Nanotechnology and malaria: Evaluation of efficacy and toxicity of green nanoparticles.

Malaria is a global health problem that causes 1.5-2.7 million deaths worldwide each year. Resistance to antimalarial drugs in malaria parasites and to insecticides in vectors is one of the most serious issues in the fight against the disease. Moreover, the lack of an effective vaccine against malaria is still a major concern. Recent developments in nanotechnology have resulted in new prospects for the fight against malaria, especially by obtaining metal nanoparticles (NPs) that are less toxic, highly biocompatible, environmentally friendly, and less expensive. Numerous studies have been conducted on the synthesis of green NPs using plants and microorganisms (bacteria, fungi, algae, actinomycetes, and viruses). To our knowledge, there is no literature review that compares toxicities and antimalarial effects of some existing metallic nanoparticles revealing their advantages and disadvantages. The purpose of this review is to assess the metal NPs obtained through various green synthesis processes, to display the worth of future malaria research, and to determine future strategies. The literature review revealed that there are very limited studies on green NPs covering all stages of malaria parasites. Additionally, green metal nanoparticles have yet to be studied for their possible toxic effects on infected as well as healthy erythrocytes. Moreover, the toxicities of green metal NPs obtained from various sources differed according to concentration, size, shape, synthesis method, and surface charge, indicating the necessity of optimizing the methods used in future studies. This work has investigated the effectiveness of green metal nanoparticles synthesized from different sources against malaria, as well as their advantages and disadvantages. It was concluded that studies on the toxic properties of green nanoparticles would be very important for future stages.

Resuscitation of the Helicobacter pylori Coccoid Forms by Resuscitation Promoter Factor Obtained from Micrococcus Luteus.

Helicobacter pylori is a gram negative, spiral-shaped, and microaerophilic bacteria which can cause life-threatening diseases. It is known that more than 55% of the human population in the world is already infected by this bacterium. The traditional treatment of H. pylori infection consists of a combination of two or more antibiotics. However, H. pylori has evolved to turning its shape from spiral to coccoid form in the presence of antibiotics and this decreases the therapeutic efficacies of conventional antibiotic applications. Resuscitation promoter factor (RPF) is a protein secreted by Micrococcus luteus have significant resuscitation effects on some bacteria especially in the group of viable but non-culturable (VBNC) pathogens. However, there is no study in the literature investigating the resuscitation effects of RPF derived from M. luteus on H. pylori in order to change its form from coccoid to spiral. The purpose of this study is to investigate the resuscitation effect of RPF-containing metabolites isolated from M. luteus on the morphological transformation of H. pylori coccoid forms to spiral forms in order to increase their susceptibilities to antibiotic treatments. Rpf-containing metabolites were primarily obtained from M. luteus culture supernatants. H. pylori was exposed to five different conditions such as prolonged culture, incubation at + 4 °C, incubation at + 22 °C, cultivation in PBS and treatment with kanamycin in order to induce transformations of bacteria to coccoid forms. Induced H.pylori coccoids were characterized by inverted microscope, UV spectrophotometer, SEM imaging, and flow-cytometer. As a result, it was found that the most suitable condition for inducing coccoid forms was cultivation of bacteria with kanamycin. Followingly, different concentrations of RPF-containing metabolites were applied on H. pylori coccoids induced by kanamycin. For the first time in this study, it was determined that the Rpf-containing metabolites obtained from M. luteus demonstrated very high resuscitation effect on kanamycin-induced H. pylori coccoid forms. This new approach for resuscitation of H. pylori coccoids is thought to play an important role in increasing the treatment effectiveness of the conventional antibiotics against the infection.

Investigation of antileshmanial activities of Cuminum cyminum based green silver nanoparticles on L. tropica promastigotes and amastigotes in vitro.

Leishmaniasis is one of the most important parasitic diseases, which is caused by Leishmania species. Nowadays; although pentavalent antimonials are used as the basic treatment option for Cutaneous Leishmaniasis, high cost, toxicity and resistance of the parasites to the medication over time are some important limitations causing several problems in the treatment. In recent years, the progress in the field of green nanotechnology provides the development of green nanoparticle-based treatment methods for Cutaneous Leishmaniasis. The importance of green nanoparticles has gradually increased due to their special reductive, stabilizing, antioxidant and non-toxic properties. Although there are many studies based on green nanoparticles against Leishmania parasites, we have not found any research about antileishmanial activities of biosynthesized silver nanoparticles (Bio-AgNPs) using Cuminum cyminum L (Cumin) seed extract. Therefore for the first time in this study in vitro antileishmanial effects of Bio-AgNPs prepared from Cumin seed extract were examined on L. tropica promastigote and amastigote forms and their efficacies were compared with chemically synthetized AgNPs. During the experiments, antileishmanial effects of synthetized nanoparticles were determined on both promastigote and amastigote forms of Leishmania parasites by detecting different parameters such as proliferation, infection index and produced nitric oxide (NO) amounts from macrophages. According to the results, it was shown that Bio-AgNPs and AgNPs excessively inhibited L. tropica promastigotes and amastigotes by significantly decreasing proliferation rates of promastigotes and metabolic activities of amastigotes, as well. Moreover, infection index rates of macrophages revealed remarkable anti-amastigote performances of Bio-AgNPs. Besides, Bio-AgNPs stimulated macrophages to release NO to kill Leishmania parasites. Consequently, for the first time, Bio-AgNPs were found to be more effective on both forms of Leishmania parasites than AgNPs. Obtained results indicated high antileishmanial potential of green nanoparticles on L. tropica parasites, causative agents of Cutaneous Leishmaniasis. Thus, obtained results demonstrated that green nanoparticles can lead to the development of new, safer, stable and more effective antileishmanial formulations against Cutaneous Leishmaniasis.

A nanotechnology-based new approach in the treatment of breast cancer: Biosynthesized silver nanoparticles using Cuminum cyminum L. seed extract.

The present study reports the anticancer activities of Cuminum cyminum L. (Cumin) seed extract, chemically synthetized silver nanoparticles (AgNPs) and biosynthesized silver nanoparticles (Bio-AgNPs) from Cumin seeds on human breast adenocarcinoma cell line (MCF-7) and human breast adenocarcinoma metastatic cell line (AU565). The synthetized nanoparticles were characterized by dynamic light scattering (DLS), UV-visible spectroscopy (UV-Vis), X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR) and Scanning electron microscopy (SEM). The cytotoxic and anticancer effects of AgNPs and Bio-AgNPs were determined by MTT assay. According to the cytotoxicity analysis, Bio-AgNPs appears to be less toxic against J774 macrophage cells than AgNPs since IC50 values were measured as 0.75 and 1.25 µg/ml for AgNPs and Bio-AgNPs, respectively. On the other hand, Bio-AgNPs demonstrated significant inhibitory effects on human breast cancer cells at non-toxic concentrations such as 0.25 and 0.5 µg/ml. However, at increased concentrations, the lethal effects of AgNPs on breast cancer cells were higher than Bio-AgNPs. When cytotoxic and anticancer characteristics of Cumin extract were investigated, it was established that it did not show any inhibitory effect on J774 cells, while killing the half of MCF-7 cells at investigated concentrations. Interestingly, Cumin extract gave rise to no inhibitory effects against AU565 cells. On the other hand, AgNPs and Bio-AgNPs exhibited considerable anticancer activities on both cell lines. The inhibition percentages of AgNPs on MCF-7 and AU565 cell lines were respectively evaluated as 95% and 97% at the highest concentrations applied (12.5 µg/ml). Similarly, we determined that 87.5% and 96% of MCF-7 and AU565 cells were respectively inhibited when they were exposed to the highest concentrations of Bio-AgNPs. Considering relatively toxic-free features of Bio-AgNPs prepared from Cuminum cyminum L. seed extracts, it can be thought that this formulation will be a pioneer in development of nanotechnology-based new anticancer drug for the treatment of breast cancer in near future.

An overview of nanotechnology-based treatment approaches against Helicobacter Pylori.

Introduction: Helicobacter Pylori (H.Pylori) is a pathogen that infects about 50% of the world's population and is known to be responsible for gastroduodenal diseases such as atrophic gastritis, peptic ulcer and stomach cancer. Nowadays, there is no treatment that ensures complete eradication. In addition, resistance to antibiotics used in the current treatment adversely affects the success rates in the fight against infection. Areas covered: This article take attention to treatment approaches using nanoparticles as an alternative to H.Pylori treatment to cope with increased antibiotic resistance. The purpose of this review is to provide an overview of the current limitations and new promising altenatives in treatment of H.Pylori, to highlight the location of nanotechnology to overcome treatment failures, and to emphasize the advantages of using membrane-coated nanoparticles for the first time. Expert opinion: Because of the current problems in the treatment of H.Pylori, there is increasing interest in alternative approaches including nanotechnology. The strong antibacterial effects of metallic nanoparticles, the advantages of polymeric nanoparticles in drug delivery and drug protection, and the prominent properties of membrane-coated nanoparticles in direct targeting demonstrate the significance of nanotechnology in developing new approaches for treatment of H.Pylori infection.

Current aspects in treatment of breast cancer based of nanodrug delivery systems and future prospects.

Breast cancer is one of the most common diseases worldwide. The risk of getting this disease in female is 30% and the mortality rate is 14%. The breast cancer treatment is based on surgery, chemotherapy and radiotherapy. However, an effective treatment method has not been developed. The main cause of failure in the treatment is cancer stem cells metastasis and chemo-resistance. The use of nanocarrier systems against breast cancer stem cells has great importance. Not only advantages of polymeric drug delivery systems are increasing the stability and reduce the side effects of drugs, but also they have disadvantages such as biocompatibility and long-term potential safety. However, in recent years, studies on exosomes provide several advantages. Exosomes usage as nanocarrier do not cause immunological reactions also the drug effectively transport into the cytosol of targeted cell and have more stability characteristics. Although there are studies about various nanocarrier systems in literature against breast cancer but in general, we have not found any review that brings them together and develops a systematic approach to solving the problem. This review mentions prospective new strategies based on various nanocarrier systems and emphasize the importance of exosome based on drug delivery systems in the treatment of breast cancer.

Problems in Stem Cell Therapy for Cardiac Repair and Tissue Engineering Approaches Based on Graphene and Its Derivatives.

BACKGROUND: Today, coronary artery disease is still one of the most important causes of mortality despite advanced surgical methods, pharmacotherapies and organ transplantation. These treatment modalities are intended to prevent further progression of myocardial infarction and do not involve the repair of the damaged part. Therefore, stem cell therapy has emerged as a new approach for the treatment of coronary artery disease. However, there are some restrictions that limit the use of these cells for desired repair. The leading limitation is that newly formed cardiomyocytes do not provide electrical integrity with local cells. OBJECTIVE: In this paper, we review the difficulties that limit the use of stem cell therapy in cardiac repair and emphasize the importance of the integration of stem cell with tissue scaffolds with conductivity. Furthermore, significance of using graphene scaffolds in cardiac tissue engineering is highlighted due to its conductivity features. RESULT: Recently, the fabrication of tissue scaffoldings has made it possible to create a biomimetic cellular environment while providing a new approach to solving these problems in treatment. Especially, the integration of stem cell therapy with graphene-based tissue scaffolds with electrical conductivity, is one of the promising new strategies to turn the success of two approaches of tissue engineering into synergistic effect in cardiac repair. CONCLUSION: Literature analysis has demonstrated that there are some limitations in use of stem cell therapy for successful treatment of cardiac repair and graphene-based tissue engineering approaches which are promising to solve these problems in the near future.

Comparative study on the antileishmanial activities of chemically and biologically synthesized silver nanoparticles (AgNPs).

The present study was conducted to investigate the antileishmanial activity of biogenic silver nanoparticles (AgNPs) compared to chemically synthesized AgNPs. A nano dimension size (10-15 nm) biogenic AgNPs was produced and characterized by UV-Vis spectroscopy and X-rays diffraction. The chemically synthesized AgNPs was recovering from our previous study with a nanoparticle (NP) size in the range of 10-40 nm. The antileishmanial activities were investigated through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell viability assay. The infectivity was determined by Giemsa staining of the infected macrophages cells. Nitric oxide (NO) accumulation was measured by Griess reagent using NaNO2 as a positive control. After 24 h of exposure with nanoparticles (NPs), a concentration-dependent growth inhibition was observed. The IC50 values were determined against promastigotes of L. infantum as 19.42 ± 2.76 µg/ml for leaves aqueous extract mediated AgNPs, 30.71 ± 1.91 µg/ml for stem mediated AgNPs and 51.23 ± 2.20 µg/ml for chemically synthesized AgNPs. It was also detected that all types of NPs produced NO at a significant level. However, the production of a high-level of NO in the biologically synthesized NPs activated macrophage cells, infected with L. infantum promastigotes indicates that NO radicals are mainly responsible for induced cell death and a decrease in the pathogenicity of the parasites. Since, biogenic nanoparticles are cost-effective, eco-friendly, simple to synthesize, and more effective than chemically synthesized silver nanoparticles, therefore, it could be used as a potential alternative for the development of antileishmanial drugs.

A new approach for development of vaccine against visceral leishmaniasis: Lipophosphoglycan and polyacrylic acid conjugates.

OBJECTIVE: To determine the antileishmanial vaccine effectiveness of lipophosphoglycan (LPG) and polyacrylic acids (PAA) conjugates on in vivo mice models. METHODS: LPG molecule was isolated and purified from large-scale Leishmania donovani parasite culture. Protection efficacies of LPG alone, in combination with Freund's adjuvant, in a physical mixture and in conjugate (consisting of various LPG concentrations) with PAA, were comparatively determined by various techniques, such as cultivation with the micro-culture method, assessment of in vitro infection rates of peritoneal macrophages, determination of parasite load in liver with Leishman-Donovan Units, and detection of cytokine responses. RESULTS: Obtained results demonstrated that the highest vaccine-mediated immune protection was provided by LPG-PAA conjugate due to all parameters investigated. According to the Leishman-Donovan Units results, the sharpest decline in parasite load was seen with a ratio of 81.17% when 35 µg LPG containing conjugate was applied. This value was 44.93% for the control group immunized only with LPG. Moreover, decreases in parasite load were 53.37%, 55.2% and 65.8% for the groups immunized with 10 µg LPG containing LPG-PAA conjugate, a physical mixture of the LPG-PAA, and a mixture of LPG + Freund's adjuvant, respectively. Furthermore, cytokine results supported that Th1 mediated protection occurred when mice were immunized with LPG-PAA conjugate. CONCLUSIONS: It has been demonstrated in this study that conjugate of LPG and PAA has an antileishmanial vaccine effect against visceral leishmaniasis. In this respect, the present study may lead to new vaccine approaches based on high immunogenic LPG molecule and adjuvant polymers in fighting against Leishmania infection.

Meglumine antimoniate-TiO2@Ag nanoparticle combinations reduce toxicity of the drug while enhancing its antileishmanial effect.

Currently, the treatment of leishmaniasis is increasingly insufficient as current antileishmanial drugs have many disadvantages such as toxic side effects, high cost, and growing drug resistance. In order to overcome these disadvantages, researchers have recently focused on combination therapy by using pentavalent antimonials in conjunction with other antileihmanial compounds. Our previous study found that TiO2@Ag nanoparticles (TiAgNps) demonstrated significant antileishmanial effects. However, a lethal dose of TiAgNps on L. topica promastigotes was found to be toxic for macrophage cells. Moreover, non-toxic concentrations of TiAgNps were ineffective in inhibiting L. topica promastigotes and amastigotes. Thus, we propose the use of TiAgNps in combination with other antileishmanial compounds like meglumine antimoniate (MA) at non-toxic concentrations, which may increase the efficacies of both agents and decrease their toxicities. Therefore, the aim of this study was to determine in vitro and in vivo antileishmanial efficacies of TiAgNps-MA combinations at non-toxic concentrations and develop a new approach for treatment that lowers the toxicities of pentavalent antimonials to minimal levels and enhances their effectiveness. In vitro screening was performed on L. topica promastigote and amastigote-macropage culture by using MTT assay to determine proliferation, perform infection index analysis, and to conduct a Griess reaction for nitric oxide production, while in vivo antileishmanial assays were applied on Balb/c mice with CL models. The results demonstrated that combinations including TiAgNps and MA at non-toxic concentrations were highly efficacious against both promastigotes and amastigotes, while MA application alone did not show any inhibitory effects. It was determined that combination applications decreased the proliferation of L. topica promastigotes 2- to 5-fold in contrast to use of MA alone, and was dependent on concentrations. Moreover, the use of combinations led to inhibition of L. topica amastigotes at rates ranging between 80% and 95%. Additionally, combinations were found to decrease metabolic activities of each form of the parasite at ranges between 7- to 20-fold, causing programmed-cell death and stimulation of macrophages for intensive production of nitric oxide, which is accepted as an important antileishmanial agent (p<0.05). Furthermore, Σ FIC analysis demonstrated that the tested combinations composed little additive, but mostly synergistic effects for inhibition of promastigotes and amastigotes. According to in vivo screening results, the combinations displayed high antileishmanial activities by successfully healing lesions and significantly reducing parasite burdens. Combined, these results show that TiAgNps-MA combinations were much more effective than use of MA alone at non-toxic concentrations and they possess high potential for development of new antileishmanial drugs to fight against leishmaniasis.

A nanotechnology based new approach for chemotherapy of Cutaneous Leishmaniasis: TIO2@AG nanoparticles - Nigella sativa oil combinations.

Since toxicity and resistance are the major drawbacks of current antileishmanial drugs, studies have been recently focused on combination therapy in fight against leishmaniasis. Combination therapy generally provides opportunity to decrease toxicity of applied agents and enhance their antimicrobial performance. Moreover, this method can be effective in preventing drug resistance. Highly antileishmanial effects of silver doped titanium dioxide nanoparticles (TiAgNps) and Nigella sativa oil were demonstrated in previous studies. However, toxicity is still an important factor preventing use of these molecules in clinic. By considering high antileishmanial potential of each agent and basic principles of combination therapy, we propose that use of combinations including non-toxic concentrations of TiAgNps and N. sativa oil may compose more effective and safer formulations against Leishmania parasites. Therefore, the main goal of the present study was to investigate antileishmanial effects of non-toxic concentrations of TiAgNps and Nigella sativa oil combinations on promastigote and amastigote-macrophage culture systems and also to develop nanotechnology based new antileishmanial strategies against Cutaneous Leishmaniasis. Numerous parameters such as proliferation, metabolic activity, apoptosis, amastigote-promastigote conversion, infection index analysis and nitric oxide production were used to detect antileishmanial efficacies of combinations. Investigated all parameters demonstrated that TiAgNps-N. sativa oil combinations had significant antileishmanial effect on each life forms of parasites. Tested combinations were found to decrease proliferation rates of Leishmania tropica promastigotes in a range between 1,5-25 folds and metabolic activity values between 2 and 4 folds indicating that combination applications lead to virtually inhibition of promastigotes and elimination of parasites were directly related to apoptosis manner. TiAgNps-N. sativa combinations also demonstrated killing effects on L. tropica amastigotes by decreasing infection index values of macrophages 5-20 folds, inhibiting their metabolic activities up to 5 fold, preventing amastigote-promastigote conversion and producing high amounts of nitric oxide. All these results emphasize high potential of TiAgNps-N. sativa oil combinations as new, safer and effective antileishmanial formulations against Cutaneous Leishmaniasis.

The effect of CO2 laser beam welded AISI 316L austenitic stainless steel on the viability of fibroblast cells, in vitro.

It has been determined by the literature research that there is no clinical study on the in vivo and in vitro interaction of the cells with the laser beam welded joints of AISI 316L biomaterial. It is used as a prosthesis and implant material and that has adequate mechanical properties and corrosion resistance characteristics. Therefore, the interaction of the CO2 laser beam welded samples and samples of the base metal of AISI 316L austenitic stainless steel with L929 fibroblast cells as an element of connective tissue under in vitro conditions has been studied. To study the effect of the base metal and the laser welded test specimens on the viability of the fibroblast cells that act as an element of connective tissues in the body, they were kept in DMEMF-12 medium for 7, 14, 28 days and 18 months. The viability study was experimentally studied using the MTT method for 7, 14, 28 days. In addition, the direct interaction of the fibroblast cells seeded on 6 different plates with the samples was examined with an inverted microscope. The MTT cell viability experiment was repeated on the cells that were in contact with the samples. The statistical relationship was analyzed using a Tukey test for the variance with the GraphPad statistics software. The data regarding metallic ion release were identified with the ICP-MS method after the laser welded and main material samples were kept in cell culture medium for 18 months. The cell viability of the laser welded sample has been detected to be higher than that of the base metal and the control based on 7th day data. However, the laser welded sample's viability of the fibroblast cells has diminished by time during the test period of 14 and 28 days and base metal shows better viability when compared to the laser welded samples. On the other hand, the base metal and the laser welded sample show better cell viability effect when compared to the control group. According to the ICP-MS results of the main material and laser welded samples which were kept in the cell culture medium for 18 months, it was determined that the Fe, Ni and Cr ion concentration released to the cell culture medium from the laser welded test sample was less than that of the main material.

Isolation and diagnosis of Helicobacter pylori by a new method: microcapillary culture.

AIM: To investigate the performance of the microcapillary culture method (MCM) in Helicobacter pylori (H. pylori) isolation and diagnosis. METHODS: Microcapillary culture (MC), classical culture (CC), rapid urease (CLO) test, and histopathologic examination (HE) were performed with biopsy samples. Homogenized biopsy samples were loaded into capillary tubes and incubated for 48 h at 37 °C without providing a microaerophilic environment. Additionally, three or four loops of the homogenized sample were inoculated in a ready-to-use selective medium (Becton Dickinson, Helicobacter Agar, Modified) specific for the isolation of H. pylori and incubated at 37 °C in a microaerophilic atmosphere provided by CampyGen (Becton Dickinson, GasPack). Bacteria reproducing in microcapillary tubes were evaluated in an inverted microscope and also were evaluated after performing a CC with the content. Results obtained by CC, CLO test, and HE were compared with those of MC. The diagnostic performances of the methods used in this study were evaluated for specificity, sensitivity, positive predictive value (PPV), negative predictive value (NPV), and CI. RESULTS: H. pylori was found positive by CLO test + HE and/or CC culture in 26 patient antrum and corpus biopsy samples. In 25 (25/26) patient biopsy samples, H. pylori was isolated by MCM, whereas in only 14 (14/26) patient biopsy samples, H. pylori was isolated by CC. CLO test and HE were found positive in 17 (17/26) patient biopsy samples. Comparing the results of the isolation of H. pylori by MCM, CC, CLO test, and HE, the sensitivity of the MCM was found as 96%, the specificity as 80%, the PPV as 83%, the NPV as 95%, and the 95%CI as 0.76 (χ (2) = 31.51, P < 0.01) whereas the sensitivity of the CC was found as 54% (χ (2) = 19.15, P < 0.01), and the sensitivity of the CLO test and HE were found as 65% (χ (2) = 25.26, P < 0.01). CONCLUSION: This new microcapillary cultivation method for H. pylori has high diagnostic sensitivity compared with CC, HE, and CLO tests.

Investigation of metal-polyelectrolyte complex toxicity.

Water-soluble binary and ternary copper complexes of polyelectrolytes were synthesized, and the toxicity of these complexes was tested in mouse fibroblast cell line (L929) in vitro. Both the binary and ternary complexes were prepared at the ratio of 0.4 mole copper(II) ions per monomer of acrylic acid and 0.5 mole copper(II) ions per monomer of methyl vinyl ether maleic anhydride, furthermore at the ratio of 1 and 2 mole bovine serum albumin per mole of polyacrylic acid and poly(methyl vinyl ether-co-maleic anhydride), respectively. Compared to binary copper(II)-polyelectrolyte complexes, these ternary complexes have been determined to be of least toxicity.

The use of platensimycin and platencin to fight antibiotic resistance.

Infectious diseases are known as one of the most life-threatening disabilities worldwide. Approximately 13 million deaths related to infectious diseases are reported each year. The only way to combat infectious diseases is by chemotherapy using antimicrobial agents and antibiotics. However, due to uncontrolled and unnecessary use of antibiotics in particular, surviving bacteria have evolved resistance against several antibiotics. Emergence of multidrug resistance in bacteria over the past several decades has resulted in one of the most important clinical health problems in modern medicine. For instance, approximately 440,000 new cases of multidrug-resistant tuberculosis are reported every year leading to the deaths of 150,000 people worldwide. Management of multidrug resistance requires understanding its molecular basis and the evolution and dissemination of resistance; development of new antibiotic compounds in place of traditional antibiotics; and innovative strategies for extending the life of antibiotic molecules. Researchers have begun to develop new antimicrobials for overcoming this important problem. Recently, platensimycin - isolated from extracts of Streptomyces platensis - and its analog platencin have been defined as promising agents for fighting multidrug resistance. In vitro and in vivo studies have shown that these new antimicrobials have great potential to inhibit methicillin-resistant Staphylococcus aureus, vancomycin-resistant enterococci, and penicillin-resistant Streptococcus pneumoniae by targeting type II fatty acid synthesis in bacteria. Showing strong efficacy without any observed in vivo toxicity increases the significance of these antimicrobial agents for their use in humans. However, at the present time, clinical trials are insufficient and require more research. The strong antibacterial efficacies of platensimycin and platencin may be established in clinical trials and their use in humans for coping with multidrug resistance may be allowed in the foreseeable future.

Conjugation, characterization and toxicity of lipophosphoglycan-polyacrylic acid conjugate for vaccination against leishmaniasis.

Research on the conjugates of synthetic polyelectrolytes with antigenic molecules, such as proteins, peptides, or carbohydrates, is an attractive area due to their highly immunogenic character in comparison to classical adjuvants. For example, polyacrylic acid (PAA) is a weak polyelectrolyte and has been used in several biomedical applications such as immunological studies, drug delivery, and enzyme immobilization. However, to our knowledge, there are no studies that document immune-stimulant properties of PAA in Leishmania infection. Therefore, we aimed to develop a potential vaccine candidate against leishmaniasis by covalently conjugating PAA with an immunologically vital molecule of lipophosphoglycan (LPG) found in Leishmania parasites. In the study, LPG and PAA were conjugated by a multi-step procedure, and final products were analyzed with GPC and MALDI-TOF MS techniques. In cytotoxicity experiments, LPG-PAA conjugates did not indicate toxic effects on L929 and J774 murine macrophage cells. We assume that LPG-PAA conjugate can be a potential vaccine candidate, and will be immunologically characterized in further studies to prove its potential.

Utility of the microculture method for Leishmania detection in non-invasive samples obtained from a blood bank.

In recent years, the role of donor blood has taken an important place in epidemiology of Leishmaniasis. According to the WHO, the numbers of patients considered as symptomatic are only 5-20% of individuals with asymptomatic leishmaniasis. In this study for detection of Leishmania infection in donor blood samples, 343 samples from the Capa Red Crescent Blood Center were obtained and primarily analyzed by microscopic and serological methods. Subsequently, the traditional culture (NNN), Immuno-chromatographic test (ICT) and Polymerase Chain Reaction (PCR) methods were applied to 21 samples which of them were found positive with at least one method. Buffy coat (BC) samples from 343 blood donors were analyzed: 15 (4.3%) were positive by a microculture method (MCM); and 4 (1.1%) by smear. The sera of these 343 samples included 9 (2.6%) determined positive by ELISA and 7 (2%) positive by IFAT. Thus, 21 of (6.1%) the 343 subjects studied by smear, MCM, IFAT and ELISA techniques were identified as positive for leishmaniasis at least one of the techniques and the sensitivity assessed. According to our data, the sensitivity of the methods are identified as MCM (71%), smear (19%), IFAT (33%), ELISA (42%), NNN (4%), PCR (14%) and ICT (4%). Thus, with this study for the first time, the sensitivity of a MCM was examined in blood donors by comparing MCM with the methods used in the diagnosis of leishmaniasis. As a result, MCM was found the most sensitive method for detection of Leishmania parasites in samples obtained from a blood bank. In addition, the presence of Leishmania parasites was detected in donor bloods in Istanbul, a non-endemic region of Turkey, and these results is a vital importance for the health of blood recipients.

Investigation of antileishmanial activities of Tio2@Ag nanoparticles on biological properties of L. tropica and L. infantum parasites, in vitro.

Leishmaniasis is a public health problem which is caused by protozoon parasites belonging to Leishmania species. The disease threatens approximately 350 million people in 98 countries all over the world. Cutaneous Leishmaniasis (CL) and Visceral Leishmaniasis (VL) are the mostly commonly seen forms of the disease. Treatment of the disease has remained insufficient since current antileishmanial drugs have several disadvantages such as toxicity, costliness and drug-resistance. Therefore, there is an immediate need to search for new antileishmanial compounds. TiO2@Ag nanoparticles (TiAg-Nps) have been demonstrated as promising antimicrobial agents since they provide inhibition of several types of bacteria. The basic antimicrobial mechanism of TiAg-Nps is the generation of reactive oxygen species (ROS). Even though Leishmania parasites are sensitive to ROS, there is no study in literature indicating antileishmanial activities of TiAg-Nps. Herein, in this study, TiAg-Nps are shown to possess antileishmanial effects on Leishmania tropica and Leishmania infantum parasites by inhibiting their biological properties such as viability, metabolic activity, and survival within host cells both in the dark and under visible light. The results indicate that TiAg-Nps decreased viability values of L. tropica, and L. infantum promastigotes 3- and 10-fold, respectively, in the dark, while these rates diminished approximately 20-fold for each species in the presence of visible light, in contrast to control. On the other hand, non-visible light-exposed TiAg-Nps inhibited survival of amastigotes nearly 2- and 2.5-fold; while visible light-exposed TiAg-Nps inhibited 4- and 4.5-fold for L. tropica and L. infantum parasites, respectively. Consequently, it was determined that non-visible light-exposed TiAg-Nps were more effective against L. infantum parasites while visible light-exposed TiAg-Nps exhibited nearly the same antileishmanial effect against both species. Therefore, we think that a combination of TiAg-Nps and visible light can be further used for treatment of CL, while application of TiAg-Nps alone can be a promising alternative in VL treatment.

Generation of avirulent Leishmania parasites and induction of nitric oxide production in macrophages by using polyacrylic acid.

Polyacrylic acid (PAA) is one of the anionic synthetic polyelectrolytes and is used in various immunological and pharmaceutical applications. PAA has been used as adjuvant in veterinary vaccines, in particular. However, to our knowledge, there are no studies that document immunostimulant properties of PAA in Leishmania infection. The main aim of this study was to investigate the interaction of Leishmania parasites with PAA: the possible effects on the infectivity of Leishmania promastigotes; and, induction of nitric oxide (NO) production in macrophages in vitro. The cytotoxicity of PAA on both macrophages and Leishmania infantum promastigotes were determined by MTT assay. NO production in the macrophage culture supernatant was measured by the Griess method. A significant, dose-dependent and time-dependent decrease in the infection index was observed after PAA exposure. The value of this decrease was found to be between 93% and 100% for all concentration and time points. PAA (molecular weight (MW) 30, 100 kDa at 1mg/1h)-exposed parasites stimulate NO production significantly at 48 h post-infection (PI), when compared to the control. This study demonstrates that Leishmania parasites lost their virulence upon interaction with PAA, and this interaction induced NO production in infected macrophages. These results may have important implications in the development of anti-leishmanial vaccines and amelioration of immune response.