Antimicrobial effects of laurel extract, laurel essential oil, zahter extract, and zahter essential oil on chicken wings contaminated with Salmonella Typhimurium.

BACKGROUND: This study aimed to evaluate the antimicrobial effects of zahter extract, zahter essential oil, laurel extract, and laurel essential oil on Salmonella Typhimurium inoculated on chicken wings. METHODS: A total of 10 groups, including eight study groups and two control groups were formed, consisting of zahter extract and zahter essential oil and laurel extract and laurel essential oil in different proportions. In the study, laurel extract at 6.4% and 12.8% concentrations, laurel essential oil at 0.2% and 0.4% concentrations, zahter extract at 0.2% and 0.4% concentrations, and zahter essential oil at 0.2% and 0.4% concentrations were used. RESULTS: The broth microdilution method was used to evaluate the antimicrobial activity of the extract and essential oils on the S. Typhimurium. Minimum inhibitory concentrations of the extracts and essential oils used in the study against S. Typhimurium were determined. The highest inhibitory effect on S. Typhimurium was observed in the 0.4% laurel essential oil group. It was determined that the inhibitory effect increased as the concentration of laurel essential oil increased. In addition, the antimicrobial activity of zahter essential oil is less inhibitory than the laurel extract, laurel essential oil, and zahter extract. CONCLUSION: According to the results of this study, it has been revealed that extracts and essential oils obtained from zahter and laurel plants, which have been shown to be natural antimicrobial, can be used in foods as an alternative to chemical additives. To develop research results, the applicability of these extracts and essential oils in different foodstuffs should be examined using different ingredients and concentrations.

Effects of the protective and therapeutic effect of yogurt enriched with Citrus sinensis essential oil against ibuprofen-induced gastric ulcer in elderly rats.

The aim of this study was to evaluate the effect of Citrus sinensis essential oil (EO) on the proximate composition of yogurt over a 28-day shelf life and to investigate the therapeutic and prophylactic effects of functional yogurt on ibuprofen-induced gastric ulcers in a rat model. It was observed that the yogurt group containing C. sinensis EO had higher acidity, total solids, and ash values. Histologic evaluation of the stomachs of rats with gastric ulcers revealed that rats fed with functional yogurt had fewer lesions compared to the control group. The treatment group had fewer lesions than the positive control (p > 0.05). Lesions in the glandular mucosa of the prophylactic group were significantly lower than those in the positive control group (p < 0.05). Yogurt with C. sinensis EO may be beneficial in reducing the severity of ulcers and improving overall health.

Combination of infra-red light with nanogold targeting macrophages in the treatment of Leishmania major infected BALB/C mice.

PURPOSE: In the treatment of cutaneous leishmaniasis (CL), developing drug resistance, existing toxic effects of drugs and failure respond to treatment cause the need to try different treatment methods. We investigated the effect of gold-conjugated macrophage-specific antibody on amastigotes under infra-red light for the treatment of CL. METHODS: Female BALB/c (4-8 weeks old, 20 ± 5 g weight) mice were used in the study. The L. major strain was inoculated on the soles of mice in amastigote form and subpassed. Nanogold (Au), Au + macrophage-specific antibody (MSA) modification and near infra-red (NIR) (5 seconds) were applied to mice groups that developed cutaneous leishmaniasis on their soles. On the 5th and 10th days of the treatment, the lesions were examined clinically and pathologically. RESULTS: When the erythema values were examined, the highest decrease was calculated in the Au + MSA + NIR group in the measurements made on the 10th day (p < 0.014). The best improvement in 10th day measurements is in the NIR and Au + MSA + NIR groups when area values were examined (p = 0.011, p = 0.001). There was a statistically significant difference between the groups in terms of parasite load (PL) (p < 0.005) in pathological evaluation. According to PL grouping, the best result is NIR (p = 0.002). When both main titles (clinical and pathological) are examined, the Au + MSA + NIR group is thought to have an optimal therapeutical feature. CONCLUSIONS: Au + MSA + NIR combination could be a new treatment approach for CL treatment.

Effects of neuromuscular blocking drugs on viability of human umbilical vein endothelial cells (HUVECs).

OBJECTIVE: This study aimed to investigate the effects of neuromuscular blocking drugs on the viability of human umbilical vein endothelial cells (HUVECs) and to investigate whether they cause vascular complications due to cell proliferation. METHODS: HUVECs were cultivated with 5% CO2 at 37°C in a predefined supplemented medium over 7 days until confluence of cell monolayers. Assays were conducted during the exponential growth phase. Suxamethonium chloride, vecuronium bromide, atracurium besylate, and rocuronium bromide were used at concentrations of 10-5, 10-6, and 10-7 M in proliferation assays in which cells were incubated with these drugs for 24, 48, and 72 hours. All experiments were performed in four replicates. RESULTS: The neuromuscular blocking drugs used had comparable effects on the survivability of HUVECs. Overall, no significant difference was observed in the survivability of HUVECs in a dose-dependent manner after exposure to the study drugs. However, some significant differences in the viability of HUVECs were found among the different measurement times. CONCLUSIONS: The findings of the current study support the safety of the studied neuromuscular blocking drugs in clinically relevant concentrations regarding their effects on endothelial cell proliferation.

In Vitro Evaluation of Adhesion of Two Acanthamoeba Strains to Cosmetic Contact Lenses.

PURPOSE: To evaluate the factors affecting the adhesion of Acanthamoeba trophozoites to the surface of cosmetic contact lenses (CCLs). METHODS: Acanthamoeba castellanii and A. hatchetti trophozoites were inoculated onto CCLs (hema copolymer [HM] [38.5% H2O], phemfilcon [PF] [55% H2O], polymacon [PM] [38% H2O], polyhema [PH] [%42 H2O], and hema [HM55] [55% H2O]), and the number of trophozoites adhered to the lens surfaces was assessed over time, that is, at 15 min, 1, and 24 hr. In addition, scanning electron microscopy (SEM) analysis of the lens surfaces was performed to evaluate the effect of lens surface topology on adhesion. RESULTS: The number of amoeba adhered to the contact lens surface was found lower with PF and PH production materials, than lenses with HM, PM, and HM55 production materials (P<0.05). No significant difference was detected in amoebic strains adhered in all the contact lens types (P>0.05). No significant difference was found on average amoeba adhesion between contact lenses with hema production material but with different water contents (45%, 55%), to see the effect of water content on amoebic adhesion (P>0.05). As a result of SEM analysis, surface topology showed no effect on adhesion. CONCLUSION: (1) Chemical composition of lenses seemed to be mostly responsible for the adhesion of Acanthamoeba. (2) Different numbers of trophozoites, obtained after the adhesion experiment, could also indicate that adherence capacity can also differ among Acanthamoeba species.

Evaluation of the in vitro activity of ceragenins against Trichomonas vaginalis.

Trichomonosis, caused by the protozoan parasite Trichomonas vaginalis, is a curable sexually transmitted disease that is most commonly encountered worldwide. Increasing importance of trichomoniasis and emerging of resistance against metronidazole lead to search for alternative drugs with different mode of activity. The purpose of this study was to determine in vitro activity of ceragenins (CSA-13, CSA-44, CSA-13, and CSA-138) against the metronidazole-susceptible (ATCC 30001) and metronidazole-resistant (ATCC 50138) strains of T. vaginalis. The effective concentrations were evaluated using two strains of T. vaginalis with different metronidazole susceptibilities (ATCC 30001 and ATCC 50138) in the presence of dilution series of ceragenins in 24-well microtitre assays. Overall, all the ceragenins killed the metronidazole-susceptible (ATCC 30001) and metronidazole-resistant (ATCC 50138) strains of T. vaginalis (p>0.05). With regard to the their effects against the studied strains of T. vaginalis, in order of effectiveness, overall, the ceragenins ordered as CSA-13 (the most effective), CSA-131 and CSA-138 (effective similarly), and CSA-44 (the least effective) (p<0.05). All of the ceragenins reduced the trophozoite numbers of both of studied strains of T. vaginalis with a time- and dose- dependent manner (p<0.05). Although all of the study ceragenins, CSA-13, CSA-44, CSA-13, and CSA-138, killed the studied strains of T. vaginalis. CSA-13 is the leading ceragenin as the most effective anti-trichomonas compound, followed by CSA-131 and CSA-138. They have a potential to have a place in the armemantarium of gynecologic and urologic practice for the management of sexually transmitted diseases.

In vitro comparison of the cytotoxicity and water sorption of two different denture base systems.

PURPOSE: Denture base resins have the potential to cause cytotoxicity in vivo, and the mechanical properties of resins are affected by water sorption. There is a correlation between residual monomer and water sorption. Thus, the purpose of this study was to evaluate water sorption and cytotoxicity of light-activated urethane dimethacrylate (UDMA) denture base resin compared to a conventional heat-activated polymethyl methacrylate (PMMA) resin. MATERIALS AND METHODS: Two denture base resins, heat-activated PMMA (Meliodent) and light-activated UDMA (Eclipse), were used in this study. Cytotoxicity (5 × 1 mm(2) ) and water sorption (1 × 1 mm(2) ) specimens were made following the manufacturers' instructions (n = 10). Cytotoxicity tests of denture base resins were performed according to ISO10993-5:1999, and water sorption was evaluated according to ISO 1567:1997. ANOVA tests were employed for evaluating data (α = 0.05). RESULTS: There was no cytotoxic effect in either the PMMA or UDMA group. In addition, contrary to short-term water storage, a significantly lower water sorption value was shown for UDMA resins compared to PMMA resins in both 3- and 6-month storage periods (p = 0.043 and p = 0.002, respectively). CONCLUSION: The tested denture base materials adhered to the ISO standards for both cytotoxicity and water sorption. The cytotoxicity of the light-activated UDMA resin tested was statistically similar to that of the heat-activated PMMA resin; however, the UDMA resin exhibited decreased water sorption in long-term water storage.

Sequential determination of serum viral titers, virus-specific IgG antibodies, and TNF-α, IL-6, IL-10, and IFN-γ levels in patients with Crimean-Congo hemorrhagic fever.

BACKGROUND: Although there have been a number of studies on the pathogenesis of Crimean-Congo hemorrhagic fever (CCHF) recently, knowledge on this topic is still insufficient. This study aims to reveal the kinetics of serum CCHF virus (CCHFV) titers, serum levels of anti-CCHFV immunoglobulin (Ig)G, tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-10, and interferon (IFN)-γ in CCHF patients. METHODS: In total, 31 CCHF cases (11 fatal) were studied. Serum samples were obtained daily from all patients from the time of admission and continued for a 7-day hospitalization period for serologic (ELISA), virologic (real-time PCR), and cytokine (ELISA) analysis. RESULTS: The mean serum CCHFV titer at admission was 5.5E + 09 copies/mL in fatal cases and 5.7E + 08 copies/mL in survivors (p < 0.001). Compared to survivors, both the mean serum levels of IL-6 and TNF-α at admission were found to be significantly increased in fatal cases. The serum levels of IL-6, TNF-α and serum CCHFV titer at admission were significantly and positively correlated with disseminated intravascular coagulation (DIC) scores (r = 0.626, p = 0.0002; r = 0.461, p = 0.009; and r = 0.625, p = 0.003, respectively). When the data obtained from the sequential determination of CCHFV titer and levels of anti-CCHFV IgG, IL-6, TNF-α, IL-10 and IFN-γ were grouped according to the days of illness, the initial serum CCHFV titer of a fatal patient was 5.5E + 09 (copies/mL) and it was 6.1E + 09 (copies/mL) in a survivor on the 2 day of illness. While significant alterations were observed in all cytokines during the monitoring period, IL-6 levels remained consistently higher in fatal cases and TNF-α levels increased in both in fatal and non-fatal CCHF cases. CONCLUSIONS: The increased CCHFV load and higher concentrations of IL-6 and TNF-α, the presence of DIC, and the absence of CCHFV specific immunity are strongly associated with death in CCHF.

Miltefosine and polyhexamethylene biguanide: a new drug combination for the treatment of Acanthamoeba keratitis.

BACKGROUND: In this study, a series of compounds - miltefosine, polyhexamethylene biguanide, chlorhexidine and propamidine isethionate - and combinations of the latter three agents with miltefosine were prepared and used in a rat model for the topical treatment of Acanthamoeba keratitis. METHODS: The corneas of rats were infected with Acanthamoeba hatchetti. On the fifth day, all corneas were microscopically examined in order to determine the grade of infections. Nine groups were then prepared: miltefosine (65.12 µg/mL); chlorhexidine (0.02%); polyhexamethylene biguanide (0.02%), propamidine isethionate (0.1%), miltefosine plus chlorhexidine, miltefosine plus polyhexamethylene biguanide; miltefosine plus propamidine isethionate; infected control; and a non-infected control group. The treatment was continued for 28 days. After the treatment, the corneas were excised and used for Acanthamoeba culture to investigate the presence of Acanthamoeba growth. For the determination of cytotoxicity of the drugs on L929 cells, colorimetric assays were performed. RESULTS: The best treatment results were obtained from the polyhexamethylene biguanide plus miltefosine group; the ratio of fully recovered eyes was 28.4%. It was proven that the miltefosine-polyhexamethylene biguanide combination yielded the highest anti-acanthamoebal activity in that approximately 86% of the eyes were cleared from amoebae. The cytotoxicity values of the miltefosine and the control groups were compared with other groups and found to be statistically different (P < 0.05). CONCLUSION: This in vivo study demonstrates that a miltefosine-polyhexamethylene biguanide combination is highly effective for the treatment of Acanthamoeba keratitis.

Effects of tyrosine kinase inhibitor E7080 and eNOS inhibitor L-NIO on colorectal cancer alone and in combination.

OBJECTIVE: To investigate the effects of E7080 and N (5)-(1-iminoethyl)-L-ornithine dihydrochloride (L-NIO) on colorectal cancer alone and in combination. METHODS: HT29 colorectal cancer cell line from Sap Institute was used. Real-time cell analysis (xCELLigence system) was performed to determine the effects of E7080 and L-NIO on colorectal cell proliferation. While apoptosis was determined with Annexin V staining, and the effect of agents on angiogenesis was determined with chorioallantoic membrane (CAM) model. RESULTS: We found that E7080 has a strong antiproliferative effect with an half maximum inhibition of concentration (IC50) value of 5.60×10(-8) mol/L. Also it has been observed that E7080 showed antiangiogenic and apoptotic effects on HT29 colorectal cancer cells. Antiangiogenic scores of E7080 were 1.2, 1.0 and 0.6 for 100, 10 and 1 nmol/L E7080 concentrations, respectively. Furthermore, apoptosis has been detected in 71% of HT29 colorectal cancer cells after administration of 100 nmol/L E7080 which may indicate strong apoptotic effect. Meanwhile administration of L-NIO alone did not show any effect, but the combination of E7080 with L-NIO increased the antiproliferative, antiangiogenic and apoptotic effects of E7080. CONCLUSIONS: Results of this study indicate that E7080 may be a good choice in treatment of colorectal tumors. Furthermore the increased effects of E7080 when combined with L-NIO raise the possibility to use a lower dose of E7080 and therefore avoid/minimize the side effects observed with E7080.

Synthesis, characterization, and assessment of cytotoxic, antiproliferative, and antiangiogenic effects of a novel procainamide hydrochloride-poly(maleic anhydride-co-styrene) conjugate.

Poly(maleic anhydride-co-styrene) (MAST) was synthesized by a free-radical polymerization reaction. A bioactive molecule, procainamide hydrochloride (PH), was then conjugated to MAST. The conjugation product was named as MAST/PH. Structural characterization of MAST and MAST/PH was carried out by Fourier Transform Infrared and Nuclear Magnetic Resonance spectroscopy. Their molecular weights were determined by size-exclusion chromatography. A mechanism was then suggested for the conjugation reaction. The results of the cytotoxicity assay, employing a mouse fibroblast cell line (L929), indicated that MAST/PH had no cytotoxicity at concentrations [Formula: see text] 62 µg mL(-1) (p > 0.05). Antiproliferative activities of MAST/PH and PH were determined by the BrdU cell proliferation ELISA assay, using C6 and HeLa cell lines. In the experiment, two anticancer chemotherapy drugs, cisplatin and 5-fluorouracil, were included as positive control. Antiproliferative activity results demonstrated that MAST/PH yielded the highest suppression profile (approximately 42%) at 20 µg/ml, while free PH exerted the same activity at 100 µg/ml. Interestingly, both MAST/PH and PH suppressed the proliferation of only one of the cell lines, C6 cells. Both cisplatin and 5-fluorouracil yielded approximately 60% antiproliferative activity on C6 cells at 20 and 100 µg/ml concentrations. Antiangiogenic capacity of both MAST and MAST/PH was also investigated by using the chicken chorioallantoic membrane assay. Results obtained indicated that while MAST/PH could be included into the category of good antiangiogenic substances, the activity score of MAST was within the weak category.

Effects of low molecular weight heparins and unfractionated heparin on viability of human umbilical vein endothelial cells.

AIM: The aim of this study was to investigate the effects of bemiparin, nadroparin, enoxaparin, and heparin on viability of human umbilical vein endothelial cells (HUVEC). METHODS: Cultivation of HUVEC was performed in an incubator having 5 % CO(2) at 37 °C and with predefined supplemented medium, until cell monolayers attained confluence which occurred after 7 days. The assays were performed in the exponential growth phase of the cells. The cell viability was assessed using the cleavage of tetrazolium salts added to the culture medium. Heparin sodium, enoxaparin sodium, bemiparin sodium, and nadroparin calcium with concentrations of 100, 10, and 1 IU/100 µL were used for the proliferation assay in which cells were incubated for 24, 48, and 72 h with these drugs. The experiments were conducted in four replicates. RESULTS: Among the study drugs with maximal concentration used in the experiments (100 IU/100 µL), heparin was found to be associated with the lowest viability score in 24 and 48 h, while bemiparin showed the lowest at 72 h. Bemiparin 100 IU/100 µL was significantly associated with lower viability score than that of bemiparin 10 IU/100 µL and bemiparin 1 IU/100 µL at every time interval. Among gradual concentrations of enoxaparin, however, concentration of 1 IU/100 µL was associated with the lowest viability scores at every time point. Heparin 1 IU/100 µL, nadroparin 100 IU/100 µL, and enoxaparin 100 IU/100 µL groups had the highest viability score after 72 h of incubation. CONCLUSION(S): Among low molecular weight heparins (LMWHs), 100 IU/100 µL concentration of bemiparin was associated with a more pronounced effect on reducing viability of HUVEC after 72 h of incubation, while nadroparin 100 IU/100 µL and enoxaparin 100 IU/100 µL showed the least effects. LMWHs differ both from each other and heparin with respect to their effects on cellular viability of HUVEC in dose- and time-dependent manner.

Antitumoral effects of Allium sivasicum on breast cancer in vitro and in vivo.

This work aims to investigate the antiproliferative properties of Allium sivasicum (AS) on breast cancer. AS extracts were studied for cytotoxicity against the breast cancer cell lines. In vitro apoptosis studies of breast cancer cells were performed by annexin V staining in flow cytometry analyses. AS showed cytotoxicity to three cancer cell lines. Annexin-positive cells level in AS treated cell lines were higher than the untreated control cells. The expressions of caspase-7 protein and TUNEL positive cells were much higher for the rats treated by AS, compared with the untreated control group. The expressions of the Ki-67 decreased in treatment groups compared with the control group. In vivo studies showed that mean tumor volume inhibition ratio in AS treated group was 38 % compared with the untreated rats. These results indicate that A. sivasicum has antitumoral potential against breast cancer.

Antitumoral effects of Melissa officinalis on breast cancer in vitro and in vivo.

BACKGROUND: There is a long standing interest in the identification of medicinal plants and derived natural products for developing cancer therapeutics. Here we investigated the antiproliferative properties of Melissa officinalis (MO) from Turkey on breast cancer. METHODS: MO extracts were studied for cytotoxicity against breast cancer cell lines (MCF-7, MDA-MB-468 and MDA-MB-231). In vitro apoptosis studies were performed by annexin V staining and flow cytometry analyses. Immunohistochemistry for Ki-67 and caspase 7 in the tumoral tissue sections of DMBA-induced mammary tumors in rats was also performed, along with TUNEL assays to detect apoptotic cells. In vivo anticancer activity testing was carried out with reference to inhibition of growth of DMBA induced mammary tumors in rats. RESULTS: MO showed cytotoxicity against three cancer cell lines, inducing increase in Annexin-positive cells. Expression of caspase-7 protein and TUNEL positive cells were much higher in rats treated by MO, compared with the untreated control group, while expression of Ki-67 was decreased. Furthermore, in vivo studies showed that mean tumor volume inhibition ratio in MO treated group was 40% compared with the untreated rats. CONCLUSION: These results indicated that MO extrcts have antitumoral potential against breast cancer.

Comparison of the antiangiogenic effects of heparin sodium, enoxaparin sodium, and tinzaparin sodium by using chorioallantoic membrane assay.

Unfractionated heparin (UFH) and low molecular weight heparins have been used as anticoagulation agents in cardiovascular clinics for decades. However, these molecules also have potent antiangiogenic effects. Whereas, angiogenesis may be the most crucial determinant of the prognosis of cardiovascular diseases, and except some special situation, antiangiogenic effect is not desirable in the most of the cardiovascular disease. In this study, we aimed to compare the antiangiogenic potency of UFH, enoxaparin, and tinzaparin. The antiangiogenic efficacies of UFH, enoxaparin, and tinzaparin were examined in vivo by using the chick chorioallantoic membrane (CAM) model. Twenty fertilized eggs were used for each studied drug. Drug solutions were prepared in 10 and 1 IU/10 µl concentrations. Decreases in the density of the capillaries were assessed and scored. All three drugs showed antiangiogenic effects on the chick CAM at the 10 IU/10 µl concentration. However, the antiangiogenic score of the UFH was significantly higher than that of enoxaparin and tinzaparin at 1 and 10 IU/10 µl concentrations. UFH had stronger and antiangiogenic potential than enoxaparin and tinzaparin. However, tinzaparin showed dose-dependent antiangiogenic effects. We think that an anticoagulant molecule with a less and dose-dependent antiangiogenic effect, as in the case of tinzaparin, may be more desirable in case of cardiovascular disease related with insufficient angiogenesis.

The antiangiogenic effects of levosimendan in a CAM assay.

BACKGROUND: There is a physiological balance between the stimulatory and inhibitory signals for blood vessel growth. In many symptomatic patients with peripheral artery disease, coronary artery disease, and ischemic chronic wounds, there is a pathological insufficiency of angiogenesis. Therefore, determining the angiogenic or antiangiogenic effects of molecules currently used in cardiovascular treatment is crucial. Although levosimendan is the most well studied calcium sensitizer in preclinical and clinical practice, to the best of our knowledge, there are no previous studies investigating its angiogenic or antiangiogenic effects. In the present study, we aimed to investigate the effects of levosimendan on angiogenesis. METHODS: The antiangiogenic efficacy of levosimendan was examined in vivo in the chick chorioallantoic membrane (CAM) model by using 20 fertilized eggs and drug solutions of 1 and 10 µmol/L concentrations. Decreases in the density of the capillaries were assessed and scored. RESULTS: Significant antiangiogenic effects were observed at 1 and 10 µmol/L concentrations of levosimendan. The antiangiogenic scores of levosimendan at 1 and 10 µmol/L concentrations were 0.6 and 1.10, respectively. The antiangiogenic score of bevacizumab, used as a positive control, was 0.95 at 1.0 µmol/L concentration. No significant difference was found between the antiangiogenic scores of levosimendan and bevacizumab (p=0.54). CONCLUSIONS: Our results indicate that levosimendan has antiangiogenic effects on the chorioallantoic membrane. However, these findings must be confirmed in future studies on humans.

Efficacy of miltefosine for topical treatment of Acanthamoeba keratitis in Syrian hamsters.

Acanthamoeba keratitis is a painful corneal infection and difficult to treat because no sufficiently efficient drug has yet been available. The aim of the study therefore was to assess the therapeutic potential of miltefosine on Acanthamoeba keratitis-infected hamster eyes. The cornea of hamsters were infected with Acanthamoeba hatchetti, a human corneal isolate. On the fifth day, all the cornea were microscopically examined in order to determine the degree of infections (G, from 0 to 3). Four groups were then prepared: miltefosine (160 µM); 0.1% propamidine isetionate plus 0.02% polyhexnide; infected control (0.05% ethanol in PBS) and a non-infected control (0.05% ethanol in PBS) groups. The treatment was continued for 28 days. After the treatment, the cornea were excised and used for Acanthamoeba culture to investigate the presence of Acanthamoeba growth. Miltefosine treatment yielded much higher cure scores than propamidine isetionate plus polyhexanide. On the last day of treatment, 85% of the miltefosine-treated eyes were graded as G0; no changes were observed in the uninfected control group eyes; G3 eyes showed only a partial improvement. Furthermore, no Acanthamoeba cells could be recovered from the miltefosine-treated eye samples. Miltefosine appeared to hold necessary therapeutic properties for the treatment of Acanthamoeba keratitis.

Effect of combined chlorhexidine gluconate and neosporin on experimental keratitis with two pathogenic strains of Acanthamoeba.

Acanthamoeba keratitis (AK) is a painful, sight-threatening, and difficult-to-treat corneal infection caused by the ubiquitous free-living amoebae Acanthamoeba species. The aim of the present study was to compare the severity of keratitis, caused by Acanthamoeba hatchetii and Acanthamoeba castellanii infections, and to assess the therapeutic effects of combined chlorhexidine (CHX) and Neosporin® treatment in rats. The rats were first divided into two groups, in which the eyes of the animals were infected with A. hatchetii or A. castellanii trophozoites. On day 5, all corneas were examined in order to determine the degree of infection (grade 0 to 3), and animals were divided into two new groups, treatment and infected control groups. The treatment was continued for 28 days, followed by excision and histological evaluation of the corneas. In conclusion, the clinical picture progressed more rapidly and severely in eyes infected by A. castellanii, while it was non-invasive and slower to progress with A. hatchetii. Moreover, eyes infected by A. hatchetii responded quicker and more positively to therapy, consistent with its clinical course, while a longer recovery was seen with A. castellanii. Histological examinations revealed the presence of A. castellanii and A. hatchetii trophozoites in the stroma of eyes of the treatment and control groups. As a result, our findings suggest that a combination of Neosporin with lower doses of CHX may be beneficial to treat patients with early diagnosis of AK.

Antiangiogenic activities of bemiparin sodium, enoxaparin sodium, nadroparin calcium and tinzaparin sodium.

INTRODUCTION: The low-molecular-weight heparins have been demonstrated to have antiangiogenic effects in various assays. We aimed to demonstrate and compare the antiangiogenic effects of four types of commercially available low-molecular weight heparins in the chick embryo chorioallantoic membrane model. MATERIALS AND METHODS: The antiangiogenic efficacies of bemiparin, enoxaparin, nadroparin, and tinzaparin were examined in vivo in the chick chorioallantoic membrane model. Drug solutions are prepared in three different concentrations (100 IU, 10 IU, or 1 IU/10 µl). For each set of experiment twenty fertilized eggs were used. The decrease of vessel formation is examined and scored according to previous literature. RESULTS: Bemiparin, enoxaparin, nadroparin, and tinzaparin sodium all have antiangiogenic effects on chick chorioallantoic membrane at the concentration of 100 IU/10 µl. This effect was also observed in 10 IU/10 µl concentrations of nadroparin and tinzaparin. CONCLUSIONS: The low molecular weight heparins studied have obvious antiangiogenic effects. There may be a difference in the potency of the drugs that could have a significant implication for further clinical research.

In vitro amoebicidal activity of a ceragenin, cationic steroid antibiotic-13, against Acanthamoeba castellanii and its cytotoxic potential.

Acanthamoeba is a free-living amoeba causing a potentially blinding infection of the cornea. Acanthamoeba keratitis is difficult to treat, without total efficacy in some patients because of cysts that are less susceptible than trophozoites to the usual treatments. Contact lens wearers are most at risk and account for some 95% of cases. Cationic steroid antibiotic (CSA)-13 is a small molecule aminosterol that has been shown to mimic the activity of endogenous antimicrobial peptides and has bactericidal activity based on membrane disruption. We investigated here the in vitro effectiveness of CSA-13 with a concentration of 100, 75, 50, and 25 mg/mL on proliferation of Acanthamoeba castellanii trophozoites and cysts and cytotoxic potential. CSA-13 was evaluated for its amoebicidal activity using an inverted light microscope at 1, 2, 4, 8, 12, and 24 h. For the determination of cytotoxicity of the CSA-13 on L929 cells, agar diffusion tests were performed. CSA-13 inhibited trophozoite growth in dose- and time-dependent ways. At 1 h, no viable trophozoites were observed in the presence of CSA-13 solution in a concentration 100 mg/mL in phosphate-buffered saline. Results of cytotoxicity experiments demonstrated that CSA-13 solution had mild toxicity at 100 mg/mL concentration on cells, whereas it had no toxicity at 75 mg/mL concentration. The findings of this experiment as in vitro ameboebicidal activity for Acanthamoeba suggest that CSA-13 has a potential to be used as a new agent in lens solutions to prevent Acanthamoeba growth and infections.