset of 3087 clinically relevant bacterial genomes with pertaining metadata, collected from 59 diagnostic units in 35 countries around the world during 2020. A relational database is available with metadata and summary data from selected bioinformatic analysis, such as species prediction and identification of acquired resistance genes.

Background Surveillance of antimicrobial resistance (AMR) and antimicrobial use (AMU) in Europe is currently annual. Aim To study the feasibility and scalability of a quarterly AMR/AMU surveillance system in the European Union/European Economic Area (EU/EEA). Methods We conducted a longitudinal study within the scope of the EU-JAMRAI project. Seventeen partners from 11 EU/EEA countries prospectively collected 41 AMU and AMR indicators quarterly from September 2017 to May 2020 for the hospital sector (HS) and primary care (PC). Descriptive statistics and coefficients of variation (CV) analysis were performed. Results Data from 8 million hospital stays and 45 million inhabitants per quarter were collected at national (n = 4), regional (n = 6) and local (n = 7) levels. Of all partners, five were able to provide data within 3 months after each preceding quarter, and eight within 3–6 months. A high variability in AMU was found between partners. Colistin was the antibiotic that showed the highest CV in HS (1.40; p < 0.0001). Extended-spectrum beta-lactamase-producing Escherichia coli presented the highest incidence in HS (0.568 ± 0.045 cases/1,000 bed-days per quarter), whereas ciprofloxacin-resistant E. coli showed the highest incidence in PC (0.448 ± 0.027 cases/1,000 inhabitants per quarter). Barriers and needs for implementation were identified. Conclusion This pilot study could be a first step towards the development of a quarterly surveillance system for AMU and AMR in both HS and PC in the EU/EEA. However, committed institutional support, dedicated human resources, coordination of data sources, homogeneous indicators and modern integrated IT systems are needed first to implement a sustainable quarterly surveillance system.