Establishment of the first stool bank in an Eastern European country and the first series of successful fecal microbiota transplantations in Bulgaria.

OBJECTIVE: For safe implementation and broader application of fecal microbiota transplantation (FMT), quality controlled stool banking is a must. Establishing a stool bank is a complex, time-consuming, and expensive process, making it a real challenge in an Eastern European country. We aimed to establish the first stool bank in Eastern Europe - in Bulgaria. SUBJECTS AND METHODS: A multidisciplinary team of gastroenterologists, microbiologists, infectionists, and geneticists was set up. We used a questionnaire based on the First European FMT Consensus in order to recruit possible stool donors. Laboratory blood and stool tests were performed on all potential donors. RESULTS: Between October 2018 and April 2019, 112 donor volunteers completed a questionnaire; 70 (62.5%) were excluded, mainly because of age above 50, an unhealthy BMI, and risk behavior. Fourty-two (37.5%) donor candidates were invited for laboratory testing of blood and feces, of which 12 (28.6%) passed this screening. Of 12 donors, 4 (33%) failed at the following screening test, which is performed every 3-6 months. Finally, 8 (7.14%) active donors were enrolled. Ten successful FMTs were performed on patients with recurrent Clostridium difficile infection. CONCLUSIONS: Even though we found many healthy volunteers, only a low percentage (7.14%) of them were suitable to become feces donors. Establishing a stool bank in an Eastern European country is essential for making FMT safe and more popular as a treatment method, finding further implementation and regulation of FMT and supporting physicians offering this treatment to their patients.

Proangiogenic signature in advanced laryngeal carcinoma after microRNA expression profiling.

The aim of this study is to evaluate which dysregulated angiomiRs compose the specific proangiogenic microRNA signature of advanced laryngeal cancer and review the literature. Thirty-six samples from twelve patients with advanced laryngeal carcinoma were collected. Total RNA was extracted and microRNA global profiling was performed using Agilent Technologies Microarray Kit. Fifty-nine microRNAs were found to have significantly different expression levels. Eleven microRNAs from the whole group were sorted as regulators of tumor angiogenesis (angiomiRs): seven were up-regulated-miR-1246, miR-181b 5p, miR-18a 5p, miR-21 3p, miR-210 3p, miR-503 5p, miR-93 5p and four were down-regulated-miR148a 5p, miR-145 5p, miR-204 5p, miR-125b 5p. For none of those microRNAs we found heterogeneity in tumor tissue. We are the first to report the specific proangiogenic microRNA signature in advanced laryngeal carcinoma and we confirm and amplify findings from previous studies that expand our perception of a specific "molecular state" of angiogenesis that is distinctive only for laryngeal cancer.

Analytical studies of the Alexandrovo Thracian tomb wall paintings.

A profound study of samples obtained from Thracian tomb wall paintings at Alexandrovo, Bulgaria (dating back to the fourth century BC) were carried out by differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FTIR) and Attenuated Total Reflectance Fourier transform infrared spectroscopy (ATR FTIR), high-resolution transmission electron microscopy (HRTEM) and energy dispersive X-ray spectroscopy (EDS). The current work provides a glimpse of the ingenious construction and painting techniques used in Thracian tomb at Alexandrovo. The results suggest that beeswax was used as a paint binder and also revealed presence of various nano-materials.

Prostaglandin E2 promotes survival of naive UCB T cells via the Wnt/ß-catenin pathway and alters immune reconstitution after UCBT.

The outcome of umbilical cord blood transplantation (UCBT) is compromised by low hematopoietic stem cell (HSC) doses leading to prolonged time to engraftment, delayed immunological reconstitution and late memory T-cell skewing. Exposure of UCB to dimethyl-prostaglandin E2 (dmPGE2) increases HSC in vivo. We determined that exposure of UCB T lymphocytes to dmPGE2 modified Wnt signaling resulting in T cell factor (TCF)-mediated transcription. Wnt signaling upregulated interleukin (IL)-7R and IL-2Rß, resulting in enhanced survival mediated by the homeostatic cytokines IL-7 and IL-15. dmPGE2 also induced components of the Wnt pathway and Wnt receptors, thereby priming UCB T cells to receive signals via Wnt ligands in vivo. We observed that the Wnt transcription factor TCF7 and its target EOMES were elevated in the T cells of patients who received PGE2-treated UCBs. Consistent with the role of Wnt/ß-catenin signaling to induce and maintain naive, memory precursors and long-lived central memory CD8(+) cells, these patients also had increased fractions of CD8(+)CD45RO(-)CD62L(+) plus CD8(+)CD45RO(+)CD62L(+) subsets encompassing these T-cell populations. These effects of the PGE2/Wnt/ß-catenin axis may have significant implications for harnessing immunity in the context of UCBT, where impaired immune reconstitution is associated with late memory T-cell skewing.

Context-dependent differences in miR-10b breast oncogenesis can be targeted for the prevention and arrest of lymph node metastasis.

Metastases, and not the primary tumor from which they originate, are the main reason for mortality from carcinoma. Although the molecular mechanisms behind metastasis are poorly understood, it is clear that epigenetic dysregulation at the level of microRNA expression is a key characteristic of the metastatic process that can be exploited for therapy. Here, we describe an miRNA-targeted therapeutic approach for the prevention and arrest of lymph node metastasis. Therapy relies on the inhibition of the pro-metastatic microRNA-10b. It is delivered to primary and lymph node metastatic tumor cells using an imaging-capable nanodrug that is designed to specifically home to these tissues. Treatment of invasive human breast tumor cells (MDA-MB-231) with the nanodrug in vitro downregulates miR-10b and abolishes the invasion and migration of the tumor cells. After intravenous delivery to mice bearing orthotopic MDA-MB-231-luc-D3H2LN tumors, the nanodrug accumulates in the primary tumor and lymph nodes. When treatment is initiated before metastasis to lymph nodes, metastasis is prevented. Treatment after the formation of lymph node metastases arrests the metastatic process without a concomitant effect on primary tumor growth raising the possibility of a context-dependent variation in miR-10b breast oncogenesis.

A pharmacoeconomical model for choice of a treatment for pregnant women with gestational diabetes.

This study discusses two main questions: the direct medical costs and the clinical effectiveness of the hospital treatment with insulin of pregnant women with gestational diabetes (GD). A prospective study that includes 50 women with GD is performed. The pregnant women are divided into 2 groups: Group I (n=30) - pregnant women treated only with a diet; and Group II (n=20) - pregnant women treated with diet and insulin. We found that the metabolite compensation degree is improved after the applied treatment with insulin. The coefficient cost/effectiveness is 6954 lv./100 women. The analysis decision tree confirms in a very convenient way the fact that insulin treatment is a clinically more effective and financially more profitable strategy.

Pilot project for education of patients with type 2 diabetes by pharmacists.

The objective was to adapt and test an educational programme for type 2 diabetes patients. The sample consisted of 24 individuals with type 2 diabetes between 31 and 75 years of age that did not require insulin, without severe complications of diabetes. A patient satisfaction questionnaire was applied in the beginning and at the end of the survey. A five-month education was conducted on: hypo- and hyperglycaemia, nourishing and physical activity, feet care, eye care and possible adverse drug reaction. The costs of the education and the pharmacological treatment were calculated. Improvement was observed in patients' diabetes knowledge and there was a decrease in the frequency of hypo- and hyperglycaemic incidents (about 60% less). At the end of the programme no incidents were matched, saving Euro 10/patient. The benefit to cost ratio is about 1:1 (Euro 140:Euro 142.80). The patients' quality of life improved by about 5% according to the Diabetes Questionnaire (IMG). The educational approach has the potential to decrease diabetes complications and therefore the economic cost of type 2 diabetes. Introduction of the programme will benefit diabetes patients, promote pharmacies as a source of independent drug information and recognise pharmacists as competent healthcare providers.

The role of information for improvement of patients' treatment in Bulgaria.

PURPOSE: This issue is an attempt to assess patients' role as regards their compliance, respectively their non-compliance. The authors try to elucidate the factors concerning the patient-physician interrelation and patient-pharmacist interrelation that influence the non-compliance in order to increase the compliance rate. METHODS: Standard individual questionnaires and interviews were developed and applied to study an attitude to compliance and non-compliance among different categories of patients, suffering from chronic diseases. RESULTS: The obtained results show that the main reasons, influencing the patients' non-adherence to physicians' advise are: the high price of the drugs, the overburdened daily regimen, negligence, distrust towards the treatment and others. The insufficient information in the patient's leaflet together with the inappropriate drug form appears to be the main factors that cause non-compliance. This happens especially in the cases, when the patients are treated with more than one drug in different drug forms and particularly it concerns elderly people. Even though the compliance among the patients investigated was comparatively high (50% to 92%), possibility for improvement still exists if the treatment that have to be initiated could be individualised. Thus the level of compliance will be improved and increased. DISCUSSION: The results from the data analysis show a great variation in the level of compliance among the chronic patients in Bulgaria. It ranges from 50 to 92%, depending on the type of the chronic disease and on the age of the patients. This fact shows the necessity for improvement of the different factors that has to begin as soon as possible.

RNA leaching of transcription factors disrupts transcription in myotonic dystrophy.

Myotonic dystrophy type 1 (DM1) is caused by a CUGn expansion (n approximately 50 to 5000) in the 3' untranslated region of the mRNA of the DM protein kinase gene. We show that mutant RNA binds and sequesters transcription factors (TFs), with up to 90% depletion of selected TFs from active chromatin. Diverse genes are consequently reduced in expression, including the ion transporter CIC-1, which has been implicated in myotonia. When TF specificity protein 1 (Sp1) was overexpressed in DM1-affected cells, low levels of messenger RNA for CIC-1 were restored to normal. Transcription factor leaching from chromatin by mutant RNA provides a potentially unifying pathomechanistic explanation for this disease.

Assessment of pharmacy patients' compliance in Bulgaria (2001--2002).

The goal of this issue is to assess the level of compliance, respectively of non-compliance among pharmacY patients between the years 2001 and 2002 in Sofia, Bulgaria. The authors study and emphasize on the factors that influence the non-compliance of the patients and also try to find different ways to increase the rate of compliance in Bulgaria. Two types of standard individual questionnaires are applied for assessment of the state of compliance and non-compliance among the pharmacy patients. The obtained results show that the main reasons, influencing the non-adherence of the patients are: the high average price of the drugs, the overload daily regimen, inattention, distrust to the treatment and others. The drug form appears to be one of the main factors that cause non-compliance, especially when the patients receive more than one drug in different drug forms few times a day and especially if elderly patients are addressed. Despite the presented comparatively high level of compliance among the patients (from 53% for 2001, to 82% for 2002), there are strategies to enhance compliance that have to be initiated. They incorporate communication tactics, patient's education, and proper dosage of the drugs and scheduling of the drug application. The results of the analysis of the received data show an increase in the level of compliance for the pharmacy patients in Bulgaria. For 2001 it was 53%, while for 2002 it has increased to 82%. This fact shows the influence of strategies for improvement of compliance.

Comparison of the compliance of pharmacy patients.

This issue is an attempt to be compared the compliance, respectively non-compliance among pharmacy patients between the years 1998 and 2001. The authors make an attempt to elucidate the factors that influence the non-compliance of the patients in order to improve it and to increase the rate of compliance among them. A questionnaire is applied for assessment the state of compliance and non-compliance among the pharmacy patients. The results of the analysis of the received data show that the level of compliance was 69% in 1998 and has decreased to 53% in 2001. The main reasons that influence the non-adherence of the patients are shown.

Phospholipid black foam films: dynamic contact angles and gas permeability of DMPC+DMPG black films.

The behavior of black foam films from aqueous dispersions of dimyristoylphosphatidyl-choline (DMPC) with addition of the soluble phospholipid dimyristoylphosphatidylglycerol (DMPG) has been studied in dynamic conditions. The dynamic contact angles theta and the gas permeability coefficient K have been measured using the diminishing bubble method. The DMPC vesicle suspension in water is obtained through sonication and DMPG is dissolved in it. Two solutions with different NaCl concentrations (0.1 M and 0.5 M) have been studied. The behavior of the dynamic contact angles is very different for DMPC black films with, and without DMPG, respectively. They follow very different time dependence during spontaneous or forced variations of the bubble size. The gas permeability coefficient is significantly reduced by the DMPG addition. The NaCl concentration also influences this specific behavior. It seems that the electrically charged DMPG anions, which determine a significant electrostatic disjoining pressure, play an important role for this specific behavior. The results are discussed in connection with data regarding the thickness and structure of these black foam films. Films from DMPC+DMPG solutions in ethanol plus water mixed solvent have been studied as well, but no quantitative results could be obtained.

Phospholipid black foam films: dynamic contact angles and gas permeability of DMPC bilayer films.

The behavior of bilayer Newton Black Films (NBF) from aqueous dispersions of dimyristoylphosphatidylcholine (DMPC) have been studied in dynamic conditions. The dynamic contact angles theta and the gas permeability coefficient K have been measured using the diminishing bubble method. Two different solutions have been used: (i) DMPC vesicle suspension in water obtained through sonication and (ii) DMPC dissolved in ethanol plus water mixed solvent. Both solutions contain 0.1 M NaCl. The behavior of the dynamic contact angles is very different for NBF from the two types of solutions. In the case (i) the initially constant theta(t) sharply increase after approximately 2 h of the spontaneous diminishing of the bubble, they follow the gas pressure variation in the cell and depend on the film area. On the contrary in case (ii) the theta(t) values are almost constant during the spontaneous diminishing of the bubble as well as during the gas pressure variation in the cell and they do not depend on the film area. The gas permeability coefficient is larger in case (ii). The results are discussed in connection with the thickness and structure of the NBF from the two types of solutions, taking into account the solubility (or insolubility) and the hydration of the adsorption layers of the DMPC molecules.

Asthma, drug medication and noncompliance.

UNLABELLED: According to the new concepts, asthma is a chronic disease of the upper respiratory tracts, caused by different cells, including the mastocits and eozinofils. The patients that are suffering from episodic, frequent, or chronic asthma are participating very energetic in the treatment--they have to know and control the symptoms of their disease and also to possess technical skills to perform the treatment. METHODOLOGY: Between March 1 and July 31, 1999, 30 patients [19 women and 11 men, mean age 35 years (range 2.50 to 65)], followed at the Clinic for Allergic Diseases of the University hospital "Aleksandrovska", Medical University of Sofia, Bulgaria, were interviewed blindly in order to assess their compliance with pharmacologic therapy. All patients were suffering from different types of asthma. RESULTS: The level of compliance among the studied group of patients is 86%. In comparison with the literary data where the level of non-compliance is 20% the obtained level is lower--14%. The reasons for non-compliance according to the patients are: the appearance of ADRs, the high prize of the drugs, the hard application of the drug form.