[CONGENITAL UNILATERAL HYDROTHORAX--A CASE REPORT].

A case of isolated unilateral congenital hydrothorax with unknown etiology, graduated abortion for medical reasons in 23 weeks. The patient was later covered by antenatal care and during the examination one foetus with the massive right-sided hydrothorax was found, (diagnosed by ultrasonography and NMR) The incidence of congenital fetal hydrothorax is estimated at approximately 1 in 15,000 pregnancies. Early ultrasound diagnosis is of paramount importance for the adequate monitoring of the pregnancy and timely proof of deviation in normal fetal development.

[Diabetes and pregnancy].

UNLABELLED: We tried to summarize our and foreign experience in the field of diabetes and pregnancy AIM: Our aim was to detect pregnant women with diabetes mellitus, as well as to introduce screening method for early detection and apply a suitable diet and treatment for these patients. MATERIAL AND METHODS: Patients with diabetes and pregnancy were researched for the period of 2007-2014 (70 patients). RESULTS: Introducing a screening method in the second trimester between 24-28 gestational week, applying a suitable diet and treatment lead to better outcome of this disease and to more safe delivery outcome. CONCLUSION: Future research work will give additional light in this field.

Coordinated evolution of the hepatitis B virus polymerase.

The detection of compensatory mutations that abrogate negative fitness effects of drug-resistance and vaccine-escape mutations indicates the important role of epistatic connectivity in evolution of viruses, especially under the strong selection pressures. Mapping of epistatic connectivity in the form of coordinated substitutions should help to characterize molecular mechanisms shaping viral evolution and provides a tool for the development of novel anti-viral drugs and vaccines. We analyzed coordinated variation among amino acid sites in 370 the hepatitis B virus (HBV) polymerase sequences using Bayesian networks. Among the HBV polymerase domains the spacer domain separating terminal protein from the reverse-transcriptase domain, showed the highest network centrality. Coordinated substitutions preserve the hydrophobicity and charge of Spacer. Maximum likelihood estimates of codon selection showed that Spacer contains the highest number of positively selected sites. Identification of 67% of the domain lacking an ordered structure suggests that Spacer belongs to the class of intrinsically disordered domains and proteins whose crucial functional role in the regulation of transcription, translation and cellular signal transduction has only recently been recognized. Spacer plays a central role in the epistatic network associating substitutions across the HBV genome, including those conferring viral virulence, drug resistance and vaccine escape. The data suggest that Spacer is extensively involved in coordination of HBV evolution.

Evaluation of viral heterogeneity using next-generation sequencing, end-point limiting-dilution and mass spectrometry.

Hepatitis C Virus sequence studies mainly focus on the viral amplicon containing the Hypervariable region 1 (HVR1) to obtain a sample of sequences from which several population genetics parameters can be calculated. Recent advances in sequencing methods allow for analyzing an unprecedented number of viral variants from infected patients and present a novel opportunity for understanding viral evolution, drug resistance and immune escape. In the present paper, we compared three recent technologies for amplicon analysis: (i) Next-Generation Sequencing; (ii) Clonal sequencing using End-point Limiting-dilution for isolation of individual sequence variants followed by Real-Time PCR and sequencing; and (iii) Mass spectrometry of base-specific cleavage reactions of a target sequence. These three technologies were used to assess intra-host diversity and inter-host genetic relatedness in HVR1 amplicons obtained from 38 patients (subgenotypes 1a and 1b). Assessments of intra-host diversity varied greatly between sequence-based and mass-spectrometry-based data. However, assessments of inter-host variability by all three technologies were equally accurate in identification of genetic relatedness among viral strains. These results support the application of all three technologies for molecular epidemiology and population genetics studies. Mass spectrometry is especially promising given its high throughput, low cost and comparable results with sequence-based methods.

Coordinated evolution of the hepatitis C virus.

Hepatitis C virus is a genetically heterogeneous RNA virus that is a major cause of liver disease worldwide. Here, we show that, despite its extensive heterogeneity, the evolution of hepatitis C virus is primarily shaped by negative selection and that numerous coordinated substitutions in the polyprotein can be organized into a scale-free network whose degree of connections between sites follows a power-law distribution. This network shares all major properties with many complex biological and technological networks. The topological structure and hierarchical organization of this network suggest that a small number of amino acid sites exert extensive impact on hepatitis C virus evolution. Nonstructural proteins are enriched for negatively selected sites of high centrality, whereas structural proteins are enriched for positively selected sites located in the periphery of the network. The complex network of coordinated substitutions is an emergent property of genetic systems with implications for evolution, vaccine research, and drug development. In addition to such properties as polymorphism or strength of selection, the epistatic connectivity mapped in the network is important for typing individual sites, proteins, or entire genetic systems. The network topology may help devise molecular intervention strategies for disrupting viral functions or impeding compensatory changes for vaccine escape or drug resistance mutations. Also, it may be used to find new therapeutic targets, as suggested in this study for the NS4A protein, which plays an important role in the network.

Pharmacovigilance: characteristics of the most widely used drugs in Bulgaria.

In the present study we analysed registered adverse drug reactions caused by 19 most used drugs in Bulgaria during three-year period. The cases were assembled from the spontaneous reporting system operated in our country. We described demographic characteristics of the patients, clinical diagnosis and reporting rates per 100,000 inhabitants for each drug. The data for antibiotics were compared with the results of our previous research. We analysed 188 adverse drug reactions i.e. 21% of all reports received in the Centre for Drug Safety of the National Drug Institute. Adverse drug reactions type B such as oedema, allergy, coma, etc. predominate. The number of cases with perforated ulcer and melaena caused by acetylsalicylic acid is considerable. The frequency of adverse drug reaction for the investigated drugs in "rare" according to WHO standards. It is necessary to assemble more information on adverse drug reactions and every serious case and inform medical doctors and pharmacists as a first step for improving the pharmacovigilance system in Bulgaria.

Reimbursed treatment, regulated in the national frame contract.

The investigation is an analysis of the main elements, included in the reimbursement system for the drug insurance. The aim is directly dependent on the valuation for the adequate service of the arise health needs of the citizens that are obligatory insured. The used methods are connected with an analysis of the regulations and of the statistical information. As result of the investigation are characterized the seize of the disease and of the drug nomenclature with a forecast reimburse therapy. The provide valuation of the objective state of the system shows that the criteria and rules are not clear and precise in respect of: the include of the drugs in the positive list, the determination of the level of reimbursement, the selection of the disease with the reimbursement treatment and especially of those with totally reimbursed therapy. The investigation shows that the accepted principles in this area are appropriate with the modern concepts and tendencies, but for their real realization are necessary additional attempts for overcoming of the negatives in the process of developments through adequate administrative decisions and approaches.

The compliance of the community and private pharmacies' patients.

This issue is an attempt to expose the extent of compliance, respectively of non-compliance among the patients of the community and private pharmacies in Sofia. The authors make an attempt to elucidate the factors that influence upon the non-compliance of the patients in order to increase the rate of compliance among them. The results of the analysis of the received data show that the level of the compliance is 69% among the studied patients. Despite the presented adequate level of compliance among the patients, there are possibilities for the individualisation of the treatment, for supply with more information and for improvement of the patient-physician and patient-pharmacist interrelations.

Antibacterials: spontaneous report analysis for a six years period in Bulgaria.

The pharmacovigilance started in Bulgaria in 1975. The doctors had to send the filled so called "yellow cards" for the suspected Adverse Drug Reactions to the Centre for Drug Safety at the National Drug Institute. In Bulgaria a spontaneous reporting on the adverse drug reactions has been studied during the period 1991-1996 for Antibacterials. The total number of the reports was 2409 and 18% of this number related to the Antibacterials. The reporting rate varied from 0.3 to 1.8 per 100,000 inhabitancy. As a whole 29% of the adverse drug reaction reports concerned the patients in the age groups of 0-5 year old. Amoxicillin is a drug with a majority of reports for the period. This article reveals the limits of the operating spontaneous reporting system in Bulgaria and suggests some possible measures to improve and refine it in a connection to the WHO and EMEA requirements.

An attempt for complex assessment of the elderly patients' compliance.

Compliance is taking the correct amount of the correct medicine at the correct time. Today, the problem of prescription medicine noncompliance is gaining considerably more attention within the pharmacy community as a part of the pharmaceutical care and by the other health professionals. Sometimes, compliance is defined "objectively" by how much medication a patient takes, such as taking a certain percentage of prescribed medication or taking a certain number of pills over a specific time period. Undoubtedly the care for the elderly patients and especially for the elderly patients that are bedridden can be an especially difficult challenge. The extent of non-compliance among them is estimated to be about 40% to 50%. The aim of this research is to assess the level of compliance and the factors, that influence this process among a group of elderly patients. We applied a complex methodology in order to avoid the mistakes that are connected with the use of a single method separately.

Cancer, drug medication and noncompliance.

This issue is an attempt to expose the extent of compliance, respectively of non-compliance among the oncological patients of the Central Oncological centre in Sofia. There are several factors that influence upon the degree of nonadherence. The authors make an attempt to elucidate them in order to increase the rate of compliance among this group of patients. The results of the analysis of the received data show that the level of the compliance is 92% among the studied cancerous patients. Despite the shown high level of compliance there are possibilities for the individualization of their treatment.

Low dose treatment with methotrexate-adverse drug reactions survey.

Antineoplastic drugs caused various and frequent adverse drug reactions (ADR) in connection with their pharmacodynamics. Methotrexate (MTX) ADRs are preferably gastrointestinal disorders and hepatotoxicity (hepatic enzyme abnormalities). The aim of this study was to detect and analyse ADR induced by low-dose MTX treatment in rheumatology. We observed 94 patients, 63 with rheumatoid arthritis and 31 with psoriatic arthritis. All patients were co-medicated with nonsteroidal anti-inflammatory drugs (NSAID) as Diclofenacum, Indomethacinum, Piroxicamum and 51% with glycocorticosteroides. During the follow-up study we collected 18 case-reports with ADR for 17% of the patients. From the patients with registered ADR, 11 was treated with standard dose of 7.5 mg MTX for a week and 7 patients received from 10 to 15 mg for a week. The distribution of the cases according patients' gender was 9 females and 7 males. Prevail individuals in age groups' 41-50 and over 61 years. The most frequent adverse drug reactions were leucopenia, trombocytopenia, skin reactions and gastrointestinal disorders as vomiting, melaena, epigastrial pain, etc. The primary risk connected with long therapy of low doses MTX is hepatotoxicity that diagnose and treatment are painful and expensive. As a result of the appearance of ADR in 5 patients the therapy with MTX was not changed, in two cases MTX is stopped timely or the dosage is changed and in the rest 11 patients MTX was excluded from the therapeutic scheme.

Psychotropic drugs in Bulgaria-frequency and risk of adverse drug reactions.

The aim of the study is to determine the frequency and risk of appearance of adverse drug reactions/ADRs/during the treatment with psychotropic drugs. The first part of the study is an analysis of the use of the psychotropic drugs for one-year period of time in our country, performed by DDD methodology. An attempt is made to equalize the Bulgarian list of the psychotropic drugs with the ATC classification and to estimate the DDD/1000/day. The data for Bulgaria is compared with that of the other countries. The collected data for the psychotropic drug use is divided into 2 groups: Psycholeptics and Psychoanaleptics. It is made an attempt to explain the main differences between them. The number of the standard therapeutic courses/NT/is used for assessment of the frequency and risk of ADRs. The results from the study show that the determined frequency of appearance of ADR for the different drugs is within "rare" and "very rare' for 100,000 inhabitants, according to the WHO terminology. Only for the drug Tardyl (EGIS Pharmaceuticals, Hungary) with INN Aminobarbitalum + Glutethimidum + Promethazini hydrochloridum the frequency is above 1%. That fact makes us to recommend a limitation of the prescription and usage of this drug to the Bulgarian Ministry of Health and to the National Drug Agency.

Trends in registered psychiatric morbidity and forecasting psychotropic drug needs in Bulgarian hospitals.

Registered psychiatric morbidity in Bulgaria as a whole and particularly in Plovdiv, the second largest region of the country, was assessed. Three aspects of psychotropic drug usage were analysed, namely, changes in registered psychotropic drugs, the prescribed daily dose (PDD) values for 2 years, and the preferred therapeutic schemes, and drug usage and needs in a psychiatric hospital with 365 beds. This was done by time series analysis for evaluation of psychiatric morbidity and drug consumption data, calculation of PDD for psychotropic medicines, and, based on a modification of the World Health Organization's morbidity method, assessment and prediction of drug use and needs in Plovdiv hospital. The results indicated that the registered morbidity had increased by 4% over the period 1989-93 to 2,427 psychiatric patients per 100,000 people. The increased consumption of especially benzodiazepines and sedative medicines was analyzed. Diazepam was prescribed the most often (91.1%), followed by levomepromazine (86.4%), haloperidol (82.7%), etc. Future drug consumption in Plovdiv hospital is expected to decrease because therapeutic practice in hospitals has been revised and improved on the basis of the World Health Organization's recommendations.

Drug resistance of a viral population and its individual intrahost variants during the first 48 hours of therapy.

Using hepatitis C virus (HCV) and interferon (IFN) resistance as a proof of concept, we have devised a new method for calculating the effect of a drug on a viral population, as well as the resistance of the population's individual intrahost variants. By means of next-generation sequencing, HCV variants were obtained from sera collected at nine time points from 16 patients during the first 48 h after injection of IFN-α. IFN-resistance coefficients were calculated for individual variants using changes in their relative frequencies, and for the entire intrahost viral population using changes in viral titer. Population-wide resistance and presence of IFN-resistant variants were highly associated with pegylated IFN-α2a/ribavirin treatment outcome at week 12 (P = 3.78 × 10(-5) and 0.0114, respectively). This new method allows an accurate measurement of resistance based solely on changes in viral titer or the relative frequency of intrahost viral variants during a short observation time.

[Not Available]. / [Potentialities for the fulfilment of the pharmacists]

The pharmacists are highly qualified specialists that possess knowledge in the field of biology, chemistry and special medical disciplines. This defines their potential for a realization in different scientific fields. The article points out the place and the role of pharmacists with higher education as leaders in the drug security policy of the country. Here are seen the different fields of realization of the pharmaceutical staff in the pharmacy system, chemical - pharmaceutical and food industry, laboratories for a drug analysis, scientific research institutes, medical control of the social security, and army. The main functions of the pharmacy are formulated as follows: drug delivery, pharmaceutical services and a function connected with health policy.

[Production of labelled gamma globulin for the detection of Chlamydia]. / Poluchavane na beliazan gamaglobulin za dokazvana ne khlamidii.

A method was worked out for the production of an antichlamydial fluorescent serum to be used in the direct immunofluorescence test. Used were the methods of culturing Chlamydia in chick embryos and cell cultures, light and electron microscopy, differential centrifugation and supercentrifugation, gel chromatography in Sepharose, ultracentrifugation in saccharose, hyperimmunization of guinea pigs, conjugation of gammaglobulin with fluorescein isothiocyanate, complement fixation, and the direct immunofluorescence itself. Applied was a new immunologic scheme for the production of fluorescent sera. The guinea pigs were hyperimmunized twice and three times at 30-day intervals with highly purified, highly concentrated and adjuvant-treated chlamydial suspensions. It was concluded that the main condition for the production of high titer labelled gammaglobulins was to obtain purified and concentrated chlamydial antigens and use these with an adjuvant. To indicate Chlamydia in testing the conjugates it is essential to prepare standard test lamellas infected at high multiplicity of purified and concentrated Chlamydia.

[Demonstration of the M-protein of the Newcastle disease virus by current immunochemical methods]. / Dokazvane M-proteina na virusa na Niukiasulskata bolest chrez suvremenni imunokhimichni metodi.

The immunosorbent micromethod with the use of nitrocellulose paper was employed for the first time to demonstrate subviral antigens from animal origin, such as the M-protein of the Newcastle Disease virus. The method proved to be highly effective. It was useful in discovering amounts as high as 10-20 ng of the antigen, while with the use of the enzyme-linked immunosorbent assay (ELISA) amounts of about 200 ng could be evaluated. The immunosorbent micromethod performed on nitrocellulose paper was readily applicable as compared to all previously employed highly sensitive methods, and could be used successfully in immunochemical investigations.

[Survival of pathogenic microorganisms in the Black Sea mussel with an experimental infection]. / Prezhiviaemost na niakoi patogenni mikroorganizmi v chernomorskata mida pri eksperimentalno zaraziavane.

The survival rate was followed up of S. typhimurium, S. enteritidis, S. aureus, and E. coli in the organs of M. galloprovincialis L. in experimental conditions. A vat was used with 30 mussels in 10 l of sea water contaminated with 10 cm3 broth culture in conc. of 2.10(4) bacterial cells per cub. cm. The water was changed at 24-hour intervals. Microbiologic studies were carried out of the gills, liver, and meat of the mussels and of the sampled sea water. Lowest survival rate showed S. aureus--up to the second day, S. typhimurium and S. orientalis--up to the 4th days (in the gills only), and E. coli--up to the sixth day (in the gills and liver). None of the experimental strains to the sixth day (in the gills and liver). None of the experimental strains was isolated from the meat. With regard to the periods of investigation cited M. galloprovincialis L. could play the role of a carrier, resp. vector of the infection and could bring about a disease after consumption. Stated is the important part played by mussels as biofilter, and, at the same time, as source of pathogenic bacterial microflora.

Antiherpes effect of Melissa officinalis L. extracts.

Studies were carried out on the virucidal and antiviral effects of Melissa officinalis L. extracts (M1, M3, M3 and M4) with respect to Herpes simplex virus type 1 (HSV-1). Virucidal effect was registered within 3 and 6 hours of treatment using M4 administered in MTC. The remaining extracts inactivate the virus at the 12th and 24th hour. No significant values of inhibiting activity of M1, M2, and M3 on the same virus in vitro or in vivo were demonstrated. The presence of caffeic, rosmarinic and ferulic acids was demonstrated by thin-layer chromatography. Their role in the antiviral activity of M. officinalis L. is discussed.