MSP CLINICAL RESEARCH | 13:30

Endometriosis and NK cells

Authors: Zuzana Prášilová (1), Barbora Sedláčková (1), Lukáš Prajka (2), Martina Grundmanová (3),
Monika Holubová (1, 4)
Supervisor: Miroslava Čedíková (1, 3)
(1) Biomedical Center, Faculty of Medicine in Pilsen, Charles University (2) Department of Gy
naecology and Obstetrics, Faculty of Medicine in Pilsen, Charles University (3) Department of
Physiology, Faculty of Medicine in Pilsen, Charles University (4) Department of Haematology and
Oncology, University Hospital Pilsen, Pilsen, Czech Republic
State-of-the-Art: Natural killer (NK) cells play a crucial role in immune surveillance and clear
ance of ectopic endometrial cells. Dysregulation of inhibitory killer cell immunoglobulin-like
receptors (KIRs) has been implicated in endometriosis, but data on their surface expression in
well-defined disease phenotypes remain limited.
Objective: The aim of this study was to characterise the expression of KIRs on peripheral blood
NK cells in women with endometrioma and to compare their distribution with that in healthy
controls.
Material and Methods: This case–control observational study included 23 treatment-naïve
women with surgically and histologically confirmed endometrioma and 23 age-matched healthy
controls. Peripheral blood mononuclear cells were analysed by multiparametric flow cytometry
using two KIR-specific antibody panels. NK cells were defined as CD45⁺CD3⁻CD56⁺ lymphocytes,
and the expression of inhibitory KIR receptors was quantified. Correlations between clinical pa
rameters and KIR expression were assessed to evaluate potential confounding effects.
Results & Discussion: The proportion of NK cells within the lymphocyte compartment did not
differ between groups. However, women with endometrioma exhibited a significantly reduced
proportion of lymphocytes and a selective enrichment of KIR2DL2⁺ and KIR2DL5⁺ NK-cell sub
sets compared with controls. No significant associations were identified between demographic
or gynaecologic variables and KIR receptor expression.
Conclusion: Women with endometrioma exhibit a distinct inhibitory KIR profile on peripheral
NK cells, with increased KIR2DL2⁺ and KIR2DL5⁺ subsets. These findings suggest altered NK-cell
inhibition contributes to systemic immune dysregulation in endometriosis and identify KIR2DL5
as a potential research target.
Funding: This work was supported by the Ministry of Health of the Czech Republic through the
Conceptual Development of Research Organization (FNPl, 00669806) and by the Czech Health
Research Council under project No. NW25-03-00338

ECHO findings in intermediate-high risk PE

Authors: Filip Lukačík (1)
Supervisor: Štěpán Jirouš (1)
(1) Department of Cardiology, Faculty of Medicine in Pilsen, Charles University and University
Hospital, Pilsen
State-of-the-Art: Acute pulmonary embolism (PE) is a potentially life-threatening condition
and represents one of the leading causes of cardiovascular mortality. Intermediate-high risk
pulmonary embolism is characterised by hemodynamic stability in the presence of right ven
tricular dysfunction, elevated cardiac biomarkers and class III-V according to the PESI (Pulmo
nary Embolism Severity Index) score or sPESI (simplified PESI) ≥ 1. The first-line therapy is antico
agulation. Among newer therapeutic approaches is catheter-directed thrombolysis (CDT), which
enables local administration of a thrombolytic agent.
Objective: To compare the effect of CDT versus standard anticoagulation therapy by assessing
selected echocardiographic parameters at 24 hours after randomisation.
Material and Methods: A retrospective analysis was performed on 30 patients hospitalised for
acute intermediate-high risk PE. This analysis constitutes part of the larger PRAGUE-26 study.
PRAGUE-26 is a multicentre, randomised, open-label, actively controlled trial designed to evalu
ate clinical outcomes in patients with acute intermediate-high risk PE undergoing CDT vs. stand
ard anticoagulation therapy. The CDT was performed using the Cragg-McNamara valved infu
sion catheter. Follow-up focused on the development of selected echocardiographic parameters
(RV diameter, RV/LV ratio, TAPSE, s′TDI, and sPAP) at 24 hours after randomisation to either the
standard anticoagulation or the CDT. Statistical analysis was conducted using the nonparametric
Mann-Whitney U test.
Results & Discussion: Baseline echocardiographic parameters did not differ significantly be
tween the groups. At 24 hours, comparison of outcomes demonstrated significant improvement
in TAPSE (+4.6 ± 3.6 mm vs. +1.8 ± 2.4 mm, p = 0.014) and sPAP (−14.3 ± 8.2 mmHg vs. −3.8
± 7.9 mmHg, p = 0,004) in the CDT group compared with the standard anticoagulation. Other
parameters (RV diameter and RV/LV ratio) showed a trend toward greater improvement in the
CDT group; however, these differences did not reach statistical significance (all p > 0.05). No
statistically significant improvement or trend was observed for s′TDI in the CDT group. No signif
icant correlation was found between the BMI or the selected comorbidities and the treatment
response in either group.
Conclusion: Significant improvement in TAPSE and sPAP, along with a trend toward greater im
provement in RV diameter and RV/LV ratio, suggests a promising potential for the routine use
of CDT therapy in acute intermediate-high risk PE. Further large-scale prospective research is
required

Authors: Aneta Švecová (1)
Supervisor: Jan Lejčko (1)
(1) Department of Cardiology, Faculty of Medicine in Pilsen, Charles University and University
Hospital, Pilsen
State-of-the-Art: Atrial fibrillation (AF) is the most common sustained arrhythmia and is as
sociated with atrial remodeling and left atrial (LA) enlargement. LA volume (LAV) and indexed
LA volume (LAVI) are clinically relevant markers used in patient characteristics and outcome
assessment in rhythm-control strategies, including catheter ablation. Three-dimensional (3D)
echocardiography is often considered more accurate than conventional two-dimensional (2D)
methods; however, real-world measurements may vary depending on acquisition quality and
rhythm during imaging.
Objective: To compare LAV and LAVI obtained by 2D TTE using Simpson’s method and by 3D
TTE in AF patients evaluated prior to pulsed-field ablation (PFA; catheter ablation using pulsed
electric fields), and to assess whether left ventricular ejection fraction (LVEF) differs between 2D
and 3D measurements.
Material and Methods: We performed a single-center observational analysis of 58 consecutive
AF patients undergoing TTE as part of the pre-procedural work-up before PFA between 03/2023
and 09/2025. Examinations were performed using a Philips EPIQ 7 ultrasound system. LAV and
LAVI were measured by both 2D Simpson’s method and 3D methods during the same exam. For
categorical comparison, an absolute difference of ≤5 mL between 2D and 3D LAV was considered
clinically negligible (“agreement”); differences >5 mL were categorized as “2D higher” or “3D
higher”. Descriptive statistics were used. Paired 2D vs 3D comparisons were performed using a
two-sided Wilcoxon signed-rank test.
Results & Discussion: The cohort included 58 patients (62.1% male) with a mean age of 64.8 ±
9.6 years. Hypertension was present in 57.9%, dyslipidemia in 49.1%, type 2 diabetes in 10.5%,
and dysglycemia in 10.5%. Mean 2D LAV was 84.3 ± 23.4 mL, whereas mean 3D LAV was 71.7 ±
23.3 mL, with an average difference (3D − 2D) of −12.6 ± 16.0 mL; this 2D vs 3D difference was
statistically significant (p < 0.001). In relative terms, 2D LAV was 17.6% higher than 3D LAV. Using
the predefined 5 mL threshold, 2D exceeded 3D in 67.2%, agreement was observed in 20.7%,
and 3D exceeded 2D in 12.1%. Contrary to the commonly expected tendency toward larger LA
volumes with 3D TTE, our cohort demonstrated a reverse trend. LVEF assessed by 2D and 3D
methods did not differ significantly.
Conclusion: In AF patients assessed prior to PFA, 2D TTE using Simpson’s method yielded high
er LAV estimates than 3D in the majority of cases. These findings highlight the importance of
standardized acquisition and analysis when comparing 2D and 3D LAV. LVEF values derived from
2D and 3D TTE were comparable

Risk Stratification and BMI Paradox in PE

Authors: Lukáš Kaun (1)
Supervisor: Milan Hromádka (1)
(1) Department of Cardiology, Faculty of Medicine in Pilsen, Charles University and University
Hospital, Pilsen
State-of-the-Art: Risk stratification of patients with pulmonary embolism (PE) is crucial for
selecting the optimal therapeutic strategy. The Pulmonary Embolism Severity Index (PESI) is
an established tool for assessing 30-day mortality risk across different patient populations.
However, clinical stability defined by PESI may not always reflect the underlying hemodynamic
stress. Integration of cardiac biomarkers and imaging methods is therefore essential to refine
risk assessment, particularly in hemodynamically stable patients with potential right ventricular
(RV) dysfunction.
Objective: The aim of this study was to evaluate the demographic, clinical, and laboratory char
acteristics of the patient group and to analyse the relationships between Pulmonary Embolism
Severity Index (PESI) scores, biomarkers, and the presence of RV dilatation.
Material and Methods: A retrospective analysis included 203 patients with PE hospitalised at
the Department of Cardiology, University Hospital Pilsen. We evaluated PESI scores, cardiac bio
marker levels (hsTnT, NT-proBNP, D-dimer), and RV dilatation on CT and echocardiography. Sta
tistical analysis was performed using non-parametric tests (Spearman correlation, Mann-Whit
ney U test, Kruskal-Wallis analysis) and Cohen‘s kappa (κ) to assess the agreement between
imaging modalities.
Results & Discussion: Mean age was 65.9±17.9 years, with a mean BMI of 30.8±7.1 kg/m². The
largest group consisted of intermediate-high risk patients (56.2%). A significant negative cor
relation was found between BMI and PESI score (ρ=−0.252; p<0.001), being strongest in low
(ρ=−0.670; p=0.004) and intermediate-low (ρ=−0.625; p<0.001) groups. hsTnT levels were signif
icantly associated with RV dilatation (median 72 vs 19.5 ng/l; p<0.001) and served as a predic
tor of in-hospital mortality (p=0.033). High agreement for RV dilatation was found between CT
and echocardiography (κ=0.893). Notably, despite comparable PESI scores (mean 101.6±23.2
vs 100.1±31.0), a fundamental difference in RV dilatation prevalence exists between intermedi
ate-high and intermediate-low groups (99.0% vs 2.3%; p<0.001).
Conclusion: The registry confirms the current ESC scheme‘s validity. A negative BMI-PESI cor
relation suggests an obesity paradox. Significant PESI limitations in intermediate-risk groups
show necessity of biomarker and echo assessment for the accurate identification of patients at
higher risk of mortality

Methods for CSF Leukocyte Differentiation

Authors: Veronika Pechová (1), Pavel Brož (1), Simona Kukrálová (1), Klára Koldušková (1), Eliška
Naušová (1), Daniel Rajdl (1)
Supervisor: Pavel Brož (1)
(1) Department of Clinical Biochemistry and Haematology, Faculty of Medicine in Pilsen, Charles
University and University Hospital, Pilsen
State-of-the-Art: Cerebrospinal fluid (CSF) examination plays an indispensable role in diag
nosing diseases of the central nervous system. Accurate quantitative cytology is essential for
appropriate clinical decision-making; however, this examination remains highly dependent on
the expertise of trained personnel. Furthermore, routine laboratory practice lacks rigorously
validated procedures for standardizing comparisons between basic leukocyte differentiation
performed in a Fuchs-Rosenthal (F–R) chamber and differentiation using permanent cytological
preparations (CP). Addressing this methodological gap is important for improving diagnostic
reliability.
Objective: The primary aim was to compare leukocyte differentiation in a F-R chamber with that
obtained from stained permanent CP. The secondary aim was to assess inter-rater agreement
between an experienced laboratory professional and a trained medical student.
Material and Methods: The study included CSF samples from 145 patients aged 10 days to 84
years at the Faculty Hospital in Pilsen. Manual cell counting and differentiation were performed
using the F–R chamber for all samples per routine protocols. Subsequently, permanent CP were
prepared from the same samples and stained using May–Grünwald–Giemsa staining. Leuko
cyte differentiation was then evaluated from the CP.
In a subset of 83 samples, leukocyte differentiation was independently evaluated by an expe
rienced professional and a trained medical student. Statistical analysis was performed using
MedCalc software (Shapiro–Wilk, Wilcoxon signed-rank test, and Bland–Altman plots). Cohen’s
kappa was used to evaluate the agreement for oligocytosis vs. pleocytosis classification. Signif
icance was set at p < 0.05.
Results & Discussion: Statistically significant differences were observed between leukocyte
differentiation in the F–R chamber and permanent CP. For mononuclear cells (MNc), median
(min–max) was 4 (1–724)/µL in the F–R chamber vs. 4 (1–1356)/µL in CP (p = 0.004). For poly
morphonuclear cells (PMNc), the values were 0 (0–6400)/µL in the F–R chamber vs. 0 (0–5471)/
µL in CP (p < 0.001).
In the subset evaluated by both raters, significant differences were found between the profes
sional and the student: MNc 7 (1–1356)/µL vs. 7 (1–793)/µL (p = 0.016) and PMNc 0 (0–5471)/µL
vs. 0 (0–6034)/µL (p = 0.002).
Cohen’s kappa showed excellent agreement in the classification of oligocytosis vs. pleocytosis
between the F–R chamber and the student (κ = 0.95) as well as between the professional and
the student (κ = 0.95).
Conclusion: Leukocyte differentiation by the student significantly differed from both the F–R
chamber and the professional. However, from a clinical perspective, agreement between raters
remained excellent, and the classification of oligocytosis vs. pleocytosis was highly consistent.
Funding: This work was supported by the Ministry of Health of the Czech Republic – Conceptual Development of Research Organization (FNPI – 00669806) and by the Cooperatio Program, re
search area Medical Diagnostics and Basic Medical Sciences

β-Blocker administration in Septic Shock

Authors: Olga Kotzamani (1)
Supervisor: Marek Nalos (1)
(1) Department of Internal Medicine I, University Hospital Pilsen, Czech Republic
State-of-the-Art: Septic shock is a life-threatening complication of sepsis characterized by
microcirculatory dysfunction and hemodynamic instability, including persistent hypotension,
tachycardia, and sympathetic overactivation despite adequate fluid resuscitation. β-blocker
therapy has been proposed as an adjunctive treatment to counteract the hyperadrenergic state.
β₁-receptor blockade reduces heart rate, myocardial contractility, and oxygen consumption, im
proving cardiac efficiency. Studies also suggest β-blockers may reduce cytokine release, limit
hypermetabolism, and protect against catecholamine-induced myocardial toxicity. However,
evidence regarding their immunomodulatory effects and safety in septic shock remains incon
clusive, requiring further investigation in critically ill patients.
Objective: To evaluate the hemodynamic safety of β-blocker therapy in septic shock and assess
its potential immunomodulatory effects through analysis of inflammatory markers.
Material and Methods: A retrospective observational study included 50 adult patients with sep
tic shock admitted to the ICU in 2024. Patients were stratified by β-blockers exposure to those
receiving parenteral cardio selective B-blockers (BB group) and those without, β-blockers (NB
group). Cardio selective agents included atenolol, acebutolol, bisoprolol, esmolol, and metopro
lol, used in patients with persistent tachycardia despite standard management. Clinical and lab
oratory data were obtained from electronic medical records. Hemodynamic parameters were
recorded daily during the first seven days after diagnosis.
Comorbidities, interventions, and 30- and 90-day mortality were also documented. Continuous
variables were expressed as mean ± SEM or median with quartiles, and group differences were
presented in graphs.
Results & Discussion: Heart rate decreased progressively during the 7-day observation pe
riod. In the NB group, mean HR declined from approximately 90 bpm on day 1 to about 72
bpm by days 6-7, while patients receiving. β-blockers maintained more stable values ranging
between 78-87 bpm without clinically significant bradycardia. Systolic blood pressure increased
modestly in both groups, while diastolic pressure remained relatively stable around 58-61
mmHg throughout the observation period. No episodes of treatment-related hypotension were
observed. Electrolyte levels and markers of organ function remained within acceptable ranges.
These findings suggest that the benefits of B-Blocker therapy primarily relate to cardiovascular
and hemodynamic modulation rather than direct suppression of systemic inflammatory activity.
Conclusion: β-blocker therapy in septic shock was well tolerated and effectively reduced heart
rate without compromising arterial pressure or systemic perfusion. These findings support
β-blockers as a safe adjunctive therapy to improve cardiovascular stability in critically ill pa
tients

Genetic pedigrees in hereditary angioedema

Authors: Eva Štuksová (1)
Supervisor: Martina Vachová (1)
(1) Department of Immunology and Allergology, Faculty of Medicine in Pilsen, Charles University
and University Hospital, Pilsen
State-of-the-Art: Hereditary angioedema (HAE) is a rare genetic disorder characterized by re
current attacks of pale, non-itchy swelling of subcutaneous tissue and mucous membranes. The
most common cause of hereditary angioedema is deficiency or dysfunction of C1 inhibitor (C1
INH), leading to excessive bradykinin production. Bradykinin contributes to increased vascular
permeability and angioedema development by acting on B2 receptors on surface of endothelial
cells and smooth muscle cells of blood vessels. Angioedema attacks cause significant discomfort
to patients and, depending on location, may have a serious clinical course. The Department
of Immunology and Allergology at the University Hospital in Pilsen is one of four specialized
centers for the diagnosis and treatment of HAE in the Czech Republic.
Objective: The aim of this study is to create family trees of patients with HAE using retrospective
data and medical history. It tracks disease in family lines and summarizes the diagnostic and
therapeutic approach, including patient care organization at the Pilsen center.
Material and Methods: The diagnosis of hereditary angioedema is based on a combination of
typical clinical presentation and laboratory findings (changes in C4 complement levels, C1 inhib
itor levels and functional activity). Molecular genetic testing focused primarily on the SERPING1
gene (and other genes in rare forms) is used to confirm the diagnosis and specify the genetic
basis.
The work is presented retrospectively in the form of case studies based on the analysis of med
ical records and targeted family history. The data obtained were used to compile family trees
in the draw.io tool, which graphically illustrate the occurrence of the disease in family lines and
describe the diagnostic and therapeutic procedure for selected patients at the University Hos
pital in Pilsen.
Results & Discussion: Based on targeted family history, detailed multigenerational family trees
were compiled in draw.io for two families of patients with HAE. The graphical representation
confirmed autosomal dominant inheritance with both sexes affected across generations. Anal
ysis of the family trees showed that the disease occurs in at least three consecutive generations
in both families.
Conclusion: The occurrence of HAE was mapped in two families from analysis of medical history
and available laboratory data. This visualization of family lines can serve as a basis for further
analysis of clinical data from medical records and for observed families to streamline long-term
dispensary care

Obesity and Birth Injury

Authors: Lucie Macháčková (1), Nela Fiedorová (2)
Supervisor: Zdeněk Rušavý (1)
(1) Department of Gynaecology and Obstetrics, Faculty of Medicine in Pilsen, Charles University
and University Hospital, Pilsen
State-of-the-Art: Birth injury refers to tissue damage in the perineal, vaginal, or cervical areas
caused by extreme pressure and stretching during the passage of the fetus through the birth
canal. This may involve spontaneous tearing of the skin and muscles or a controlled surgical inci
sion performed by a physician. The number of obese parturients is rising continually, hence the
increased interested in the effect of BMI on labor outcome. Large cohort studies have suggested
obesity as a protective factor for maternal perineal injury. As this conclusion is rather surprising
and comes from countries with a different obstetric care, we wanted to find out whether it is
valid also among Czech parturients that delivered in our institution.
Objective: The objective of this study is to test the hypothesis that women with a higher BMI
have a significantly reduced risk of sustaining severe birth injuries compared to ones with nor
mal weight. This study focuses on whether BMI can be considered a protective factor for child
birth trauma in our setting.
Material and Methods: The methodology employed a retrospective cohort analysis, compar
ing a group of parturient women divided by BMI into a normal weight group (BMI under 25) and
an obese group (BMI over 30). The study included all women who gave birth at term in the de
partment of Gynecology and Obstetrics, Faculty of Medicine in Pilsen between 1/2023-12/2025
the necessary data was extracted from clinical records and the hospital information system.
Monitored parameters included age, parity, delivery mechanism, and BMI. Furthermore, the
analysis accounted for the duration of labor, neonatal gender, and specific interventions, such
as oxytocin administration for labor augmentation, mode of labor induction, vaginal birth after
cesarean (VBAC), or operative deliveries using vacuum extraction (VEX) and forceps.
Results & Discussion: The results of the analysis demonstrated that parturient women with a
normal body weight (BMI under 25) show a statistically higher probability of delivery without
perineal injury compared to obese patients (39,4 vs 32,3%, p=0,0002). While a higher overall in
cidence of ruptures was recorded in the group with a BMI over 30, the data did not demonstrate
an increased risk of the most severe injuries involving the anal sphincter (rpt. per III.st. 1,5 vs
1,2 %, p= 0,5677).
Conclusion: The final analysis shows that a normal BMI represents a significant protective fac
tor in reducing the risk of birth injuries. In obese women, a higher incidence of ruptures was
demonstrated. These results are in contradiction to previously published data from different
countries