Výpočetní predikce vedlejších účinků nově vyvíjených léčiv Fibigar Jakub

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Výpočetní predikce vedlejších účinků nově vyvíjených léčiv

Fibigar Jakub

Pracoviště řešitele: Katedra toxikologie a vojenské farmacie, FVZ UO Brno

Školitel a pracoviště školitele: kpt. Mgr. et Mgr. T. Kučera, Ph.D., Katedra toxikologie a vojenské farmacie, FVZ UO Brno

a vojenské farmacie, FVZ UO Brno Abstrakt:

Cílem práce je vytvoření algoritmu a softwarového řešení pro predikci možných vedlejších účinků nových léčiv in silico na základě interakce účinných látek léčiv (ligandy) se známými biomakromolekulami a poskytnutí tohoto řešení odborné veřejnosti.

Pro predikci interakcí léčiv a biomakromolekul je použita metoda reverzního molekulového dokování. Použití proteinové databáze sc-PDB umožňuje identifikaci vazebných míst vhodných pro zadané ligandy na více než 16.000 cílových strukturách.

Za použití software AutoDock Vina a statistického software R je molekulovým dockingem zjištěna interakce všech zadaných ligandů vůči všem biomakromolekulám z databáze proteinů sc-PDB, a dále jsou vybrány relevantní cílové struktury.

Pro vybrané biomakromolekuly platí, že zadané ligandy mohou být jejich substrátem, inhibitore či induktorem a jsou proto vhodné k testování in vitro k analýze jejich případných toxických účinků, jejich nových terapeutických účinků, příp. vysvětlení mechanismů jejich známých účinků.

Přínosem práce je možnost urychlení, usnadnění a zhospodárnění vývoje nových léčiv.

Samozřejmostí je pokračování vývoje, oprav a zdokonalování algoritmů a celého řešení.

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Vztah ligamentum transversum acetabuli k rovině acetabula hodnocený na 3D modelech pánevních kostí

Lejsek Jakub

Pracoviště řešitele: Ústav anatomie LF HK

Školitel a pracoviště školitele: Mgr. M. Kuchař, Ústav anatomie LF HK Abstrakt:

Správná pozice pacienta při totální endoprotéze kyčelního kloubu je nezbytná pro prostorově přesné umístění acetabulární komponenty. Pro zajištění nejpřirozenějšího pohybu by měla osa acetabula směřovat laterálně, dolů a dopředu a ideálně svírat úhel 45° s mediánní rovinou a 15° s rovinou frontální. Díky konstantnímu výskytu, dobré přístupnosti a jednoznačnému průběhu ligamentum transversum acetabuli je jednou z možností polohování pacienta podle průběhu tohoto vazu vzhledem k vertikále. V rámci našeho projektu chceme popsat anatomickou souvislost roviny hrany acetabula s průběhem ligamentum transversum acetabuli a definovat variabilitu tohoto vztahu v rámci současné české populace i mezi oběma pohlavími.

Ve spolupráci s Radiologickou klinikou FN HK jsme získali anonymní CT data více než 100 pacientů Fakultní nemocnice, ze kterých jsme pomocí polo-automatické segmentace vytvořili přes 200 virtuálních modelů pánevních kostí. Na každý z modelů byl namodelován průběh ligamentum transversum acetabuli a ukotven do vertikály, dále proložena rovina uzavírající acetabulum a automaticky změřen vzájemný úhel.

I přes zjištěnou variabilitu se orientace podle průběhu ligamentum transversum acetabuli jeví jako vhodná metodika pro správné umístění acetabulární složky totální endoprotézy kyčelního kloubu.

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Faktory ovlivňující výsledek imunohistochemického vyšetření u historických parafínových bloků

Macháčková Lucie

Pracoviště řešitele: Fingerlandův ústav patologie LF a FN HK

Školitel a pracoviště školitele: MUDr. P. Kašparová, Ph.D., Fingerlandův ústav patologie LF a FN HK

Fingerlandův ústav patologie, Fakultní nemocnice Hradec Králové Abstrakt:

Imunohistochemie představuje nedílnou součást moderní patologie. Slouží nejen k přesnému určení diagnózy, ale často i k přímému ovlivnění léčby konkrétního pacienta (prediktivní markery). V případech, kdy je potřeba revidovat starší materiál a provést nové imunohistochemické vyšetření, je nezbytné mít jistotu, že stáří materiálu významným způsobem neovlivnilo výsledky vyšetření. Cílem naší studie bylo ověřit, zda lze imunohistochemické vyšetření archivních bloků považovat za validní.

Do studie jsme použili archivní parafínové bloky z let 1968, 1978, 1988, 1998, 2008, 2018, obsahující vzorky plicní tkáně s nemalobuněčnými plicními karcinomy. Každý rok byl zastoupen 6 případy. Vyšetřované markery byly zvoleny jednak s ohledem na hledisko klinické (markery diagnostické a prediktivní), jednak podle typu pozitivity barvení (cytoplazmatická, membránová, nukleární). Celkem bylo vyšetřeno 9 markerů. U všech zvolených případů byl nejprve z původního parafínového bloku zhotoven jeden povrchový řez k výběru vhodných oblastí k vyšetření. Vždy byly zvoleny dva vzorky ze samotného karcinomu a dva vzorky z okolní plicní tkáně, které byly použity jako vnitřní kontrola.

Z vybraných oblastí bylo zhotoveno celkem 6 TMA bloků. Každý blok obsahoval 6 případů (po jednom případu z každého roku). Ze složených bloků byly zhotoveny povrchové řezy a řezy v co největší dosažitelné hloubce vzorku. Následně bylo provedeno imunohistochemické vyšetření vzorků (řezy z různých hloubek bloků). Vyhodnocení bylo provedeno porovnáním intenzity barvení v jednotlivých vzorcích, tedy obdobně, jako je imunohistochemické vyšetření hodnoceno v praxi. Bylo pozorováno, zda a jakým způsobem je intenzita barvení u jednotlivých markerů ovlivněna: 1) stářím bloku a 2) hloubkou řezu v rámci bloku.

Z námi získaných výsledků je patrné, že vliv zkoumaných faktorů se lišil nejen u různých typů pozitivity, ale v některých případech i u jednotlivých protilátek. Největší stabilitu barvení jsme pozorovali u cytoplazmatických markerů, kde byla pozitivita uchována téměř beze ztráty, stáří bloku ani hloubka řezu výsledek neovlivnily. Membránové markery vykazovaly větší variabilitu, v některých případech byl výsledek prakticky srovnatelný s cytoplazmatickými markery, u jiných došlo k mírné ztrátě pozitivity vlivem stáří bloku;

v povrchových řezech byla pozitivita slabší než v řezech hlubších. Největší variabilita barvení byla pozorována u nukleárních markerů – od mírného zeslabení intenzity barvení ve starších vzorcích a povrchových řezech, až po téměř úplné vymizení pozitivity jednoho markeru ve starších blocích a jen mírné obnovení barvení v hlubokých řezech.

Z výsledků naší studie vyplynulo, že imunohistochemické vyšetření použitých cytoplazmatických a membránových markerů lze v archivních blocích považovat za validní.

U nukleárních markerů je výsledek variabilní, ovlivněný především konkrétní použitou protilátkou.

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Viral infections in patients after kidney transplantation (5 years retrospective study)

Hoonjan Pryia Kaur, Afshari Moeina

Department of the origin: Department of Clinical Microbiology, Faculty of Medicine and University Hospital, Hradec Kralove

Tutor and Tutor’s Dept.: MUDr. M. Fajfr, Ph.D., Department of Clinical Microbiology, Faculty of Medicine and University Hospital, Hradec Kralove

Abstract:

Aim of study: This study finished the long-term research in topic of infections in patients after kidney transplantation. Previous study was focused on bacteriology and therefore we concentrate on virology. The aim of the following conducted research is to have better understanding of viral infections (and bacterial in previous research) and colonisation during first year after kidney transplantation. The knowledge acquired should lead to more precise prophylaxis and empiric therapy.

Material and methods: The patients’ cohort is made by every patient transplanted at Transplantation Centre (University Hospital in Hradec Kralove) during period 2013 – 2017, for total of 213 patients. Females create 34.3 % (n=73) and males creates 65.7 % (n=140). The viruses associated with complications in immunocompromised patients chosen for research are – herpesviruses (cytomegalovirus -CMV, herpes simplex virus - HSV, varicella zoster virus - VZV and Epstein Barr virus - EBV), hepatitis viruses (HBV, HCV) and polyomaviruses (BK polyomavirus - BKPyV). The study has two parts – direct prove by PCR and indirect diagnosis by serology. For PCR evaluation the quantity criterion is very important. Three levels were used: low (up to 10³), medium (10³-10⁵) and high (over 10⁵).

In serology, we evaluate the serological status before transplantation, reactivations after transplantation and number of days before transplantation and reactivations.

Results: The data from 2013 had shown the discrepancy between PCR and serology. The PCR positivity was lower in all viruses than in serology. The PCR positivity was mostly seen in blood samples (20.0%) but rarely in urine (3.3%). All PCR positivity were in low or medium level. The CMV was reactivated in 20.0% and the BKPyV in 13.3% of cases.

In CMV reactivation, the first diagnosis was made equally by PCR or serology. while CMV reactivation level was found by PCR in 20.0%, the finding in serology was 46.7%. The biggest discrepancies were in EBV reactivation. No PCR prove was made, but according to serology the reactivation was in 46.7% of cases.

Discussion: In the review research, the seroprevalence in patients after renal transplant was the highest for HSV (in 53%), followed by CMV (in 8-32%) and VZV (in 4-12%) (Cukuranovic et al., 2012). According to the recent studies from Germany, the BKPyV had the highest reactivation rate detected by PCR (48.1%), followed by EBV (20.2%) and CMV (17.0%) (Navarro et al., 2018). The Spanish studies has showed the serological reactivation of EBV in 33.5%, but only 12.7% of the reactivation were symptomatic (Lauzurica et al., 2005).

Conclusion: In general, serology showed higher seroprevalence rate than PCR results. This was also shown in our study.

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Novel device for measurement of VEP for clinical practice – Part I

Chua Huey Shin, Miller Katrin Ji-In, Akhtar Sayyeda Niha

Department of the origin: Department of Pathological Physiology, Faculty of Medicine, Hradec Kralove

Tutor and Tutor’s Dept.: prof. MUDr. M. Kuba, CSc., Department of Pathological Physiology, Faculty of Medicine, Hradec Kralove

Abstract:

Two groups of international students worked independently from each other in the Electrophysiological lab to achieve normative data of visual evoked potentials (VEPs) with the use of a new set of visual stimuli for the novel portable VEP device.

This device should enable to extend objective diagnostics of neuro-ophthalmological and psychiatric disorders (outside standard labs) and its certification by International Society for Clinical Electrophysiology of Vision (ISCEV) requires providing of precise age-related norms of the recorded VEP parameters.

The first group has examined 27 volunteers (20 females, 7 males 18 – 27 years old).

Monocular VEPs (dominant eye with normal visual acuity – with correction if needed) were unipolarly recorded from Oz and Pz skin electrodes. The following visual stimulations were used:

- Checkerboard pattern-reversal stimulations with two check sizes 60´ and 15´ (according to ISCEV standards) activating the parvocellular system of the visual pathway

- Radial motion (expansion/contraction of low contrast circular rings) activating the magnocellular pathway

- Visual stimulus with a cognitive task (recognition of red and green patterns in the odd-ball paradigm) activating associative/cognitive parietal visual areas

For each monocular VEP, 20 artefact free epochs were recorded twice. The VEP from the lead with maximum amplitude was selected in each stimulation for statistical evaluation of main peak latencies and interpeak amplitudes.

Results:

- All kinds of the recorded VEPs displayed good reliability/reproducibility – there were no significant differences between results of the repeated measurements, nor between results of both groups.

- There was quite low inter-individual variability of all evaluated VEP parameters which was comparable with the variability of standard VEP examinations (variation coefficient was in the range 6 – 14%).

- After merging of the data from both groups (total number of subjects = 51), norms of VEP latencies were obtained for diagnostics in Ophthalmology (Pattern reversal 60´: 113±7 ms), Neurology (motion VEPs: 167±23 ms) and Psychiatry (cognitive VEPs: 354±37 ms).

Acknowledgements: Financially supported by the project PROGRES Q40/07 of Charles University and by the grant of AZV agency NV18-08-00314.

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Novel device for measurement of VEP for clinical practice – Part II

Babikir Eithar Osama Eltayeb, AttaAlla Marten Reda Youssef Ibrahim, Lasheen Noor Hasan Ahmed Ahmed

Department of the origin: Department of Pathological Physiology, Faculty of Medicine, Hradec Kralove

Školitel a pracoviště školitele: prof. MUDr. M. Kuba, CSc., Department of Pathological Physiology, Faculty of Medicine, Hradec Kralove

Abstract:

Aim of study: A continuance of part I focusing on validity of data within both groups and prospective clinical implications, validation of use of the device in extending the objective diagnosis of neuro-ophthalmological and psychiatric disorders (outside standard labs) for its certification by the International Society for Clinical Electrophysiology of Vision (ISCEV).

Materials and methods:

Two groups of students worked independently of each other in the Electrophysiological lab to achieve normative data of visual evoked potentials (VEPs) with the use of a new set of visual stimuli for the novel portable VEP device. The method of collection of raw data from subjects was identical as to that of group I.

In group II 13 females and 11 males ranging from 18-29 years of age were tested. For each monocular VEP, 20 artefacts free epochs were recorder twice. The VEP from the lead with maximum amplitude was selected in each stimulation for statistical evaluation of mean peak latencies and inter-peak amplitudes.

The Anderson-Darling test of normality was used to verify Gaussian distribution of our result. Thereafter, the statistical significance of the difference was tested using the t-test in the case of a normal distribution of values. A non-parametric test, such as the Wilcoxon test was used in the case, of values not in a normal distribution. As the result of both tests, expressed as p value is over 0.05, we confirm that the compared groups do not differ significantly.

Conclusions:

The following results were yielded:

- No significant difference between the first and second readings within eachgroup

and no significant difference between the results of groups I and II prove reliability of our data.

- Upper limit of latencies (the main diagnostic criterion – M + 2 SD) in all tested VEPs is comparable with standard lab norms and thus the device (stimuli) are well usable in clinical diagnostics.

- Evaluation of sex differences presented significantly shorter latencies in females, with exception of the P300 latency in which females presented a non-significant prolongation of 19 ms compared to men (matter for further research).

Acknowledgements: Financially supported by the project PROGRES Q40/07 of Charles University and by the grant of AZV agency NV18-08-00314.

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Bacterial profile of ocular infections

De Araújo Jorann Attracta

Department of the origin: Department of Clinical Microbiology, Faculty of Medicine and University Hospital, Hradec Kralove

Tutor and Tutor’s Dept.: Mgr. R. Kukla, Ph.D., Department of Clinical Microbiology, Faculty of Medicine and University Hospital, Hradec Kralove

Abstract:

Background: Bacterial ocular infections can lead to conjunctivitis, keratitis and other infections which could progress to severe visual impairment and ultimately even blindness.

Common pathogens causing ocular infections are Staphylococcus aureus, Streptococcus pneumoniae & Haemophilus influenzae. In our study, we focused on bacteria isolated from ocular samples with acute & chronic eye infections in order to analyze trends among species.

Materials and methods: A 4-year (2015-2018) retrospective analysis of bacterial findings was conducted in acute and chronic ocular infections at University Hospital, Hradec Kralove.

2002 ocular samples with culture positivity were tested, out of which 1362 samples were classified as acute and 640 into chronic infections. Conjunctiva or corneal swabs were cultivated on Blood agar, MacConkey agar, Chocolate & Sabouraud agar and in liver broth form multiplication. Bacteria were identified based on morphology of colonies and MALDI- TOF.

Results & Discussion: Coagulase-negative staphylococci were the major group of cultured bacteria- with 830 isolates (60.9%) in acute infections & 354 isolates (55.3%) in chronic infections, with Staphylococcus epidermidis being the most predominantly present.

Nevertheless, they are classified among contaminants, so their presence is necessary to evaluate, in low quantities. Amongst known pathogens, Staphylococcus aureus is predominantly seen in comparison with others, i.e. Streptococcus pneumoniae and Haemophilus influenzae. Upon focusing on Corynebacterium, studies suggest Corynebacterium macginleyi as a possible conjunctiva specific pathogen. It was identified that among corynebacteria this species was identified in 18 (31%) of acute infections and in 17 (50%) of chronic infections. Our results suggest that it’s necessary to consider the importance of Corynebacterium macginleyi and to perhaps evaluate its antibiotic susceptibility during its analysis.

Conclusion: Results indicate that Staphylococcus aureus is the main pathogen isolated from both acute & chronic infections. We also determined a significantly increased number of Corynebacterium macginleyi among other corynebacteria isolated from eye infections.

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Intra-abdominal infections - spectrum of bacteria, retrospective study

Hegazy Hend Mohamed

Tutor and Tutor’s Dept.: Mgr. K. Vaverková, Department of Clinical Microbiology, Faculty of Medicine and University Hospital, Hradec Kralove

Abstract:

Introduction:

Intraabdominal infections is a broad term that includes a number of infectious processes, caused by polymicrobial bacteria. The gut microflora has been found to be a recurrent etiology, such as E.coli & Enterococcus species. Two classifications can be used to subdivide IAIs: uncomplicated or complicated, depending on the extent of the infection;

and community-acquired or hospital-acquired, considering the place of acquisition. IAIs represent a wide range of pathological conditions that include both inflammation of single organs and any type of peritonitis (primary, secondary, tertiary). Moreover, IAIs management generally involves antimicrobial therapy directed against the causative microorganisms and can sometimes require invasive procedure to control the source of the infection.

However, antimicrobial therapy is becoming more difficult due to the arising multi-resistant strains. As a result, IAIs are associated with significant morbidity and mortality rates.

Aim of study:

To evaluate the microbiological findings in patients with intraabdominal infection.

To compare the main pathogens involved in infections from our hospital with general data.

Materials and methods:

Retrospective data gained from the evaluation of cultivated samples obtained from patients treated for intraabdominal infections in University Hospital Hradec Králové. The data were analyzed from a time period of six years from 2014 to 2019. The isolated strains were identified by MALDI – TOF mass spectrometer.

Conclusion:

This retrospective study yielded that the most common pathogens in intrabdominal infections, in descending order are E.coli, Enterococcus species and Bacteriodes species. Which correlates with the worldwide most common intrabdominal pathogens.

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Addition of Hyaluronic Acid in Dental Pulp Stem Cell Cryopreservation and Cultivation Media

Hiebert Emma Yoshiko

Department of the origin: : Department of Histology and Embryology, Faculty of Medicine, Hradec Kralove

Tutor and Tutor’s Dept.: MUDr. T. Soukup, Ph.D., Department of Histology and Embryology, Faculty of Medicine, Hradec Kralove

Abstract:

Aim: The aim of this project was to investigate whether the addition of hyaluronic acid to dental pulp stem cell cryopreservation and cultivation media would change stemness of these cells over time.

Materials & Method: To investigate these changes, addition of hyaluronic acid to the cultivation media was done in two different concentrations: 1 mg/ml and 2 mg/ml. These cells were cryopreserved in 3 different cryopreservation medias: (1) dimethylsulfoxide 10 % + FBS containing media, (2) dimethylsulfoxide 5% + hyaluronic acid 0.1 mg/ml + FBS containing media and (3) dimethylsulfoxide 5% + hyaluronic acid 0.2 mg/ml + FBS containing media.

After defrosting, the cells were further cultivated in the above mentioned media. The hyaluronic acid used in this project was 0.8 MDa.

At each passage, cells were counted and flow cytometry was done to check for stemness markers such as CD49f. A pilot study was conducted on previously cryopreserved cells and then a further study was conducted on 3 freshly isolated independent dental pulp stem cell lines.

Results: Cells that were cryopresyerved in media with hyaluronic acid showed higher stemness after defrosting of cells. Addition of hyaluronic acid to the cultivation media also showed an inital increase in stemness makers.

Conclusion: An intial postive effect on stemness of stem cells was seen when hyaluronic acid was added. The results of this project provide a starting point for further investigation to confirm the effect of hyaluronic acid on the stemness and viability of dental pulp stem cells.

Different stemness and stem cell markers as well as other methods such as immunostaining and qPCR can be used to get more detailed information on how the cells are changing in the new media. We hope that this can increase the amount of time stem cells stay viable after cryopreservation so that they can be used more easily in labs and in medical settings.

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Urinary tract bacteria characterization: retrospective study (age, gender distribution, susceptibility to antibiotics)

Shanker Dhivyaa Bommi

Tutor and Tutor’s Dept.: MUDr. M. Fajfr, Ph.D., : Department of Clinical Microbiology, Faculty of Medicine and University Hospital, Hradec Kralove

Abstract:

Aim: the main aim for this study is to evaluate the bacteria distribution cultured from the urinary tract. We evaluated the possible dependency of isolated bacteria to age and gender of patients and also to the sample type.

Material and Methods: the study cohort is made using all samples from the urinary tract examined in the Department of Clinical Microbiology, University Hospital in Hradec Kralove during one year. The accepted samples were: spontaneously micturated urine, urine from single catheterization, urine from permanent catheterization, urine from nephrostomy. During the study time period 20806 samples were evaluated from 7947 patients. The samples were dominantly from hospitalized patients or hospital out-patients; limited samples were from general practitioners. Males made 48.8% (n=3382) and females made 51.2% (n=4065) of patients. In total 54.9% (n=4364) patients only have single samples and the rest have multiple samples.

Results: The most frequent bacteria found in urine samples were Escherichia coli, Klebsiella pneumoniae and Enterococcus species. The evaluation according to ward type had shown relatively high percentage of negative results in ICU patients. The findings of Candida, Klebsiella and Pseudomonas were highly associated with hospitalization. The samples from elder populations (age 60+) had more positive cultivations (average 66%) than younger population (37%). The single catheterised urine was 47% negative and positive sample were mostly associated with E. coli (50% of positive samples). The urine samples from permanent catheters were highly associated with Candia (82.5% of all Candida positivity) and Providentia, Proteus and Pseudomonas (up to 60% of all cultured strains). The samples from permanent catheterisation were highly associated with resistance (at about 50%

of betalactams, 60% of colistin resistance and 79% of multidrug resistant strains).

Discussion: The results give some interesting findings - high negativity rate in samples from ICU. This founding is probably because these patients are periodically screened without urinary tract infection symptoms. The permanent catheters positivity was made by bacteria with high affinity to foreign material and with the ability to create bio-films. The data from foreign countries shown comparable results. The recent study from India shown the most frequent bacteria both in nosocomial catheter and non-catheter related urinary tract infections as E. coli (Karishetti et al., 2019). The recent study from European countries suggested use of Carbapenems or combined antibiotic therapy for empiric treatment of health care associated urinary tract infection due to high resistance level of causative agents (Tandogdu et al.,2019) Conclusion: The results will be used for more accurate empiric therapy according to age, gender or sample type.

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Resistance patterns of selected Enterobacterales isolated from rectal swabs of patients on ICU

Khalil Omar Ashraf Khalil

Tutor and Tutor’s Dept.: Mgr. R. Kukla, Ph.D., Department of Clinical Microbiology, Faculty of Medicine and University Hospital, Hradec Kralove

Abstract:

Purpose of the study is to assess the prevalence, types and trends of resistance of selected Enterobacterales isolated from rectal swabs of patients hospitalized in Haematology, Metabolic, Pediatric or Intensive Medicine intensitive care units during years 2013-2018. Data for analysis was collected from 4186 bacterial strains out of total 2255 patients. Bacterial species selected for resistance analysis were as follows: Citrobacter spp., Enterobacter spp., Escherichia coli, Klebsiella spp., Morganella morganii, Proteus spp., Providencia rettgeri and Serratia spp. Study aimed on evaluation of specific resistance patterns in above mentioned bacteria including carbapenemases, extended-spectrum beta-lactamases – types ESBL, AmpC and K1 and also on resistance against selected antibiotics such as fluoroquinolones, carbapenems, colistin and aminoglycosides. Analyzed data are being processed continuously and complete results will be presented in SVOC conference.

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Evaluation of the Response of Cells to Royal Jelly Using Cell Proliferation Assays

Haidar Heba M. K.

Department of the origin: Department of Histoloy and Embryology, Faculty of Medicine, Hradec Kralove

Tutor and Tutor’s Dept.: prof. MUDr. J. Mokrý, Ph.D., Department of Histoloy and Embryology, Faculty of Medicine, Hradec Kralove

Abstract:

Background:

Royal jelly is a viscous, whitish to yellow jelly substance that is secreted from the hypopharyngeal gland of the honeybee to nourish the queen bee. It is a bioactive substance that increases the growth and the metabolic processes of bee larvae. Hence why it is added as an active ingredient to skin care products. The aim of this study is to investigate the effect of royal jelly at different concentrations on animal cell cultures using cell proliferation assays.

Materials and Methods:

We evaluated the influence of royal jelly on human embryonic C2C12 myoblasts and primary mouse embryonic fibroblasts. In regards to the C2C12 cells, they were cultured in Dulbecco’s Modified Eagle’s Medium supplemented with 10% foetal bovine serum and 1% penicillin and streptomycin. This was done in a 96 well plate were 1500 cells were cultured per well.

In contrast, the mouse embryonic fibroblasts were cultured 2500 cells per well in a similar medium with exception of 15% foetal bovine serum instead of 10%. Following that, cells were incubated in a CO2 incubator at 37°C for 48 hours. Then we added royal jelly at decreasing concentrations starting with 10 mg/ml and ending with 0.5mg/ml. The positive control cells were cultured in the same medium without royal jelly. The negative control cells were cultured in the same medium with the addition of dimethyl sulfoxide ”DMSO”. I used 60% concentration of DMSO because it is toxic to the cells at this concentration. Afterwards the cells were incubated for another 24 hours. Then 10 μL per well of Cell Proliferation Reagent WST-1 were added and the cells were incubated for another 4 hours. The proliferative activity was measured using spectrophotometry at measurement wavelength of 405 nm, and a reference wavelength of 620 nm. Finally, the data was collected and reordered into an Excel spread sheet.

Results:

So far, we have received only preliminary data that support evidence about a positive effect of royal jelly on both C2C12 myoblasts and the fibroblasts. We have found that the addition of royal jelly has increased the proliferative activity of cells in comparison to the control group in both experiments. For example, data from MEF cells showed 140% proliferation at 10 mg/ml concentration of royal jelly compared to the control cells.As data were received from one experiment only, they need to be verified in repeated experiments. We also plan to expand the spectrum of cells to be tested such as neural stem cells using the neurosphere assay. Unfortunately, due to restrictions associated with the COVID-19 pandemic, the experiments could not be resumed.

Conclusion:

Despite the fact that royal jelly has shown promising results, further testing is needed in order to solidify our conclusions.

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Hyperlipidémie – porovnání atorvastatinu a rosuvastatinu

Chalupová Michaela

Pracoviště řešitele: II. interní gastroenterologická klinika LF a FN HK

Školitel a pracoviště školitele: doc. MUDr. M. Brndiar, CSc.; II. interní gastroenterologická klinika LF a FN HK

Abstrakt:

Ve své práci SVOČ jsem se zabývala porovnáváním účinků dvou preparátů z řad statinů používaných k terapii různých forem hyper-/dyslipidémie. Zvýšený LDL-cholesterol (a pokles HDL) iniciuje proaterogenní patologie ve stěně cév. Léčba statiny by měla vést k výraznému poklesu hladiny LDL-cholesterolu v plazmě, mírně klesají i triacylglyceroly a lehce stoupá HDL-cholesterol. Účinkem statinů tak dochází ke stabilizaci aterosklerotických plátů, optimalizaci endoteliálních funkcí a jiným protektivním dějům. Vztah mezi snížením LDL- cholesterolu a poklesem rizika aterosklerotických kardiovaskulárních onemocnění je brán jako kauzální.

Na vybraném náhodném vzorku šedesáti pacientů s průměrným věkem 68 let jsem porovnávala zvolené naměřené laboratorní biochemické parametry a jejich změnu v čase, kdy pacient daný preparát užíval. Konkrétně se jednalo o transaminázy (ALT a AST), kreatinfosfokinázu (CK), celkový cholesterol (CHOL), LDL-cholesterolu (LDL-C), HDL- cholesterolu (HDL-C) a triacylglyceroly (TAG). K tomu jsem u každého z pacientů zaznamenávala přítomné ovlivnitelné i neovlivnitelné rizikové faktory vzniku aterosklerózy (věk, pohlaví, kouření, arteriální hypertenze, diabetes mellitus, obezita), symptomy možné přítomnosti aterosklerotických změn (např. ischemická choroba srdeční, prodělaný infarkt myokardu) i možné nežádoucí účinky terapie (dyspepsie, myalgie aj.).

Jaterní enzymy je nutné během terapie monitorovat, zvláště u pacientů s poruchou funkce jater, případně ledvin. Mírné přechodné zvýšení transamináz lze při terapii statiny očekávat. U žádného z pacientů našeho souboru nedošlo při kontrolním měření po nasazení terapie k trojnásobnému nárůstu sérových transamináz, jež by bylo klinicky významné. Zvýšením CK by se mj. projevily myositidy či myopatie jako nežádoucí účinek léčby statiny. Desetinásobný nárůst sérové CK svědčící o závažné rhabdomyolýze se v souboru nevyskytl. Získaná data potvrdila, že statiny jsou velmi bezpečným léčivem.

Soubor pacientů jsem rozdělila na dvě hlavní skupiny, 38 pacientů užívajících rosuvastatin a 18 jedinců s terapií atorvastatinem (4 užívali pouze ezetimib), a ty ve vybraných parametrech porovnávala. Pokles celkového cholesterolu i LDL-C byl v průměru vyšší u skupiny s rosuvastatinem, zato HDL-C vyšel příznivěji u atorvastatinu.

K tomu jsem vytvořila další podskupiny, pacienty ženského a mužského pohlaví, kuřáky a nekuřáky, a i u nich výsledky porovnávala. Například u žen došlo k výrazně vyššímu poklesu LDL-C, než tomu bylo u mužů, a u nekuřáků oproti kuřákům nedošlo v průběhu terapie průměrně k navýšení hladiny TAG.

Získané hodnoty, další srovnání a jejich možný přínos uvádím ve své práci SVOČ.

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Výskyt abnormalit EKG u pacientů s mnohočetným myelomem léčených transplantací kostní dřeně

Lejsková Lucie

Pracoviště řešitele: I. interní kardioangiologická klinika LF a FN HK

Školitel a pracoviště školitele: prof. MUDr. R. Pudil, Ph.D., I. interní kardioangiologická klinika LF a FN HK

Abstrakt:

Výskyt nežádoucích kardiovaskulárních účinků protinádorové terapie může významně ovlivnit kvalitu života a prognózu léčených pacientů, u pacientů s mnohočetným myelomem se může v patogenezi výsledného poškození myokardu uplatnit navíc ukládání AL amyloidu do myokardu.

Cíle studie: analyzovat výskyt EKG abnormit u vysoce rizikových pacientů s diagnózou mnohočetného myelomu léčených transplantací kostní dřeně, kteří podstoupili protinádorovou potenciálně kardiotoxickou terapii potenciálně prodlužující QT interval.

Soubor pacientů a metodika: do studie bylo zařazeno 121 pacientů (59 žen), průměrný věk 69±8 let. Pro vyhodnocení byl použit EKG záznam pořízený před transplantací kostní dřeně, analýza digitalizovaných EKG záznamů byla provedena s využitím Image J software.

Výsledky: fibrilace síní byla přítomna u 4 pacientů (3,3%), klidová srdeční frekvence byla 76,5 (±17) tepů/min., průměrná délka PQ intervalu byla 161 (±27,3) ms. Průměrná délka QRS intervalu byla 88,5 (±13,8) ms. Průměrná délka QT/QTc intervalu byla 377,1 (±47,2)/ 419 (±34,6) ms. Prodloužený QTc interval byl nalezen u 1 ženy (1,7%) a 15 (27%) mužů. Poruchy nitrokomorového vedení byly zastoupeny následovně: LAH 10 (8,1%) pacientů, iRBBB u 19 (15,5%) pacientů, RBBB byl zjištěn u 2 (1,6%) pacientů, LBBB u 1 (0,8%) pacientů. Nízkou voltáž v končetinových svodech (amplituda QRS všech svodech ≤ 0,5mV) měli 3 (2,4%) pacientů, nízkou voltáž v prekordiálních svodech (amplituda QRS ve všech svodech ≤ 1,0 mV) mělo 7 (5,7%) pacientů, poruchu progrese R kmitu mělo 30 (24%) pacientů, pseudoinfarktový typ EKG mělo 24 (19,7%) pacientů. Hodnota Sokolowa indexu v prekordiálních svodech byla 2,24 (±0,89) mV.

Závěr: EKG abnormality pacientů s mnohočetným myelomem jsou relativně časté. Mohou odhalit pacienty s preexistující patologií kardiovaskulárního systému či postižení myokardu při samotném onemocnění.

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Výsledky rekonstrukčních operací ušního bubínku

Mrázová Kristýna

Pracoviště řešitele: Klinika otorinolaryngologie a chirurgie hlavy a krku LF a FN HK Školitel a pracoviště školitele: MUDr. L. Školoudík, Ph.D., Klinika otorinolaryngologie a chirurgie hlavy a krku LF a FN HK

Abstrakt:

Úvod: Myringoplastika je rekonstrukční chirurgický výkon, jehož cílem je obnovení mechanické bariéry mezi zevním a středním uchem a zlepšení sluchu.

Cíle: Hlavním cílem práce je posouzení sluchu u pacientů 1 rok po rekonstrukční operaci perforace bubínku. Vedlejším cílem je analýza faktorů ovlivňujících zacelení bubínku po rekonstrukční operaci.

Metodika: Práce vychází z retrospektivního zpracování dat u pacientů operovaných na Klinice otorinolaryngologie a chirurgie hlavy a krku FNHK v období let 2013-2017. Do studie byli zařazeni pacienti operovaní pro perforaci bubínku. Vyřazeni byli pacienti s neúplnou dokumentací. U každého pacienta byl zaznamenán tónový audiogram před operací, 1 měsíc a 1 rok po operaci, získán aritmetický průměr ze 4 hlavních frekvencí (0,5, 1, 2 a 4 kHz) pro vzdušné vedení (AC) a kostně vzdušný rozdíl (ABG). Ze zdravotnické dokumentace byly získány informace o stavu bubínku před operací a 1 rok po operaci, anamnestické údaje a komorbidity. Pro analýzu faktorů ovlivňujících zhojení bubínku jsme sledovali následující faktory: věk, pohlaví, velikost perforace bubínku, lokalizace perforace, materiál použitý k rekonstrukci bubínku, středoušní záněty v anamnéze, jiné středoušní patologie přítomné v době operace, diabetes mellitus, arteriální hypertenze, ICHS.

Výsledky: Ve sledovaném období bylo operováno 84 pacientů pro perforaci bubínku, z toho 49 žen, 35 mužů, věkový průměr 42 let, medián 44 let. Průměrné sluchové ztráty před operací byly 38 dB, ABG 22 dB, ABG ≤ 10dB byl u 8 (10 %) pacientů. Měsíc po operaci byly průměrné sluchové ztráty 30 dB, ABG 11 dB, ke snížení ABG došlo u 74 (88 %) pacientů, ABG ≤ 10dB byl u 40 (48 %) pacientů. Rok po operaci byly průměrné sluchové ztráty 29 dB, ABG 11 dB, k pooperačnímu snížení ABG došlo u 70 (83 %) pacientů, ABG ≤ 10dB byl u 39 (46 %) pacientů. Ke zhojení bubínku po 1 měsíci došlo u 75 (89 %) pacientů, po 1 roce u 78 pacientů (93 %) pacientů. Při analýze faktorů ovlivňujících zhojení bubínku byly významné následující: traumatické perforace, velikost perforace a přidružená recidivující/chronická onemocnění ucha.

Závěr: Práce potvrdila přínos rekonstrukčních operací bubínku pro pacienty s postižením převodního systému ucha. Zlepšení sluchu jsme prokázali u 85 % pacientů. Zacelení bubínku je ovlivněno mechanismem vzniku perforace, velikostí perforace a přidruženými onemocněními středního ucha.

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Relevance stanovení trabekulárního kostního skóre u mužů s osteoporózou na Královéhradecku

Pařízek Vojtěch

Pracoviště řešitele: Ústav klinické biochemie a diagnostiky LF a FN HK, Osteocentrum Školitel a pracoviště školitele: MUDr. L. Pavlíková, MUDr. R. Hyšpler, Ph.D., Ústav klinické biochemie a diagnostiky LF a FN HK, Osteocentrum

Abstrakt:

Úvod: Trabekulární kostní skóre (trabecular bone score, TBS) je relativně nově stanovovaný ukazatel kvality kostní hmoty. V Osteocentru ÚKBD FNHK je rutinně vyšetřován od roku 2018. Principem stanovení je matematický odhad trabekulární struktury lumbálních obratlů pomocí analýzy šedoúrovňové struktury AP DEXA snímku páteře matematickými algoritmy.

Cíle: Hlavním cílem práce je ověřit klinický význam stanovovaní TBS jako samostatného ukazatele rizika osteoporotické fraktury (dále OPF). Vedlejší cíl práce je pokusit se nalézt klinicky významné korelace mezi hodnotou TBS a vybranými parametry (viz Metodika).

Metodika: Do studie byli vybráni muži trpící osteoporózou s určenou hodnotou TBS, kteří navštívili Osteocentrum v roce 2018 nebo 2019. Za hodnocené parametry byly zvoleny věk, TBS, BMD, T-skóre, PINP, CTX-1, přítomnost OPF, základní osteologická diagnóza, užívání glukokortikoidů, nikotinismus a abusus alkoholu. Statistické vyhodnocení bylo provedeno pomocí software SigmaPlot 12.0 (Systat Software, USA) a p < 0,05 bylo považováno za statisticky signifikantní. K vyhodnocení byly použity metody jednocestné analýzy variance a lineární regrese. Data jsou prezentována jako průměr ± směrodatná odchylka.

Výsledky: K vytvoření retrospektivní analýzy dat byl vytvořen soubor 196 mužů, byli vyřazeni pacienti s jinou osteologickou diagnózou než osteoporózou (např. m. Paget, osteogenesis imperfecta, vitamin D rezistentní rachitis a jiné). Zpracován byl tedy soubor 179 pacientů.

Nejmladšímu pacientovi bylo 24 let, nejstaršímu 91 let, věkový průměr byl 65 let a medián 66 let. Práce potvrdila závislost mezi hodnotou TBS a přítomností OPF (průměr TBS u mužů bez OPF byl 1,21 ± 0,118 a u mužů s ní 1,17 ± 0,105, p = 0,039). Práce nepotvrdila statisticky významný rozdíl mezi hodnotou BMD a přítomností OPF (p = 0,709). Dále byla nalezena negativní korelace mezi hodnotou TBS a věkem (TBS = 1,379 – (0,0028×věk), R² = 0,112, p

< 0,001) a pozitivní korelace mezi hodnotou BMD a věkem (BMD = 0,76 + (0,00244×věk), R² = 0,056, p = 0,001). Prokázána byla i souvislost hodnoty TBS a abusu alkoholu (průměr TBS u abstinentů byl 1,23 ± 0,088 a u abuserů alkoholu 1,15 ± 0,116, p < 0,001), zatímco vazba mezi hodnotou BMD a abusem alkoholu prokázána nebyla (p = 0,782).

Závěr: Práce potvrdila význam klinického stanovení hodnoty TBS v osteologické ambulanci.

Pacienti s nižším TBS vykazovali vyšší riziko OPF než pacienti s kvalitnější strukturou kosti.

Dále práce ukázala, že kvalita kosti s věkem klesala, zatímco její hustota vlivem léčby stoupala. Prokázán byl též patologický dopad abusu alkoholu na kvalitu kosti, ne na hustotu.

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Analýza bezpečnosti inhibitorů B-buněčného receptoru v léčbě relabované chronické lymfocytární leukemie v běžné praxi

Ponteš Martin

Pracoviště řešitele: IV. interní hematologická klinika LF a FN HK

Školitel a pracoviště školitele: MUDr. M. Šimkovič, Ph.D., IV. interní hematologická klinika LF a FN HK

Abstrakt:

Úvod: Chronická lymfocytární leukemie (CLL) je nejčastější leukemií dospělých v euroamerické populaci. V posledních letech došlo k revolučním změnám v léčbě této lymfoproliferace. Nově máme možnost ovlivnit signalizaci přes B-buněčný receptor (BCR) pomocí ibrutinibu a idelalisibu. Jedná se o vysoce účinné perorální cílené léky, které jsou obecně velmi dobře snášeny. Nicméně, jejich podávání je ve srovnání s cytostatiky spojeno s odlišným profilem nežádoucích účinků. Informace o toxicitě máme dominantně z randomizovaných studií. Naopak data z běžné praxe jsou omezená. Cíl a metody:

Retrospektivní analýza bezpečnosti léčby relabované CLL inhibitory BCR v období 2015- 2019 ve FNHK. Závažnost toxicity byla určena dle kritérii CTCAE. K určení nezávislých prognostických faktorů doby do sledované události byla použita Coxova regresní analýza.

Výsledky: Celkově bylo zařazeno 88 konsekutivních nemocných s relabovanou CLL. Ibrutinib byl podáván N=60 (mužů 39, 65%, věkový medián 68 let, rozmezí 46-83). Medián počtu předchozích léčebných linií byl 2, rozmezí 1-6. Nepříznivé prognostické znaky: pokročilé stádium dle Raie 37/60 (62%), nemutované IGHV 41/52 (79%), mutace TP53 a/nebo del17p 21/58 (36%), komplexní karyotyp 21/55 (38%). ORR/CR bylo dosaženo u 85/9% pacientů.

Medián sledování byl 12,4 měsíců, odhadovaný medián období do progrese (PFS) byl 32,6 měsíců. Přežití po progresi bylo velmi krátké (medián ≈ 15 dní). Nežádoucí účinky se vyskytly u 92% nemocných; krvácení 42%, průjem 25%, neutropenie 22%, fibrilace síni 20%, exantém 17% a respirační infekce 15%. Výskyt závažné toxicity (stupeň III-V): infekce 27%, neutropenie 22% nemocných. Léčba byla ukončena 18/60 (30%).; pro toxicitu 50% a progresi 50%. Rituximab + idelalisib byl podáván N=28 (mužů 16, 57%, věkový medián 71 let, rozmezí 54-83). Medián počtu předchozích léčebných linií byl 2, rozmezí 1-5. Nepříznivé prognostické znaky: pokročilé stádium dle Raie 18/28 (64%), nemutované IGHV 19/24 (79%), mutace TP53 a/nebo del17p 10/27 (37%), komplexní karyotyp 10/24 (42%). ORR/CR bylo 81/9%. Medián sledování byl 17,4 měsíců, odhadovaný medián PFS byl 13,9 měsíců.

Přežití nemocných po ukončení léčby bylo velmi krátké. Nežádoucí účinky se vyskytly u 93%

nemocných: neutropenie 39%, průjem 32%, exantém 29%, respirační infekce 18%, myoskeletální bolest 18% transaminitida 18%. Výskyt toxicity stupně III-V: infekce 25%, neutropenie 39% nemocných. Léčba byla ukončena 17/28 pacientů (68%). Důvody ukončení léčby: toxicita 47% a progrese CLL 53%. Bude prezentována analýza prediktivního významu výskytu nežádoucích účinků pro dosažení ORR a PFS. Závěr: Naše výsledky ukazují, že výskyt závažných komplikací při léčbě relabované CLL pomocí inhibitorů BCR je v běžné praxi vyšší, než udávala data z registračních studií. Nežádoucí účinky byly častou příčinou předčasného ukončení léčby ibrutinibem či idelalisibem.

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Melanoma survival rates in the East Bohemia region; the relation between the time of diagnosis, histology, sex and survival

Litvina Olha

Department of the origin: Department of Dermatology and Venereology, Faculty of Medicine and University Hospital, Hradec Kralove

Tutor and Tutor’s Dept.: doc. MUDr. J. Čelakovská, Ph.D., Department of Dermatology and Venereology, Faculty of Medicine and University Hospital, Hradec Kralove

Abstract:

Introduction: The incidence of melanoma skin cancer has been increasing worldwide in recent decades. Thus, it raises many questions concerning the survival of the patients and factors influencing the prognosis.

Aim: To evaluate the survival rates of patients diagnosed with melanoma skin cancer in the East Bohemia region of the Czech Republic, and determine, whether the time (year) of diagnosis, sex of the patient and histology of the lesion all have an impact on the survival rates.

Methods: This is an epidemiological study based on the examination of patients with a new histopathologically confirmed primary melanoma (lentigo maligna, melanoma in situ, superficial spreading melanoma or nodular melanoma) in the period 1996-2017. Our study pool is 2887 patients, including the deceased patients that died from melanoma skin cancer.

All patients were examined and diagnosed at the Department of Dermatology and Venereology, University Hospital, Hradec Králové.

Results: The cumulative survival decreases with time (5-year cumulative survival - 89.68%).

However, higher rates of survival (5-year survival) were observed among the patients diagnosed in the more recent years, 2006-2010 (92.65%) and 2011-2015 (94.88%), rather than in 1996-2000 (81.18%) and 2001-2005 (84.74%). The lowest 5-year survival rates were in patients with nodular melanoma (84.42%), the highest – in the cases of lentigo maligna diagnosis (99.45%). The research has shown no significant impact of sex on survival (p=0.0576).

Conclusion: This study has confirmed the impact of the time (year) of diagnosis and histology of the lesion on the survival rates. The results of our study are comparable with studies from other countries.

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Gene-Environment interactions in Schizophrenia, Bipolar Disorder, Depression and Anxiety Disorder

Raj Divyam

Department of the origin: Department of Psychiatry, Faculty of Medicine and University Hospital, Hradec Kralove

Tutor and Tutor’s Dept.: prof. MUDr. L. Hosák, Ph.D, Department of Psychiatry, Faculty of Medicine and University Hospital, Hradec Kralove

Abstract:

A GxE interaction occurs when a genetically predisposed individual is hit by a strong environmental factor. This alters the individual’s gene expression, making them more susceptible to develop an illness. The diseases mentioned (Schizophrenia, Anxiety Disorder, Depression and Bipolar Disorder) all are disabling psychiatric disorders with long-term treatment and disruption of an individual’s life. Concerns have been raised as the diseases are now more prevalent than ever. Increased occurence can be seen in large cities, drug abusers and traumatised individuals proving an environmental link. In developed nations with longer life spans, the number of patients with major psychiatric disorders is expected to rise in the future. This presentation aims to provide insight on research being done right now to combat this trend. It also aims to provide some examples of gene-environment interactions and how large scale studies have proven this “Two-Hit” hypothesis and that we need to consider both factors when we risk stratify individuals for these conditions.

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The role of wireless video capsule enteroscopy in the screening of colorectal cancer in the average-risk Czech population

Sazzad Sadman Bin

Department of the origin: 2nd Department of Internal Medicine-Gastroenterology, Faculty of Medicine and University Hospital, Hradec Kralove

Tutor and Tutor’s Dept.: doc. MUDr. I. Tachecí, Ph.D., 2nd Department of Internal Medicine-Gastroenterology, Faculty of Medicine and University Hospital, Hradec Kralove Abstract:

Background & Aim: The Czech National colorectal cancer (CRC) screening program (launched in 2000, focusing on the average-risk population over 50 years) has undoubtedly a positive effect on the mortality and incidence of this disease in the past years. The main disadvantage of this program is low adherence of the target population (fear of procedures, especially a standard flexible high-resolution video-colonoscopy (FVC)). Wireless colon capsule endoscopy (CCE) can be a non-invasive solution acting as a pre-selection before FVC. FVC still remains the therapeutic method for cases with confirmed premalignant or malignant colorectal lesion only. The main aim of our project was to verify the feasibility of CCE and its diagnostic yield compared with FVC. Another aim was to evaluate the possible role of students in the pre-assessment of CCE.

Methods: Both CCE and FVC were performed after fortified bowel-cleansing in asymptomatic subjects (aged 50 – 75 years, with no history of colorectal neoplasia and family history of CRC). FVC was performed by an experienced endoscopist. CCE was blindly evaluated by an endoscopist and a medical student (after sufficient training). All pathological findings were classified as non-neoplastic or neoplastic lesions (including adenomas, neoplastic serrated lesions and carcinomas). The findings of CCE were evaluated in comparison to FVC as a gold standard for colon examination.

Results: Twenty asymptomatic subjects were enrolled in the study (11 men and 9 women, mean age 59. No complication was observed at any time. 5 CCE were terminated before reaching the rectum. A total of 63 neoplastic lesions between 4 and 30 mm were diagnosed by FVC in 14 subjects. The CCE evaluated by an endoscopist diagnosed a total of 53 lesions in 15 patients (1 was false positive). Evaluated by a student, 45 lesions in 18 patients were found (6 false positive cases). Diagnostic yield of the student (adjusted for false positive findings) reached 62 %. Lesion missed by the student were flat-lesions (4), serrated-lesions (3) and lesions with diameter < 5 mm (15), right-sided (16) and cases with bad mucosal visibility due to poor bowel-cleansing (14). One cancer diagnosed by FVC, was correctly identified both by the endoscopist and student during the CCE.

Conclusions: The CCE as a non-invasive procedure has a high potential to become a pre- selective investigation before FVC. So far, major limits for the involvement of students in the evaluation of CCE were difficulties in proper recognition of some lesions (flat, serrated or right-sided lesions). It could be overcome by further training.

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Prophylaxis Indomethacin treatment for prevention of Post ERCP Pancreatitis

Wisal Maham

Department of the origin: 2nd Department of Internal Medicine - Gastroenterology, Faculty of Medicine and University Hospital, Hradec Kralove

Tutor and Tutor’s Dept.: MUDr. Š. Šembera, MUDr. K. Zhychko, prof. MUDr. S. Rejchrt, Ph.D., 2nd Department of Internal Medicine - Gastroenterology, LF and FN HK

Abstract:

Endoscopic retrograde cholangiography (ERCP) has become a widely used procedure for therapy of pancreaticobiliary diseases. Acute Post ERCP pancreatitis(PEP) remains one of the concerning complications.The incidence of PEP varies from 3.5-9.7%.varying depending on the procedure related factors.Rectal Indomethacin suppository has been supported as prophylactic prevention of PEP, explained by its ability to inhibit pathways involved in the pathogenesis of PEP. The study aimed to evaluate the effect of the standard use of Indomethacin to prevent PEP in routine clinical practice. Methods: A retrospective cohort study conducted by collecting data from patients hospitalised in FNHK,undergoing ERCP from 3/2013 to 12/2015.666 patients belonged to the group before routine use of prophylactic Indomethacin was recommended and 606 after. Furthermore, two subgroups of 100 consecutive patients were randomly selected from each group to asses in detail the procedure risk factors for PEP. We identified patients with PEP and assessed them for these risk factor as well. The criteria for pancreatitis were amylase level 3x upper normal limit 24 hrs after procedure and abdominal pain.Results: A total of 1272 patients were analysed. In the first group before standard use of Indomethacin, the average of patients was 68.6(SD 13.9), 47% of the patients were male. Procedure-related PEP risk factors were pre-cut sphincterotomy in 16% and wirsung cannulation in 34%. In the second group average age of patients was 66.7 years (SD 14.7), and 49% were male, pre-cut sphincterotomy in 23% and wirsung cannulation in 44%. In the first group no Indomethacin was used, whereas in the second group 100 mg Indomethacin was administered to 84% of the patients immediately after the procedure. PEP was observed in 2 patients (3.15%) in the first group and in first group and in 17patients (2.81%)in the second group with Indomethacin prophylaxis.Procedure-related risk factors for PEP in patients with PEP were in the first and second group respectively: wirsung cannulation 38.1%/70.6% and pre-cut sphincterotomy 23.8/58.8. Second group with PEP received Indomethacin prophylaxis only in 43%. The risk ratio of standard Indomethacin use was calculated to be 0.89 (p-value 0.7). Conclusion:

Effectiveness of Indomethacin was not proved in our study. There was a low incidence of PEP with no statistically significant change. Secondly, the adherence to recommended guideline was not optimal which should be improved.required to demonstrate statistically significant protective effect. Secondly, the adherence to recommended guidelines was not optimal which should be improved.

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Lawsonella clevelandensis; case report and literature review

Ramesh Rommel

Department of the origin: Department of Infectious Diseases, Mayo Clinic, Rochester, USA Tutor and Tutor’s Dept.: Zerelda Esquer Garrigos MD., Department of Infectious Diseases, Mayo Clinic, Rochester, USA

Abstract:

Background: Vascular graft infections (VGI) pose a significant risk of morbidity and mortality. In up to 50% of VGI cases, the causative organism is not established by conventional culture. With the incorporation of 16S ribosomal (r) RNA gene PCR methods, a causative agent may be identified from clinical samples of extracted vascular grafts. We report the first case of VGI due to Lawsonella clevelandensis, an anaerobic Gram- positive, partially acid-fast, fastidious organism identified using 16S ribosomal rRNA gene PCR approach.

Materials/methods: We reviewed medical records of a 65-year old patient with inferior abdominal aortic aneurysm, status post endovascular aneurysm repair with stent graft who presented with a one-month history of progressive bilateral inguinal pain, fever, chills, night sweats and unintentional weight loss of 30 pounds.

Results: Labs revealed leukocytosis of 17.5 x10⁹ /L, elevated C-reactive protein at 132.8 mg/L, erythrocyte sedimentation rate at 100mm/hr, and thrombocytosis of 758x10⁹ /L.

Positron emission tomography showed high fluorodeoxyglucose avidity within the aneurysm wall and graft, along with bilateral psoas abscesses. Vascular graft was removed and replaced with cryopreserved aortobi-iliac graft. Operative specimens including aortic aneurysm sac fluid and aortic tissue were obtained. Aerobic and anaerobic bacterial cultures, fungal culture, and mycobacterial cultures, along with corresponding direct smears, were negative.

Histopathology revealed mild to moderate acute inflammatory infiltrate and a cluster of weakly acid-fast, Gram-variable bacilli. A 16S rRNA gene PCR identified L.

clevelandensis. As the organism did not grow on cultures for susceptibility testing, the patient was treated with a six-week course of intravenous vancomycin and oral doxycycline therapy, followed by chronic suppression with cefadroxil. The patient’s symptoms resolved, and he recovered wel since surgery.

Conclusions: L. clevelandensis is a highly fastidious anaerobic organism, primarily detected by molecular methods. Clinical infection due to this organism appears to be characterized by localized abscess formation. Cases of disseminated infection have not been reported in the literature. Our case highlights the importance of incorporating molecular methods in cases of vascular graft infection, where conventional cultures are negative.

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The importance of wireless video capsule enteroscopy in obscure gastrointestinal bleeding

Thirumushi Sairaj Revathi

Department of the origin: 2nd Department of Internal Medicine - Gastroenterology, Faculty of Medicine and University Hospital, Hradec Kralove

Tutor and Tutor’s Dept.: doc. MUDr. I. Tachecí, Ph.D., 2nd Department of Internal Medicine - Gastroenterology, Faculty of Medicine and University Hospital, Hradec Kralove Abstract:

Background and Aim: Wireless video capsule Enteroscopy (CE) is a non-invasive endoscopic investigation indicated especially in the small bowel diseases. It is the first choice diagnostic method in patients with obscure occult gastrointestinal bleeding (OGIB) after negative standard endoscopic procedures (gastroscopy and colonoscopy). Many studies were published with different diagnostic yield of CE in this indication. The main problem is different interpretation of pathological findings as a potential source of bleeding. The aim of our study was to evaluate the importance of CE in OGIB and to set possible role of medical students in pre-assessment of the CE recordings.

Methods: We reviewed medical records and CE of patients investigated for OGIB, who underwent CE at our University Hospital. Examinations were performed according to a standard CE protocol (12 hours of fasting without special small bowel cleansing). CE records were evaluated by a medical student, who was blinded to medical data. Her evaluations were exported as thumbnails, classified according to bleeding potential and compared with final diagnosis set by an experienced endoscopist. The main output of the study was diagnostic yield of CE in OGIB and the agreement between student’s and endoscopist’s findings.

Results: Twenty patients were enrolled (14 men, 6 women, mean age 61 years). Eleven subjects underwent CE for overt obscure (melaena) and 9 for occult obscure ( iron-deficiency anaemia) gastrointestinal bleeding. No complications were observed during all CE procedures.

Pathological findings were diagnosed in 16 cases (80 %) (by an endoscopist) and in 14 cases (70 %) (by a student). Lesions with a high bleeding potential (diagnostic yield of CE in OGIB) were observed in 8 cases (40 %, by an endoscopist) and in 13 cases (65 %, by a student). The CE evaluated by a student was false positive (the pathological finding was misinterpreted as a source of bleeding) in 5 cases (25 %), we did not observe any false negative finding.

Conclusion: CE is a diagnostic method with sufficient diagnostic yield in patients with OGIB.

Pre-assessment of CE by a trained medical student can be beneficial. High proportion of false- positive findings can be corrected during the second reading by an experienced endoscopist.

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Objective measures of muscle strength and fatigue using a new portable device

Bhorkar Aprajita Milind

Department of the origin: Department of Neurology, Faculty of Medicine and University Hospital, Hradec Kralove

Tutor and Tutor’s Dept.: MUDr. O. Vyšata, Ph.D., Department of Neurology, Faculty of Medicine and University Hospital, Hradec Kralove

Abstract:

The degree of muscle weakness is difficult to quantify using existing methods, as both manual testing and dynamometric evaluation require patient cooperation.

Ultrasound examinations can be used to assess muscle thickness, however this does not directly indicate the degree of muscle strength.

Similarly, conductive studies may indicate the amount of active motor units, while needle EMG indicates the pathology of the neuromuscular unit.

However, even these methods do not directly indicate loss of muscle strength.

The aim of this work is to estimate the variability of repeated measurements using a device for the objective measurement of muscle strength, in addition to finding predictors of muscle strength in a healthy population

Our device measures the muscle strength and fatigue of the tibialis anterior muscle using increasing current intensity stimulation at the motor point of the muscle.

The 2 measurements were performed on 27 healthy volunteers, and the association between the values, sex, weight, height, calf circumference and degree of training was measured by the Pearson product-moment correlation coefficient.

This method was found to possess excellent reproducibility with physical endurance being the only significant predictive factor for the strength curve. There were no differences found between men and women

Unlike manual methods, this device is capable of measuring strength in:

- Unconscious patients

- Non-cooperating patients (with cognitive deficiency, sensoric aphasia, apraxia, neglect syndrome, depression, catatonic, simulating)

- Patients with central motoneuron lesions

- Patients with peripheral nerve conduction blocks, demyelinating or polyneuropathies

Currently there is no alternative to measuring muscle strength and fatigue in the above- mentioned disorders.

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New Approaches to Motion Analysis in Multiple Sclerosis Patients

Ibrahim Mohamed Hagar

Tutor and Tutor’s Dept.: MUDr. O. Dostál, Faculty of Medicine and University Hospital, Hradec Kralove

Abstract:

In clinical practice, the classification of physical disability in patients with Multiple Sclerosis (MS) is achieved by using the clinical scale „EDSS“ -Expanded Disability Status Scale.

However, EDSS has limited sensitivity and large variability in grading, increasing the need for precise and independent measurement tools for movement impairment.

Our patients and healthy control subjects were geared with Perception Neuron Suit- a Motion Sensor Suit containing 32 sensors each composed of gyroscope, accelerometer and magnetometer- and an EEG cap, the experimental data sets were acquired while performing a neurological exam on the participants. We used the data for complex motion analysis regarding the physical disability of the patients, which is showing good potential of use in early diagnostics and further patient monitoring.

The entire study is based on observations in the clinical environment, and suggests the importance of augmented reality to decision making and diagnostics in neurology.

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Dependency of Recanalization times on Presenting Symptoms in Acute Ischaemic Stroke

Hamza Abduljabar

Tutor and Tutor’s Dept.: MUDr. S. Halušková, Department of Neurology, Faculty of Medicine and University Hospital, Hradec Kralove

Abstract:

Anterior circulation stroke (ACS) is associated with typical symptoms, while posterior circulation stroke may cause a wide spectrum of less specific symptoms. A consecutive series of patients treated with intravenous thrombolysis (IVT) and endovascular treatment (EVT) (±

IVT) at a single-center included in a prospective registry was retrospectively reviewed to investigate 1) the impact of initial presenting symptoms and affected vascular territory on recanalization times and 2) the dependency of 90-day clinical outcome on the initial symptoms, achieved recanalization times (in IVT and EVT [± IVT] groups) and on the degree of achieved recanalization (in EVT [± IVT] group). In the IVT group, severe neurological deficit on admission and speech difficulties were associated with shorter and nausea/vomiting with longer onset-to-needle time and, vertigo with longer door-to-needle time (DNT). In the EVT (± IVT) group, coma was associated with longer DNT, ACS with shorter onset-to-groin time and, drooping of the mouth corner with shorter door-to-groin time. Thus, treatment was initiated later in stroke with less specific symptoms than in ACS with typical symptoms.

In both groups, age and neurological deficit on admission were independent negative predictors of good 90-day outcome and independent positive predictors of 90-day mortality.

Hemiparesis was another independent negative predictor of good 90-day outcome in the IVT group. In the EVT (± IVT) group, successful recanalization was another independent positive predictor of good 90-day outcome, speech difficulties an independent negative predictor and the occlusion of the intracranial internal carotid artery an independent positive predictor of 90-day mortality.