Metin Korkmaz

Tissue Parasites in Patients with Chronic Urticaria

Chronic urticaria is an important diagnostic and therapeutic problem. We aimed to investigate the sero‐prevalence of tissue parasites causing toxocariasis and fasciolosis in patients with chronic urticaria. All cases were analyzed for antibodies against Toxocara canis and Fasciola hepatica by modified (homemade) ELISA. The excretory/secretory products of Toxocara and Fasciola were used as antigens (ES‐ELISA) in the test. In this study, the highest toxocariasis seropositivity (29.0%) rate and the highest fasciolosis seropositivity (14.5%) rate were found in patients with chronic urticaria. Fasciolosis seropositivity and total seropositivity of toxocariasis and fasciolosis in patients with chronic urticaria was significantly higher than in healthy controls (p<0.05). Toxocariasis seropositivity in patients with chronic urticaria was not significantly higher than that in healthy controls (p<0.05). We suggest that parasitic infections should be considered as an important cause of chronic urticaria. Serological methods should be used to expose the diagnosis of tissue parasites in such cases.

Identification of potential serodiagnostic and subunit vaccine antigens by antibody profiling of toxoplasmosis cases in turkey

Toxoplasmosis, caused by infection of the protozoan parasite Toxoplasma gondii, is associated with mild disease in healthy individuals, whereas individuals with depressed immunity may develop encephalitis, neurologic disorders, and other organ diseases. Women who develop acute toxoplasmosis during pregnancy are at risk of transmitting the infection to the fetus, which may lead to fetal damage. A diagnosis is usually confirmed by measuring IgG, or IgM where it is important to determine the onset of infection. A negative IgM result essentially excludes acute infection, whereas a positive IgM test is largely uninterpretable because IgM can persist for up to 18 months after infection. To identify antigens for improved diagnosis of acute infection, we probed protein microarrays displaying the polypeptide products of 1357 Toxoplasma exons with well-characterized sera from Turkey. The sera were classified according to conventional assays into (1) seronegative individuals with no history of T. gondii infection; (2) acute infections defined by clinical symptoms, high IgM titers, and low avidity IgG; (3) chronic/convalescent cases with high avidity IgG but persisting IgM; (iv) true chronic infections, defined by high avidity IgG and no IgM. We have identified 38 IgG target antigens and 108 IgM target antigens that can discriminate infected patients from healthy controls, one or more of which could form the basis of a ‘tier-1′ test to determine current or previous exposure. Of these, three IgG antigens and five IgM antigens have the potential to discriminate chronic/IgM persisting or true chronics from recent acutely infected patients (a ‘tier-2′ test). Our analysis of the antigens revealed several enriched features relative to the whole proteome, which include transmembrane domains, signal peptides, or predicted localization at the outer membrane. This is the first protein microarray survey of the antibody response to T. gondii, and will help in the development of improved serodiagnostics and vaccines.

Fascioliasis in eosinophilic patients in the Isparta region of Turkey

Abstract.Background: The aim of this study was to investigate fascioliasis in a group of eosinophilic and non-eosinophilic patients in the Isparta region of Turkey. Patients and Methods: All cases were examined for antibodies against Fasciola hepatica by the modified ES-ELISA method. Seropositive patients with fascioliasis were investigated by radiological and laboratory evaluations. Results: Of the 756 eosinophilic patients, 6.1% were found to be positive for fascioliasis by ES-ELISA, while the rate was only 0.9% among the non-eosinophilic group (statistically significant, p < 0.001). Among the seropositives, 19 cases were evaluated further. Ultrasonographic findings were found in 14 of the 19 cases. In six of nine cases, computed tomography findings were related to fascioliasis. F. hepatica eggs were found in ine of the 19 cases. Conclusion: Fascioliasis was demonstrated in the Isparta region of Turkey for the first time. The higher prevalence of fascioliasis among patients with eosinophilia indicates the importance of this sign for this infection.

Use of Two Sensitive and Specific Immunoblot Markers, Em70 and Em90, for Diagnosis of Alveolar Echinococcosis

ABSTRACT Antibodies against Echinococcus multilocularis metacestodes were screened by immunoblotting sera from patients with alveolar echinococcosis (n = 39), cystic echinococcosis (n = 109), or other parasitic infections (n = 66) and healthy individuals (n = 32). Two antigens, approximately 70 and 90 kDa, are found to be valuable for confirmatory diagnosis, with a sensitivity and specificity of 100 and 99.51%, respectively.

Isolation of Toxoplasma gondii strains similar to Africa 1 genotype in Turkey

INTRODUCTION Toxoplasma gondii is a protozoon parasite that has a worldwide dissemination. It can cause serious clinical problems such as congenital toxoplasmosis, retinochoroiditis, and encephalitis. Currently, T. gondii genotypes are being associated with these clinical presentations which may help clinicians design their treatment strategy. CASE REPORTS Two T. gondii strains named Ankara and Ege-1 were isolated from newborns with congenital toxoplasmosis in Central and Western Anatolia, respectively. Ankara and Ege-1 strains were isolated from the cerebrospinal fluid of newborns. According to microsatellite analysis, Ankara and Ege-1 strains were sorted as Africa 1 genotype. CONCLUSION T. gondii strains isolated in Turkey were first time genotyped in this study. Africa 1 genotype has previously been isolated in immunosuppressed patients originating from sub-Saharan Africa. The reason of detecting a strain mainly detected in Africa can be associated with Turkeys specific geographical location. Turkey is like a bridge between Asia, Europe and Africa. Historically, Anatolia was on the Silk Road and other trading routes that ended in Europe. Thus, detecting Africa 1 strain in Anatolia can be anticipated. Consequently, strains detected mainly in Europe and Asia may also be detected in Anatolia and vice versa. Therefore, further studies are required to isolate more strains from Turkey.

Behaviour of Toxoplasma gondii RH Ankara strain tachyzoites during continuous production in various cell lines.

Toxoplasma gondii is an obligate intracellular protozoan parasite. The objective of the present study was to examine the behaviour of Toxoplasma gondii RH Ankara strain tachyzoites in a cell culture environment. The study represents the first step in determining whether T. gondii RH Ankara strain tachyzoites, grown in cell culture, are of sufficient quality to allow cessation of in vivo tachyzoite production for diagnostic assays. In the present study, T. gondii RH Ankara strain tachyzoites were continuously produced in myeloma X63.Ag8.653, HeLa, Hep-2, and Vero cell cultures for 2 months. The average size of the tachyzoites was 3 x 5.7 microm prior to the first inoculation but after continuous production, a marked decrease was noted in average tachyzoite size. The smallest tachyzoite size, was 1 x 2.1 microm after 2 months, in myeloma cell cultures even though the yield of tachyzoites increased. With other cell cultures, tachyzoite yields were not as high as myeloma cell culture although decrease in size was less. The smallest decrease in tachyzoite size, averaging 2 x 3.8 microm after 2 months, was observed in tachyzoites produced in HeLa cell cultures. A virulence assay in small groups of BALB/c mice, using tachyzoites derived from cell cultures, was also conducted. The preliminary results of the virulence assay suggest that as the size of the tachyzoites decreased, the virulence in mice decreased. Future research will focus on the effect of the size of cell culture-derived T. gondii RH Ankara strain tachyzoites on the virulence, protein expression, and the reliability of diagnostic assays. Ultimately, the behaviour of tachyzoites from various T. gondii strains will be observed in cell culture to determine if size is altered.

A mass screening survey of cystic echinococcosis by ultrasonography, Western blotting, and ELISA among university students in Manisa, Turkey.

Cystic echinococcosis (CE) is one of the most important zoonotic diseases in a wide geographic area, including Turkey. In the present project, a total of 4275 students from Celal Bayar University, Manisa, Turkey, were screened by ultrasonography (US) and specific antibodies for CE were examined by Western blotting (WB) and ELISA in finger prick blood samples of 2034 of 4275 volunteered students. We aimed to report the apparent prevalence of CE based on different diagnostic procedures and to compare WB and ELISA with US in diagnosis of CE in a mass screening setting. Six new cases were diagnosed as CE by US during the survey. In addition to these cases, three students were also detected to have been previously operated and pathologically confirmed for hepatic CE. US revealed parenchymal changes in these cases in concordance with their operation history; so, the prevalence of CE by US was calculated as 0.21% (9/4275) (95%CI, 0.11-0.39%) among university students in Manisa. Bands were detected at 8, 28, 32, 38, 42, 47, 70 and 90kDa by WB and the cases were considered to be positive for CE when at least three of the bands were seen together. Apparent prevalence of CE by ELISA and WB were found to be 2.11% (43/2034) (95%CI, 1.57-2.83%) and 0.25% (5/2034) (95%CI, 0.10-0.57%), respectively. Of the six US positive cases, WB was positive in only one case with two cysts in the liver. All of four cases with liver involvement were positive by ELISA. The high prevalence of CE among university students in Manisa indicated that CE is a major health problem in this area of Turkey. Our results supported that WB is rather difficult and not feasible as a mass screening test and may not be effective for confirmation especially in asymptomatic cases. As a result, we recommend US to be used initially in mass screening surveys for CE followed by confirmation by ELISA for suspected cases. Further examination primarily by chest X-ray followed by computed tomography and/or magnetic resonance imaging, if needed, should be recommended for US negative, ELISA and WB positive individuals who may have non-abdominal cysts.

Fasciola hepatica infection in echinococcosis suspected cases.

OBJECTIVE Fasciola hepatica, a liver fluke of sheep and cattle, may accidentally infect humans. The main signs and symptoms of the fasciolosis are eosinophilia, abdominal pain and hepatomegaly and may also be attributed to Cystic Echinococcosis (CE) caused by Echinococcus granulosus, which is a prevalent infection in Turkey. METHODS Sera samples of 226 CE suspected cases were tested for anti-F. hepatica antibodies by an excretory secretory ELISA (ES-ELISA) and for anti-E. granulosus antibodies by ELISA and indirect hemagglutination (IHA) tests. Cases which were seropositive for fasciolosis were further evaluated radiologically and examined for F. hepatica eggs. RESULTS Five (2.2%) and 96 (42.4%) of the 226 CE suspected cases were found seropositive for fasciolosis and CE, respectively. Although the radiological findings strongly suggested that there was fasciolosis in three cases, F. hepatica eggs were detected in two patients only. CONCLUSION These data suggest that human fasciolosis is not as rare as previously reported in Turkey. F. hepatica infection should be suspected especially in the presence of eosinophilia, abdominal pain and liver lesions.

The ability of 67Ga scintigraphy to detect the lesions of Echinococcus multilocularis infection: Preliminary results

Abstract]AimTo assess the ability of67Ga scintigraphy to detect the lesions ofEchinococcus multilocularis (EM) infection.Materials and MethodsAn animal model of EM infection was developed. The infected tissues taken from stock infection were placed into the abdominal cavity of uninfected animals operatively. The success of implantation was controlled 20–25 days after implantation. Five infected and 2 healthy animals were studied. All of the animals were examined by ultrasound before the scintigraphic evaluation. After the injection of 7.4 MBq (200 µCi)67Ga citrate intravenously, static images from the whole anterior thorax and abdomen were obtained at 24, 48 and 72 hours. Visual and semiquantitative analyses were performed. In semiquantitative analysis, an irregular region of interest was drawn over the thorax as the background, excluding the heart and a second region of interest was drawn over the abdomen, excluding the liver and spleen. Abdomen/ background ratios were calculated using the mean counts.ResultsIn the visual evaluation, it was noticed that there was considerably increased67Ga uptake in the abdomens of the infected animals. In infected animals, mean abdomen/background ratios at 48 and 72 hours (3.76 ± 1.04, 4.13 ± 0.72, respectively) were increased compared with mean abdomen/background ratios at 24 hours (2.94 ± 0.77). These increases in abdomen/background ratios were statistically significant at 72 hours (p = 0.04). Between the infected animals and control group, mean abdomen/background ratios were compared, and statistically significant differences were found in the images obtained at 48 and 72 hours.ConclusionImaging at 72 hours seems to be more suitable imaging time for the diagnosis of alveolar echinococcosis.67Ga scintigraphy may successfully demonstrate the lesions of EM infection localized intraperitoneally. The method of67Ga scintigraphy is useful because it is simple, non-invasive and relatively safe.

Eosinophil cationic protein in patients with fascioliasis: Its probable effects on symptoms and signs

Eosinophilia is 1 of the most common signs of fascioliasis especially during acute stage. In this study, our aim was to determine eosinophil cationic protein (ECP) in the sera of patients with fascioliasis and to investigate the likely relation between ECP and symptoms and signs of fascioliasis. Presence of high liver enzyme levels, eosinophilia, abdominal pain or urticaria was not found to be significantly associated with ECP positivity (p>0.05), but statistical analyses revealed that ECP positivity was significantly related to weight loss status of patients (p<0.005). In conclusion, ECP may be 1 of the likely causes of the symptoms and signs of fascioliasis.