Influence of CD34+ cells transenocardial transplantation on diastolic function of the left ventricle in patients with dilative cardiomyopathy

IZVLEČEK Izhodišča in hipoteze. Diastolična disfunkcija levega prekata je napovednik naravnega poteka srčnega popuščanja z ohranjenim ali zmanjšanim iztisnim deležem levega prekata. Učinek transplantacije matičnih celic, ki se uporablja za zdravljenje napredovalega srčnega popuščanja, na diastolično funkcijo levega prekata ni jasen. Zato smo z doktor¬skim delom ocenili, ali i) so ehokardiografski parametri diastolične disfunkcije povezani s parametri, ki jih pridobimo z elektroanatomsko kartografijo pred transplantacijo matičnih celic, ter ii) ali presaditev CD34+ matičnih celic izboljša diastolično funkcijo bolnikov s srčnim popuščanjem. Metode dela. V raziskavo smo vključili zaporedne bolnike z neishemično in ishemično dilatacijsko kardiomiopatijo, ki so bili predvideni za zdravljenje s CD34+ matičnimi celicami. Bolniki so imeli iztisni delež levega prekata manj kot 40 %, bili so v sinusnem ritmu, v klinično stabilnem stanju (funkcijski razred III po klasifikaciji newyorškega kardiološkega združenja) vsaj tri mesece pred vključitvijo. Bolnike smo razdelili v skupine glede na etiologijo kardiomiopatije in izhodiščno vrednost ehokardiografskega znaka povišanega polnitvenega tlaka levega prekata (E/e\u27 ? 15) v podskupine. Pri vseh bolnikih smo iz periferne krvi mobilizirali CD34+ celice s pomočjo stimulacije kostnega mozga s filgrastimom in jih izolirali z aferezo. Z elektroanatomsko kartografijo smo določili glede na velikost unipolarnega endokardnega potenciala in linearno skraj¬šanje srčne mišice (LLS) predele zdrave (povprečen unipolarni potencial ? 8,27 mV in LLS ? 6 %) in hibernirane (povprečen unipolarni potencial ? 8,27 mV in LLS < 6 %) srčne mišice in predele brazgotine (povprečen unipolarni potencial < 8,27 mV in LLS < 6 %). Pri bolnikih z neishemično kardiomiopatijo smo izmerili tudi globalni čas relaksacije levega prekata in ga primerjali z ehokardiografskimi parametri diastolične disfunkcije. Matične celice smo vbrizgali transendokardno v predele hibernirane srčne mišice. Ob vključitvi v študijo in nato 3, 6 in 12 mesecev po transplantaciji CD34+ celic smo opravili ehokardiografsko preiskavo z oceno diastolične disfunkcije in spremljali klinične (klinični pregled in 6MWT) in laboratorijski pokazatelj srčnega popuščanja (nivo NT-proBNP v krvi). Diastolično disfunkcijo smo ocenjevali z ehokardiografskimi parametri E/A, DT, TR in E/e\u27 in skladno s priporočili ASE/EACVI opredelili stopnjo diastolične disfunkcije. Rezultati. Vključili smo 58 bolnikov (povprečna starost vseh bolnikov je bila 54 ± 8,4 let20,8 % je bilo žensk). Neishemično kardiomiopatijo je imelo 38 bolnikov – 18 s povišanim polnilnim tlakom (skupina DKMP+), 20 z normalnim polnilnim tlakom (skupina DKMP?). Ishemično kardiomiopatijo je imelo 20 bolnikovvsi so imeli povišan polnilni tlak (skupina IKMP+). Z endomiokardnim beleženjem unipolarnih potencialov smo v skupini DKMP+ ugotovili večji obseg hiberniranega miokarda kot v skupini DKMP? (4,9 ± 2,7 mV in 2,7 ± 2,9 mVp = 0,03) in večji obseg brazgotin (2,2 ± 1,6 mV in 0,9 ± 1,1 mVp = 0,02). Globalni čas relaksacije pri elektroanatomski meritvi ni koreliral s parametri diastolične funkcije (E/A, DT in stopnjo diastolične disfunkcije po ASE/EACVI), je pa koreliral s parametrom polnilnega tlaka E/e\u27. Transplantacija CD34+ ni bila povezana z značilno spremembo parametrov diastolične disfunkcije ali koncentracijo NT-proBNP v nobeni izmed skupin, bila pa je povezana z izboljšanjem 6MWT (DKMP+: 48 ± 58 mp = 0,026DKMP?: 52 ± 63 m, p = 0,05IKMP+: 96 ± 62 mp < 0,001). Transplantacija CD34+ celic je bila povezana z značilnim upadom v parametru polnilnega tlaka levega prekata E/e\u27 le v skupini DKMP+ (?8,0 ± 10,3 mmHgp = 0,008), ne pa v skupinah DKMP– in IKMP+. Zaključki. Podaljšan globalni čas relaksacije miokarda, izmerjen s pomočjo elektro¬ana-tomske mape, korelira s parametrom zvišanega polnilnega tlaka E/e\u27, ne pa z ostalimi parametri diastolične disfunkcije. Zdravljenje s CD34+ matičnimi celicami zniža polnilni tlak levega prekata (E/e\u27) le pri bolnikih z DKMP+ (ki imajo že zvišan izhodiščni polnilni tlak), ne pa pri bolnikih z DKMP? (kjer je parameter E/e\u27 izhodiščno normalen) in z IKMP+ (kjer je obseg brazgotin večji). Izsledki raziskave nakazujejo novo možnost ocene zvišanega polnilnega tlaka levega prekata s pomočjo elektroanatomske mape in naka¬zujejo možnost ustreznejšega izbora kandidatov za zdravljenje s CD34+ matičnimi celicami. Ključne besede. Kardiomiopatija, CD34+ celice, diastolična, ehokardiografija, elektro-anatomska mapaABSTRACT Background and hypotheses. Left ventricular diastolic dysfunction is a predictor of the natural course of heart failure with a preserved or reduced left ventricular ejection fraction. The effect of stem cell transplantation used to treat advanced heart failure due to left ventricular diastolic dysfunction is unclear. The doctoral thesis evaluated whether i) echocardiographic parameters of diastolic dysfunction are related to parameters obtained by electroanatomical mapping prior to stem cell transplantation, and ii) whether CD34+ stem cell transplantation improves the left ventricular diastolic function of patients with advanced heart failure. Methods. We included consecutive patients with non-ischemic and ischemic dilated cardiomyopathy who were enrolled for a treatment with CD34+ stem cells. Patients had less than 40 % left ventricular ejection (LVEF), were in sinus rhythm, clinically stable (NYHA class III according to the New York Cardiac Association) for at least three months prior to enrollment. Patients were divided into groups according to the etiology of cardiomyopathy and baseline echocardiographic sign of elevated filling pressure (E/e\u27 ⡥ 15) into subgroups. In all patients, CD34+ cells were mobilized from peripheral blood by bone marrow stimu-lation with filgrastim and isolated with apheresis. Electroanatomical mapping determined, according to the size of the unipolar endocardial potential and linear shortening of the heart muscle (LLS), areas of healthy (average unipolar potential ⡥ 8.27 mV and LLS ⡥ 6 %) and hibernated (average unipolar potential ⡥ 8.27 mV and LLS < 6 %) myo¬cardi-um, and scars (average unipolar potential < 8.27 mV and LLS < 6 %). We also measured global left ventricular relaxation time in patients with non-ischemic cardiomyopathy and compared it with echocardiographic parameters of diastolic dysfunction. Stem cells were injected transendocardially into areas of hibernated cardiac muscle. When enrolled in the study and 3, 6 and 12 months after CD34+ cell transplantation, an echocardiographic examination was performed to assess diastolic dysfunction and to monitor clinical (clinical examination and 6MWT) and laboratory parameters of heart failure (NT-proBNP blood level). Diastolic dysfunction was assessed by echo¬cardio-graphic parameters E/A, DT, TR and E/e\u27, and the degree of diastolic dysfunction was determined in accordance with the ASE/EACVI recommendations. Results. We included 58 patients (mean age of all patients was 54 ± 8.4 years, 20.8 % were women). Non-ischemic cardiomyopathy had 38 patients ⠒ 18 with elevated filling pressure (DKMP+ group), 20 with normal filling pressure (DKMP⠒ group). Twenty patients had ischemic cardiomyopathyall had elevated filling pressure (IKMP+ group). Endomyocardial recording of unipolar potentials resulted in a greater area of hibernated myocardium in the DKMP+ group than in the DKMP⠒ group (4.9 ± 2.7 mV and 2.7 ± 2.9 mVp = 0.03) and a greater area of scars (2.2 ± 1.6 mV and 0.9 ± 1.1 mVp = 0.02). Global relaxation time in electroanatomical measurement did not correlate with diastolic function indicators (E/A, DT and ASE/EACVI diastolic dysfunction rate), but correlated with E/e\u27. CD34+ transplantation was not associated with a significant change in diastolic dysfunction indicators or NT-proBNP concentration in any of the groups, but was associ-ated with improvement in 6MWT (DKMP+: 48 ± 58 mp = 0.026: DKMP⠒: 52 ± 63 mp = 0.05: ICMP+: 96 ± 62 mp <0.001). CD34+ cell transplantation was associated with a significant decrease in the left ventricular E/e\u27 pressure indicator only in the DKMP+ group (⠒8.0 ± 10.3p = 0.008) and not in the DKMP– and IKMP+ groups. Conclusions. Prolonged global myocardial relaxation time, measured using an electro-anatomical map, correlates with the E/e\u27 elevated filling pressure indicator but not with other indicators of diastolic dysfunction. Treatment with CD34+ stem cells lowers left ventricular filling pressure (E/e\u27) only in DKMP+ patients (who have baseline elevated filling pressure), but not in DKMP– patients (where the E/e\u27 indicator is baseline) or IKMP+ (where the extent of the scars is larger). The results of the study indicate a new possibility of assessment of elevated left ventricular filling pressure using an electro-anatomical map and indicate the possibility of a more appropriate selection of candidates for treatment with CD34+ stem cells

[Cisteinski katepsini, stefini in razgradnja izvenceličnega matriksa med invazijo človeških transformiranih celičnih linij raka dojke]

Background. Human breast cellular model, comprising four cell lines originating from spontaneously immortalized human breast epithelial MCF10A cell line, its c-Ha-ras transfectant, MCF10AT, and two tumourigenic derivatives, cultured from two sequential mouse xenographs, MCF10AT-Ca1a and MCF10AT-Ca1d, were used to compare the relative protein concentration of cathepsins and stefins in single cells. Methods. The relative protein concentration of cathepsins and stefins in single cells was analysed by confocal microscopy, and compared to their protein expression in cell homogenates. Results. The most invasive, MCF10AT cell line contained several fold higher protein concentration of cathepsin B and increased levels of stefins, but similar levels of cathepsin L, compared with the parental MCF10A cells. This was associated with five fold higher endocytosis of Matrigel-DQ-collagen IV (DQC) and a simultaneous increase in signal overlap between DQC and cathepsin L as well as DQC and stefin B, but a decrease in that of DQC and cathepsin B overlap in the MCF10AT cells. Simultaneously, increased signal overlaps between both cathepsins and between cathepsins-stefins pairs, were observed in this cell line. Conclusions. These results suggest that the increased collagen endocytosis and degradation in theinvasive phenotype significantly affect also the subcellular localization of cysteine cathepsins and stefins. Based on these and the reports of other authors, we hypothesize that the intracellular degradation may also be assoeiated with cathepsin L, whereas cathepsin B in the ras transformed breastcells is involved in both, the intracellular and pericellular degradation of extracellular matrix during cell migration and invasion

ANALYSIS OF THE EXPOSURE TO STRESS AT THE WORKPLACE IN THE SELECTED ORGANIZATION

Stres je stalni spremljevalec našega vsakdanjega poklicnega in zasebnega življenja. Vsakdo se je že vsaj enkrat v življenju srečal s stresom, bodisi v pozitivnem bodisi v negativnem smislu. V preteklosti je bilo zaradi stresa ogroženo telesno zdravje ljudi, danes pa se vse bolj odraža prek psiholoških učinkov. Današnji način življenja se hitro spreminja in razvija, od posameznika zahteva nenehno spremljanje in sprejemanje sprememb ter hitro prilagajanje nanje. Posameznik se je prisiljen s spremembami soočiti, jih sprejeti ter vključiti v svoje življenje in način dela. Spremembe v načinu življenja in razvoj tehnologije pa morajo upoštevati tudi organizacije, saj jim to omogoča nadaljevanje svoje poslovne poti in s tem preživetje. V diplomskem delu je najprej predstavljeno področje stresa, nato pa so podani njegovi vzroki in posledice – tako za posameznika kot za organizacijo. Namen raziskave, ki je bila izvedena v farmacevtski organizaciji, je bilo ugotoviti prisotnost stresa glede na spol, delovno mesto, izobrazbo in starost. Rezultati raziskave, ki je bila izvedena z metodo anketiranja, so pokazali zmerno raven stresne ogroženosti v organizaciji. Glede na rezultate so stresu bolj izpostavljene ženske, zaposleni na nevodstvenih delovnih mestih, zaposleni z višješolsko/visokošolsko izobrazbo in najstarejša zaposlena oseba v organizaciji. Rezultati kažejo, da se organizacija zaveda prisotnosti stresa in izvaja aktivnosti v smeri njegovega zmanjševanja. Kljub temu se kot pomoč pri zmanjševanju stresa predlaga še dodatne aktivnosti – izobraževanja, srečanja, komunikacijo in merjenje vzdušja v organizaciji.Stress is our constant everyday companion in the professional as well as in the private life. Everyone has to deal with stress, be it in a positive or negative way, at least once. In the past, stress has heavily affected people’s physical health, but today it is increasingly reflected through psychological effects. The way of life is rapidly changing and developing. It demands constant monitoring, acceptance of changes and fast adjustments. An individual is thus forced to confront, accept and incorporate quick changes into his everyday life and work. Organizations on the other hand, have to consider the change in lifestyle and developments in technology as this enables them to carry out their business plans and ensure their survival. The diploma thesis presents the present-day stress field, and shows its causes and consequences - for the individual and for the organization. The purpose of the research, conducted in a pharmaceutical organization, was to determine the presence of stress, considering the gender, job position, education and age of its employees. The survey that was carried out revealed a moderate level of stress in the organization. According to the results, women, non-managerial employees, higher-educated employees and the oldest personnel have the highest risk of stress. The results also show that the organization is aware of the presence of stress and does carry out activities towards its reduction. Nevertheless, additional activities such as education, meetings, communication and measuring the atmosphere in the organization are proposed as additional aids in combating stress

Cardiac myxoma

BACKGROUND: Although cardiac myxoma (CM) are rare and benign, they can cause life-threatening complications, such as hemodynamic disturbances or embolization. Surgical excision of the tumour is the treatment of choice. The aim of the study was to evaluate the epidemiological characteristics, clinical presentation, imaging findings, and outcomes of surgical treatment of patients with CM treated in the largest tertiary care centre in Slovenia. PATIENTS AND METHODS: We retrospectively analysed the medical records of all patients referred to our institution between January 2005 and December 2020 and identified 39 consecutive adult patients with pathologically confirmed CM. RESULTS: The average annual incidence of CM in the study was 3 per 2 million population per year. Patients were more often female (n = 25, 64%). The mean age at diagnosis was 63.1 ± 13.6 years. Dyspnoea was the most common presenting symptom (31%). CM was an incidental finding in 11 patients (28%). Seven patients presented with thromboembolic event (18%). Transthoracic echocardiography (TTE) was performed in all patients, however additional imaging was required in 22 patients (56%). All patients in our series were successfully treated surgically without in-hospital mortality. During the follow-up period (6 months to 16 years) three patients (8%) died, and all deaths were unrelated to CM. There was no recurrence of CM during the follow-up. CONCLUSIONS: Our single-centre study confirms that CM is rare cardiac tumour with diverse clinical presentation. Our data shows data that CM might be more prevalent than considered before. Surgical resection of the tumour is safe with excellent short- and long-term outcomes

Can echocardiographic assessment of interatrial septum shape and motion improve the accuracy of the BLUE protocol?

Acute respiratory failure is one of the most challenging critical conditions due to a wide variety of differential diagnosis. Bedside lung ultrasound in emergency (BLUE) protocol allows accurate differentiation between the most common underlying causes of acute respiratory failure in up to 90% of the cases. The assessment of left atrial pressure affecting left ventricular filling is essential in critically ill patients guiding volume substitution, optimization of left ventricular function and prevention of pulmonary congestion, thus ensuing haemodynamic stability. A simple, non-invasive method of left atrial pressure evaluation is the echocardiographic assessment of interatrial septum shape and motion, which is affected by interatrial pressure gradient. Aiming to improve the accuracy of the BLUE protocol, we propose the simple, non-invasive echocardiographic assessment of interatrial septum shape and motion as an upgrade, providing additional information of the loading of left and right atrium thus distinguishing the most common causes of acute respiratory failure