Androgens in women: Establishing reference intervals for dehydroepiandrostenedione sulphate and androstenedione on the Roche Cobas.

Introduction: Immunoassays are the most common method in routine practice for measuring androgens in women. Study's aim was to establish new population specific indirect reference intervals (RI) for dehydroepiandrostenedione sulphate (DHEAS) and for new androstenedione test available on automated Roche Cobas electrochemiluminescent immunoassay method. Materials and methods: From extracted laboratory records, testosterone, sex hormone binding globulin and follicle-stimulating hormone were used as reference tests to exclude possibly diseased women. After the data selection steps, the study included 3500 subjects for DHEAS and 520 for androstenedione aged 20-45 years. To evaluate the need for age partitioning, we calculated standard deviation ratio and bias ratio. For each hormone, 90% and 95% RIs were calculated with appropriate statistical method. Results: Total age group (20-45 years) 95% RIs were: 2.77-11.50 µmol/L for DHEAS and 2.48-8.89 nmol/L for androstenedione. Age-stratified 95% RIs for DHEAS were: 3.65-12.76 µmol/L (20-25 years); 2.97-11.50 µmol/L (25-35 years) and 2.30-9.83 µmol/L (35-45 years). Age-stratified 95% RIs for androstenedione were: 3.02-9.43 nmol/L (20-30 years) and 2.23-7.75 nmol/L (30-45 years). Conclusion: New RIs for DHEAS were slightly wider for age group 20-25 and 35-45, while the differences in the age group 25-35 years were more pronounced. Androstenedione RI showed significantly higher concentrations than the manufacturer's. Age-related decrease of androgens should be considered when calculating RIs. We propose population specific, age-stratified RIs for DHEAS and androstenedione on electrochemiluminescent method, which should improve test interpretation in women of reproductive age.

Laboratory policies and practices for thyroid function tests in Croatia: survey on behalf of Working Group for Laboratory Endocrinology of the Croatian Society of Medical Biochemistry and Laboratory Medicine.

Introduction: Laboratory plays important part in screening, diagnosis, and management of thyroid disorders. The aim of this study was to estimate current laboratory preanalytical, analytical and postanalytical practices and policies in Croatia. Materials and methods: Working Group for Laboratory Endocrinology of the Croatian Society of Medical Biochemistry and Laboratory Medicine designed a questionnaire with 27 questions and statements regarding practices and protocols in measuring thyroid function tests. The survey was sent to 111 medical biochemistry laboratories participating in external quality assurance scheme for thyroid hormones organized by Croatian Centre for Quality Assessment in Laboratory Medicine. Data is presented as absolute numbers and proportions. Results: Fifty-three participants returned the questionnaire. Response rate varied depending on question, yielding a total survey response rate of 46-48%. All respondents perform thyroid stimulating hormone (TSH). From all other thyroid tests, most performed is free thyroxine (37/53) and least TSH-stimulating immunoglobulin (1/53). Laboratories are using nine different immunoassay methods. One tenth of laboratories is verifying manufacturer's declared limit of quantification for TSH and one third is verifying implemented reference intervals for all performed tests. Most of laboratories (91%) adopt the manufacturer's reference interval for adult population. Reference intervals for TSH are reported with different percentiles (90, 95 or 99 percentiles). Conclusion: This survey showed current practices and policies in Croatian laboratories regarding thyroid testing. The results identified some critical spots and will serve as a foundation in creating national guidelines in order to harmonize laboratory procedures in thyroid testing in Croatia.

Oxidative homeostasis in follicular fluid and reproductive outcomes - from bench to bedside.

Free radicals and oxidant molecules are part of our organism in a stable balance. However, when addressing female infertility, questions about their role in oocyte quality arise. This review outlines the major alterations of redox homeostasis in the follicular fluid through pathophysiological conditions in female reproduction and its potential effect on IVF outcome. A review of the literature was accurately performed. Manuscripts investigating follicular fluid biomarkers, especially related to oxidant molecules, were screened and used in this review. Studies assessing the follicular reactive species were found and screened. Moreover, studies assessing the IVF outcomes related to biomarkers were considered. The results are provided in an analytical pathway. The study of biomarkers confirms the shift to enhanced oxidizing modification of macromolecules and antioxidative consumption in the follicular fluid of women undergoing IVF treatment. A lack of congruency in methods appears to be marked in the design of scientific studies. However, it is not clear whether redox disbalance has a disruptive effect on the oocyte competence or whether it plays a role in the oocyte maturation process. Red-ox balance plays a questionable role in IVF outcomes. Possible further insights may consider the antioxidant role of adjuvants during controlled ovarian stimulation cycles.

Establishing paediatric reference intervals for thyroid function tests in Croatian population on the Abbott Architect i2000.

INTRODUCTION: Evaluation of thyroid function is often requested and therefore defining paediatric reference intervals (RIs) is of vital importance. Currently, there is a distinct lack of paediatric RIs for thyroid function tests in Croatia. Thus, we established RIs for thyroid stimulating hormone (TSH), total triiodothyronine (TT3), total thyroxine (TT4), free triiodothyronine (FT3) and free thyroxine (FT4) in the Croatian paediatric population. MATERIALS AND METHODS: Reference intervals were calculated from 397 apparently healthy children, aged from 2 days to < 19 years. Serum samples were analysed for thyroid function tests on the Abbott Architect i2000. Age- and sex-specific 95% RIs with 90% confidence intervals were established according to Clinical and Laboratory Standards Institute guidelines. To express the magnitude of sex and age variation, standard deviation ratio (SDR) was calculated using two-level nested ANOVA. The criterion for considering partitioning reference values was set to SDR > 0.3. RESULTS: All thyroid function tests required age partitioning, confirmed by SDR above 0.3. There was no need for sex partitioning, confirmed by SDR below 0.3. Still, FT3 was partitioned due to visually noticeable sex related difference for the oldest group (12 years to < 19 years). CONCLUSION: This is the first study to establish RIs for thyroid function tests in the Croatian paediatric population. We propose RIs for widely used Abbott platform, thus giving laboratories method- and population-specific paediatric RIs for thyroid function tests that should improve clinical test interpretation.

Optimizing laboratory defined macroprolactin algorithm.

INTRODUCTION: Macroprolactinaemia is a well-known analytical problem in diagnostics of hyperprolactinaemia usually detected with polyethylene glycol (PEG) precipitation method. Since there is no harmonization in macroprolactin detection and reporting results, this study proposes and evaluates the usefulness of in-house developed algorithm. The aims were to determine the most suitable way of reporting results after PEG treatment and the possibilities of rationalizing the precipitation procedure. MATERIALS AND METHODS: This is a retrospective study based on extracted data for 1136 patients. Prolactin concentrations were measured before and after PEG precipitation on Roche cobas e601. Macroprolactinaemia was defined by percentage recovery and post-PEG prolactin concentrations. RESULTS: Prevalence of macroprolactinaemia using recovery criteria of ≤ 40%, ≤ 60%, and post-PEG prolactin concentrations was 3.3%, 8.8% and 7.8%, respectively. Raising the cut-off value from the upper limit of the manufacturer's reference interval to 32.9 µg/L does not drastically change detected macroprolactinaemia with recovery criteria. Post-PEG prolactin concentrations showed more than half of the patients with macroprolactinaemia would be overlooked. Regardless of the criteria, a cut-off of 47.0 µg/L would miss most of the macroprolactinaemic patients. Repeated recovery measurements of follow-up patients showed there is a significant difference with mean absolute bias of 9%. CONCLUSIONS: Post-PEG prolactin concentration with corresponding reference interval is the most suitable way of reporting results. All samples with prolactin concentration above the upper limit of the manufacturer's reference interval should be submitted to PEG precipitation. Follow-up period could be prolonged since the difference between the recoveries of repeated measurements is not clinically significant.

Undetectable Serum Levels of Anti-Müllerian Hormone in Women with Ovarian Hyperstimulation Syndrome During In Vitro Fertilization and Successful Pregnancy Outcome: Case Report.

We report a unique case of undetectable serum levels of anti-müllerian hormone (AMH) in women with polycystic ovary syndrome (PCOS) who developed ovarian hyperstimulation syndrome (OHSS) during in vitro fertilization (IVF). A case is described of a 28-year-old woman with clinical symptoms of PCOS and AMH serum level below analytical sensitivity (<1.0 pmol/L). After undergoing controlled ovarian stimulation, the patient developed OHSS. After follicle aspiration, seven oocytes were recovered. Three of them were used for intracytoplasmic sperm injection (ICSI) and fertilized, but with unsuccessful pregnancy outcome. A successful pregnancy was achieved in the second IVF/ICSI cycle with six oocytes retrieved and three embryos transferred. At 39 weeks of gestation, the patient delivered a healthy baby weighing 3930 g and 50 cm long. In conclusion, although AMH is considered a useful tool in ovarian reserve assessment and in predicting response to controlled ovarian hyperstimulation, the case presented shows that AMH should not be used as an independent ovarian marker.

Pregnant women's knowledge and attitudes to prenatal screening for fetal chromosomal abnormalities: Croatian multicentric survey.

We aimed to assess opinion and preferences of Croatian pregnant women regarding the prenatal screening tests for trisomies. The study was conducted in Zagreb, Split, Cakovec, Nova Gradiska, Gospic and Zlatar. It was organized in the public primary healthcare centers, among the pregnant volunteers during their first visit to an antenatal clinic (7-12 weeks pregnant). The respondents filled anonymous questionnaire reflecting their knowledge and attitudes to the screening options. In total, 437 completed questionnaires were selected. The average maternal age and the level of education differed significantly between the respondents in the respective cities (P < 0.001). Of the respondents with positive attitude towards screening, the majority would prefer the first-trimester combined test (160/219; 73.1%), while 37/219 (16.9%) opted for the second-trimester biochemical screening. The remaining 22/219 (10.0%) would accept only the ultrasound screening. Among the 224 respondents, who would accept the combined first-trimester test, 95 (42.4%) held a college and university degree, whereas among 59 women, who would choose the second-trimester biochemical screening, 14 were highly educated (23.7%). The difference was statistically significant (P = 0.016). The univariate regression analysis showed that age, level of education and previous information were significant variables predictive for the choice of the test; the level of education and previous knowledge remained significant in the multivariate model. The survey has revealed some of the points that should be improved in the future concept of screening program in Croatia. Health professionals should persist to mend women's knowledge about prenatal screening, taking into consideration women's preferences as well.

Visfatin is not associated with inflammatory markers in patients on hemodialysis.

BACKGROUND: Cardiovascular events are a major cause of death in patients with end-stage renal disease. Endothelial dysfunction represents a key event in atherosclerosis development and has been implicated in the pathophysiology of different forms of cardiovascular disease, including chronic kidney disease. In recent years, visfatin, a ubiquitous adipokine, has been described as a potent marker of endothelial inflammation and dysfunction. The aim of the study was to investigate the association of visfatin with well-known markers of inflammation and endothelial dysfunction. METHODS: Serum and plasma samples from 66 patients (40 males and 26 females) treated by hemodialysis were analysed for visfatin, fibrinogen, CRP, PAI-1 levels. Visfatin was determined by ELISA method while CRP, fibrinogen and PAL-1 were obtained by standard laboratory methods. RESULTS: We observed statistically significant correlation between visfatin level and fibrinogen (r = 0.51; p = 0.008) and the time on dialysis in female patients (r = 0.70; p < 0.001). PAI-1 and CRP did not correlate with visfatin in males nor in females. CONCLUSIONS: Visfatin is correlated with time on dialysis and with fibrinogen only in female dialysis patients. To confirm this, further studies are needed with a higher number of patients.

Macroprolactinemia: new insights in hyperprolactinemia.

Hypersecretion of prolactin by lactotroph cells of the anterior pituitary may lead to hyperprolactinemia in physiological, pathological and idiopathic conditions. Most patients with idiopathic hyperprolactinemia may have radiologically undetected microprolactinomas, but some may present other causes of hyperprolactinemia described as macroprolactinemia. This condition corresponds to the predominance of higher molecular mass prolactin forms (big-big prolactin, MW > 150 kDa), that have been postulated to represent prolactin monomer complexed with anti-prolactin immunoglobulins or autoantibodies. The prevalence of macroprolactinemia in hyperprolactinemic populations between 15-46% has been reported. In the pathophysiology of macroprolactinemia it seems that pituitary prolactin has antigenicity, leading to the production of anti-prolactin autoantibodies, and these antibodies reduce prolactin bioactivity and delay prolactin clearance. Antibody-bound prolactin is big enough to be confined to vascular spaces, and therefore macroprolactinemia develops due to the delayed clearance of prolactin rather than increased production. Although the clinical symptoms are less frequent in macroprolactinemic patients, they could not be differentiated from true hyperprolactinemic patients, on the basis of clinical features alone. Although gel filtration chromatography (GFC) is known to be the gold standard for detecting macroprolactin, the polyethylene glycol precipitation (PEG) method has offered a simple, cheap, and highly suitable alternative. In conclusion, macroprolactinemia can be considered a benign condition with low incidence of clinical symptoms and therefore hormonal and imaging investigations as well as medical or surgical treatment and prolonged follow-up are not necessary.

Reference intervals for reproductive hormones in prepubertal children on the automated Roche cobas e 411 analyzer.

OBJECTIVES: To establish reference intervals for luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol (E2), progesterone (P), total and free testosterone (T) and sex-hormone binding globulin (SHBG) in prepubertal children and to assess age- and gender-related differences. DESIGN AND METHODS: A total of 948 subjects, 480 girls and 468 boys, between 1 and 11 years of age, were included in this study. All assays were performed on a Roche cobas e 411 immunoassay analyzer. Reference intervals have been evaluated according to the most recent CLSI guidelines. RESULTS: Median values of LH, FSH and T were significantly higher in subgroups ranging from ≥ 8 to < 11 years, for both genders. In girls of that age, reference values of E2 were significantly higher than in younger ones, and in boys of the corresponding age. CONCLUSION: Established reference intervals are applicable to other laboratories that use the same instrumentation.

Biochemical screening of fetal aneuploidies and neural tube defects by "double-test" in Croatia: a 10 years' experience.

The aim of the study is to investigate the efficiency of the second-trimester biochemical screening, with maternal serum alpha-fetoprotein (MS-AFP) and free beta-subunit of human chorionic gonadotropin (free beta-hCG), during the ten-year period. The study included 11,292 of pregnant women between the 15th and 18th gestational week, who underwent screening from November 1996 to December 2006. The risk for trisomy 21 and trisomy 18 were calculated by computer software, based on a model which generated the final risk for fetal aneuploidies from the pregnant woman's a priori age risk and the likelihood ratio of the distribution of the biochemical markers, according to the second-trimester gestation. With the cut-off value of the final risk > or = 1:250, the detection rate for trisomy 21 was 75% (21/28). In women less than or equal to 35, the detection was 57.1% (8/14) and 92.9% (13/14) in those over 35 years, respectively. The detection rate of trisomy 18 was 50% (2/4). The results confirmed that the implementation of double-test, as non-invasive screening for fetal aneuploidies, should be accepted as a complementary method of antenatal care.

Anti-Müllerian hormone: a unique biochemical marker of gonadal development and fertility in humans.

Anti-Müllerian hormone (AMH) is a glycoprotein belonging to the transforming growth factors (TGF-P). AMH plays a fundamental role in the regression of Müllerian ducts in male embryo. In its absence, Müllerian ducts develop into female inner reproductive organs. In boys, it is significantly produced in Sertoli cells of testes until puberty and then slowly decreases to residual values for the rest of the men's life. AMH serves as a biochemical marker of the presence of testes in cryptorchidic males. In females, AMH is secreted by granulosa cells of small follicles in the ovary. Serum values are almost undetectable during infancy and then rapidly increase with the onset of puberty, reflecting the initial recruitment of primordial follicles. AMH is produced in growing follicles until they reach a stage when dominant follicle is detached from a cohort of antral follicles. The measurement of serum AMH levels during woman's reproductive life represents an ideal tool for the assessment of the ovarian follicular reserve. The advantage of AMH in relation to the ovarian steroid hormones is that serum levels do not fluctuate significantly during the menstrual cycle. In addition, circulating AMH strongly correlates with antral follicle count (AFC), visualized by ultrasound in the follicular phase of the cycle. As the number and quality of the oocytes diminish throughout the woman's reproductive life, serum concentrations of AMH gradually decrease and fall below detectable levels in menopause. This could be of particular interest in subfertile and infertile women undergoing assisted reproductive techniques (ART) in achieving pregnancy.

The impact of temporal variability of biochemical markers PAPP-A and free beta-hCG on the specificity of the first-trimester Down syndrome screening: a Croatian retrospective study.

BACKGROUND: The variability of maternal serum biochemical markers for Down syndrome, free beta-hCG and PAPP-A can have a different impact on false-positive rates between the 10+0 and 13+6 week of gestation. The study population comprised 2883 unaffected, singleton, spontaneously conceived pregnancies in Croatian women, who delivered apparently healthy child at term. Women were separated in 4 groups, dependently on the gestational week when the analyses of biochemical markers were performed. The concentrations of free beta-hCG and PAPP-A in maternal serum were determined by solid-phase, enzyme-labeled chemiluminiscent immunometric assay (Siemens Immulite). Concentrations were converted to MoMs, according to centre-specific weighted regression median curves for both markers in unaffected pregnancies. The individual risks for trisomies 21, 18 and 13 were computed by Prisca 4.0 software. FINDINGS: There were no significant differences between the sub-groups, regarding maternal age, maternal weight and the proportion of smokers. The difference in log10 MoM free beta-hCG values, between the 11th and 12th gestational week, was significant (p = 0.002). The difference in log10 MoM PAPP-A values between the 11th and 12th, and between 12th and 13th week of gestation was significant (p = 0.006 and p = 0.003, respectively). False-positive rates of biochemical risk for trisomies were 16.1% before the 11th week, 12.8% in week 12th, 11.9% in week 13th and 9.9% after week 13th. The differences were not statistically significant. CONCLUSIONS: Biochemical markers (log10 MoMs) showed gestation related variations in the first-trimester unaffected pregnancies, although the variations could not be attributed either to the inaccuracy of analytical procedures or to the inappropriately settled curves of median values for the first-trimester biochemical markers.

Serum levels of antimüllerian hormone in women with regular menstrual cycles.

Antimüllerian hormone (AMH) is produced by Sertolli cells of the testes and granulosa cells of the ovaries. Recent studies have indicated that AMH may be a novel measure of ovarian reserve. Also, earlier reports have presented minimal fluctuations ofAMH levels throughout the menstrual cycle. The aim of this preliminary study was to demonstrate the relation of serum AMH levels and age in women with regular menstrual cycles and normal hormonal regulation of ovarian function. The study included 35 women divided into two groups of women aged 30 or younger and those older than 30. Hormone concentrations were assessed by measurements of lutropin (LH), follitropin (FSH), estradiol (E2), testosterone (T), sex hormone binding globulin (SHBG) and AMH on cycle day 3-5 (follicular phase); and LH, FSH and E2 on cycle day 13-15 (ovulation). Progesterone level was determined on cycle day 19-23 (luteal phase). Median age differed significantly between the two groups of study subjects (P = 0.001). Study results confirmed regular ovarian response to physiological gonadotropin stimulation, which is the assumption for normo-ovulatory cycles. Some decrease in the mean serum AMH levels was recorded in women over 30 years of age, although the difference was not statistically significant (P = 0.0693). There was no statistically significant difference in serum AMH concentrations between follicular phase and ovulation in study women (P = 0.3124). Our preliminary results, although obtained in a limited number of women, support the diagnostic value of AMH as a reliable marker of ovarian reserve.

[Antifungal drug resistance: mechanisms of resistance, freqeuncy, prevention and control of resistance].

Fungi are important causes of human infections, especially systemic mycoses. Pathogenic fungi have many complex mechanisms of resistance to antifungal drugs. In this article, information about the cellular, genetic and clinical factors contributing to antifungal-drug resistance, and their diagnostic and epidemiologic characteristics are reviewed. Understanding the mechanisms of resistance should assist in developing better detection and preventive strategies. The emergence of acquired antifungal resistance, especially in immunocompromised hosts, has necessitated routine antifungal susceptibility testing. Initial antifungal screening of clinical isolates by the disk-diffusion method would be followed by confirmation of resistant strains by the broth dilution method. Also, strategies to avoid and suppress the antifungal resistance are discussed. There are several strategies to overcome antifungal resistance, including increased antifungal-dose intensity, immunomodulation, combined antifungal therapy and new antifungals.

Diagnosis of Trichomonas vaginalis infection: The sensitivities and specificities of microscopy, culture and PCR assay.

OBJECTIVES: The aim of this study was to compare wet mount-, Giemsa stain-, acridine orange fluorescent stain-, cultivation- and polymerase chain reaction (PCR)-based approaches to establish which method or combination of methods was most effective in the laboratory diagnosis of trichomoniasis. STUDY DESIGN: Out of 200 investigated patients with various gynecological complaints, Trichomonas vaginalis infection was detected in 27 (13.5%) by any of methods investigated. Among women with trichomonads, a typical clinical finding was presented in only nine. For analysis of sensitivity and specificity of the methods used, the receiver operating characteristic (ROC) curve concept with culture as a gold standard was applied. RESULTS: Infection was diagnosed by wet mount in 14 (7.0%) women, by Giemsa stain in 11 (5.5%) and by acridine orange stain in 16 (8.0%) women. In 21 (10.5%) women, it was diagnosed by culture in Diamond's medium, and in 22 (11.0%) by PCR. For the initial diagnosis of trichomoniasis, wet preparation is the test that is widely available in most STD clinics, but its sensitivity is poor (66.67%). Giemsa stain shows a low sensitivity of 52.38%. Acridine orange shows reasonable sensitivity and specificity of 71.43% and 99.44%, respectively. The sensitivity and specificity of PCR (80.95% and 97.21%) did not exceed that of culture. CONCLUSION: With regard to the fact that trichomoniasis can have an atypical or even asymptomatic course, in order to accurately diagnose this disease, microbiological investigation is necessary. Comparison of different methods showed that at least two techniques, such as culture and acridine orange staining, have the potential for better diagnosis of T. vaginalis infection. PCR detection of infection has been demonstrated to be highly specific and sensitive, but its availability and cost effectiveness are in question. PCR could provide an alternative for laboratory diagnosis of trichomoniasis by culture.

[The role of parasites and fungi in secondary infertility]. / Uloga parazita i gljiva u nastanku sekundarnog steriliteta.

INTRODUCTION: Parasite-host relationships can cause diminished or absent ability to conceive, ectopic pregnancy or pregnancy with undesired course. LITERATURE REVIEW: There are reports that some protozoa, helminths and fungi may impair women's reproductive capacity, causing deformities of genital tract, so that conception is impossible, or, if it does occur, normal implantation and development of placenta are impossible. Schistosoma haematobium may cause vulvar papule, swelling, tumors, irregular vaginal hemorrhage, tubular infertility and ectopic pregnancies. Patients with cirrhosis caused by schistosomas have gonadal dysfunction and schistosomiasis itself can lead to tubular infertility. Some authors found microfilaria of Mansonella perstans in follicular aspirates in patients with tubular adhesions. Chronic Entamoeba histolytica infection can cause pelvic pain and dyspareunia in some patients. Although Trichomonas vaginalis is a common cause of tubal inflammation, this protozoa affects semen quality and leads to secondary infertility. Soluble parasite extract of T. vaginalis can lead to impaired motility of 50% spermatozoa in vitro and affects semen quality by increased viscosity and amount of debris, or damage spermatozoid membrane. In enterobiosis, presence of adult worms and eggs in fallopian tube, can be followed by chronic salpingitis and tubal occlusion. Also in ascariosis, presence of adult forms and eggs can lead to acute colpitis. chronic endometritis, salpingitis or ovarian abscess. The consequence of fungal infections, such as colpitis and endometritis, caused by Candida albicans, may be infertility. Also, according to some reports, C. albicans leads to decreased spermatozoan motility. CONCLUSION: Hence parasites and fungi can cause infertility, we recommend examination of both partners in treatment of infertility.

[Human babesiosis--recent discoveries]. / Humana babezioza--nova saznanja.

INTRODUCTION: Babesiosis is caused by intraerythrocytic parasites of the genus Babesia, which is a common animal infection worldwide. This protozoa requires both a competent vertebrate and a nonvertebrate host (Ixodes sp. etc.) to maintain the transmission cycle. HUMAN BABESIOSIS: Human babesiosis is predominantly caused by Babesia microti (rodent-borne piroplasm, an emerging zoonosis in humans in North America) and by Babesia divergens (bovine pathogen, in Europe). Occasionally, infection in America is caused also by a newly recognized species, so-called WA1 piroplasm. The spectrum of human babesiosis in the USA is broad, and ranges from an apparently silent infection to a fulminant. In Europe, babesiosis is considerably rarer, but more lethal (42% mortality rate in Europe and 5% in the USA, for clinically apparent infections) and mostly in splenectomized patients. Various determinants are involved in the severity of infection, such as age, immunocompetence and coinfection with other pathogens (Borrelia burgdorferi). B. microti antigens can trigger specific activation of T-cells and the infection can be effectively controlled by a Th1-dominant CD4+ T-cell response. The diagnosis of babesiosis should include examination of blood smears stained by Giemsa, as well as serologic evaluation with indirect immunofluorescent antibody tests and possibly PCR. The treatment of babesiosis depends on severity of cases; if it is mild it resolves spontaneously, whereas very severe cases with B. divergens require prompt treatment that includes erythrocyte exchange transfuision along with intravenous clindamycin and oral quinine to arrest hemolysis and prevent renalfailure. This paper offers an overview of recent developments in the investigation of Babesia sp. and babesiosis.

Correlation between clinical symptoms and diagnosis of trichomoniasis in women.

The aim of this study was to investigate correlation between clinical symptoms and diagnosis of trichomoniasis in women. 200 women were included in the study. Swabs were taken from all patients from the posterior vaginal fornix. Each sample was examined using the following five methods: wet mount, Giemsa stain, acridine orange fluorescence stain, cultivation in Diamond medium and PCR method. Trichomoniasis was diagnosed in 27 women using any of the applied methods and 33.3% presented with typical frothy yellow-green discharge, characteristic for tichomoniasis and yellowish discharge characteristic for the third group of vaginal discharge. White discharge, characteristic for Candida infection, was found in 18.5% of patients with diagnosed trichomoniasis. Finally, 14.8% of positive patients had a normal discharge. Based on the results of our study we came to the conclusion that microbiological investigations are necessary for accurate diagnosis of trichomoniasis, as well as for revealing asymptomatic infections, in order to prevent spreading of this relatively common disease.

[First case of Malassezia globosa isolation in Serbia]. / Prvi slucaj isolacije Malassezia globosa kod nas.

Today is known that genus Malassezia includes seven species: M. furfur, M. sympodialis, M. obtusa, M. globosa, M. restricta, M. sloofflae and M. pachydermatis, but role of each of the species in the pathogenesis of disease has not been elucidated yet, so further laboratory isolation and identification are necessary. We report the first case of isolation of Malassezia globosa in Serbia (Belgrade), in a patient suffering from Pityriasis versicolor. Identification of M. globosa was based on macroscopic, microscopic and biochemical characteristics. Isolation was done on Leeming and Notman medium and on mDixona agar, at 350C, during 7 days in aerobic conditions. Also the yeast's biochemical phenotype was determined as catalase (+), lipase (+), esculin degradation (-), Tween (20, 40, 60 and 80) assimilation (-). M. globosa is a lipophilic yeast of the genus Malassezia and the common member of the skin flora. In concordance with some predisponing factors M. globosa is implicated in the pathogenesis of several skin diseases (pityriasis versicolor, malassezia foliculitis, seborheic dermatitis and some forms of atopic dermatitis). In immunocompromised patients and neonates this yeast can even cause fatal systemic infections. Because the role of Malassezia spp. In pathogenesis of skin disease is not still determined, we suggest laboratory diagnosis and identification of these species as a routine diagnostic procedure.