[Effects of azithromycin and Chlorella vulgaris treatment on certain cytokine values and NK cell activity in an acute murine toxoplasmosis model]. / Akut toksoplazmozlu fare modelinde Azitromisin ve Chlorella vulgaris'in tedavi etkinliginin, NK aktivasyonu ve bazi sitokin degerlerinin arastirilmasi.

Toxoplasmosis is a common infection with a complicated treatment process. Azithromycin (AZT) is a macrolide antibiotic that can be effectively used in patients with cerebral and ocular toxoplasmosis and has fewer side effects. Chlorella vulgaris (CV), a single-cell green algae that contains nutrients and has various biological effects. CV extract (CVE) has been shown to have protective effects against infections via immune enhancement by increasing the cytotoxicity of NK cells, IL-12 and IFN-γ levels. The aim of this study was to investigate the protective effects of AZT and CV, individually and in combination, against acute toxoplasmosis in mice, and their effects on NK cell cytotoxixity, IL-12, IFN-γ, and IL-2 levels. Six groups of mice (Balb/c) were formed. With the exception of the healthy control (HC) group, all other groups were infected with 1 ml (11 x 104 trofozoit/ml) Toxoplasma gondii RH strain trophozoites. No further action was performed for infected control (IC) group. After 24 hours from trophozoite infection, CVE was given to CV group, AZT to azithromycin group and CVE + AZT combination to CV + AZT group by oral gavage for 6 days. All of the mice from IC, CV, AZT and CV + AZT groups were sacrified on the 8th day of the infection and serum, peritoneal fluid and spleen samples were collected. Trophozoite count of the groups were determined in all groups except HC group and the average growth inhibition activity was calculated by using the growth inhibition formula. In all groups IL-12, IFN-γ, IL-2 levels were measured with ELISA method and cytotoxicity of the NK cells were measured using Cytotox 96 Non-Radioactive Cytotoxicity Assay. The number of trophozoites were significantly lower in the CV group than the IC group (p< 0.001), and also significantly lower in CV + AZT combination group than the AZT group. According to the growth inhibition calculations CV treatment showed 88.6%, AZT treatment 98.46%, AZT + CV combination treatment 99.4% antiprotozoal activity against T.gondii compared with the IC group. NK cell cytotoxicity in the CV and the combination group were significantly higher than all the other groups (p< 0.001). IL-12 and IFN-γ levels were highest in IC group and the lowest in AZT + CV group. This situation has been linked to the fact that the severity of the infection has fallen considerably. IL-2 levels were significantly higher in CV, CV + AZT groups than in the other groups (p< 0.001). In our study, even CV administration alone caused a significant decline in infection.This may be related to the increased NK cytotoxicity, IL-2, IL-12 and IFN-γ levels. CV + AZT combination seems to be an effective treatment option than AZT alone, particularly in patients who are difficult to treat with common methods or in patients with immunosuppression.

[Effects of atovaquone and astragalus combination on the treatment and IL-2, IL-12, IFN-γ levels on mouse models of acute toxoplasmosis]. / Atovakuon ve astragalus kombinasyonunun akut toksoplazmozlu fare modeli tedavisi ve IL-2, IL-12, IFN-γ düzeyleri üzerindeki etkileri.

Reactivation of Toxoplasma gondii infections and serious clinical manifestations such as encephalitis may develop in immunocompromised subjects and AIDS patients. Different protocols are used for the treatment of toxoplasmosis in high-risk patient groups, however life-long prophylactic therapy against reactivation risk in AIDS patients may lead to several undesired results. Atovaquone is an effective antiprotozoal agent against toxoplasmosis with minor side effects. On the other hand, Astragalus membranaceus root extract (AmE) has been shown to have immunomodulatory and antimicrobial activities, empowering immunity by enhancing proliferation and activation of phagocytic cells mainly macrophages, and inducing Th1 type immune response. The aim of this study was to investigate the effectiveness of atovaquone alone and in combination with AmE, in the treatment of toxoplasmosis, and on the levels of IL-2, IL-12 and IFN-γ in experimentally infected mice with T.gondii. For this purpose, four experimental groups, each consisting of eight BALB/c mice, were set with the approval of Ethics Committee for the Animal Experiments. All the mice were infected with 0.5 ml of a suspension containing 2 x 104/ml trophozoites prepared from T.gondii RH strain by intraperitoneal injection. Twenty-four hours after the infection, atovaquone (100 mg/kg/day) was given to atovaquone group, AmE (0.075 mg/g) to astragalus group and atovaquone (100 mg/kg/day) plus AmE (0.075 mg/g) to Atovaquone + Astragalus (Ato + Astra) group by oral gavage. The mice in the fourth group, which was the control group, were all infected but untreated. The above administrations were carried out for seven days. On the 8th day peritoneal fluids of mice were collected under anaesthesia and trophozoite numbers per 1 ml were detected by counting on the Thoma slide. In addition, the heart bloods of mice were drawn and IL-2, IL-12, IFN-γ levels were determined in serum samples by using commercial ELISA kits (eBioscience, Austria). The mean number of trophozoites in Ato + Astra group was found significantly lower than the number of trophozoites in the other three groups (p< 0.05). The number of trophozoites in the atovaquone and astragalus groups were found significantly lower than the number of trophozoites in the control group (p< 0.05). There was a significant increase in IL-2 levels of astragalus group compared with the other three groups, in addition when IL-2 levels of Ato + Astra group were compared with ones in other three groups, a significant decrease was noticed (p< 0.05). There was a definite increase in IL-12 levels of atovaquone, astragalus and the control groups compared to those in Ato + Astra group (p< 0.05). A significant increase was found in IFN-γ levels in atovaquone and Ato + Astra groups compared with those in the control group (p< 0.05). Within the reach of our literature survey, this study was the first research in which the effectiveness of the combination of atovaquone and AmE was investigated in the treatment of acute toxoplasmosis. The results of our study suggested that there might be a synergy between atovaquone and AmE in the treatment of acute toxoplasmosis. In case these results are supported by further studies, atovaquone and AmE combination may have a potential to be used for therapy in immunocompromized patients such as AIDS patients who have a risk for toxoplasmosis.

[Investigation of combined effectiveness of spiramycin and beta-glucan in mice models of acute toxoplasmosis and determination of IL-10, IL-12 and TNF-α levels]. / Spiramisin ve Beta-Glukan Kombinasyonu Etkinliginin Akut Toksoplazmozlu Fare Modelinde Arastirilmasi ve IL-10, IL-12 ve TNF-α Düzeylerinin Belirlenmesi.

Toxoplasmosis which is caused by Toxoplasma gondii, has a high risk of fetal infection development if the infection occurs during pregnancy. Treatment with oral spiramycin is recommended during pregnancy in order to prevent the transmission of protozoa to fetus and development of infection. Since beta- glucan is known to stimulate the immune system and increase the phagocytic activity of the cells, it has been shown to exhibit immunomodulatory effect on many infectious diseases. The objectives of this study were to investigate the effectiveness of beta-glucan alone and in combination with spiramycin and to determinate the levels of interlökin (IL)-10, IL-12 and tumor nekrosis factor (TNF)-α in mice experimentally infected with T.gondii. For this purpose, four experimental groups each consisting of eight BALB/c mice, were formed with the approval of Ethics Committee for the Animal Experiments. All the mice were infected with 2 ml of suspension containing 2 x 102/ml of trophozoite prepared from T.gondii RH strain (Refik Saydam National Public Health Agency, Parasitology Laboratory of Communicable Diseases Research Department, Ankara, Turkey), by intraperitoneal injection. Twenty-four hours after the infection, beta-glucan (3 mg/day) was given to the beta-glucan group, spiramycin (200 mg/kg/day) to the spiramycin group, beta-glucan (3 mg/day) plus spiramycin (200 mg/kg/day) to the beta-glucan-spiramycin (BG-S) group by oral gavage. The fourth group which was the control group was infected but untreated. The above administration was carried out for seven days. On the 8th day, under anaesthesia, 1 ml normal saline was given into the peritoneum, drawn back later and the number of trophozoites in 1 ml of peritoneal fluid was determined by counting them on the Thoma slide. Moreover, by drawing the heart blood; IL-10, IL-12, TNF-α levels were determined in serum samples by ELISA method (eBioscience Platinum, Austria). The number of trophozoites in the BG-S group was found significantly lower than the number of trophozoites in control, beta-glucan and spiramycin groups (p< 0.05). There was no significant difference between the beta-glucan and spiramycin groups, however the number of trophozoites in both groups was significantly lower than the number of trophozoites in the control group (p< 0.05). There was a certain decrease in IL-10 level in spiramycin and BG-S groups, compared to the control group, in addition when IL-10 levels in spiramycin and BG-S groups were compared with BG group, a significant decrease was noticed (p< 0.05). There was no difference in IL-12 levels between the groups, while there was a certain decrease in TNF-α level in beta-glucan, spiramycin, BG-S group in comparison to the control group. Within the reach of our literature survey, this study is the first research in which the effectiveness of the combination of beta-glucan and spiramycin in the treatment of acute toxoplasmosis was investigated. The results of our study suggested that there might be synergy between beta-glucan and spiramycin in the treatment of acute toxoplasmosis.

The influence of glucose added urine on the in vitro antimicrobial activity of various antibiotics.

BACKGROUND & OBJECTIVE: Factors associated with the medium, including calcium and magnesium ion concentration and pH have been shown to affect the results of susceptibility testing but very little is known about glycosuria and the effect of glucose on the antimicrobial effect of antibiotics. In this study we assessed the influence of glucose added urine on the in vitro activities of various antibiotics by the microbroth dilution method. METHODS: Sixteen Escherichia coli isolates from patients with urinary infections were used in this study. Nine antibiotics were tested for their antimicrobial activity. Minimum inhibitory concentrations (MICs) were performed by the microbroth dilution method parallel in Mueller Hinton broth and glucose added urine. RESULTS: MICs of nearly all antibiotics were higher in glucose added urine than MICs in broth. MIC(90) against ampicillin was 32-fold higher in glucose added urine than MIC(90) in broth. MIC(90)s against ampicillin-sulbactam, cephalothin, cefuroxime, ceftriaxone and ciprofloxacin in glucose added urine were significantly (P<0.05) higher than MIC(90) in broth. Equal MIC(90) in glucose added urine and broth were obtained for amikacin, sulphamethoxazole and trimethoprime. INTERPRETATION & CONCLUSION: Our findings demonstrated that MICs of antibiotics are influenced by the glucose added urine.

The effects of streptozotocin-induced diabetes on brucellosis of rats.

It is believed that an infection is more common and runs a more protracted course in people with diabetes. In clinical practice, it is important to be aware of these associations, as the prognosis is often dependent upon prompt recognition and appropriate treatment. To show the course of brucellosis in the diabetic state, a model of Brucella melitensis infection was used in the setting of streptozotocin-induced diabetes in rat. B. melitensis infection proceeded more severely in diabetic rats and the severity of diabetes affected the prognosis. However, no association was found between B. melitensis and insulin using in vitro and in vivo experiments. Our study illustrates that B. melitensis infection in diabetes should be taken seriously.

[Investigation of intestinal parasites in AIDS patients]. / AIDS hastalarinda barsak parazitlerinin arastirilmasi.

In this study, enteric parasites were investigated in the stool samples of 38 AIDS patients (23 with chronic diarrhea and 15 without diarrhea) prospectively. At least three stool samples from each patient were investigated microscopically for ova or trophozoites. The samples were concentrated with formol-ether method and wet preparations stained with lugol were examined. In addition, the concentrated samples were stained with modified asid-fast (Kinyoun's), rhodamine-auramine, modified trichrom and calcoflor methods. Enteric parasites were detected in 18 (47%) of the 38 patients, 16 patients harbored a single parasite, and 2 patients were found to be infected with more than one parasite. Only one (7%) of 15 AIDS patients without diarrhea, were found to be infected with Giardia lamblia. On the other hand, 17 (74%) of 23 AIDS patients with chronic diarrhea were found to be infected with various enteric parasites. Cryptosporidium spp. was detected in 9 (39%) of these 23 patients, and in 2 of them Microsporidium spp. accompanied Cryptosporidium. In 2 (9%) of these 23 patients G. lamblia were detected, while Isospora belli, Blastocystis hominis, Entamoeba histolytica, Strongyloides stercoralis and Trichuris trichiura were detected in one patient each. As a result, the detection rate of emerging parasites, including Cryptosporidium spp, Microsporidium spp, I. belli, B. hominis, and S. stercoralis was significantly higher than conventional parasites (39% versus 13%; z=2.34, p=0.01), and CD4 T cell counts were found to be significantly lower among AIDS patients with chronic diarrhea than those without diarrhea (x2=34.33, p<0.001).

The influence of urine on the in vitro antimicrobial activity of various antibiotics against different Escherichia coli phenotypes.

BACKGROUND: The influence of urine on the in vitro activities of various antibiotics used in the therapy of urinary tract infections was assessed by the microbroth dilution method in this study. METHODS: Thirty Escherichia coli strains were used: 10 E. coli strains susceptible to ampicillin, 10 strains resistant to ampicillin and ampicillin+sulbactam and ten extended spectrum beta-lactamase producer strains. The minimum inhibitory concentrations (MICs) of ampicillin, ampicillin + sulbactam, cephalothin, cefuroxime, ceftriaxone, amikacin, ciprofloxacin, sulfamethoxazole and trimethoprim were performed parallel in Mueller-Hinton broth and human urine by the microbroth dilution method. RESULTS: The MIC(90) of all antibiotics except cephalothin were higher in the urine. MICs performed in the urine were found significantly higher than those performed in broth. CONCLUSIONS: This study demonstrated that MICs of antibiotics are influenced by the human urine and that MICs of some antibiotics used in the treatment of urinary tract infections may be overestimated by the standard antibiotic testing methods.