Volume 9 (2016): Issue 1 (September 2016)

Histiocytoses comprise a group of diverse diseases of unknown etiology with various clinical presentation and evolution. The underlying pathology is characterised by accumulation and infiltration of variable numbers of cells of the monocyte-macrophage line in the affected tissues and organs. Histiocytoses are divided into three major classes: Langerhans cell histiocytosis (LCH), non- Langerhans cell histiocytosis, and malignant histiocytic disorders. The term LCH (also known in the past as histiocytosis X) encompasses the following rare diseases: Eosinophilic Granuloma, Hand-Schuller-Christian disease, Letterer-Siwe disease, Hashimoto-Pritzker disease, in which accumulation of pathologic Langerhans cells (LCs) leads to tissue damage. LCs usually reside in the skin and ensure protection against infections by destroying foreign substances. LC accumulation is caused by antigen stimulation and inadequate immune response. Thus, clinical LCH manifestations range from isolated disease with mono- or multifocal bone lesions to disseminated multisystem disease. LCH is a rare disease, affecting mainly children and young smokers, aged 20-50 years. Lung involvement in LCH usually presents as a mono-system disease and is characterized by Langerhans cell granulomas (LCG) infiltrating and impairing the distal bronchioles. The definite diagnosis is based on lung biopsy of CAT selected LCG areas. So far, there is no an effective treatment, but the better understanding of the mechanisms involved in the pathogenesis of the disease would help in the development of effective therapeutic strategies in the future.

Modem biobanks should be considered as important structural platforms for sharing samples and data, providing benefits that go beyond those of a single study. To increase the impact of biobanking on feasible strategies to protect and improve peoples’ health, it could be useful to increase awareness about biobank research potential in the scientific community, as well as to promote collaboration among different fields of expertise. The basis for involvement of biobanks in public health has already been partly built up, emphasising the synergy in organising efforts and the openness to infrastructure collaboration, to ensure reliable long-term funding and resource sharing to the benefit of all. To obtain the maximum benefits possible from biobanks, which currently constitute a set of heterogeneous structures, a strategy of coordinated governance is needed, based on harmonisation of different aspects, built on the consensus of all stakeholders involved and society in general. It is important that researchers strive to understand public attitudes in this biobanking involvement so that the relationships between public trust in biobanking, donors’ behaviours and support for public-health involvement in the health-care system can be better understood.

The objective of the study was to investigate the influence of sperm DNA fragmentation index (DFI) by DNA integrity test on pregnancy outcome and pregnancy loss after assisted reproductive technology (ART) procedure: autologous intracytoplasmic sperm injection (ICSI), donation eggs ICSI, and intrauterine insemination (IUI). We investigated men from 531 couples undergoing autologous ICSI procedure (n=416), from couples undergoing donation eggs procedure (n=39) and IUI (n=76). We performed the following interventions: semen analysis, DNA integrity test, embryo scoring by Gardner and Schoolcraft grading system (1999). The study showed no statistically significant differences between the group regarding pregnancy rate (χ²=0.55; p>0.05; OR=1.25, 95% Cl 1.23-1.46; p>0.05). However, with increased levels of DFI, the number of pregnancy losses became higher (including biochemical pregnancies and spontaneous abortions) at OR=5.65 (95% Cl 4.32-7.11; p=0.05). We examined the percentage of grade I blastocysts (by Gardner and Schoolcraft, 1999) before donation eggs embryo transfer and found a statistically significant correlation with both the DFI (χ²=7.80; p<0.05) and sperm morphology (χ²=6.14; p<0.05). Analysis of the relationship between DFI and IUI output (clinical pregnancy, miscarriage) revealed significant correlations in both directions: between DFI and pregnancy rate after IUI (χ²=6.29; p<0.05) and between the DFI and pregnancy development after IUI (χ²=6.87; p<0.05). The three group categories (autologous, heterologous ICSI procedures and IUI) studied showed that sperm samples with DFI>27% were associated with increased riskofearlypregnancyloss. Men with infertility should undergo DNA fragmentation assay in addition to the standard semen analysis. When DFI exceeds 27%, ICSI should be a method of choice, even in cases where the conventional parameters of semen analysis tests are normal.

The study is a statistical analysis of the dynamics and structure of forensic autopsies performed on dead bodies and body remains at University Hospital “Dr. Georgi Stranski” in Pleven. The study was based on forensic autopsy records database and forensic autopsy reports for the period 2009-2013. A total of 976 cases, including forensic autopsies, exhumations, examination of bone remains and inspection of dead bodies at the site of death (without following forensic autopsy), were analyzed. The studied data was coded and processed with Microsoft Excel software. A detailed data analysis of the number of autopsies, frequency of death by cause, distribution of death by gender, age, residence, place of death and its forensic category is presented. According to our results, the deaths due to diseases comprised 35% of all forensic autopsies. The most common cause of violent death was damage by mechanical factors (53%), followed by asphyxia (24.7%). The most common mechanical factor was vehicle crash trauma, followed by falling (22.5%). Accidents were the most common cause of death 62%, followed by suicides (31 %) and homicides 7%. Fifty-nine percent of the suicides were by hanging. The preliminary analyses of the studied data lead to several conclusions. The number of forensic autopsies was found to be progressively decreasing, but the road traffic death rate remained unchanged. Most of the forensic autopsies were performed in July. The highest death rates by age was observed in the age group between 51 and 60 years, with predominance of males over females (3.3 males: 1 females).

It is well known that some psychological features of adolescents are positively associated with alcohol drinking but less attention is paid to the psychological features of their significant others. The aim of the study was to describe a common psychological profile of significant others to regularly alcohol drinking adolescents. A survey was conducted using an original questionnaire, which classifying alcohol drinking as regular, social (incidental) and abstinence. Of 903 students (aged 15-19), 169 identified themselves as regular drinkers (RDA) and 279 - as abstainers (NDA). The significant others of these two groups were compared statistically in terms of a wide range of psychological features. It was found that RDAs were significantly more likely to have: fathers (OR=1.94; 95% Cl 1.04-3.62) and friends (OR=l .55; 95% Cl 1.02-2.36) that tended to avoid obligations in favor of pleasure; extravert friends (OR=1.55; 95% Cl 1.06-2.28) and lovers (OR=1.72; 95% Cl 1.14-2.59); impulsive lovers (OR=l .76; 95% Cl 2.86- 1.08), and obeying (OR=l .95; 95% Cl 1.01-3.80) friends; conventional fathers (OR=2.17; 95% Cl 1.27-3.72) and less likely to have independent mothers (OR=0.57; 95% Cl 0.32-0.99) and hardworking friends (OR=0.58; 0.35- 0.95). It seems that significant others of RDAs are less likely to exhibit models of rational reactions in everyday life that prevent adolescents from developing rational personality, thus making it possible to increase the probability of regular drinking in adolescence.

Recent scientific evidence has shown that risk behaviors tend to form a risk cluster, but less attention is paid to clustering ability of light forms of risky behavior like social drinking. The objective of the study was to analyze the risk clustering potential of social drinking. We conducted a survey using a self-completed questionnaire. Of 903 students (aged 15-19), 279 (30.9%) were found to be abstainers (NDA), and 455 (50.39%) were social drinkers (SDA). These two groups were compared statistically, concerning smoking and drugs use among them and their significant others. SDAs were more likely to smoke (OR=3.29; 95% Cl 2.35-4.59) than NDAs but not more likely to use soft drugs. Their fathers (OR=1.4; 95% Cl 1.04-1.89), friends (OR=1.78; 95% Cl 1.31-2.42) and lovers (OR=2.01; 95% Cl 1.39-2.89) were also significantly more likely to smoke, but only friends were more likely to use soft drugs (OR=1.75; 95% Cl 1.19- 2.58). SDAs were also more likely to start smoking in order to be closer to their peers (OR=l .84; 95% Cl 1.01- 3.37) and to smoke when communicate with peers (OR=2.12; 95% Cl 1.40-3.21). Risk clustering in social drinking adolescents is limited to smoking and does not expand to drug use, irrespective of provocation by friends. Heavy smoking among SDAs’ significant others might contribute for SDA smoking. So SDAs seem to be resistant to the psycho-social mechanisms adding drug use to drinking but sensitive to psycho-social mechanisms adding smoking to drinking.

The aim of the study was to carry out a toxicological chemical analysis of methanol in detectable quantities in the blood of patients with acute alcohol intoxication. Blood samples from 85 patients with acute alcohol intoxication were analysed for the presence of methanol. All patients with acute methanol intoxication were excluded from the study. The methods of gas chromatography with vapor-phase analysis (head-space) and flame ionization detection (FID) were used. The limit of detection (LOD=0.015 g/L) and the limit of quantification (LOQ=0.025 g/L) of methanol in whole blood were evaluated. In 30% of the cases, methanol was found in the blood in detectable quantities. The levels of methanol were on the average 5 to 6 times lower than the toxic methanol level (0200 g/L) and they were not due to natural metabolic processes (ingestion of fruit, fruit juices or vegetables). No reliable statistically linear correlation between the concentration of ethanol and methanol was found. Methanol subintoxications are major factors in alcohol intoxications, in which the quantity of the alcohol ingested is not as important as its quality. Chronic methanol subintoxication of people who often consume alcohol of poor quality is discussed.

We present a case of 58-year-old female patent with a long-term history of paranoid schizophrenia treated with different antipsychotics. She died with sign and symptoms of hypothermia. Autopsy revealed an ischemic stroke. We could not exclude involvement of antipsychotic medications as culprits.

Degenerative aortic stenosis is the second most common acquired valvular heart disease in adults (after mitral insufficiency) and the second most common cause for cardiac surgery (after coronary heart disease). The reasons for the occurrence of these diseases (congenital abnormality of the valve: bicuspid aortic valve disease, advanced renal failure, impaired calcium-phosphorus metabolism) have been established only in a small portion of these patients. The absence of a specific reason, causing calcification and narrowing of the aortic valve in recent years has challenged researchers to start investigating genetic factors that may correlate with the development of degenerative aortic stenosis. Regardless of the conducted studies, knowledge and identification of predictive genetic factors in the occurrence and progression of aortic stenosis are still insufficient. It is assumed that a specific genetic variant in the Lipoprotein (a) locus (LPA locus), reflected by the Lipoprotein (a) [Lp(a)] plasma levels, is connected to the pathology of aortic stenosis in multiethnic groups. The study of the genetic nature of aortic stenosis and significance of Lp( a) plasma levels and genetically determined variations of its structure associated with the manifestation and progression of valvular calcification in the future might provide predictive intervention. Similar studies relating to genetic polymorphisms in LPA locus, plasma concentrations of Lp(a) and their correlation with aortic stenosis have not beenconducted in Bulgaria so far.

Administration of antiplatelet therapy Aspirin and Clopidogrel (CLP) is a corner stone inpatients with Acute Coronary Syndrome (ACS) undergoing Percutaneous Coronary Intervention (PCI) with/without stent implantation. The CYP2C19*2 allele is the most important genetic variant determining response to CLP. We aim to investigate frequency of CYP2C19*2 polymorphism in patients with ACS and significance for the individual response to CLP therapy. The preliminary data of a study including a total of 120 patients with ACS undergoing PCI with stent placement and treated with dual antiplatelet therapy (CLP and Aspirin) are presented. So far 18 patients (41-81 year age) are tested for CYP2C19*l/*2 polymorphisms. The genotype CYP2C19*1/*1; CYP2C19*l/*2 and CYP2C19*2/*2 is demonstrated in 50%, 33%, 17% respectively, of the patients. The established frequency of CYP2C 19*2 allele (33%) is significantly higher (x²=5.220; p=0.022) than in healthy Bulgarian individuals (16%). In-stent thrombosis have developed 3 (17%) of patients: 2 are C YP2C19* l/*2 carriers, and 1 - homozygous CYP2C19*2/*2. The preliminary data demonstrate high prevalence of CYP2C19*2 polymorphism in patients with ACS and point to significance of the variant for CLP therapy. Further extension of the study with larger samples and monitoring of the patients are required to determine the effects of the polymorphism on the prognosis for major adverse cardiovascular events.

Platelet activation is a complex process in which platelet reorganization takes place associated with changes in the cell shape, topology, membrane elasticity and microparticle production. The aim of this study was to investigate the changes/aberrations in the platelet activity, elasticity and morphology in healthy subjects, carriers of A allele of prothrombin G20210A polymorphism. Blood samples from 18 healthy subjects were used for platelet analysis by force-mode atomic force microscopy. Restriction analysis was used to investigate the carriage of G20210A polymorphism in the prothrombin gene. Flow- cytometry was applied to evaluate platelet activation. Young’s modulus of the plasma membranes of platelets derived from healthy subjects, carriers of variant A allele of prothrombin 20210G>A polymorphism (407±69 kPa) is two times higher than the one determined for non­carriers (195.4±48.7 kPa; p<0.05). The background activity of platelets measured as an interrelation of Cd41/Cd61 and CD62 by flow cytometry was also higher in carriers of variant A allele of prothrombin 20210G>A polymorphism (5.0%) than in non-carriers (1.3%). Platelets isolated from healthy carriers of variant A allele of prothrombin 20210G>A polymorphism exhibited a higher level of activity and a higher degree of stiffness at the stage of spreading as compared to platelets from non­carriers.

We present a case of a 75-year-old female who underwent a total abdominal hysterectomy with bilateral salpingooophorectomy for an ovarian tumor. Hematoxylin-eosin- stained slides from tumor specimen revealed 1) focci of benign Brenner tumor; 2) mucinous cysts and 3) intracystic papillary projections resembling low-grade papillary transitional cell carcinoma (Grade 1-2) with squamous differentiation and comedo-type necrosis; 4) focal areas resembling noninvasive papillary transitional cell carcinoma Grade 3. Immunohistochemical investigation with a panel of antibodies (p63, p53, Ki-67, Wilms Tumor 1 - WT1, pi 6) was initiated. Areas resembling urothelial carcinoma showed diffuse nuclear positive reaction for p63 and wild-type expression of p53. Ki-67-nuclear positivity varied from less than 5% up to 30% in areas resembling high-grade urothelial carcinoma. WT1 expression was not seen. Weak but still exceeding background staining was observed in predominantly cytoplasmic fashion with few scattered positive nuclei in transitional cell nest of the benign component. No reactivity, however, was seen within the proliferative component. The histopathological diagnosis was a borderline/atypical proliferative Brenner tumor.

The patient has been regularly followed up and is at present disease-free 5 years after diagnosis. In this paper, the authors describe the morphological characteristics of Brenner tumors and address some debatable issues in the light of recent immunohistochemical and molecular studies.

Liposarcoma is one of the most common soft tissue sarcomas in adults with a relative incidence amongst other sarcomas ranging from 9.8% to 16%. It usually locates in the limbs and retroperitoneum. Primary liposarcomas of the larynx and hypopharynx are rare, comprising less than 20% of all head and neck liposarcomas. According to World Health Organization, these tumors are divided into four histologic types, and well-differentiated liposarcoma is the most common one. It is a tumor of low-grade malignancy that may recur locally, but does not metastasize. We present a case of laryngopharyngeal well- differentiated liposarcoma in an old patient with two previous removals. We also discuss recently published cases with this unusual location of liposarcoma.