Molecular genetic study of congenital adrenal hyperplasia in Serbia: novel p.Leu129Pro and p.Ser165Pro CYP21A2 gene mutations.

PURPOSE: Congenital adrenal hyperplasia (CAH) is an autosomal recessive disease characterized by impaired adrenal steroidogenesis and most often caused by CYP21A2 gene mutations. For the first time, we reported complete spectrum and frequency of CYP21A2 gene mutations in 61 unrelated patients with classical and non-classical CAH from Serbia. METHODS: Direct DNA sequencing of whole CYP21A2 gene and polymerase chain reaction with sequence-specific primers for detection of CYP21A1P/CYP21A2 chimeras were combined. RESULTS: We identified 18 different pathogenic alleles-two of them novel. Mutation detection rate was highest in patients with salt-wasting form of CAH (94.7%). The most prevalent mutation was intron 2 splice site mutation, c.290-13A/C>G (18.5%). Other mutation frequencies were: CYP21A1P/CYP21A2 chimeras (13%), p.P30L (13%), p.R356W (11.1%), p.G110fs (7.4%), p.Q318X (4.6%), p.V281L (4.6%), p.I172N (2.8%), p.L307fs (2.8%), p.P453S (1.9%), etc. Mainly, frequencies were similar to those in Slavic populations and bordering countries. However, we found 6.5% of alleles with multiple mutations, frequently including p.P453S. Effects of novel mutations, c.386T>C (p.Leu129Pro) and c.493T>C (p.Ser165Pro), were characterized in silico as deleterious. The effect of well-known mutations on Serbian patients' phenotype was as expected. CONCLUSIONS: The first comprehensive molecular genetic study of Serbian CAH patients revealed two novel CYP21A2 mutations. This study will enable genetic counseling in our population and contribute to better understanding of molecular landscape of CAH in Europe.

Influence of acetylsalicylic acid (aspirin) on biofilm production by Candida species.

Candida spp. are important causative agents of infections associated with biofilm formation. Management of biofilm-related infections is extremely difficult and therefore new therapeutic solutions are needed. This study for the first time explored the possible effect of aspirin on Candida spp. Biofilm-producing capacity. Two strains of C. guilliermondii, and one strain per species of C. kefyr, C. glabrata, C. albicans, and C. parapsilosis were included in the study. The antifungal property of aspirin was tested by the broth microdilution method, while effect of aspirin on biofilm formation was determined by the microtiter-plate test. The minimal inhibitory concentrations of aspirin obtained ranged from 2.17 to 8.67 mM and minimal fungicidal concentrations were from 4.33 to 8.67 mM. The concentrations of aspirin which induced statistically significant decrease in biofilm formation ranged from 0.43 mM to 1.73 mM of aspirin, depending on the tested yeast strain. Therefore, the significant effects of aspirin on growth and biofilm formation of Candida spp. were achieved only with suprapharmacological concentrations of the drug. The influence of the inoculum size on the effect of aspirin on biofilm formation was determined for C. albicans only and a significant decrease was observed also at suprapharmacological concentrations of aspirin, irrespective of the inoculum size. The results obtained in the present study show aspirin to be a drug with the potential to affect and suppress biofilm formation by Candida spp., and provide support for further investigation.

Meropenem therapy failure in Listeria monocytogenes infection.

Listeria monocytogenes is highly susceptible to meropenem in vitro, but data on the efficacy of meropenem in clinical cases of listeriosis are scarce. Described here is the case of a child with aplastic anemia who acquired nosocomial listeriosis and failed to respond to initial meropenem therapy. Resolution of fever was not noted after 5 days of therapy with meropenem and, more importantly, clinical worsening was observed during this period. The patient began to improve after ampicillin was introduced to the therapeutic regimen. In total, meropenem was administered for 15 days and ampicillin for 10 days.

Autopsy material analysis at the Cardiology Clinic in 1999.

Clinico-pathological studies serve as a valuable source of information in everyday practice of most medical institutions. The aim of this study was to correlate the clinical and pathological diagnoses of principal disease and cause of death after autopsy in patients who died during 1999 at the Cardiology Clinic, Institute of Cardiovascular Diseases, Sremska Kamenica, Yugoslavia. Medical histories and autopsy reports of dead patients were analyzed: 255 (8.46%) patients died, but only 72 (28%) underwent autopsy. In 53 patients (73%) clinical cause of death was cardiac. Among these patients, consequences of coronary (atherosclerotic) artery disease prevailed in 41 patients (77%). Noncardiac cause of death was established in 19 patients (27%) and vascular causes prevailed with 17 (23%). The comparison of clinical and principal disease established by autopsy revealed a complete concordance in 62 cases, partial concordance in 1 and no concordance in 9 cases. Thus, concordance was found in 86-87.5%. The correlation coefficient was 0.48. Comparison of clinical and autopsy diagnoses of causes of death revealed concordance in 53 patients, with correlation coefficient 0.26.

[The Crigler-Najjar syndrome, type I. Therapeutic dilemmas]. / Krigler-Najarov sindrom, tip jedan. Terapijske dileme.

Crigler-Najjar syndrome type I is a rare hereditary disease that is manifested by high concentration of unconjugated bilirubin in serum, which may lead to neurologic disorders. Management consists of conservative therapy and partial liver transplantation and, since recently, hepatocyte transplantation. Both therapeutic procedures have been continually improved and are yielding better results. We present a girl, aged three and a half years, who has been living at the Neonatology Department of the University Childrens Hospital in Belgrade since birth due to the permanent need for phototherapy and other measures so as to keep her bilirubin at minimal level. We present numerous management dilemmas, having in mind the controversial attitudes of physicians from other world centres, starting from those who consider that surgery should be done before the age of six years to those who consider that the patient should be treated conservatively only. Our attempts to avoid neurological disorders developing in our little patient surpasses professional interest only, because this girl is an exceptionally lovely and smart child with IQ 128.

[Electrocardiography and echocardiography in the diagnosis of chronic cor pulmonale]. / Elektrokardiografija i ehokardiografija u dijagnostici hronicnog plucnog srca.

The study was aimed at evaluating the possibilities of electrocardiography (ECG) and echocardiography in establishing the diagnosis of chronic pulmonary heart disease (CPHD). Sixty patients with a chronic obstructive pulmonary disease (COPD) and clinical symptoms of cardiac decompensation were observed. The examined patients ranged from 26 to 74 years of age, the mean age being 59.25 years. The diagnosis of CPHD was established by electrocardiography in 43 patients (78.18%) while in 57 patients (95%) it was done by ultrasound cardiography. Morphologic changes of the right ventricle (RV) were quantified, i.e. the diagnosis of chronic pulmonary heart disease was confirmed by echocardiography parameters such as right ventricle free wall thickness--RVFWT (0.82 +/- 0.09 cm), enddiastolic right ventricle dimension--ERVD (3.78 +/- 0.73 cm) and tricuspid regurgitation--TR (2.9 +/- 0.59, i.e. 2.58 +/- 0.55 after therapy). The information on the right ventricle function was provided by the values of echocardiography parameters, right ventricle ejection fraction--RVEF (30.98 +/- 5.88%, i.e. 34.1 +/- 5.64 after therapy) and right ventricle systolic pressure--RVSP (60.41 +/- 16.98 mmHG, i.e. 51.50 +/- 12.76 after therapy). Monitoring of these values after therapy, as well as of the RTO changes gave a good insight into the therapy effects. Echocardiography has been concluded to be of better sensitivity than ECG in diagnosing CPHD and both methods, noninvasive and easily applicable, have an important role in examining cardiac changes in patients with COPD.

[Evaluation of ejection fraction and end-diastolic volume using one-dimensional and two-dimensional echocardiography]. / Procena istisne frakcije i enddijastolnog volumena pomocu jednodimenzionalne i dvodimenzionalne ehokardiografije.

A group of 30 healthy persons and 120 coronary patients were investigated by one and two-dimensional echocardiography and digital subtraction angiography. End-diastolic volume and ejection fraction were estimated. Patients with coronary disease had greater volumes than healthy persons and smaller ejection fraction as well. The correlation of the given values showed that data gathered by two-dimensional echocardiography have greater coefficient in comparison to digital subtraction angiography (end-diastolic volume index r = 0.86, ejectional fraction r = 0.88, p < 0.01) than the same data estimated by one-dimensional technique. Two-dimensional echocardiography is a reliable noninvasive method for evaluation of these significant indicators of left ventricle function.