Rapid differential diagnosis of vaginal infections using gold nanoparticles coated with specific antibodies.

Vaginal infections caused by bacteria, Candida and Trichomonas vaginalis, affect millions of women annually worldwide. Symptoms and signs have limited value in differential diagnosis of three causes of vaginitis. Current laboratory methods for differential diagnosis are either expensive or time consuming. Therefore, in this work, development of a method based on gold nanoparticles has been investigated for rapid diagnosis of vaginal infections. Specific antibodies against three main causes of vaginal infections were raised in rabbits. The antibodies were then purified and conjugated to gold nanoparticles and used in an agglutination test for detection of vaginal infections. Finally, sensitivity and specificity of this test for diagnosis of vaginal infections were estimated using culture method as gold standard. Purification of antibodies from sera was confirmed by electrophoresis. Construction of nanoparticles was proved by TEM and FT-IR methods. Conjugation of antibodies to gold nanoparticles was confirmed using XPS method. Sensitivity and specificity of gold nanoparticles for diagnosis of Candida species were 100%, for Gardnerella were 100% and 93%, and for T. vaginalis was 53.3% and 100%, respectively. Gold nanoparticle-based method is a simple, rapid, accurate, and cost-effective test for differential laboratory diagnosis of vaginal infections.

Scolicidal agents for protoscolices of Echinococcus granulosus hydatid cyst: Review of literature.

Hydatid cyst is the larval stage of dog tape worm Echinococcus granulosus. Protoscolices are parasite larvae that develop into adult worms in the final host intestine. During surgical treatment of human hydatidsosis spillage of live protoscolices is the major cause of hydatidosis recurrence. To prevent this problem scolicidal agent such as hypertonic salt are used to kill the protoscolices that may disseminate into the patient's tissues during surgery. However, they may have some unacceptable side effects. To find scolicidal agents with high efficacy, the effect of different compounds on protoscolices of hydatid cyst in vitro has been reviewed. Using PubMed, Scopus, Google Scholar, and SID databases articles about scolicidal effects of different agents on protoscolices of hydatid cyst in vitro were collected. Foeniculum vulgare after 5 min, metalonic extracts of Allium sativum and hypertonic saline after 10 min, and warm water after 2 min kill all alive protoscolices. The above agents that in minimum time and minimum concentration have 100% scolicidal activity, could be good candidates for further investigations.

Immunological diagnosis of human hydatid cyst using Western immunoblotting technique.

BACKGROUND: Echinococcosis is a parasitic disease with worldwide distribution which is caused by the tapeworms Echinococcus granulosus. Diagnosis of the disease relies on imaging techniques, but the techniques are not able to differentiate the cyst from benign or malignant tumors; hence, appropriate serologic methods are required for the differential diagnosis of the infection. MATERIALS AND METHODS: In this investigation, different sheep hydatid cyst antigens probed with thirty sera of patients with hydatid cyst and also thirty human normal sera using Western immunoblotting technique. Considering results of surgery as gold standard, sensitivity and specificity of Western blotting was estimated. RESULTS: Sera of 29, 26, and 16 patients with hydatid cyst reacted with specific bands of hydatid cyst fluid (HCF), protoscolex crude antigen, and cyst wall crude antigen, respectively. However, none of the normal human sera reacted with those specific bands. CONCLUSION: A 20 kDa band of sheep HCF is an appropriate antigen for serodiagnosis of hydatid cyst infection.

Mutual action of anticancer and antiparasitic drugs: are there any shared targets?

Parasites and cancers have some common features. It has been shown that some parasites interfere with tumor growth. In addition, they both have common antigens such as the Tn antigen. A communal action of anticancer and antiparasitic drugs has been reported. This shared action may be related to common targets for these drugs in cancers and parasites. In this paper, mutual action of anticancer and antiparasitic drugs, with the aim of discussing shared targets of these drugs, has been reviewed.

Retraction notice to "The therapeutic potential of human adipose-derived mesenchymal stem cells producing CXCL10 in a mouse melanoma lung metastasis model" [Cancer Lett. 419 (2018) 30-39].

This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the Editor. After publication, the Editors were contacted by a concerned reader regarding alleged image duplication. These allegations are in regard to Fig. 3a being duplicated from a previously published paper in the journal Stem Cells (Stem Cells. 2008 Sep;26 (9):2332-8. doi: 10.1634/stemcells.2008-0084) and Fig. 8a being duplicated from a previously published paper in the journal Molecular Cancer (Mol Cancer 13, 255 (2014). https://doi.org/10.1186/1476-4598-13-255). After a thorough investigation by the editorial team, the Editors determined that there are multiple identical details between Fig. 5A (Cancer Letters) and Fig. 3A (Stem Cells) and the authors did not produce satisfactory evidence that the published images in Cancer Letters were original. Due to this, the Editor does not have confidence in the results and conclusions presented and has made the decision to retract.

Bio-Membrane SELEX as a New Approach for Selecting ss-DNA Aptamers that Bind to the Hydatid Cyst Laminated Layer.

BACKGROUND: Hydatid cyst (HC) is the larval stage of the canine intestinal tapeworm (cestode), Echinococcus granulosus. In addition to the high global economic cost of livestock farming, the infection can lead to dangerous problems for human health. Therefore, research into new diagnosis and treatment approaches is valuable. This study is set out to explore aptamers that bind to HC antigens. METHODS: The similarity between HC genotype in sheep and humans was that sheep HCs were collected and used as a biological membrane for aptamer selection. Four Bio- Membrane SELEX rounds were conducted, and ssDNA aptamers were selected. Selected aptamers' affinity and specificity to the laminated layer antigens were evaluated using membrane staining by fluorescein primer as a probe. Biotinylated primer was used as a probe for aptahistochemistry and dot blot techniques. Subsequently, cloning and plasmid extraction was conducted. The affinity and specificity of sequenced aptamers were examined with the dot blot method. RESULTS: Selected aptamers reacted with HC wall in aptahistochemistry, aptahistofluorescent, and dot blot experiments. Following cloning and sequencing, 20 sequences were achieved. A strong reaction between HC total antigens and sequenced aptamers has emerged in the dot blot method. CONCLUSION: We propose a novel method to determine specific aptamers in this investigation. Bio-Membrane SELEX could be assumed as a practical and sensitive method for aptamer selection. Selected aptamers in this study possibly may be used for specific HC antigens detection.

Sensitive detection of specific cell-free DNA in serum samples from sheep with cystic echinococcosis.

BACKGROUND: Developing more sensitive methods for the diagnosis of echinococcosis is essential. In this study PCR assay for sensitive detection of specific cell-free DNA (cfDNA) of Echinococcus granulosus sensu lato in the sera of the sheep naturally infected with echinococcosis was investigated. METHODS: To extract cfDNA from 35 infected sheep, the modified phenol-chloroform method was used for two different volumes (0.5 and 2 ml) of serum samples. From each extracted sample, two DNA volumes (5 and 10 µl) were amplified using both standard PCR and semi-nested PCR targeting NADH dehydrogenase subunit I. RESULTS: Standard and semi-nested PCR on 0.5 ml of serum samples detected Echinococcus DNA in 8 and 12 out of 35 sheep, respectively; however, using 2 ml of serum samples, they detected 24 and 27 samples. By increasing the volume of template DNA, the PCRs could detect 29 and 33 out of 35 samples. The results were confirmed by sequencing of randomly selected PCR amplicons and comparing them with GenBank databases. CONCLUSIONS: Larger volumes of serum for DNA extraction, greater volumes of DNA template for PCR, and employing a semi-nested PCR protocol, increased the sensitivity of PCR to 95%. This approach can also be applied to the diagnosis of echinococcosis in humans.

Parasites as potential targets for cancer immunotherapy.

Parasites and cancers have some common antigens. Much scientific evidence in the human population, animal models, and in vitro experiments exhibit that parasites have significant anti-cancer effects. The larval stage of the tapeworm Echinococcus granulosus, Toxoplasma gondii, Trypanosoma cruzy, Plasmodium's, and Trichinella spiralis are among the parasites that have been subjects of anti-cancer research in the last decades. Anti-tumor effects of parasites may be due to the direct impact of the parasites per se or indirectly due to the immune response raised against common antigens between malignant cells and parasites. This manuscript reviews the anti-cancer effects of parasites and possible mechanisms of these effects. Options for using parasites or their antigens for cancer treatment in the future have been discussed.

Parasites and cancers: parasite antigens as possible targets for cancer immunotherapy.

An adverse relationship between some parasite infections and cancer in the human population has been reported by different research groups. Anticancer activity of some parasites such as Trypanosoma cruzi, Toxoplasma gondii, Toxocara canis, Acantamoeba castellani and Plasmodium yoelii has been shown in experimental animals. Moreover, it has been shown that cancer-associated mucin-type O-glycan compositions are made by parasites, therefore cancers and parasites have common antigens. In this report anticancer activities of some parasites have been reviewed and the possible mechanisms of these actions have also been discussed.

Molecular diagnosis of Old World leishmaniasis: real-time PCR based on tryparedoxin peroxidase gene for the detection and identification of Leishmania spp.

BACKGROUND & OBJECTIVES: Rapid and accurate diagnosis and identification of Leishmania sp causing cutaneous leishmaniasis is crucial in control and therapeutic programs. The problem of diagnosis with traditional methods is that they have a low sensitivity or time consuming but molecular techniques would be an alternative method for rapid and accurate diagnosis. In this work, tryparedoxine peroxidase gene-based real-time PCR was used for accurate identification of Leishmania spp causing Old-World cutaneous leishmaniasis. METHODS: In this study, biopsies of specimens were taken from the ulcerative sites in 100 patients and used for direct microscopy, culture in NNN or fixed in alcohol for identification of Leishmania spp using tryparedoxin peroxidase gene-based realtime PCR (qPCR). RESULTS: Using direct microscopy and culture method, Leishmania parasites were isolated from 68 out of 100 patient samples. However, 13 patients with negative finding on traditional tests, had positive results on RT-PCR test. After melting curve analysis of PCR product, Leishmania major in 75 and L. tropica in 4 cases were identified. The sensitivity and specificity of RT-PCR for diagnosis of cutaneous leishmaniasis was 98.7 and 59.8%, respectively. CONCLUSION: Results of this study showed that RT-PCR was the most sensitive diagnostic test for cutaneous leishmaniasis and represents a tool for rapid species identification.

Environmental Contamination of Different Areas of Isfahan Province of Iran with Toxocara spp. Eggs using Molecular Methods.

Background: Toxocariasis is a parasitic disease caused by the larval stage of Toxocara canis and Toxocara cati. Infective stage of this parasite for human develops on soil. So, in this work contamination of the soil of public environments in five geographical areas of Isfahan province of Iran has been investigated. Materials and Methods: In this descriptive study, 355 soil samples were collected from parks, children's playgrounds, student dormitories, and university environments, and examined by Flotation method. The samples were then inspected using microscopic and molecular methods. Results: From the 355 examined soil samples in 77 (21.69%), and 87 (24.50%) cases Toxocara eggs were detected by microscopic and molecular methods, respectively. In the molecular method, 31 (8.70%) cases of T. cati and 44 (12.39%) cases of T. canis were identified. Conclusion: Toxocara eggs were identified in all areas of Isfahan province, although contamination rate was higher in Fereydun Shahr and Semirum counties.

Evaluation of Th17 immune responses of recombinant DNA vaccine encoding GRA14 and ROP13 genes against Toxoplasma gondii in BALB/c mice.

Toxoplasma gondii, a worldwide opportunistic parasite, causes serious diseases in both humans and fetuses with defective immune systems. The development of an effective vaccine is urgently required to prevent and control the spread of toxoplasmosis, caused by the apicomplexan parasite Toxoplasma gondii which is one of the most damaging zoonotic diseases of global importance. Plasmid DNA vaccination is a promising procedure for vaccine development and following the previous studies, pcROP13 + pcGRA14 cocktail DNA vaccine was evaluated for Th17 immune responses. Four groups of BALB/c mice were immunized intramuscularly three times at 2-week intervals. Subsequently, the production of anti- T. gondii antibodies and serum levels of cytokines IL-17, and IL-22 were evaluated against the RH strain of T. gondii. In addition, both the reactive oxygen species (ROS) and parasite load were assessed using ELISA and Q-PCR, respectively. The results of this study showed that high levels of IgG were found in mice immunized with cocktail DNA vaccine (p < 0.05). The cytokines level of Th17, IL-17, and IL-22, increased remarkably in the immunized mice (p < 0.05). Also, significant induction (p < 0.05) was observed in ROS. In addition, immunization with pcROP13 + GRA14 resulted in a considerable decrease in parasite load compared to the control groups (p < 0.05). Based on the results, the pcROP13 + GRA14 cocktail DNA vaccine induced Th17 related cytokines and decreased the parasite load in spleen and brain tissues. Hence, pcGRA14 + pcROP13 cocktails are suitable candidates for DNA-based vaccines and due to the development of protective immune responses against T. gondii infection, future studies may yield promising results using these antigens in vaccine design.

Effects of Toxoplasma gondii and Toxocara canis antigens on WEHI-164 fibrosarcoma growth in a mouse model.

Cancer is the main cause of death in developed countries. However, in underdeveloped countries infections and parasitic diseases are the main causes of death. There are raising scientific evidences indicating that parasitic infections induce antitumor activity against certain types of cancers. In this study, the effects of Toxoplasma gondii and Toxocara canis egg antigens in comparison with Bacillus Calmette Guerin (BCG) (known to have anticancer distinctive) on WEHI-164 fibosarcoma transplanted to BALB/c mice was investigated. Groups of 6 male BALB/c mice injected with T. gondii antigen, BCG, or T. canis egg antigen as case groups and alum alone as control groups. All mice were then challenged with WEHI-164 fibrosarcoma cells. The mice were examined for growth of the solid tumor and the tumor sizes were measured every other day up to 4 wk. The mean tumor area in T. gondii, BCG, or alum alone injected mice in 4 different days of measurements was 25 mm(2), 23 mm(2), and 186 mm(2) respectively. Also the mean tumor area in T. canis injected mice in 4 different days was 25.5 mm(2) compared to the control group (alum treated) which was 155 mm(2). T. gondii parasites and T. canis egg antigens induced inhibition of the tumor growth in the fibrosarcoma mouse model. We need further study to clarify the mechanisms of anti-cancer effects.

Withdrawn: Effect of Ursodeoxycholic Acid, Lawsonia Inermis and Achil-lea Millefolium Extracts on Hydatid Cyst Protoscolices

The article has been withdrawn on the request of the authors and the editor of the journal Infectious Disorders - Drug Targets.Bentham Science apologizes to the readers of the journal for any inconvenience this may have caused. Bentham Science Disclaimer: It is a condition of publication that manuscripts submitted to this journal have not been published and will not be simultaneous-ly submitted or published elsewhere. Furthermore, any data, illustration, structure or table that has been published elsewhere must be reported, and copyright permission for reproduction must be obtained. Plagiarism is strictly forbidden, and by submit-ting the article for publication the authors agree that the publishers have the legal right to take appropriate action against the authors, if plagiarism or fabricated information is discovered. By submitting a manuscript the authors agree that the copyright of their article is transferred to the publishers, if and when the article is accepted for publication.

Identifying the Most Effective Hydatid Cyst Fluid Fraction for Anticancer Vaccination of 4T1 Breast Tumor-Bearing Mice.

BACKGROUND: The hydatid cyst fluid antigens have high homology with cancer cell antigens and also exhibit considerable immunogenicity. Therefore, their utilization for cancer immunization can cause an effective antitumor immune response. However, the main challenge is identifying the most effective antigens for this purpose. METHODS: Hydatid cyst fluid fractions including the glycolipid fraction, glycoprotein fraction, 78 kDa fraction, and antigen B fraction were prepared. Then, the BALB/c mice were immunized against different antigens and, subsequently, 4T1 cells were subcutaneously implanted. The tumors' growth, metastasis, and tumor-bearing mice survival were assessed in different immunized groups. In addition, IL-2, IL-4, IFN-γ, and TNF-α serum levels were estimated to evaluate the immune system response. RESULTS: BALB/c mice immunization against the complete hydatid cyst fluid antigens exhibited more significant inhibition of the tumors' growth and metastasis and increase of tumor-bearing mice survival in comparison with its derived fractions. However, the 78 kDa fraction exhibited the best results according to the same factors in comparison with all the prepared fractions. CONCLUSIONS: The 78 kDa fraction of the hydatid cyst fluid was the most effective fraction of hydatid cyst fluid for immunization against 4T1 breast tumors.

Anti-Toxoplasma gondii antibodies attach to mouse cancer cell lines but not normal mouse lymphocytes.

Toxoplasma gondii (T. gondii) is prevalent intracellular parasite and a cause of worldwide infection in the human population. An inhibitory effect of this parasite on cancer growth has been demonstrated in cell culture and animal models. To determine whether the anticancer activities of T. gondii are associated with host immune response, in the current study the reactivity of anti-T. gondii antiserum with the surface of cancer cell lines was investigated. Anti-T. gondii antibodies were raised in rabbit and the reaction of this antiserum in comparison with other anti-parasite antisera (anti-T. vaginalis, anti-hydatid cyst fluid, anti-protoscolices antigens) with mouse melanoma or breast cancer cells lines was investigated using flow cytometry. Anti-T. gondii antiserum reacted markedly with the surface of mouse melanoma and breast cancer cells, and less so with the normal mouse spleen lymphocytes. Meanwhile, the other anti-parasite antisera did not react strongly with the surface of cancer cells compared with normal mouse spleen lymphocytes. In summary, it has been demonstrated herein that anti-T. gondii antiserum may selectively react with the surface of mouse cancer cells but not with normal mouse spleen lymphocytes. Therefore, further study on anti-Toxoplasma antibodies may be useful for directing the application of selective drug delivery in cancer treatment.

Monoclonal Antibodies Production Against a 40KDa Band of Hydatid Cyst Fluid.

BACKGROUND AND OBJECTIVES: Hydatid cyst is the larval stage of the tapeworm Echinococcus granulosus. Hydatid cyst fluid, cyst membrane and Protoscolices, contain a complex mixture of antigens that can induce immune responses in the host. Anti-cancer properties of Protoscolices and hydatid cyst fluid has been shown. In order to identify antigens of hydatid cyst fluid that have anti-cancer effect, in this study production of monoclonal antibodies against one of the hydatid cyst fluid band (40KDa) has been investigated. There are many published patents about applications of monoclonal antibodies. METHODS: In this experimental study, 40KDa band of hydatid cyst fluid that has cross reaction with sera of patients with breast cancer was used as antigen. A group of mice were immunized with this antigen, and then their spleen cells were extracted and fused with SP2 cells. Monoclonal antibodies production was checked in wells with signs of cell growth using ELISA and western blotting. The reaction of the produced monoclonal antibodies with breast cancer cells was tested using flow cytometry method. Finally, effect of the monoclonal antibodies on growth of breast cancer cells was investigated in vitro. RESULTS: The results of this study showed that in the first plate antibody against 40KDa was detected in several wells. In the second plate monoclonal antibodies with high titer was detected in one well. The produced monoclonal antibodies reacted with the surface of breast cancer cells. However, they had no significant effect on growth of breast cancer cells in culture medium. CONCLUSION: Monoclonal antibodies against hydatid cyst fluid 40KDa band were produced. These antibodies reacted with the surface of breast cancer cells but had no significant effect on growth of these cells.

A 53 KDa Glycan Antigen of Hydatid Cyst Wall May Involve in Evasion from Host Immune System.

BACKGROUND: Recent studies have shown that similar host glycan antigens are expressed by helminths such as Echinococcus granulosus hydatid cysts to evade from host immune system. In this work to investigate these antigens further, immunological cross-reactivity between human sera and hydatid cyst wall antigens has been investigated. MATERIALS AND METHODS: Hydatid cyst wall antigens were used in enzyme-linked immunosorbent assay and Western immunoblotting and probed with pooled sera of hydatidosis patients and healthy controls. Sodium metaperiodate treatment was used to investigate glycan antigens. RESULTS: A band with molecular weight about 53 KDa reacted with both hydatid patients' sera and also normal human sera. It has been shown that this band was a glycan antigen. CONCLUSIONS: A 53 KDa glycan antigen of hydatid cyst wall that reacted with all human sera may have an important role for evasion from host immune system.

The therapeutic potential of human adipose-derived mesenchymal stem cells producing CXCL10 in a mouse melanoma lung metastasis model.

Interferon γ-induced protein 10 kDa (IP-10) is a potent chemoattractant and has been suggested to enhance antitumor activity and mediate tumor regression through multiple mechanisms of action. Multiple lines of evidence have indicated that genetically-modified adult stem cells represent a potential source for cell-based cancer therapy. In the current study, we assessed therapeutic potential of human adipose derived mesenchymal stem cells (hADSC) genetically-modified to express IP-10 for the treatment of lung metastasis in an immunocompetent mouse model of metastatic melanoma. A Piggybac vector encoding IP-10 was employed to transfect hADSC ex vivo. Expression and bioactivity of the transgenic protein from hADSCs expressing IP-10 were confirmed prior to in vivo studies. Our results indicated that hADSCs expressing IP-10 could inhibit the growth of B16F10 melanoma cells and significantly prolonged survival. Immunohistochemistry analysis, TUNEL assay and western blot analysis indicated that hADSCs expressing IP-10 inhibited tumor cell growth, hindered tumor infiltration of Tregs, restricted angiogenesis and significantly prolonged survival. In conclusion, our results demonstrated that targeting metastatic tumor sites by hADSC expressing IP-10 could reduce melanoma tumor growth and lung metastasis.

Diagnosis of Giardia duodenalis Infection using Dot Blot in Comparison with Microscopy.

BACKGROUND AND PURPOSE: Giardia duodenalis is an intestinal flagellate parasite which spreads all over the world and is considered as a health problem in the most rural and low sanitation areas. Many diagnostic tests have been developed for the detection of Giardia parasite in stool samples but all of them have some disadvantages such as lack of sensitivity and specificity. In search for a simple and accurate test, diagnosis of Giardia infection using dot blot method has been investigated in this work. MATERIALS AND METHODS: In this descriptive study, 30 stool samples which their infection with Giardia were confirmed by direct examination and formalin ether considered as case group. Thirty stool samples without Giardia infection according to formalin ether examination were also considered as a control group. Giardia cysts were isolated from the stool samples using sucrose method. In order to raise antiserum against Giardia cysts, the purified cysts were then sonicated and injected to a rabbit. A mono specific antiserum against the 66KDa band of Giardia cyst antigen was also prepared. The two antisera were used in the dot blot test. Finally, the sensitivity and specificity of the dot-blot method were estimated by considering formalin ether as the gold standard. RESULTS: When Poly specific antiserum was used, the sensitivity and specificity of the dot blot for detection of Giardia infection were 77% and 64% respectively. However the sensitivity and specificity of this assay were 97% and 64% respectively when monospecific antiserum was used. CONCLUSION: It seems that dot blot is an easy method for the diagnosis of Giardia especially in the rural areas. However more work is recommended for further development of this test.