Chitosan-titanium dioxide-glucantime nanoassemblies effects on promastigote and amastigote of Leishmania major

The purpose of the present study was to design nanoassemblies of chitosan-titanium dioxide (TiO2) nanoparticles (NPs) loaded with glucantime for using their synergistic effects and enhancing the toxic effects of glucantime on Leishmania parasites. The nanoassemblies were prepared by electrostatic interactions and optimized by a response surface central composite design. The effects of glucantime, chitosan and TiO2 NPs amounts were studied on the particle size, zeta potential, loading efficiency, and release efficiency of drug from nanoassemblies. The conjugation of TiO2/chitosan-glucantime was verified by UV spectroscopy and changes in surface charge of NPs. The anti-promastigots effect of glucantime loaded in TiO2/chitosan nanoassemblies was studied by tripan blue dye test and their anti-amastigotes effect by counting the average number of parasites per infected J774 macrophages in 100 cells. The optimized formulation obtained by using 12.5 mg glucantime, 25 mg chitosan and 6 mg TiO2 NPs. Although TiO2 NPs alone were effective more than negative control in reduction of promastigots and amastigotes but they didn't show significant difference compared with free glucantime (p > 0.05). However, at the concentration of 50 μg/mL and after 72 h exposure nanoassemblies decreased the proliferation of L. major promastigotes and amastigotes 13 and 4-fold, respectively compared with glucantime alone. © 2017 Elsevier B.V

Prevalence of Intestinal Parasitic Infection Among Inhabitants and Tribes of Chelgerd, Iran, 2008-2009.

INTRODUCTION Although a notable development in treating and controlling of parasitic infections in recent years has occurred but, these infections are still counted as important problems in many countries. AIM The aim of this study is to determine the prevalence of intestinal parasitic infections in the inhabitant and tribe populations who were referred to central health care of Chelgerd, Iran. MATERIALS AND METHODS This descriptive cross-sectional study was carried out from April 2008 to October 2009 in Chelgerd, Iran. A total of 655 samples of feces from inhabitants and tribes were collected and each sample was examined by Direct smear, formol- ethyl acetate concentration and Trichorom staining. RESULTS Out of 655 stool samples, 367(56%) patients revealed at least one intestinal parasite (pathogenic /non-pathogenic protozoa/helminth), 233(67.7%) in tribes and 134(43%) in inhabitants. There was significant difference between infected inhabitants and infected tribes (p=0.001). Although the intestinal parasitic infections were more in female than male it was not statistically significant (p=0.52). There was no significant difference in various age groups. Common intestinal parasitic infections which were detected in both the populations were Giardia intestinalis (28.2%) and Blastocystis hominis (27.5%). CONCLUSIONS We found that the prevalence of intestinal parasitic infections was higher in the tribe than inhabitant populations. Prevalence of intestinal protozoa infections was much higher than the helminthic infections. These findings reflect poor sanitary conditions in this region. They should be educated and provided better facilities to get rid of intestinal parasitic infections

Chitosan-titanium dioxide-glucantime nanoassemblies effects on promastigote and amastigote of Leishmania major

The purpose of the present study was to design nanoassemblies of chitosan-titanium dioxide (TiO2) nanoparticles (NPs) loaded with glucantime for using their synergistic effects and enhancing the toxic effects of glucantime on Leishmania parasites. The nanoassemblies were prepared by electrostatic interactions and optimized by a response surface central composite design. The effects of glucantime, chitosan and TiO2 NPs amounts were studied on the particle size, zeta potential, loading efficiency, and release efficiency of drug from nanoassemblies. The conjugation of TiO2/chitosan-glucantime was verified by UV spectroscopy and changes in surface charge of NPs. The anti-promastigots effect of glucantime loaded in TiO2/chitosan nanoassemblies was studied by tripan blue dye test and their anti-amastigotes effect by counting the average number of parasites per infected J774 macrophages in 100 cells. The optimized formulation obtained by using 12.5 mg glucantime, 25 mg chitosan and 6 mg TiO2 NPs. Although TiO2 NPs alone were effective more than negative control in reduction of promastigots and amastigotes but they didn't show significant difference compared with free glucantime (p > 0.05). However, at the concentration of 50 μg/mL and after 72 h exposure nanoassemblies decreased the proliferation of L. major promastigotes and amastigotes 13 and 4-fold, respectively compared with glucantime alone. © 2017 Elsevier B.V

Induction of Apoptosis in Toxoplasma gondii Infected Hela Cells by Cisplatin and Sodium Azide and Isolation of Apoptotic Bodies and Potential Use for Vaccination against Toxoplasma gondii

Background: Toxoplasma gondii can infect a wide range of mammalians, especially humans. It controls several intracellular signals for the inhibition of apoptosis. This study aimed to investigate the apoptogenic effect of cisplatin and sodium azide on T. gondii infected HeLa cells and isolate apoptotic bodies (blebs) as a potent stimulator of the immune system. Methods: The cytotoxic properties of cisplatin and sodium azide (NaN3) on HeLa cells were evaluated by MTT assay. Moreover, the apoptogenic activity of cisplatin and NaN3 was studied using flow cytometry (Annexin V/PI double staining) and scanning electron microscopy (SEM). Finally, apoptotic bodies were separated by centrifugation. Results: MTT assay data showed that the survival rate of cells treated with different concentration of NaN3 was significantly reduced, compared to negative control groups. Concerning cisplatin, only concentration of 20 μM had not a significant impact on the cell viability; however, the other concentration of cisplatin significantly reduced cell viability, compared to negative control groups. The level of early apoptosis in uninfected HeLa cells was higher compared to infected HeLa cells treated with cisplatin and NaN3. Finally, apoptotic bodies were separated from T. gondii infected HeLa cells treated with cisplatin. Conclusion: Apoptosis was induced in both uninfected and infected HeLa cells with T. gondii and apoptotic bodies were isolated from infected cells. Therefore, further studies on apoptotic bodies are required in order to find a proper candidate for vaccine preparation against T. gondii infections

The prevalence of human trichuriasis in Asia: a systematic review and meta‑analysis

Trichuriasis is one of the most common soil-transmitted helminth (STH) infections, affecting populations globally. The condition is particularly prevalent in tropical and subtropical areas with low levels of sanitation and poor living conditions. The objective of this systematic review and meta-analysis was to evaluate the prevalence of Trichuris trichiura infection in Asia at the country and region level. Multiple databases/academic search engines (Web of Science, PubMed, ProQuest, Scopus, and Google Scholar) were searched for literature on T. trichiura prevalence in Asia published through January 2021. Pooled prevalence was determined using the meta-package in R (version 3.6.1). Out of 13,836 articles, 226 studies (5,439,500 individuals) from 26 countries met the inclusion criteria. Of the 226 studies, 151 were community-based studies that included individuals across the age spectrum, while 75 studies focused on school children (typically in the 5–16 years age range). The overall T. trichiura pooled prevalence was 15.3% (95% CI: 12.4–19.1%), with a pooled prevalence of 13.3% (95% CI: 10.0–17.1%) for the community studies and 20.9% (95% CI: 14.7–27.9%) for the studies only including school children. For studies including all age groups, individuals in the 1–15 years age group had the highest pooled prevalence at 23.4% (95% CI: 1.7–49.4%). There was a significant difference found in overall pooled prevalence by sex (p < 0.001) and community type (rural versus urban) (p < 0.001). Although prevalence appears to be decreasing, study findings suggest that T. trichiura infection continues to be a public health problem in Asia. Therefore, control programs focused on at-risk individuals in endemic areas are needed

Study on four trichostrongylus species in isfahan by PCR-RFLP

Background : Trichostrongyliases as zoonotic diseases are of major public health and socio-economic importance. Accurate identification of these parasites is the first step in strategic planning for the prevention,control and treatment of the parasitic disease. The aim of this study is using PCR-RFLP technique for genotyping. Materials and methods : samples were collected from elementary tract content of 70 sheep in Isfahan and Khorasgan slaughterhouses. Identification of Trichostrongylus was determined based on morphological character and key identification after direct and flotation methods . For genotyping studies, DNA was extracted and the ITS-2 of ribosomal DNA of each species with specific primers was amplified by PCR and restricted with the endonucleases DraI, RsaI, HinfI. The profile was visualized in agarose gel under ultraviolet transillumination. Results: The eggs of most species are morphologically indistinguishable at the generic level .rDNA-ITS2 fragment size of all species were the same size about 330bp, however, there were differences among species in their PCR-RFLP patterns. The two fragments produced with RsaI in all species were the same size about 138bp,190bp. The PCR product T.probulurus with DraI remained unrestricted. Restriction with DraI produced two fragments in the PCR product of T.axei and T.colubriformis about 110bp,215bp, however, the larger of the two fragments in T.vitrinus is smaller in size than in T.axei and T.colubriformis had a size about145bp,185bp.The two fragments produced with HinfI in T.colubriformis had a size about 90bp,238bp, By contrast other Trichostrongylus remained unrestricted. Conclusion: Based of our results from genotyping studies using PCR-RFLP technique, it can be concluded that four species had been identified .This study provides a pattern to distinguish Trichostrongylus species in Isfahan region using the DraI, RsaI, HinfI restriction endonucleases

Molecular identification of Leishmania isolates obtained from patients suspected as having cutaneous leishmaniasis referred to reference laboratories from Yazd province in central Iran

Background: Cutaneous leishmaniasis (CL) continues to be an increasing public health problem in Iran. The dominant etiologic agents of CL in the Old World are Leishmania major and Leishmania tropica. One of the important endemic foci of CL in Iran is Yazd. Recently, previous studies showed the equal prevalence of L. major and L. tropica as the agents of cutaneous leishmaniasis in this area. This prompted us to identify the genotype of L. major isolates obtained from patients with cutaneous leishmaniasis. Materials and Methods: After completing a clinical/epidemiologic data questionnaire for 218 patients with suspected skin lesions, scraping samples were collected, and each specimen was examined using both direct microscopy and molecular assay of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results : Results showed that of the 218 samples, Leishman body was observed in 77 by direct smear and 104 by PCR assay. Molecular assay indicated 50 cases as L. major, 52 cases as L. tropica, and two cases as unknown. Molecular characterization of L. major isolates showed four patterns, named LmA1, LmA2, LmA3, and LmA4. Conclusion: Our study is the first report for molecular characterization of L. major from one of the important central province of Iran that could affect the control strategies in this field

Frequency of Entamoeba histolytica and Entamoeba dispar prevalence among patients with gastrointestinal complaints in Chelgerd city, southwest of Iran

BACKGROUND: Differentiation between Entamoeba histolytica and Entamoeba dispar is very important for both clinical therapy and epidemiological studies. Although these two species are morphologically identical, they have differences in genetic, chemical specifications and pathogenicity. This study was carried out to differentiate E. histolytica from E. dispar and also to find out frequency of the two species. METHODS: Fecal samples were collected three times from 655 patients with gastrointestinal complaints (47.3% male and 52.7% female), who were referred to the primary health care centers of Chelgerd, Chaharmahal and Bakhtiary province. Samples were examined microscopically with direct smear, formalin-ethyl-acetate concentration and trichrom staining methods to distinguish E. histolytica from E. dispar complex and differentiate them from non-pathogenic intestinal amoeba. Genomic DNA was extracted from microscopy positive isolates and polymerase chain reaction (PCR) was carried out to different the two morphologically identical Entamoeba isolates. RESULTS: Among the 655 recruited patients, eleven subjects with E. histolytica I E. dispar isolates (1.7%) were identified by microscopy methods. Ten of the positive isolates (90.9%) were identified as E. histolytica by PCR and one isolate (9.09 %) was positive for E. dispar. CONCLUSIONS: This study revealed that E. histolytica was more prevalent than E. dispar in the studied area. This result was different from the previously reported data in other parts of Iran

Frequency of blood-tissue parasitic infections in patients with multiple sclerosis, as compared to their family members

Background: Multiple Sclerosis (MS) is a chronic demyelinating disease of the central nervous system which has been identifies more prevalent in economically developed countries than in the developing countries. Low prevalence of parasitic infections (which can activate immune response and prevent or modulate damage to host antigens) in these areas is among the possible responsible factors for such a difference. In this study we aimed to compare frequency of blood-tissue parasitic infections in patients with MS, as compared to their healthy family members. Methods: This study was conducted on 50 relapsing remitting MS patients and 50 family members attending MS clinic at Alzahra Hospital. IgM and IgG anti-Toxoplasma gondii were measured. Given the high prevalence of cutaneous leishmaniasis in Isfahan, all the participants were also examined for protozoan leishmania microscopically. Furthermore malaria parasite was investigated. Results: Eighteen patients and 24 healthy family members had positive test in IgG Toxoplasma gondii(P = 0.09). In both groups, there was no positive IgM Toxoplasma gondii. In investigating leishmania, only 3 participants in the case group and 2 in the control tested positive (P = 0.25). No case of malaria was found among the participants. Conclusion: Our results showed a mismatch with hygiene hypotheses examined. However, considering that the prevalence of parasites varies with time, and depends on numerous epidemiological factors; these results do not discredit the theory investigated

A new effective antiplasmodial compound: Nanoformulated pyrimethamine

Objectives: The aim of this study was to evaluate the efficacy of pyrimethamine-loaded poloxamer 407 nanomicelles on Plasmodium berghei strain NICD in vivo. Methods: Pyrimethamine-loaded nanomicelles were prepared and their zeta potential, particle size and polydispersity index were measured. For antiplasmodial assessment, 54 mice were randomly divided into six groups. Four groups were infected intraperitoneally with P. berghei, whereas the two remaining groups did not receive the parasite (negative controls). Three of the P. berghei-infected groups received treatment with either pyrimethamine-loaded nanomicelles (2 mg/kg), pyrimethamine (2 mg/kg) or empty nanomicelles (2 mg/kg); the fourth group remained untreated (positive control). The parasitaemia rate, survival rate and histopathological changes in the liver, spleen and kidneys were examined and were compared with the negative and positive control groups. Results: The mean parasitaemia rate differed significantly between the nanoformulated pyrimethamine group and each of the other groups (P < 0.05). Moreover, the survival rate of mice in the nanoformulated pyrimethamine group (7/9; 78) was significantly higher compared with each of the other groups (P < 0.01). The main histopathological changes, including hepatic necrosis in the liver, lymphoid hypoplasia in the spleen, and tubular nephrosis and perivascular and interstitial lymphocytic infiltration in the kidneys, were considerably lower in the nanoformulated pyrimethamine group than in the pyrimethamine and positive control groups. Conclusion: Pyrimethamine-loaded nanomicelles showed potent antimalarial activity and can be considered as a potential candidate for further examination of their suitability as an antimalarial drug. © 2019 International Society for Antimicrobial Chemotherap