Novel chloroquine loaded curcumin based anionic linear globular dendrimer G2: a metabolomics study on Plasmodium falciparum in vitro using 1H NMR spectroscopy.

Due to side-effects and inefficiency of the drugs used in malaria treatment, finding alternative medicine with less side-effects has attracted much attention. In this regard, in the present study, nanocomposite synthesized and its effects on the metabolites of P. falciparum were investigated. Subsequent to synthesis of nanocomposites, characterization was carried out using nuclear magnetic resonance (NMR), liquid chromatography-mass spectrometry (LC-MS), scanning electron microscopy, dynamic light scattering and Fourier-transform infrared tests. Solubility and drug release were measured and its toxicity on Vero cell was assessed using the MTT assay. The antiparasitic effect of the nanocomposite on the metabolites of P. falciparum was investigated by 1H NMR spectroscopy. Among synthesized nanocomposites, the average size of 239 nm showed suitable solubility in water as well as slow drug release. The MTT assay showed no toxicity for Vero cell lines. Concentrations of 2.5 µg mL-1 of nanocomposite eliminated 82.6% of the total parasites. The most effected metabolic cycles were glyoxylate and dicarboxylate metabolism. In this study, 1H NMR spectroscopy was used with untargeted metabolomics to study the effect of the nanocomposite on P. falciparum. Playing an essential role in understanding drug-target interactions and characterization of mechanism of action or resistance exhibited by novel antiprotozoal drugs, can be achieved by targeting metabolic using LC-MS.

Effects of some natural products from fungal and herbal sources on Giardia lamblia in vivo.

Giardia lamblia (G. lamblia) is the most widely known protozoan parasite that causes human gastrointestinal infection worldwide. Some natural compounds exhibited pivotal effects against different infectious diseases. In this research, the antigiardial activity and cytotoxicity of fungal chitosan, nano-chitosan, Rhamnus cathartica (R. cathartica) and emodin were evaluated in Balb/c mice. Genotyping of G. lamblia was assessed by PCR-RFLP technique. Different concentrations of mentioned compounds were used to check their antigiardial and cytotoxicity effects on human intestinal epithelial cells (HT-29) after 24, 48 and 72 h. The G. lamblia strain used in the current work was genotyped and revealed as an AII assemblage. All the concentration showed acceptable activity against G. lamblia cysts and trophozoites in comparison to the negative and positive controls (furazolidone and metronidazole) in vitro (P 0.05). The maximum mortality rate (100%) was achieved at 100 and 50 µg kg-1 concentrations after 48 and 72 h of exposure time, respectively. Our results provide significant information about the new antigiardial agent and proposed the nano-chitosan and emodin for the development of new drugs against G. lamblia in the future.

Antimalarial Effects of Nano Chloroquine Loaded Curcumin In vivo.

BACKGROUND: Malaria is still the deadliest parasitic disease caused by Plasmodium spp. Due to drug resistance and their unpleasant side effects, of conventional researchers are enormously seeking to achieve antimalarial drugs with more curative effective, less toxic and cost-affordable drugs using more advanced technology such as nanodrugs. PURPOSE: The present study aimed to examine the antimalarial effects of a novel synthesized nonochloroquine-loaded curcumin relying on dendrimer G2 in susceptible mice. METHODS: Antimalarial activity and toxicity of the nanocomposite were examined on BALB/C mice with microscopy, checking RBCs morphology and related enzymatic activity rate. RESULTS: The maximum inhibitory effect of the nanocomposite was seen at 10 mg/kg, killing 98% of P. berghei compared to sole chloroquine, whereas ED50 was reported at 5.5 mg/kg. The safety of the synthesized nanocomposite was confirmed with biochemical tests with no detrimental effects on mice. The sustainability and longevity of the nanodrug increased significantly with the NDC-CQ assay compared to the control groups. CONCLUSION: The study showed that nonochloroquine-loaded curcumin had a promising inhibitory effect on P. berghei growth in infected mice compared to standard drugs. However, further studies and clinical trials with large samples are recommended to study different aspects of using nanodrug.

A review on Wickerhamomyces anomalus yeast: a promising eco-friendly approach to biological controlof malaria.

Malaria is a deadly parasitic disease transmitted by female Anopheles mosquitoes. One of the most extensive malaria control measures proposed by the World Health Organization (WHO), which has received better attention in recent years, is the biological control of Anopheles mosquitoes. In this concept, Wickerhamomyces anomalus yeast has received more attention from researchers in this field. In the present review, we have investigated the anti-malaria effect of Wickerhamomyces anomalous. In the present review, we searched PubMed, ProQuest, Scopus, Embase, Google Scholar, Science Direct, and Wiley databases for relevant articles. Keywords used in the inquiries were biological control, yeast, Wickerhamomyces anomalus, malaria, Anopheles mosquito, and Plasmodium. Wickerhamomyces anomalus has a wide range of anti-microbial activity. By producing killer toxins (KT), this yeast can kill microorganisms, so it has called killer yeast. This was investigated and proven using monoclonal antibody, western blot analysis and immunofluorescence (IFA) technique. It has also been used in various studies regarding the biological control of malaria by killing Anopheles mosquito larvae. Considering the proven lethal effect of toxins produced by Wickerhamomyces anomalus, the results could be a big step forward towards ending the life cycle of malaria parasites in the body of vector mosquitos.

Antiplasmodial Effect of Nano Dendrimer G2 Loaded with Chloroquine in Mice Infected with Plasmodium berghei.

BACKGROUND: Malaria is a parasitic lethal disease caused by Plasmodium protozoa. The resistance and drugs' side effects have led to numerous researches for alternative suitable drugs with better efficiency and lower toxicity PURPOSE: In the present study, we investigated in vivo antimalarial effects of G2 linear dendrimer-based nano-chloroquine. METHODS: After the preparation of nano dendrimer G2, chloroquine loading was done. Determine the characterization of particles were specified by DLS, SLS and SEM. The LC-MS and FTIR were used for verifying the nano dendrimer G2 and the loading of chloroquine into the compound. The Solubility N-chloroquine and measurement of drug release rate were done. Antiplasmodial activity of N-chloroquine on BALB/c mice was performed by the microscope and enzymatic methods. At the end, In vivo toxicity of N-chloroquine on tissues was assayed. The RBC morphology and enzyme levels were identified. RESULTS: The results showed the synthesized N-chloroquine had suitable size and solubility. Highest inhibitory effect on Plasmodium parasitic growth was observed at 16 mg/kg dose of N-chloroquine, which eliminated 95% of the parasites (p > 0.05). ED50 is observed at 7.7 mg/kg of N-chloroquine dose. Biochemical findings showed the synthesized N-chloroquine was safer than chloroquine. The N-chloroquine no adverse effects were observed in examined tissues. CONCLUSION: Due to the better effect of the synthesized N-chloroquine on Plasmodium berghei in mice compared to chloroquine, this nanoparticle can be considered as an effective anti-plasmodium compound while more comprehensive research is recommended.

The Metabolomic Profiles of Sera of Mice Infected with Plasmodium berghei and Treated by Effective Fraction of Naja naja oxiana Using 1H Nuclear Magnetic Resonance Spectroscopy.

BACKGROUND: The use of venom fractions from the Iranian cobra could be useful adjunct treatments of malaria with chloroquine. A metabolomic investigation with 1HNMR spectroscopy was conducted on an effective fraction tested earlier using Plasmodium berghei as an experimental murine model. PURPOSE: We sought to ascertain both safety and anti-parasitic effects of experimental therapies. METHODS: After purification of the venom fractions, 25 mice were infected, then treated for 4 days with 0.2 ml of 5 mg/kg, 2.5 mg/kg and 1 mg/kg of the effective fraction, chloroquine, and a drug vehicle. An ED50 was obtained using Giemsa staining and real-time PCR analysis. The toxicity tests inspecting both liver and kidney tissues were performed. RESULTS: A clear inhibitory effect on parasitaemia was observed (with 75% inhibition with 5 mg/kg and 50% reduction when 2.5 mg/kg dosage used). ED50 obtained 2.5 mg/kg. The metabolomics were identified as differentiation of aminoacyl-t-RNA biosynthesis, valine, leucine, isoleucine biosynthesis and degradation pathways were observed. CONCLUSION: Upon therapeutic effects of cobra venom fraction, further optimization of dose-dependent response of pharmacokinetics would be worthwhile for further exploration in adjunct experimental venom therapies.

Antimalarial effects of the hydroalcoholic extract of Allium paradoxum in vitro and in vivo.

Malaria still is the most fatal parasitic disease affecting 50% of the world's population. Although annual deaths attributed to malaria has reduced, crucial importance of its prevention and treatment remains a priority for health care systems and researchers. The worldwide increase in resistance to most common antimalarial drugs such as chloroquine, their unpleasant side effects and low efficiencies persuade researchers to prioritize finding alternative drugs including herbal medication from plant roots. The present study aimed to examine in vitro and in vivo effects of hydroalcoholic extract of herbal medicinal plant, Allium paradoxum, on growth rate in Plasmodium falciparum and Plasmodium berghei. The cytotoxicity assay was performed for hydroalcoholic extract of A. paradoxum. The 3D7 strain of P. falciparum was cultured. The IC50 assay and enzymatic activity of lactate dehydrogenase were performed. BALB/c mice were infected with P. berghei in vivo. Toxicity and histopathological changes in the tissues of liver and kidney were also examined. The highest efficacy of A. paradoxum extract was observed at 80 µg/mL in P. falciparum culture resulting in 60.43% growth inhibition compared to control groups. The significantly highest parasite growth inhibition with 88.71% was seen in the mice infected with P. berghei when administered with 400 mg/kg extract compared to control groups. No significant changes in the liver and kidney cells were observed between experimental and control groups. The study showed that A. paradoxum extract exhibited significant antimalarial properties in vitro on P. falciparum and in vivo in mice infected with P. berghei. There was no significant toxicity in the liver and kidney of the treated mice.

Different manifestation of Echinococcus granulosus immunogenic antigens in the liver and lungs of intermediate host.

Hydatidosis is one of the most important zoonotic diseases with a worldwide distribution and it seems that the survival of Echinococcus granulosus in nature for many years, is due to having different mechanisms to escape from the host immune systems. One of these efficient mechanisms is the production of various antigens and proteins by the larva of the parasite and the main purpose of this study is evaluation of manifestation of various antigens in different parts of intermediate host. The hepatic and pulmonary hydatid cysts were gathered from sheep and the antigens of different parts of the cysts (laminated layer, protoscolices and cyst fluid) were separated and analyzed by SDS-PAGE and then transferred to nitrocellulose paper and finally, Western blot analysis was evaluated the immunogenicity of proteins. The antigens of laminated layer, protoscolices and hydatid cyst fluid, in different tissues of the liver and lungs, manifest various proteins and also these antigens are immunogenically different. Also, it is found more immunogenic proteins in the laminated layer than the other parts of the cysts. The various proteins are generated by Echinococcus granulosus larva depending on the type of tissues attacked by the parasite. Increasing the chance of survival may be the main cause of manifestation various antigens in different parts of cysts and host tissues. These antigenic variations might have made diagnostic serologic test unreliable.

In Vitro Antiprotozoal Effects of Nano-chitosan on Plasmodium falciparum, Giardia lamblia and Trichomonas vaginalis.

BACKGROUND: Treatment of parasitic infections with conventional drugs is associated with high toxicity, and undesirable side effects require cogent substitutions. Nanotechnology has provided novel approaches to synthesize nano-drugs to improve efficient antipathetic treatment. PURPOSE: Nano-chitosan as a nontoxic antimicrobial agent was examined against three most prevalent protozoa in humans, Plasmodium falciparum, Giardia lamblia and Trichomonas vaginalis. METHODS: Chitosan extracted from Penicillium fungi was converted to nanoparticles to maximize its therapeutic properties. Safety of nano-chitosan was examined by determining its hemolytic property and toxicity on PC12 cells. The studied parasites were identified with RFLP-PCR and cultivation in relevant media. Characteristics of nano-chitosan as an useful and valuable curative compound was evaluated by FTIR, DLS and SEM. Dose dependent anti-parasitic effect of nano-chitosan was evaluated. RESULTS: The highest anti-parasitic activity of the nano-chitosan was observed at 50 µg/mL by which growth rates of cultivated P. falciparum, T. vaginalis and G. lamblia were inhibited by 59.5%, 99.4%, and 31.3%, respectively. The study demonstrated that nano-chitosan with the least toxicity, low side effects, and substantial efficacy deserved to be considered as an anti-parasitic nano-compound. CONCLUSION: Nano-chitosan significantly inhibited protozoan growth in vitro promising to explore its use to combat parasitic infections. Further investigations covering extended sample size, in vivo experiments and optimizing the concentration used may lead to efficient treatment of protozoan diseases.

Serological Evaluation of Anti-Toxoplasma gondii Antibodies in Patients with Acute Leukemia and Lymphoma through Chemotherapy.

BACKGROUND: Toxoplasma gondii is a protozoan parasite that belongs to the family Coccidae. We aimed to evaluate IgG avidity and the changes of anti-Toxoplasma immunoglobulins M (IgM) and G (IgG) in patients with acute leukemia and lymphoma. METHODS: Ninety eight patients with Acute myeloid leukemia (AML), Acute Lymphoblastic Leukemia (ALL) and lymphoma, selected from patients referring to Imam Reza Hospital of Tabriz (38°04'N 46°18'E), in terms of the presence of anti-Toxoplasma IgM, IgG, IgG avidity antibodies and the major risk factors were evaluated. RESULTS: The results of pre-chemotherapy evaluation showed that of the examined patients, only two cases, one patient with ALL and another patient with lymphoma, had a positive IgM titer. Overall, 46 cases had positive IgG titers, including 20 patients with AML, 15 patients with ALL and 11 patients with lymphoma. Three (3.06%) patients were positive for anti-T. gondii IgM and one of them was with new infection of toxoplasmosis in lymphoma patients. The post-chemotherapy IgG titer evaluation showed 46 [46.9% (95% CI 37.4-56.7)] positive IgG cases that this result was similar to the result of pre-treatment phase. One [1% (95% CI 0.2-5.6)] positive IgG avidity case was detected using ELISA method, in a patient with lymphoma whose IgM was also positive. There was no significant difference between the type of leukemia and the history of contact with cat. CONCLUSION: Performing specialized tests to diagnose toxoplasma infection before starting treatment, in immunodeficiency patients who undergo chemotherapy, is necessary; therefore, these tests should be considered in therapeutic protocols.

Analysis of the active fraction of Iranian Naja naja oxiana snake venom on the metabolite profiles of the malaria parasite by 1HNMR in vitro.

OBJECTIVES: Malaria is an important parasitic disease with high morbidity and mortality in tropical areas. Resistance to most antimalarial drugs has encouraged the development of new drugs including natural products. Venom is a complex mixture of active pharmaceutical ingredients. The purpose of this study was to investigate the antimalarial activity of purified fractions of Naja naja oxiana. MATERIALS AND METHODS: Lyophilized venom was purified with a Sephacryl S-200 HR column and the fractions lyophilized and inhibitory concentration 50% against Plasmodium falciparum 3D7 in vitro obtained. The 4th fraction was run on a Mono Q column, and activity against P. falciparum was detected by lactate dehydrogenase assay and purity by SDS PAGE. Large scale culture of the parasite was carried out with and without the active fraction on the ring stage for 48 hr. The parasites were collected and lyophilized and analyzed by 1HNMR. Chemometrics studies were performed using MATLAB, differentiating metabolites were identified by Human Metabolic Database, and metabolic pathways by the Metaboanalyst online package. RESULTS: The active fraction from the ion exchange column had a 50% inhibitory concentration of 0.026 µg/ml on P. falciparum in vitro (P<0.001) with molecular weight of 63 kDa by SDS-PAGE and no hemolytic activity. Metabolomics studies on the two groups with and without the fraction identified 5 differentiating metabolites and a number of related pathways. CONCLUSION: The metabolites were succinic acid, l-glutamic acid, pyruvic acid, cholesterol, and NAD. The changes in the Krebs cycle and metabolism pathways of nicotinamide and pyruvate were noticeable.

Induction of Immune Responses by DNA Vaccines Formulated with Dendrimer and Poly (Methyl Methacrylate) (PMMA) Nano-Adjuvants in BALB/c Mice Infected with Leishmania major.

BACKGROUND: Leishmaniasis is a parasitic disease induced by a protozoan from the genus Leishmania. No effective vaccine has yet been developed against the disease. AIM: In this work, two nano-vaccines, TSA recombinant plasmid and dendrimer and poly (methyl methacrylate) (PMMA) nanoparticles (as adjuvants), were designed and tested for their immunogenicity in BALB/c mice. METHODS: After the plasmid construction and preparation of adjuvants, three intramuscular injections of the nano-vaccines (100 µg) and the recombinant TSA protein (20 µg) were subcutaneously performed. Eventually, the challenged animals were infected with the parasites (1*106 promastigotes). After the last injections of the nano-vaccines, the responses of their antibody subclasses and cytokines were assessed via ELISA method before and after the challenge. RESULTS: This study revealed that the new nano-vaccines were strong and effective in inducing specific antibody and cellular responses and reducing the parasite burden in the spleen compared to the control groups of Leishmania major-infected BALB/c mice. CONCLUSION: Based on the results, we can suggest that the formulated vaccines are suitable candidates for further studies in the field of leishmaniasis control.