Determination of Parasitic Contamination in Vegetables Collected from Local Markets in Izmir Province, Türkiye

OBJECTIVE: Fresh vegetables are an important part of a healthy and nutrient-rich diet but the consumption of raw vegetables without proper washing is the main way for transmission of parasites. This study was aimed at determining the rate of parasitic contamination in prewashed fresh vegetables sold at randomly selected 10 retail markets which is the last step to reach the consumer in Izmir, Türkiye. METHODS: A total of 80 samples selected from eight types of vegetables including tomato, spinach, lettuce, rocket, mint, parsley, dill, and cucumber were examined for parasitic agents microscopically by sedimentation method after washing samples with normal saline. Statistical analysis was performed using SPSS software version 20.0. RESULTS: Protozoan cysts, helminth eggs, and larvae were detected in 21 (26.2%) of 80 samples from eight different vegetable species. Rhabditidiform larvae 18.7%, Blastocystis spp. 5%, Toxocara spp. 2.5%; Ascaris spp., Fasciola spp., Entamoeba histolytica/ Entamoeba dispar and hooked worms were found in 1.2%. Spinach and mint samples were contaminated with parasites significantly more than other fresh vegetable samples (p<0.008, odds ratio =80.0; p<0.017, odds ratio =46.6 respectively). Cruznema spp., a plant nematode, was found at the highest rate according to the results of culture, polymerase chain reaction and sequencing, respectively. CONCLUSION: In this study, the parasitic contamination was found in approximately one of the four vegetables sold in randomly selected markets in Izmir. These findings show that vegetables sold in local markets can cause parasitic infections if they are consumed without adequate washing and awareness should be raised on this issue. In addition, it was concluded that morphological examinations should be confirmed by molecular studies and sequencing as much as possible in order to avoid misdiagnosis of rhabditidiform larvae.

Liquid Biopsy for Promising Non-invasive Diagnostic Biomarkers in Parasitic Infections.

BACKGROUND: Liquid biopsy refers to the sampling and molecular analysis of body fluids such as blood, saliva, and urine in contrast to conventional tissue biopsies. Liquid biopsy approach can offer powerful non-invasive biomarkers (circulating markers) for diagnosis and monitoring treatment response of a variety of diseases, including parasitic infections. METHODS: In this review, we concentrate on cell-free DNA (cfDNA), microRNA (miRNA), and exosomes in the published literature. RESULTS: Considering the high prevalence and severity of parasitic infections worldwide, circulating biomarkers can provide a new insight into the diagnosis and prognosis of parasites in the near future. Moreover, identifying and characterizing parasite- or host-derived circulating markers are important for a better understanding of the pathogenesis of parasite infection and host-parasite relationship at the molecular level. Profiling of biomarkers for parasitic diseases is a promising potential field, though further studies and optimization strategies are required, both in vitro and in vivo. CONCLUSION: In this review, we discuss three approaches in the liquid biopsy including circulating cfDNA, miRNAs, and exosomes for diagnosis and evaluation of parasites and summarize circulating biomarkers in non-invasive samples during parasitic infections.

Comparison of the multi-epitope recombinant antigen DIPOL and hydatid fluid for the diagnosis of patients with cystic echinococcosis.

The use of serological tests containing multiple immunodominant antigens rather than single antigens have the potential to improve the diagnostic performance in Cystic Echinococcoses (CE) as a complement tool to clear the inconclusive imaging data. Here, we comparatively evaluated the diagnostic value of Hydatid Fluid (HF) and the recently described recombinant multi-epitope antigen DIPOL in IgG-ELISA in a clinically defined cohort of CE patients. The serum samples from 149 CE patients were collected just before surgical or Percutaneous- Aspiration- Injection- Reaspiration (PAIR) procedures. Additionally, serum samples of patients with other parasitic infections (n=49) and healthy individuals (n=21) were also included in the study as controls. To investigate the association between the genotype of the parasite and DIPOL, cyst materials from 20 CE patients were sequenced. In terms of overall sensitivity, HF was higher than DIPOL (82.55%,78.52%, respectively). However, while the sensitivity of HF was higher than DIPOL in patients with active and transitional cysts (83.3%, 75.4%, respectively), sensitivity of DIPOL in inactive cysts was higher compared to HF (95.6%, 78.3%, respectively). The sensitivity of DIPOL depending on cyst stage was statistically significant (P= 0.041). In terms of specificity, DIPOL was found to be better than HF (97.71%, 91.43%, respectively). By genotyping, the majority of 20 patients showed G1 genotype (80%). All patients harboring G3 and G1/G3 cyst genotypes were positive with both antigens, while 87.5% of patients with G1 genotype were seropositive with HF and 75% with DIPOL. The overall sensitivity and high specificity of DIPOL suggest that this recombinant protein containing immunodominant epitopes is a potential substitute for the HF by serological tests for the diagnosis of CE.

Comparison of the New Multiepitope Recombinant Peptide Antigen with Cyst Fluid Antigen in the Follow-up of Patients with Cystic Echinococcosis

Objective: The follow-up of patients with cystic echinococcosis (CE) offers the opportunity of evaluating the prognosis of the infection as well as detecting relapse. This study aimed to evaluate the performance of the new multiepitope recombinant peptide (recDipol) antigen in the follow-up of CE patients treated by surgery or percutaneous aspiration injection respiration. Methods: A total of 137 blood samples from 28 patients were evaluated by IgG-ELISA method using recDipol and hydatid fluid (HF) antigens. The patients were simultaneously checked for recurrence by ultrasonography. Results: The seropositivity rate of the 28 patients varied considerably during the follow-up. When the first blood of the patients was evaluated, 4 (14.28%) were seronegative by HF-ELISA and 9 (32.14%) were recDipol-ELISA. During the entire follow-up, only 1 (3.5%) and 6 (21.4%) patients were seronegative by HF-ELISA and recDipol ELISA, respectively. Conclusion: We found that recDipol did not perform as expected in the follow-up due to the higher number of seronegative patients compared to HF-ELISA in the first blood and during the entire follow-up. Our results suggest that imaging methods are gold standards in the diagnosis and that, in parallel, longer-term patient follow-up is required with recombinant antigens that have an improved diagnostic performance.

The Prevalance of Blastocystosis among Patients with Gastrointestinal and Dermatologic Complaints and Effects of Blastocystis spp. Density on Symptomatology.

OBJECTIVE: Blastocystosis has been linked with non-specific symptoms, such as diarrhea, abdominal pain, and distention. In this study, we evaluated the relationship between Blastocystis spp. with urticaria and intestinal symptoms. METHODS: The results of the stool examinations of the patients who were referred to Ege University Medical Faculty Hospital's Medical Parasitology Department Direct Diagnosis Laboratory with gastrointestinal (GIS) and/or dermatologic symptoms between January 2011 and July 2016 were retrospectively scanned. RESULTS: Of the evaluated 37108 stool samples, 2573 (6.93 %) were identified to be positive for Blastocystis spp. The patients with gastrointestinal complaints comprised 68.4% of Blastocystis spp. positive samples (1.761 samples) while 30.1% of patients had dermatologic symptoms (urticaria) (776 samples). Blastocystis spp. density in the non-amplified (without using any stool concentration technique) stool samples of the patients with GIS and dermatological symptoms was as follows: 2.47%, 1.35% rare, 21.73%, 22.17% few, 49.65%, 54.29% medium, 26.27%, and 22.17% dense, respectively. CONCLUSION: 75.92% and 76.46% of the patients with GIS and dermatological complaints had medium to dense parasite densities in their stool samples respectively. This suggests a positive correlation between parasite density and GIS and dermatologic symptomatology.

Travel-Related Parasitic Infections in Travellers to Southeast Asia and Western Pacific Countries.

In the last decades, there has been a significant increase in international human mobility with increase in the prosperity, travel possibilities, and number of refugees. In the first half of 2016, the Asian continent showed the fastest growth in the number of tourists. Such increase is seen due to the interest in Asian history, culture, and cuisine. In the globalizing world, human mobility causes changes in the epidemiology of diseases and the spread of various infections across continents. Parasitic infections that may pose a risk for travellers to the Asia-Pacific are malaria, leishmaniasis, filariasis, foodborne trematode infections, schistosomiasis, soil-transmitted infections, and tourist diarrhea. Consulting a travel medical expert and using health services such as pre-travel vaccination and chemoprophylaxis will reduce the risk of infectious diseases among travelers.

Incidence of Parasitic Diarrhea in Patients with Common Variable Immune Deficiency.

OBJECTIVE: Parasites might cause atypical and severe infections in immunocompromised hosts. The prevalence of diarrhea among common variable immune deficiency (CVID) syndrome patients varies between 20% and 94%, which indicates that diarrhea and gastrointestinal system (GIS) complaints could be the second leading cause of morbidity in CVID patients after respiratory tract infections. This study aimed to assess the prevalence of intestinal parasites in CVID patients with GIS complaints and diarrhea. METHODS: In this study, all cases followed up in the Immunology and Allergy Clinic of Ege University School of Medicine from July 2008 to August 2015 with the diagnosis of CVID were reviewed retrospectively. The stool samples of patients with diarrhea were identified using direct microscopy of native (0.09% NaCl) and Lugol's iodine preparations followed by formol-ethyl acetate concentration to apply modified Kinyoun, trichrome, acid-fast trichrome, and modified trichrome stains for the presence of intestinal parasites. RESULTS: Overall, 26 of 37 CVID patients had diarrhea; white and red blood cells (WBCs and RBCs, respectively) were identified in 11 and 10 of these 26 samples, respectively. Intestinal parasites were found to be present in 7 of the 11 patients with WBCs and 3 of the 10 patients with RBCs. With the addition of patients who neither had WBCs nor RBCs in their stool, a parasitic agent was detected in 13 (50%) of the 26 patients with diarrhea. There was no significant difference between the diarrheic patients with or without intestinal parasites with respect to cramps, fever, nausea and vomiting, tenesmus, bloody feces, and presence of mucus in the stool. Only one patient had malabsorption, which was not associated with intestinal parasites. The most common parasites detected in this study were Cryptosporidium spp. (n=9; 69.2%), Giardia spp. (n=7; 53.8%), and Blastocystis spp. (n=3; 23.1%). We also identified that parasitic diarrhea in CVID patients tended to last longer (M (mean): 16.2 days) than other causes of infectious diarrhea; this is in accordance with previous studies. CONCLUSION: Cryptosporidium spp. was found be the major cause of parasitic intestinal infection in this patient population. It was concluded that parasitic infections may cause chronic diarrhea, which are major causes of morbidity in CVID patients. Therefore, special attention is necessary for the identification of intestinal parasites in CVID patients with diarrhea.

Preparation and characterization of polyethyleneglycolmethacrylate (PEGMA)-co-vinylimidazole (VI) microspheres to use in heavy metal removal.

Polyethyleneglycolmethacrylate (PEGMA) and vinylimidazole (VI) were used in order to obtain microspheres of PEGMA-VI copolymers that can be used in heavy metal removal applications. The obtained copolymers were characterized and their use as sorbents in heavy metal removal was investigated. In the first part of the study, PEGMA-VI microspheres were prepared by suspension polymerization method. The obtained swellable microspheres with 10-50 microm average diameter did not have permanent porosity according to the morphological and physicochemical determinations. The sizes of microspheres became smaller with increasing VI and cross-linker ethyleneglycoldimethacrylate (EGDMA) contents and increasing agitation rate. The VI content, EGDMA ratio, pH and ionic strength were determined as the effective parameters on the swelling behavior of PEGMA-VI microspheres. In the second part of the study, Cu(II) ions were used as a model species in order to investigate the usability of the obtained PEGMA-VI microspheres in heavy metal removal. Adsorption capacities under optimum conditions were determined. The Cu(II) ion adsorption capacity increased by increasing the initial Cu(II) ion concentration, and it reached the maximum value (i.e., 30 mg Cu(II)/g PEGMA-VI microspheres) at 400 mg Cu(II)/L initial Cu(II) ion concentration under the determined optimum conditions. Microspheres were found to be reusable after desorption for several times.

An anticancer C-Kit kinase inhibitor is reengineered to make it more active and less cardiotoxic.

Targeting kinases is central to drug-based cancer therapy but remains challenging because the drugs often lack specificity, which may cause toxic side effects. Modulating side effects is difficult because kinases are evolutionarily and hence structurally related. The lack of specificity of the anticancer drug imatinib enables it to be used to treat chronic myeloid leukemia, where its target is the Bcr-Abl kinase, as well as a proportion of gastrointestinal stromal tumors (GISTs), where its target is the C-Kit kinase. However, imatinib also has cardiotoxic effects traceable to its impact on the C-Abl kinase. Motivated by this finding, we made a modification to imatinib that hampers Bcr-Abl inhibition; refocuses the impact on the C-Kit kinase; and promotes inhibition of an additional target, JNK, a change that is required to reinforce prevention of cardiotoxicity. We established the molecular blueprint for target discrimination in vitro using spectrophotometric and colorimetric assays and through a phage-displayed kinase screening library. We demonstrated controlled inhibitory impact on C-Kit kinase in human cell lines and established the therapeutic impact of the engineered compound in a novel GIST mouse model, revealing a marked reduction of cardiotoxicity. These findings identify the reengineered imatinib as an agent to treat GISTs with curbed side effects and reveal a bottom-up approach to control drug specificity.

Rational drug redesign to overcome drug resistance in cancer therapy: imatinib moving target.

Protein kinases are central targets for drug-based cancer treatment. To avoid functional impairment, the cell develops mechanisms of drug resistance, primarily based on adaptive mutations. Redesigning a drug to target a drug-resistant mutant kinase constitutes a therapeutic challenge. We approach the problem by redesigning the anticancer drug imatinib guided by local changes in interfacial de-wetting propensities of the C-Kit kinase target introduced by an imatinib-resistant mutation. The ligand is redesigned by sculpting the shifting hydration patterns of the target. The association with the modified ligand overcomes the mutation-driven destabilization of the induced fit. Consequently, the redesigned drug inhibits both mutant and wild-type kinase. The modeling effort is validated through molecular dynamics, test tube kinetic assays of downstream phosphorylation activity, high-throughput bacteriophage-display kinase screening, cellular proliferation assays, and cellular immunoblots. The inhibitor redesign reported delineates a molecular engineering paradigm to impair routes for drug resistance.

Antimicrobial activity of cefazolin-impregnated mesh grafts.

BACKGROUND: The aim of this study is the preparation and characterization of cefazolin-impregnated meshes (Surgipro; Tyco Healthcare USSC, Norwalk, CT, USA) to be used as antimicrobial devices. METHODS: During the impregnation, poly(DL-lactide-co-glycolide) solution with cephazolin in dichloromethane was used as coating material. In vitro release experiment was carried out first; later cefazolin-impregnated meshes were evaluated for the characteristics of antimicrobial efficacy and in the last part of the study native and cefazolin-impregnated meshes were implanted in the rats. Cefazolin content was proposed as the effective parameter to control the cefazolin release rate and it was concluded that the higher amounts of initial cefazolin content caused higher release rates. In all cases (or with different cefazolin content for each mesh), the release rates were very rapid in the first 24 h and in the following periods rather slow release rates were obtained. RESULTS: Antimicrobial activity was increased in the case of cefazolin-impregnated form and this efficiency was also increased by the higher amount of cefazolin in certain mesh pieces. Similar antimicrobial activities were observed in the in vitro studies. CONCLUSION: In this study, almost all of the cefazolin-impregnated mesh grafts showed very high antimicrobial activity compared with the bare mesh (or mesh without cefazolin).

Bioadhesive drug carriers for postoperative chemotherapy in bladder cancer.

Transurethral resection (TUR) is the primary mode of therapy for both diagnosis and treatment of bladder cancer. Due to the recurrency of tumoral tissues after TUR further treatment is necessary which is usually in the form of intravesical chemotherapy or immunotherapy. But these therapies have some disadvantages such as disturbancy to patients, adjustment of the suitable dosage, loss of active agents without using. In this study, an alternative approach was proposed and pharmaco-therapeutic agent delivery systems which will supply the suitable dosage of the agent for a certain time period were designed to solve those problems. For this aim, Mitomycin-C loaded alginate and chitosan carriers were prepared to use as an alternative system in the post-operative chemotherapy in bladder cancer. The carriers were prepared in the form of cylindirical geometries to facilitate the insertion of the carrier in in vivo studies. The effects of some parameters (i.e., polymer MW, cross-linker concentration, Mitomycin-C/polymer ratio etc.) over the morphology, swelling behavior, bioadhesion and in-vitro drug release rate of the carriers were evaluated. The obtained results for chitosan and alginate carriers were concluded comparatively.

Norfloxacin-loaded chitosan sponges as wound dressing material.

The aim of this study was the preparation and characterization of chitosan sponges including a model antibiotic (i.e., norfloxacin). The chitosan sponges were prepared by a solvent evaporation method. The matrix was also cross-linked during the preparation. The results indicated that the chitosan sponges were in the fibrillar structure. The swelling behavior, norfloxacin loading, in vitro release characteristics, and antibacterial activity were determined. The effects of cross-linker concentration, norfloxacin/chitosan ratio, chitosan molecular weight, and base concentration were investigated. The most effective parameter was found to be the degree of neutralization. It was also observed that the equilibrium swelling ratio decreased with increasing cross-linking density. The norfloxacin release was found to be swelling controlled initially and diffusion controlled at the extended release periods. It was also found that the antibacterial activity was directly proportional to the release rate.