[Comparison of the incidence, treatment and prognosis of myocardial infarction in Hungary's differently developed districts]. / A szívinfarktus incidenciája, a betegek ellátásának és prognózisának összehasonlítása Magyarország különbözo fejlettségu járásaiban.

INTRODUCTION: Several international studies have already confirmed the importance of the socioeconomic status of acute myocardial infarction patients in terms of patient care and prognosis. To our knowledge, a nationwide examination of this kind has not yet taken place in Hungary. The investigation of this problem field was made possible by the fact that from January 1, 2014, all healthcare providers must record the data of patients treated with a diagnosis of acute myocardial infarction in the database of the Hungarian Myocardial Infarction Registry (HUMIR). OBJECTIVE: In this study, the authors searched for an answer to whether the complex development index (CDI) in Hungary's 174 districts and 23 capital districts influences the treatment and prognosis of acute myocardial infarction patients. METHOD: Based on the CDI worked out by the Hungarian Central Statistical Office, the authors divided the Hungarian districts into low (CDI_L), medium (CDI_M) and high (CDI_H) CDI groups according to their values. They examined the incidence, hospital treatment and prognosis of acute myocardial infarction in these administrative-territorial units. The HUMIR included 66,253 patients treated by myocardial infarction between 2015 and 2019. Their place of residence could be identified based on the zip code and in which district it was located. In the examined population, 29,101 patients with ST-elevation (STEMI) and 37,152 without ST-elevation (NSTEMI) received treatment for acute myocardial infarction. RESULTS: In the population over 15 years of age, the age-standardized incidence of STEMI was 68.8 per 100,000 inhabitants a year in the CDI_L group and 52.7 per 100,000 inhabitants a year in the CDI_H group. Almost the same values were found in all three CDI subgroups of NSTEMI incidence (69.5 and 67 per 10,000 inhabitants a year). The frequency of percutaneous coronary intervention in the case of STEMI was higher than in NSTEMI, but within the groups, CDI did not influence the performance of this treatment. In the case of STEMI, the rates of patients who underwent percutaneous coronaria intervention in all three CDI subgroups (CDI_L, CDI_M, CDI_H) were 83.5%, 83.7%, 83.5%, while in the case of NSTEMI they were 57.4%, 57.7%, 57.3%. The authors applied a Cox multivariate regression analysis to examine myocardial infarction mortality. The CDI did not affect the 30-day mortality rates in the case of any myocardial infarction: the hazard ratio (HR) values were 0.906 and 0.914 (p = 0.04659; p = 0.04686) in the case of STEMI, while 1.067 and 1.001 (p = 0.16520; p = 0.98933) in the case of NSTEMI. In the case of a STEMI diagnosis, the risk of the 30-364-day and the 1-year mortality in the subgroup of CDI_H was significantly lower (HR = 0.822 and 0.816) than in the subgroup of CDI_L (p = 0.00096 and p = 0.00001). In the case of NSTEMI diagnosis, the authors found a difference in the risk of beyond 1-year mortality by comparing the districts in the subgroup of CDI_L with the districts in CDI_H: in the latter case, the HR of the mortality was 0.876, which was significantly lower (p = 0.00029) than in the subgroup of CDI_L. CONCLUSION: The CDI has independent prognostic significance in determining the late prognosis of acute myocardial infarction patients. Orv Hetil. 2022; 163(47): 1862-1871.

Simple Formula for pH/Impedance Probe Positioning in Children-Time to Update Standard Practice?

OBJECTIVE: To develop a novel formula for pH probe placement with adequate accuracy. METHODS: Children (3-18 y) undergoing pH-metry were prospectively evaluated. Their height and corrected pH probe position under X-ray (2 vertebrae above the diaphragm) was recorded and the linear-regression analysis was performed to derive a novel formula. Its accuracy was checked on an additional group of prospectively included children. The success rate of a newly developed formula was estimated and compared to the performance of previously used formulae. The difference in the suggested placement of the probe (cm from nostrils) was calculated. RESULTS: Based on 670 children with pH probe placed under X-ray, the following formula was developed using the linear-regression analysis: L = 0.184x + 4.4 (cm) (L = probe placement depth, x = body height). Its accuracy was confirmed on additional 111 children resulting in almost 85% success rate. The formula showed significant difference in the suggested placement from formulae used previously: +4.9 ± 0.8 cm, +2.4 ± 0.1 cm, +0.7 ± 0.6 cm, +1.1 ± 0.4 cm, +1.8 ± 0.3 cm, +2.2 ± 0.5 cm from the one by the Strobel, Moreau, Wilson, Nowak, Staiano-Clouse formulae, and the GOSH table with the calculated success rates of 1.8%, 43.2%, 65.8%, 77.5%, 65.8% and 54.1%, respectively. A table suggesting placement depth based on the body height was developed. CONCLUSION: The present formula provides 85% success of pH probe placement in children ≥ 3 y suggesting its use in routine practice. More data are needed to confirm that probe adjustment under X-ray is unnecessary.

Community and environmental data-driven monitoring of waste management.

Environmental operators perform their activities in accordance with the relevant legal provisions; however, this does not mean that they operate at their technological optima using the operational information available. The possible negative effects (odor, noise, etc.) of a sub-optimal operation can be felt first and foremost by those living in the immediate vicinity of the given object. It would be important to make effective use of these citizens feedback (quickly to revealing the root causes) thus minimize negative environmental impact of operations. The solution proposed in this paper is a portal called EnviroMind, which allows citizens feedback to be recorded in an easy, immediate, and structured way via a form and on the other hand, it provides a real-time graphical odor transmission model output in a dashboard to operators. Using this portal as a monitoring system the magnitude of the odor effect could be reduced and a smaller area around the industrial object could be affected. In a landfill monitoring pilot project where this monitoring system was used the decrease in the number of indicated odor observations was 85% and the decrease in maximal distance from landfill to odor detection positions was 45%. It is proposed to use EnviroMind monitoring system for all industrial objects which have a significant odor effect on the environment, because by using it we can make the odor effect visible to operators in real time, thus, the reaction time for solving the problem can be minimized.Implications: monitoring is available online to the surrounding community, the affected population, so that quick responses and interventions are available; in the knowledge of the current technological activity carried out on the site its expected odor effect in the area can be determined, whether a protected area can be reached and what odor concentration is expected; in every 15 minutes model results to accurately track expected odor emission values; possibility of intervention, stopping or modification of the technology steps. Experience and main achievements of portal operation in a landfill monitoring pilot project from recent 3 years: the decreasing number of odor perceptions (the decrease in the number of indicated observations was 85%) and the cessation of odor effects in certain areas (and the decrease in maximal distance from landfill to odor detection positions was 45%).

Management of patients treated for myocardial infarction in different regions of Hungary and patient survival for 10 years / Szívinfarktus miatt kezelt betegek ellátása Magyarország különbözo régióiban és a betegek 10 éves túlélése.

Összefoglaló. Elozmény: A szívinfarktus miatt kezelt betegek ellátásának regionális adataira és a betegek hosszú távú kórlefolyására vonatkozó hazai kutatás eddig nem történt. Célkituzés: A vizsgálat célja a Magyar Infarktus Regiszter pilotidoszakában rögzített betegeknél az ellátás és a 10 éves túlélés elemzése a magyarországi nagyrégiókban. Módszer: A Magyar Infarktus Regiszter (késobbi neve: Nemzeti Szívinfarktus Regiszter) 2010. január 1. és 2013. december 31. között a centrumok önkéntes részvételével 23 142 beteg adatait rögzítette, akik írásban hozzájárultak egészségügyi és klinikai adataik kezeléséhez. Az adatgyujtés a Kutatásetikai Bizottság engedélyével rendelkezett. A vizsgált populációban 12 104, ST-elevációval járó myocardialis infarctuson (STEMI) és 10 768, ST-elevációval nem járó myocardialis infarctuson (NSTEMI) átesett beteg szerepelt. A feldolgozott adatok 128 220 betegévre vonatkoznak, amelyeket nagyrégiók szerint (Nyugat-, Közép- és Kelet-Magyarország) hasonlítottunk össze. Eredmények: A STEMI-betegek 78,4%-ánál, az NSTEMI-betegek 51,6%-ánál történt katéteres érmegnyitás (PCI). NSTEMI esetén a Közép-Magyarország és Nyugat-Magyarország régiókban a beavatkozás gyakoribb volt, mint a Kelet-Magyarország régióban (p<0,01). Az utánkövetés során a PCI a Nyugat-Magyarország régióban, a revascularisatiós szívmutét (CABG) a Nyugat-Magyarország és a Kelet-Magyarország régióban szignifikánsan gyakoribb volt, mint a Közép-Magyarország régióban (p<0,01). A STEMI-betegek között a 10 év alatt a férfiak 49,2%-a, a nok 46,6%-a halt meg, az NSTEMI-csoportban 63%, illetve 57,6%. Az akut szakban elvégzett PCI mindkét betegcsoportban, nemben, az utánkövetés minden idopontjában és a vizsgált régiókban csökkentette a halálozást (p<0,01). A STEMI-betegek esetén a túlélés a régiók között nem különbözött (p = 0,72), míg az NSTEMI után a 10 éves túlélés a Nyugat-Magyarország régióban jobb volt (p<0,01). Következtetés: A magyarországi nagyrégiók között az infarktusos betegek ellátásában és prognózisában regionális különbségek vannak. Orv Hetil. 2021; 162(36): 1438-1450. HISTORY: Regional data on patients' care for myocardial infarction and the long-term follow up of patients have not yet been studied in Hungary. OBJECTIVE: The study aims to analyze the care and 10-year survival of patients recorded during the Hungarian Myocardial Infarction Registry's pilot period in large regions of Hungary. METHOD: Between Jan 1, 2010 and Dec 31, 2013, the Hungarian Myocardial Infarction Registry recorded data on 23 142 patients with voluntary participation. The Research Ethics Committee approved the program. The study included 12 104 patients with ST-elevation myocardial infarction (STEMI) and 10 768 patients with non-ST-elevation myocardial infarction (NSTEMI). The data processed refer to 128 220 patient years based on large regions (West, Central and East Hungary). RESULTS: Percutaneous coronary intervention occurred in 78.4% of STEMI patients and 51.6% of NSTEMI patients. In the NSTEMI group, percutaneous coronary interventions (PCIs) in the Central-Hungary and West-Hungary regions were significantly more common than in the East-Hungary region (p<0.01). During follow-up, PCI in the West-Hungary region, revascularization surgery in the West-Hungary and East-Hungary regions were significantly more common than in the Central-Hungary region (p<0.01). Among STEMI patients, 49.2% of men and 46.6% of women died within 10 years, while in the NSTEMI group 63% and 57.6%, respectively. PCI reduced mortality in both patient groups, sex, at all times of follow-up and in the regions studied (p<0.01). As for STEMI patients, survival was similar in all regions (p = 0.72), while after NSTEMI, 10-year survival in the West-Hungary region was better (p<0.01). CONCLUSION: There are regional differences in the care and prognosis of patients with myocardial infarction. Orv Hetil. 2021; 162(36): 1438-1450.

Incidence, pre-hospital delay and prognosis of acute myocardial infarction in big regions of Hungary: Population data from the Hungarian myocardial infarction registry.

AIM: To examine the incidence and treatment of acute myocardial infarction (AMI) as well as 30-day and 1-year prognoses of patients in different regions of Hungary. According to the statistical system of the European Union, Hungary can be divided into three major socio-economic regions-west Hungary, central Hungary and east Hungary. METHODS AND RESULTS: The Hungarian Myocardial Infarction Registry (HUMIR) is a prospective comprehensive and mandatory disease registry for patients with AMI. The total population of Hungary is currently 9.8 million: 39% live in the eastern region (ER), 31% in the central region (CR) and 30% in the western region (WR). Population over 30 years, the age-standardised incidence of AMI was 177.5 (175.7-179.3) per 100 000 person-year. During hospital treatment, 82.5%-84.6% of patients with ST-elevation (STEMI) and 54.8%-58.8% without ST-elevation (NSTEMI) myocardial infarction underwent PCI. The total ischaemic time of patients with STEMI was shortest in WR (221 minutes) compared with two other regions (CR: 225 minutes and ER: 262 minutes). In the STEMI group, the 30-day mortality rates of male patients were lowest in the WR (P = .03). If PCI was performed, mortality rates for both sexes were lowest in the WR (P < .01; P = .04). The 1-year mortality rate in the male population who received PCI was lowest in the WR. In the NSTEMI group, the 30-day mortality rate exhibited no differences. Regarding 1-year mortality, those who underwent PCI in the WR showed the lowest mortality. CONCLUSION: The major regions of Hungary revealed significant differences regarding the incidence, prehospital delay, treatment and mortality of AMI. Logistic regression analysis confirmed the independent prognostic significance of the region on the 30-day mortality of patients with STEMI (hazard ratio = 0.88, P = .0114; CI: 0.80-0.97).

[Emergency care of patients with myocardial infarction: from the onset of symptoms until opening the vessel]. / Szívinfarktus miatt kezelt betegek sürgosségi ellátása: a panasz kezdetétol az ér megnyitásáig.

Introduction and aim: The authors analyse emergency care data for 6878 patients treated for acute myocardial infarction (AMI) using data from the Hungarian Myocardial Infarction Registry (HUMIR) and the National Ambulance Service (NAS). Method: Patients received treatment between 01/01/2017 and 31/12/2018, and all patients underwent percutaneous coronary intervention (PCI): 47.5% of patients had ST-elevation myocardial infarction (STEMI) and 3614 patients (52.5%) had non-ST-elevation myocardial infarction. The time between the beginning of the complaint and notification of NAS was regarded as the patient delay (PD). The time from the notification of NAS until arrival on the scene (M1), that of the on-site care (M2) and of the transport from the scene to the hospital (M3) were recorded. In-hospital care was evaluated from admission until opening the vessel ("door to balloon time"). The results were also broken down by counties. The median values and the quartiles (Q1, Q3) were given when the time was reported. Results: Patient delay in both types of infarction was unfavourably long: 101 minutes for STEMI and 687 minutes for NSTEMI. Immediate ambulance action was recorded in 58.7% for STEMI patients and 43.7% for NSTEMI patients. In both types of myocardial infarction, the median M1 time was 13 minutes, on-site care (M2) was 23 minutes, and M3 time was 30 minutes. In patients treated for STEMI, the time from hospital admission until opening the infarct-related artery was 37 minutes, and the total ischemic time was 243 minutes. In 9.5% of STEMI patients, the infarct-related artery was opened within 2 hours, in 49.1% within 4 hours, and in 88.1% within 12 hours. Significant differences were found between the counties for each of the periods examined. Conclusions: The PD is currently the biggest problem in providing optimal care timely for myocardial infarction patients. There are significant regional differences in rescue times, and further analysis is needed to investigate the causes. Orv Hetil. 2020; 161(12): 458-467.

[Analysis of pre-hospital delay time of patients with myocardial infarction]. / Szívinfarktus miatt kezelt betegek prehospitális késési idejének elemzése.

INTRODUCTION AND AIM: The authors examined the pre-hospital delay of patients with ST-elevation myocardial infarction receiving percutaneous coronary revascularization. METHOD: In the Hungarian Myocardial Infarction Registry (HUMIR), between January 1, 2014 and March 31, 2016, 7146 patients were found who had all necessary time intervals available. In the database of the National Ambulance Service (OMSZ), 3288 patients were found who had the necessary time intervals. The following time intervals were investigated: the time from the beginning of the complaint to the rescue service notification (patient delay time = PDT), the time of the rescue service to arrive at the scene (R1), the on-site care time (R2) and the time from the scene until arriving to the centre (R3). The case of care at the centre, we investigated the time from the onset of symptoms until the balloon inflation (SBI). If the first hospital had no cardiac catheterization laboratory, we measured the transfer time to the cath centre. The methodological details related to the operation of the HUMIR had been described in our earlier communication. Rescue times (R1, R2, R3) were investigated on the basis of the paper-based records of the National Emergency Service. The patients were divided into two groups based on the fact that the first admission hospital is a centre with a heart catheter facility (C) or a non-invasive hospital (H). RESULTS: 2621 patients (79.7%) were admitted to a hospital with cath lab (C) and 667 patients with secondary transport. Patients with primary transport to C were younger, but for other data, the two groups did not differ. The median of PDT for patients in group C was 114, and 121 minutes for patients in group H. There was no significant difference between R1 and R2 time between the two groups. R3 time in group C was longer than for H patients. In the case of secondary transport, the median time was 98 minutes until the centre. The median time from the beginning of the complaint to the balloon inflation (total ischemic time) was 260 minutes in group C and 356 minutes in group H. CONCLUSION: Based on the analysis of the total ischemic time and the pre-hospital delay, it is clear that the care of myocardial infarction patients can be further improved by reducing the patient's decision time and increasing the proportion of primary transport. Rescue times (R1, R2, R3) met the expectations, however, further analysis of the M2 time should be considered. Orv Hetil. 2019; 160(1): 20-25.

[Prognostic significance of the total ischemic time in patients with ST-elevation myocardial infarction]. / A teljes ischaemiás ido prognosztikus jelentosége az ST-elevációval járó szívinfarktus miatt kezelt betegekben.

INTRODUCTION: The significance of the total ischemic time (from the beginning of the complaint to the opening of the vessel) is an important factor for myocardial salvage. AIM: The aim of the study was to determine the prognostic significance of the TIT in patients with ST elevation myocardial infarction in Hungary. METHOD: From 1 January 2014 all patients with myocardial infarction were recorded by law in an on-line database of the Hungarian Myocardial Infarction Registry. Between 1 January 2014 and 31 March 2016, 27 157 patients with 28 408 myocardial infarction events were recorded. To investigate TIT, 7146 STEMI patients were selected who were treated with percutaneous coronary intervention (PCI) within 24 hours of the beginning of the complaint and all of its components were known. RESULTS: Average follow-up was 740 ± 346 days. The median time of the TIT is 260 minutes, within which the earliest prehospital time was found (median 205 minutes). The TIT influenced survival: if this time was less than 400 minutes, the 30-day and the 1-year deaths were 7.5% and 12.2%, respectively. In longer TIT, higher mortality rate was found (9.2% versus 19.7%, respectively). Multivariate analysis was performed for short (<30 days), medium (30-364 days) and long-term (≥365 days) survival. Diabetes mellitus is a short-term prognostic factor, abnormal creatinine, and severe coronary status have affected short and medium survival. PCI was significant in terms of medium and long-term survival. Previous myocardial infarction and TIT influenced the long-term survival significantly. CONCLUSIONS: In Hungary, TIT is too long, and its dominant part falls within the prehospital period. The TIT is an independent prognostic factor, so reducing this time can improve the long-term prognosis of patients with ST-elevation myocardial infarction. Orv Hetil. 2018; 159(27): 1113-1120.

Analysis of selected genes associated with cardiomyopathy by next-generation sequencing.

BACKGROUND: As the leading cause of congestive heart failure, cardiomyopathy represents a heterogenous group of heart muscle disorders. Despite considerable progress being made in the genetic diagnosis of cardiomyopathy by detection of the mutations in the most prevalent cardiomyopathy genes, the cause remains unsolved in many patients. High-throughput mutation screening in the disease genes for cardiomyopathy is now possible because of using target enrichment followed by next-generation sequencing. The aim of the study was to analyze a panel of genes associated with dilated or hypertrophic cardiomyopathy based on previously published results in order to identify the subjects at risk. METHODS: The method of next-generation sequencing by IlluminaHiSeq 2500 platform was used to detect sequence variants in 16 individuals diagnosed with dilated or hypertrophic cardiomyopathy. Detected variants were filtered and the functional impact of amino acid changes was predicted by computational programs. RESULTS: DNA samples of the 16 patients were analyzed by whole exome sequencing. We identified six nonsynonymous variants that were shown to be pathogenic in all used prediction softwares: rs3744998 (EPG5), rs11551768 (MGME1), rs148374985 (MURC), rs78461695 (PLEC), rs17158558 (RET) and rs2295190 (SYNE1). Two of the analyzed sequence variants had minor allele frequency (MAF)<0.01: rs148374985 (MURC), rs34580776 (MYBPC3). CONCLUSION: Our data support the potential role of the detected variants in pathogenesis of dilated or hypertrophic cardiomyopathy; however, the possibility that these variants might not be true disease-causing variants but are susceptibility alleles that require additional mutations or injury to cause the clinical phenotype of disease must be considered.

Serum osteocalcin is associated with improved metabolic state via adiponectin in females versus testosterone in males. Gender specific nature of the bone-energy homeostasis axis.

OBJECTIVE: The osteoblast-derived protein osteocalcin (OCN) is known to be involved in glucose metabolism by increasing adiponectin secretion from adipocytes. Recently, OCN was also found to enhance testosterone production in mouse testes, suggesting that OCN effects on energy metabolism may be mediated through testosterone. Our aim was to assess a possible gender difference in the metabolic effect of OCN in humans. METHODS: We included 135 women and 155 men exhibiting changes in glucose tolerance in our study. Oral and intravenous glucose tolerance tests (OGTT and IVGTT, respectively) and a hyperinsulinemic normoglycemic clamp were performed. For clamp indices, whole body (M1) and muscle (M2) glucose uptake values were used. Leptin, adiponectin serum lipid, lipoprotein, total serum OCN and testosterone levels, and body composition were determined. RESULTS: Higher OCN values were associated with improving metabolic state in both genders. Adiponectin and OCN correlated significantly only in females (r=+0.254, p=0.0029), while in men, testosterone and OCN values showed a significant positive correlation (r=+0.243, p=0.0023), independent of age, BMI, HbA1c and body composition. In women, adiponectin was confirmed by feature selection analysis as being an independent determinant of OCN, in addition to age and three of the IVGTT glucose values. In men, besides M1, BMI, M2, leptin, body fat percent, and the 90-minute OGTT glucose reading testosterone, but not adiponectin were identified as independent contributors for OCN. CONCLUSION: We confirmed the 'classic' adiponectin-mediated insulin-sensitising effect of OCN only in females. In men, a testosterone-mediated OCN metabolic effect is more likely.

Photodynamic Therapy with Hypericin Improved by Targeting HSP90 Associated Proteins.

In this study we have focused on the response of SKBR-3 cells to both single 17-DMAG treatment as well as its combination with photodynamic therapy with hypericin. Low concentrations of 17-DMAG without any effect on survival of SKBR-3 cells significantly reduced metabolic activity, viability and cell number when combined with photodynamic therapy with hypericin. Moreover, IC10 concentation of 17-DMAG resulted in significant increase of SKBR-3 cells in G1 phase of the cell cycle, followed by an increase of cells in G2 phase when combined with photodynamic therapy. Furthermore, 17-DMAG already decreased HER2, Akt, P-Erk1/2 and survivin protein levels in SKBR-3 cells a short time after its application. In this regard, 17-DMAG protected also SKBR-3 cells against both P-Erk1/2 as well as survivin upregulations induced by photodynamic therapy with hypericin. Interestingly, IC10 concentration of 17-DMAG led to total depletion of Akt, P-Erk1/2 proteins and to decrease of survivin level at 48 h. On the other hand, 17-DMAG did not change HER2 relative expression in SKBR-3 cells, but caused a significant decrease of HER2 mRNA in MCF-7 cells characterized by low HER2 expression. These results show that targeting HSP90 client proteins increases the efficiency of antineoplastic effect of photodynamic therapy in vitro.

Photoactivated hypericin induces downregulation of HER2 gene expression.

Photodynamic therapy is an alternative method for cancer treatment in which a photosensitizer exposed to a light source of suitable wavelength is excited and can subsequently react through free radical mechanisms. Recently, oxygen free radical-mediated changes in gene expression have been established. The present study shows the effect of photoactivated hypericin on the expression of the human epidermal growth factor receptor 2 (HER2) oncogene at both the mRNA and the protein level in SKBR-3 and MCF-7 breast adenocarcinoma cell lines. The photodynamic therapy-induced decrease in mRNA expression was reversed by the singlet oxygen scavenger trolox, which supports a role for singlet oxygen. In addition, prevention of the generation of reactive oxygen species by pretreatment with trolox effectively blocked the antiproliferation activity of photoactivated hypericin. These results may have important implications at least for recurrent breast cancer with HER2 expression alone or in combination with conventional therapies.

Impact of 2 doses of clorgyline on the rat preimplantation embryo development and the monoamine levels in urine.

We have evaluated the impact of chronic administration of clorgyline, a potent monoamine oxidase A inhibitor and a former antidepressant, on the preimplantation embryo development in Wistar rats. Females were injected intraperitoneally daily for 30 days with saline (control animals), or with a low-dose clorgyline (LDC, 0.1 mg/kg per d) or with a high-dose clorgyline (HDC, 1 mg/kg per d). Embryos were isolated on day 5 of pregnancy and urine was collected by puncture of the urinary bladder. The number of embryos per female did not differ between experimental groups and control, but we have recorded a decreased number of embryos in HDC group compared to LDC (P < .05). We have found that LDC significantly reduced the presence of healthy embryos and increased the presence of the degenerated embryos (P < .001). The administration of the LDC resulted in the lowest cell number in blastocysts. We have observed significantly increased serotonin levels in HDC group compared to both control (P < .05) and LDC animals (P < .01). Norepinephrine (NE) levels in both experimental groups were significantly elevated compared to controls. Dopamine levels did not differ between groups (P > .05). We speculate that lesser negative effect of HDC compared to LDC on the preimplantation embryo development could be the consequence of the lower NE levels and/or elevated serotonin levels. Potential mechanisms mediating clorgyline-induced impaired preimplantation embryo development are proposed.

Down-regulation of Bcl-2 and Akt induced by combination of photoactivated hypericin and genistein in human breast cancer cells.

Presented experiment considers combination of genistein and photodynamic therapy with hypericin with a view to achieve higher therapeutic outcome in human breast adenocarcinoma cell lines MCF-7 and MDA-MB-231, both identified in our conditions as photodynamic therapy resistant. Since genistein is known to suppress Bcl-2 expression, we predicted that photodynamic therapy with hypericin might benefit from mutual therapeutic combination. In line with our expectations, combined treatment led to down-regulation of Bcl-2 and up-regulation of Bax in both cell lines as well as to suppression of Akt and Erk1/2 phosphorylation induced by photoactivated hypericin in MCF-7 cells. Although Akt and Erk1/2 phosphorylation was not stimulated by photodynamic therapy with hypericin in MDA-MB-231 cells, it was effectively suppressed in combination. Variations in cell death signaling favoring apoptosis were indeed accompanied by cell cycle arrest in G(2)/M-phase, activation of caspase-7, PARP cleavage and increased occurrence of cells with apoptotic morphology of nucleus. All these events corresponded with suppression of proliferation and significantly lowered clonogenic ability of treated cells. In conclusion, our results indicate that pre-treatment with tyrosine kinase inhibitor genistein may significantly improve the effectiveness of photodynamic therapy with hypericin in MCF-7 and MDA-MB-231 breast cancer cells.