Molecular diagnosis, phylogenetic analysis, and antifungal susceptibility profiles of Candida species isolated from neutropenic oncological patients.

BACKGROUND: Neutropenia is the most important cause of life-threatening invasive fungal infections (IFIs). Here, we studied the frequency and antifungal susceptibility profiles of Candida species that colonized or caused infections among neutropenic patients with solid or hematological malignancies. METHODS: A total of 362 clinical samples were collected from 138 patients. After initial isolation using a mix of mycological methods, isolates were screened using chromogenic culture media. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was applied for molecular identification. Positive or suspected cases were confirmed using the reference method of sequencing. Antifungal susceptibility testing for voriconazole and caspofungin was carried out using the microbroth dilution method. An in-silico assay was applied for phylogenetic analysis. RESULTS: Thirty-four Candida strains were isolated. C. albicans (47.06%) and C. glabrata (29.41%) were the most frequent strains. Antifungal treatment reduced the chance of Candida colonization by almost 76% in neutropenic patients (OR: 1.759; 95% CI: 1.349 to 2.390; p value: 0.000). An unusual and non-resistant strain, C. lambica, was reported from the bloodstream of a 56-year-old man with hematologic malignancy (HM). Eight isolates were non-susceptible, and one isolate was resistant to voriconazole. Also, four isolates were non-susceptible to caspofungin. CONCLUSION: We can conclude that there is a cause-and-effect relationship between neutropenia, HM background, and Candida species separated from neutropenic patients, which can lead to possible infections. Further and repetitive studies are recommended using different molecular methods for better prediction and management of fungal infections in neutropenic patients.

Induction of TLR5, IRAK1, and NF-κB expression by Trichomonas vaginalis in cervical cancer cell (HeLa) and normal human vaginal epithelial cell (HVECs) lines.

INTRODUCTION: Trichomoniasis is the most common non-viral sexually transmitted infection that increases the risk of cervical cancer. Trichomonas vaginalis (T. vaginalis) can regulate the pro-inflammatory cytokine production in the host cells. Toll-like receptors (TLRs) are a family of the pattern recognition receptors (PRRs) of mammalian cells, expressed in various host cells and have an important role in recognizing pathogens, and pro-inflammatory responses. The aim of the present study is to investigate the role of TLR5 in cervical cancer cells (HeLa) and human vaginal epithelial cells (HVECs) exposed to T. vaginalis. METHODOLOGY: First, the cells and parasites were cultured in RPMI and trypticase yeast extract maltose (TYM), respectively. After adaption of parasite and epithelial cells by RPMI-TYM medium co-culture (9:1 vol/vol), HVECs and HeLa cells were stimulated with T. vaginalis trophozoites (24-hour incubation at 37 °C, 5% CO2). Following RNA extraction and cDNA synthesis, the gene expression levels of TLR5, IRAK1, and NF-κB were assessed using real-time PCR. Besides, the protein levels were measured using western blotting. All tests and controls were normalized using ß-actin as a housekeeping control. RESULTS: Real-time PCR results showed an increased gene expression of TLR5, IRAK1, and NF-κB in T. vaginalis exposed HVECs and HeLa cells compared to the control group (p < 0.05). Additionally, western blot analysis showed a statistically significant increase in TLR5, and NF-κB proteins in both groups after exposure to the parasite (p < 0.05). CONCLUSIONS: These findings provide insight into the host-parasite interaction, and the results indicated that T. vaginalis could stimulate TLR5 and activate related pathways.

Muscular and hepatosplenic candidiasis in a patient with acute myeloblastic leukemia: A case report and literature review.

Key Clinical Message: Muscular and subcutaneous candidiasis is a rare entity in immunocompromised patients, but it should be kept in mind when we see multiple cystic soft tissue masses in addition to target-shaped hepatosplenic lesions in neutropenic patients. US and MRI are useful imaging modalities for the diagnosis and follow-up of these patients. Abstract: Soft tissue candidiasis is an opportunistic infection in immunocompromised patients and must always be diagnosed and treated as soon as possible. In this case report, the patient is a 14-year-old boy with acute myeloid leukemia M3-type who presented with numerous soft tissue and hepatosplenic candidal abscesses.

Development of a high-resolution melt-based assay to rapidly detect the azole-resistant Candida auris isolates.

Background and Purpose: Candida auris is a multidrug-resistant yeast that rapidly spreads, making it the leading Candidate for the next pandemic. One main leading cause of emerging resistant C. auris isolates is nonsynonymous mutations. This study aimed to detect the Y132F mutation, one of the most important azole resistance-associated mutations in the ERG-11 gene of C. auris, by developing a reliable high-resolution melt (HRM)-based method. Materials and Methods: Five C. auris isolates from Iran, plus three control isolates from other Clades were used in the study. The antifungal susceptibility testing through micro broth dilution was performed to recheck their susceptibility to three azole antifungals, including fluconazole, itraconazole, and voriconazole. Moreover, the polymerase chain reaction (PCR) sequencing of the ERG-11 gene was performed. Following the bioinformatic analysis and HRM-specific primer design, an HRM-based assay was developed and evaluated to detect ERG-11 mutations. Results: The minimum inhibitory concentrations of fluconazole among Iranian C. auris isolates ranged from 8 to 64 µg/mL. The PCR-sequencing of the ERG-11 gene and bioinformatic analyses revealed the mutation of Y132F, a substitution consequence of A to T on codon 395 in one fluconazole-resistant isolate (IFRC4050). The developed HRM assay successfully differentiated the targeted single nucleotide polymorphism between mutant and wild types (temperature [Tm]: 81.79 ℃ - cycle threshold [CT]: 20.06 for suspected isolate). For both mutant and non-mutant isolates, the mean Tm range was 81.79-82.39 °C and the mean CT value was 20.06-22.93. These results were completely in accordance with the findings of DNA sequencing. Conclusion: The fast-track HRM-based method successfully detected one of the most common mechanisms of resistance in the ERG-11 gene of C. auris within 3 h. Finally, the development of more panels of HRM assays for the detection of all azole resistance mutations in C. auris ERG-11 is recommended to expand the scope of the field and facilitate the elaboration of rapid and accurate methods of antifungal resistance assessment.

Dual Blockade of PD-1 and LAG3 Immune Checkpoints Increases Dendritic Cell Vaccine Mediated T Cell Responses in Breast Cancer Model.

PURPOSE: Increasing the efficiency of unsuccessful immunotherapy methods is one of the most important research fields. Therefore, the use of combination therapy is considered as one of the ways to increase the effectiveness of the dendritic cell (DC) vaccine. In this study, the inhibition of immune checkpoint receptors such as LAG3 and PD-1 on T cells was investigated to increase the efficiency of T cells in response to the DC vaccine. METHODS: We used trimethyl chitosan-dextran sulfate-lactate (TMC-DS-L) nanoparticles (NPs) loaded with siRNA molecules to quench the PD-1 and LAG3 checkpoints' expression. RESULTS: Appropriate physicochemical characteristics of the generated NPs led to efficient inhibition of LAG3 and PD-1 on T cells, which was associated with increased survival and activity of T cells, ex vivo. Also, treating mice with established breast tumors (4T1) using NPs loaded with siRNA molecules in combination with DC vaccine pulsed with tumor lysate significantly inhibited tumor growth and increased survival in mice. These ameliorative effects were associated with increased anti-tumor T cell responses and downregulation of immunosuppressive cells in the tumor microenvironment and spleen. CONCLUSION: These findings strongly suggest that TMC-DS-L NPs loaded with siRNA could act as a novel tool in inhibiting the expression of immune checkpoints in the tumor microenvironment. Also, combination therapy based on inhibition of PD-1 and LAG3 in combination with DC vaccine is an effective method in treating cancer that needs to be further studied.

Global prevalence and subgroup analyses of coronavirus disease (COVID-19) associated Candida auris infections (CACa): A systematic review and meta-analysis.

BACKGROUND: Increased hospitalisation rates in the Coronavirus disease 19 (COVID-19) era lead to a new wave of hospital-acquired infections such as emerging multidrug-resistant Candida auris. We aimed to evaluate and estimate the global prevalence of coronavirus-associated C. auris infection (CACa). METHODS: We searched related databases between December 2019 and April 2022 for studies that reported data about CACa. Meta-analysis was performed using MedCalc software version 20.104 according to the DerSimonian and Laird method applying the random-effects model. We evaluated heterogeneity using the χ2 -based Q statistic (significant for p-value < .1) and the I2 statistic (>75% indicative of 'notable' heterogeneity). Moreover, if possible, an odds ratio (OR) analysis was performed for eligible data. RESULTS: Our meta-analysis includes ten eligible studies, including 1942 patients hospitalised with COVID-19; 129 were C. auris cases. The overall pooled prevalence of CACa was estimated at 5.7%. The mortality rate of CACa was estimated at 67.849%. Hypertension was the most prevalent comorbidity (59.374%), followed by diabetes mellitus (52.898%) and cardiovascular diseases (31.392%). Men with a prevalence rate of 80.012% were 3.27 (OR) times more prone to getting infected by C. auris. CONCLUSION: We concluded that the prevalence of C. auris infections decreased during the COVID-19 pandemic and the prevalence gradient changed from Asia to America. Unfortunately, there are many descriptive studies with duplicate content in the field of epidemiology of C. auris infections which are increasing every day. We suggest further non-descriptive studies to accurately establish the cause-and-effect relationships between C. auris and COVID-19 infections.

Nanoliposomes increases Anti-Trichomonas vaginalis and apoptotic activities of metronidazole.

Trichomoniasis, caused by Trichomonas vaginalis (T. vaginalis), is the most common non-viral sexually transmitted disease worldwide. As current trichomoniasis chemotherapies have many side effects, we examined the Anti-Trichomonas effects of nano-liposomal metronidazole (NLMTZ) compared to metronidazole (MTZ) in vitro. Liposomes were produced using the thin film hydration-sonication technique with a slight modification coated with MTZ. The average hydrodynamic diameter of monodispersed NLMTZ was evaluated by DLS and the morphological measurements were performed by scanning electron microscopy (SEM). The effects of NLMTZ and MTZ (5, 10, 20 and 40 µg/mL) on T. vaginalis trophozoites (105 cells/mL) in trypticase-yeast extract-maltose (TYM) medium were evaluated in different exposure times. Then, cell viability, IC50, SEM analysis and the expression of the metacaspase gene were assessed by qRT-PCR. Growth inhibition of MTZ in a concentration of 40 µg/mL was 39.34% after 3 h, whereas NLMTZ caused 51% growth inhibition after 3 h and lysed Trichomonas completely after 12 h. The IC50 values were estimated at 31.51 and 15.90 µg/mL after a 6 h exposure for MTZ and NLMTZ, respectively. Moreover, both T. vaginalis treated with MTZ and NLMTZ had high levels of metacaspase mRNA expression relative to the control groups (P< 0.05). A significant difference was observed between the apoptotic intensities of T. vaginalis treated with MTZ and NLMTZ (P< 0.05). This study showed that nano-liposomal MTZ is a potentially excellent approach for the treatment of trichomoniasis in vitro, although further studies are needed before consideration of clinical trials.

Antifungal Effects of Voriconazole-Loaded Nano-Liposome on Fluconazole-Resistant Clinical Isolates of Candida albicans, Biological Activity and ERG11, CDR1, and CDR2 Gene Expression.

This study aimed to assess the effect of voriconazole (VCZ)-loaded nano-liposomes on biological activity and expression of ERG11, CDR1, and CDR2 genes in fluconazole (FCZ)-resistant Candida albicans. In this study, 5 resistant isolates of C. albicans and 3 susceptible clinical isolates to FCZ were scrutinized from 60 patients suspected of candidiasis. The liposomal formulation of VCZ was produced. After that, the minimum biofilm inhibitory concentration (MBIC) testing was performed and the percentage of growth inhibition was determined. Finally, ERG11, CDR1, and CDR2 mRNA levels were amplified by the quantitative reverse transcription PCR (qRT-PCR) instrument. The obtained results unveiled that VCZ-loaded nano-liposome reduction of minimum inhibitory concentration in C. albicans isolates was remarkable. The results of the MBIC in the most optimum inhibitory concentration of VCZ-loaded nano-liposome were determined to be 4.54 and 4.88 µg/mL for susceptible isolate and resistant isolate, respectively. The ERG11 gene expression in FCZ-resistant C. albicans strains in VCZ-treated, liposomal formulation of VCZ-treated, and nontreated specimens stood at 91%, 63%, and 100%, respectively. Expression levels of CDR1 genes in FCZ-resistant C. albicans were shown to be 91%, 88%, and 100%, respectively. Concerning CDR2 genes, this rate varied to 91%, 78%, and 100% in FCZ resistant, respectively. What our study unveiled was that the use of liposomal VCZ formulation could further reduce the expression of azole-resistant genes compared to VCZ itself. In addition, thanks to more efficacious penetration of the liposomal form, the rate of growth inhibition was considerably higher.

Current applications and prospects of nanoparticles for antifungal drug delivery.

Currently, the significance of fungi as human pathogens is not medically concealed in the world. Consequently, suitable recognition and treatment of such infections are of great importance and necessitate the need for comprehensive information in this regard. The introduction of new antifungals and their use today, especially in the last two decades, have revolutionized the treatment of fungal infections. On the other hand, increasing drug resistance in the world has overshadowed such developments. The use of NPs results in the treatment of fungal infections and owing to their specific properties, these particles, unlike the pure antibiotics, can exert a greater inhibitory power although with less concentration compared with conventional drugs. Important reasons that have led to the use of antifungal drugs in delivery systems include reduced drug efficacy, limited penetration through tissue, poor aqueous solubility, decreased bioavailability, and poor drug pharmacokinetics. It is therefore hoped that unfavorable properties of antifungal drugs be mitigated via their incorporation into different types of NPs. This review summarizes the different types of NPs as delivery systems of antifungal as well as their advantages over pure drugs.

Cryptosporidiosis in HIV-positive patients and related risk factors: A systematic review and meta-analysis.

Cryptosporidium is one of the major causes of diarrhea in HIV-positive patients. The aim of this study is to systematically review and meta-analyze the prevalence of Cryptosporidium in these patients. PubMed, Science Direct, Google Scholar, Web of Science, Cochrane and Ovid databases were searched for relevant studies dating from the period of 1 January 2000 to 31 December 2017. Data extraction for the included studies was performed independently by two authors. The overall pooled prevalence was calculated and subgroup analysis was performed on diagnostic methods, geographical distribution and study population. Meta-regression was performed on the year of publication, proportion of patients with diarrhea, and proportion of patients with CD4 < 200 cells/mL. One hundred and sixty-one studies and 51,123 HIV-positive participants were included. The overall pooled prevalence of Cryptosporidium infection in HIV-positive patients was 11.2% (CI95%: 9.4%-13.0%). The pooled prevalence was estimated to be 10.0% (CI95%: 8.4%-11.8%) using staining methods, 13.5% (CI95%: 8.9%-19.8%) using molecular methods, and 26.3% (CI95%: 15.0%-42.0%) using antigen detection methods. The prevalence of Cryptosporidium in HIV patients was significantly associated with the country of study. Also, there were statistical differences between the diarrhea, CD4 < 200 cells/mL, and antiretroviral therapy risk factors with Cryptosporidiosis. Thus, Cryptosporidium is a common infection in HIV-positive patients, and safe water and hand-hygiene should be implemented to prevent cryptosporidiosis occurrence in these patients.

TITLE: Cryptosporidiose chez les patients VIH-séropositifs et facteurs de risque associés : revue systématique et méta-analyse. ABSTRACT: Cryptosporidium est l'une des principales causes de diarrhée chez les patients séropositifs pour le VIH. Le but de cette étude est de revoir et méta-analyser systématiquement la prévalence de Cryptosporidium chez ces patients. Les bases de données PubMed, Science Direct, Google Scholar, Web of Science, Cochrane et Ovid ont été recherchées pour des études pertinentes datant du 1er janvier 2000 au 31 décembre 2017. L'extraction des données pour les études incluses a été réalisée indépendamment par deux auteurs. La prévalence globale combinée a été calculée et une analyse en sous-groupes a été effectuée sur les méthodes de diagnostic, la répartition géographique et la population étudiée. Une méta-régression a été réalisée pour l'année de publication, la proportion de patients atteints de diarrhée et la proportion de patients avec CD4 < 200 cellules/mL. Cent soixante et une études et 51,123 participants séropositifs ont été inclus. La prévalence globale combinée de l'infection à Cryptosporidium chez les patients VIH-séropositifs était de 11,2 % (IC95 % : 9,4 %­13,0 %). La prévalence regroupée a été estimée à 10,0 % (IC95 % : 8,4 %­11,8 %) en utilisant des méthodes de coloration, 13,5 % (IC95 % : 8,9 %­19,8 %) en utilisant des méthodes moléculaires et 26,3 % (IC95 % : 15,0 %­42,0 %) en utilisant des méthodes de détection d'antigènes. La prévalence de Cryptosporidium chez les patients infectés par le VIH était significativement associée au pays d'étude. En outre, il existe des différences statistiques entre la diarrhée, les CD4 < 200 cellules/mL et les facteurs de risque du traitement antirétroviral avec la cryptosporidiose. Ainsi, Cryptosporidium est une infection courante chez les patients séropositifs, et une eau salubre et une hygiène des mains doivent être mises en œuvre pour prévenir la survenue de cryptosporidiose chez ces patients.

Interactions between immune response to fungal infection and microRNAs: The pioneer tuners.

Due to their physiological and biological characteristics, numerous fungi are potentially emerging pathogens. Active dynamicity of fungal pathogens causes life-threatening infections annually impose high costs to the health systems. Although immune responses play crucial roles in controlling the fate of fungal infections, immunocompromised patients are at high risk with high mortality. Tuning the immune response against fungal infections might be an effective strategy for controlling and reducing the pathological damages. MicroRNAs (miRNAs) are known as the master regulators of immune response. These single-stranded tuners (18-23 bp non-coding RNAs) are endogenously expressed by all metazoan eukaryotes and have emerged as the master gene expression controllers of at least 30% human genes. In this review article, following the review of biology and physiology (biogenesis and mechanism of actions) of miRNAs and immune response against fungal infections, the interactions between them were scrutinised. In conclusion, miRNAs might be considered as one of the potential goals in immunotherapy for fungal infections. Undoubtedly, advanced studies in this field, further identifying of miRNA roles in governing the immune response, pave the way for inclusion of miRNA-related immunotherapeutic in the treatment of life-threatening fungal infections.

Nanostructured lipid carriers of ivermectin as a novel drug delivery system in hydatidosis.

BACKGROUND: The larval stage of the tapeworm Echinococcus granulosus is the causative agent of hydatid disease in humans. This zoonotic parasitic infection remains a major health problem in certain areas of the world where is still endemic. In view of the ineffectiveness of some drug treatments, the surgical removal of cysts remains the preferred treatment option together with the administration of albendazole and mebendazole. However, severe side effects of these drugs have been reported which demands developing new scolicidal agents that confer suitable efficacy and fewer side effects during surgery. METHODS: To that purpose, in the present work we assessed the effectiveness of ivermectin (IVM), a macrocyclic lactone endectocide that has shown to be an effective nematocidal drug against other important parasitic infections. To overcome the limitations observed in some drug formulations and resistance, we used nano lipid carriers (NLCs) as a targeted and sustained drug delivery system for IVM. We evaluated the in vitro cestocidal and apoptotic effects of NLCs-loaded IVM versus IVM by quantifying the expression of caspase-3 mRNA. RESULTS: We found that after 60 and 120 min of administration, 800 µg/ml and 400 µg/ml NLCs-loaded IVM induced 100% mortality, respectively. On the other hand, the 800 µg/ml of IVM induced 100% mortality rate 150 min after administration. Additionally, we found that NLCs-loaded IVM induced higher mRNA caspase-3 expression suggesting a more potent apoptotic effect on the parasite. CONCLUSIONS: These data suggest that NLCs-loaded IVM may be a promising alternative to current treatments although in vivo studies are needed.

Assessment of the global paradigms of genetic variability in Strongyloides stercoralis infrapopulations determined by mitochondrial DNA sequences.

Microevolutionary data of Strongyloides stercoralis infrapopulations are regarded as a requirement for determining the global genetic structure and transmission paradigms of this neglected tropical nematode among the neighboring countries of the world. English databases were searched from 2010 to 2019, analyzing a total of 10 publications. The cytochrome c oxidase subunit 1 sequences of S. stercoralis isolated from Asian and African continents were subjected to calculate the diversity indices and genetic differentiation. A parsimonious haplotype network indicated a star-like trait a total of 106 (Homo sapiens) and 48 haplotypes (Canid) being grouped into four distinct geographical haplogroups. A significant genetic diversity was identified in human-derived S. stercoralis (Haplotype diversity: 0.78) and those with dog (Hd: 0.86) origins. Cladistic phylogenetic tree indicated the Japanese, Thailandish, and Myanmarese clades have a sister relationship with the Laotian clade. The statistically significant Fst values indicated that human S. stercoralis populations of Japanese-Thailandish, Japanese-Myanmarese, and Japanese-Laotian origins were genetically differentiated (Fst: 0.48430 to 0.54903). We conclude that a high gene migration of human strongyloidiasis is being unequivocally shared between the Laotion-Myanmarese and Laotion-Thailandish population pairs. The current findings enhance our knowledge to assess the transmission dynamics and the evolutionary patterns of S. stercoralis in various geographical regions of the globe; also it will serve as a basis for public health policy to control human strongyloidiasis particularly in immunocompromised individuals. Besides, the infected canids and other environmental reservoirs for zoonotic transmission of S. stercoralis to humans should be de-wormed along with their owners.

Therapeutic efficacy of nanocompounds in the treatment of cystic and alveolar echinococcoses: challenges and future prospects.

Echinococcus granulosus sensu lato and E. multilocularis are the causative agents of life-threatening cystic and alveolar echinococcoses (CE and AE), respectively, which lead to serious public health concerns across the globe. Benzimidazoles (BMZs) are the drugs of choice for the treatment of human CE and AE. Presently, the chemotherapeutic failures of BMZs against CE and AE are caused by their low aqueous solubility, poor absorption, and consequently their erratic bioavailability. Among the BMZ compounds used for CE/AE treatment, albendazole (ABZ) and mebendazole (MBZ) are the only drugs licensed for human use. Nevertheless, the administration of these BMZs for a long period of time leads to undesirable adverse effects. Therefore, there is an urgent need for designing new formulations of BMZs with increased bioavailability. To bridge these therapeutic gaps, nanoparticle enantiomers of ABZ and drug delivery systems based on nanostructured entities currently provide an interesting new formulation of already existing drugs to improve the pharmacokinetic effects of BMZs. This study provides an overview of the tested nanocompounds against E. granulosus and E. multilocularis, including their effective dose, type of nanoparticles (NPs), assay setting, and therapeutic outcomes. This review suggests that BMZ derivatives loaded in NPs can significantly improve the scolicidal and cysticidal activities compared with single BMZ. Moreover, BMZ-loaded polymeric NPs show a tendency to increase mortality rate against protoscoleces and microcysts compared with metallic formulations, nanoemulsions, lipid nanocapsules, solid lipid NPs, liposomes, and nanocrystals. In the future, the use of the newly structured entities, attained by bridging ligands to the modified surface of NPs, as well as the electromagnetically produced nanodrugs could be helpful for developing fine-tuned formulations as an alternative to the already existing drugs against these neglected parasitic infections.

Implications of exosomes as diagnostic and therapeutic strategies in cancer.

Exosomes offer a new perspective on the biology of cancer with both diagnostic and therapeutic concepts. Due to the cell-to-cell association, exosomes are involved in the progression, metastasis, and therapeutic efficacy of the tumor. They can be isolated from blood and other body fluids to determine the disease progression in the body, including cancer growth. In addition to being reservoirs of biochemical markers of cancer, exomes can be designed to restore tumor immunity. Tumor exosomes interact with different cells in the tumor microenvironment to confer beneficial modulations, responsible for stromal activity, angiogenesis, increased vascular permeability, and immune evasion. Exosomes also contribute to the metastasis with the aim of epithelial transmission to the mesenchyme and the formation of premetastatic niches. Moreover, exosomes protect cells against the cytotoxic effects of chemotherapeutic drugs and prevent the transmission of chemotherapy resistance to adjacent cells. Therefore, exosomes are essential for many fatal cancer agents, and understanding their origins and role in cancer is important. In this article, we attempted to clarify the potential of exosomes for the application in cancer diagnosis and therapy.

Molecular Phylodiagnosis of Enterocytozoon bieneusi and Encephalitozoon intestinalis in Children with Cancer: Microsporidia in Malignancies as an Emerging Opportunistic Infection.

BACKGROUND : Microsporidia may cause infection in both immunocompromised and immunocompetent populations. The best strategy to control microsporidiosis is obtaining thorough knowledge of its outbreak and pathogenicity. PURPOSE : Because of the lack of precise estimation of microsporidia prevalence among Iranian children with cancer, the current study aimed at evaluating the rate of intestinal microsporidia in children undergoing chemotherapy. METHODS:  Patients with cancer undergoing chemotherapy in a children's hospital in Northwestern Iran were studied; 132 stool samples were collected and stained by the Weber and Ryan-blue modified trichrome staining techniques. The extracted DNA samples were evaluated by the nested polymerase chain reaction (PCR) method. All positive isolates were sequenced for genotyping and phylogenetic analysis. RESULTS: A total of 17 (12.8%) samples were microscopically positive for microsporidia infection, whereas only 14 (10.6%) cases were positive based on nested PCR results. In the positive samples detected with nested PCR, the frequency of Enterocytozoon bieneusi and Encephalitozoon intestinalis infections was 71.4% (n = 10) and 28.6% (n = 4), respectively. After sequencing and phylogenetic analysis, the genotype of E. bieneusi was type D and the sequences of the isolated species were similar to those of the registered ones. CONCLUSION: E. bieneusi is a major contributor to microsporidiosis in young immunocompromised patients in Iran. Microsporidia species are well-detected when confirmatory techniques such as molecular methods are in agreement with staining. So, to ensure this, a suggestion has been made to introduce a certain diagnostic test for microsporidiosis.

Current antifungal drugs and immunotherapeutic approaches as promising strategies to treatment of fungal diseases.

Invasive candidiasis (IC) can affect individuals with various underlying diseases hospitalized in different parts of hospitals. In recent decades, IC has caused 27-55% mortality in general population. Although Candida albicans (C. albicans) is still the most common cause of IC, non-albicans infections such as C. krusei, C. glabrata, C. lusitaniae, C. tropicalis, and C. parapsilosis have been increased in recent years. Treatment of invasive fungal infections is challenging as the number of existing antifungals is limited and more problems include: toxicity, drug interactions, and drug resistance. These problems provide a clear rationale for the development of new immunotherapies to increase outcomes in patients with invasive fungal infections. Thus, the purpose of this paper is to complete review of the current and modern antifungal drugs in IC therapy and focus on the role of immunotherapy in preventing and controlling the disease. Therefore, we review the features of current research efforts directed towards devising safe and effective immunotherapeutic options for fungal infections, including work on antifungal vaccines, engineered T-cells, cytokines, monoclonal antibodies, and other agents.

Fungal vaccines, mechanism of actions and immunology: A comprehensive review.

Fungal infections include a wide range of opportunistic and invasive diseases. Two of four major fatal diseases in patients with human immunodeficiency virus (HIV) infection are related to the fungal infections, cryptococcosis, and pneumocystosis. Disseminated candidiasis and different clinical forms of aspergillosis annually impose expensive medical costs to governments and hospitalized patients and ultimately lead to high mortality rates. Therefore, urgent implementations are necessary to prevent the expansion of these diseases. Designing an effective vaccine is one of the most important approaches in this field. So far, numerous efforts have been carried out in developing an effective vaccine against fungal infections. Some of these challenges engaged in different stages of clinical trials but none of them could be approved by the United States Food and Drug Administration (FDA). Here, in addition to have a comprehensive overview on the data from studied vaccine programs, we will discuss the immunology response against fungal infections. Moreover, it will be attempted to clarify the underlying immune mechanisms of vaccines targeting different fungal infections that are crucial for designing an effective vaccination strategy.