Molecular prevalence of Pneumocystis jirovecii and Cryptosporidium in patients with asthma.

Asthma is characterized by chronic airway inflammation. In addition to allergens, microorganisms can affect the clinical course of asthma. It has been shown that some fungi play an important role in the progression of asthma. However, the effects of Pneumocystis jirovecii and Cryptosporidium spp., on the disease are little known. We investigated P. jirovecii and Cryptosporidium spp. in the sputum and stool sample of patients with asthma (n = 40) by microscopy and PCR compared to the healthy group (n = 40). P. jirovecii (12.5 %), and Cryptosporidium spp. (12.5 %) were detected in the sputum samples of only asthmatic patients (p = 0.029 and 0.029 respectively). However, Crpytosporidium spp. was detected equally in stool samples of both groups (p = 0.682). Our results indicate that P. jirovecii and Cryptosporidium spp. should be considered in patients with asthma and molecular screening of these neglected eukaryotes in respiratory tract samples may be beneficial in the clinical management of the disease.

Evaluation of the Impact of the COVID-19 Pandemic on Cystic Echinococcosis Indirect Hemagglutination Test Dynamics: A Single-center Experience.

OBJECTIVE: Cystic echinococcosis (CE) is one of the neglected tropical diseases announced by the World Health Organization. In the period entered with the Coronavirus disease-2019 pandemic, the fight against such diseases has become even more difficult. In our study, we aimed to make inferences about the effects of the pandemic on the diagnosis of the disease by evaluating the number and results of CE indirect hemagglutination test (IHA) before and during the pandemic. METHODS: The number of IHA test requests and positivity rates in the 30-month periods before and after March 11, 2020, when the first case was seen in our country, were evaluated retrospectively. Statistical analysis was made with SPSS version 23 (SPSS, Chicago, IL, USA) program. RESULTS: The results of 1444 patients before the pandemic and 870 patients during the pandemic period were examined. The difference between IHA positivity rates, which was found to be 18.49% before the pandemic and 14.6% during the pandemic, was statistically significant (p=0.016). The positivity rates of women and men were found to be statistically similar in both periods (pbefore=0.621, pafter=0.238). The age group with the highest IHA positivity rate was 20-39 in both periods, and the difference between the positivity rates of the age groups was statistically significant (p<0.001). CONCLUSION: A significant decrease was observed in the rate of IHA positivity during the pandemic period. The status of no increase in positivity rates despite a significant decrease in IHA tests makes us think that the diagnosis may be missed in some patients or that there could be disruptions in their follow-up. For this reason, in order to continue the fight successfully against CE, which is an important public health problem for our country, early diagnosis and regular follow-ups should be emphasized with educations, and the laboratory-clinician communication should be strengthened in order to use tests more efficiently.

Molecular Prevalence of Microsporidia Infection in Patients with Lung Cancer.

Infections are still among the most important causes of morbidity and mortality in patients with lung cancer, which has the highest rate of cancer-related deaths in the world. Microsporidia, which are opportunistic parasitic fungi, primarily localize to the intestine by ingestion but can disseminate to the respiratory tract or can be acquired by spore inhalation. Cancer patients are at higher risk for microsporidia, a life-threatening infection, than the normal population is. We aimed to characterize the prevalence of microsporidia infection for the first time by evaluating the intestinal and respiratory tracts of patients with lung cancer. In this study, we investigated 98 patients with lung cancer and 103 healthy individuals for microsporidia infection and evaluated the clinical findings of patients who were found to be positive. Sputum and stool samples were tested by microscopic examination, in addition to pan-microsporidia and genus-specific polymerase chain reactions. Nine patients with lung cancer had positive results for microsporidia (9.2%), which was significantly higher than the rate in healthy individuals (P = 0.008), and most of them had clinical findings. Among these positive patients, polymerase chain reaction revealed microsporidia in the sputum samples of seven patients, the stool sample of one patient, and both the sputum and stool samples of one patient. Encephalitozoon cuniculi was identified as the predominant pathogen in 87.5% (7/8) of positive sputum samples. Microsporidia infection was significantly associated with advanced stages of cancer. However, in the control group, Encephalitozoon intestinalis was detected in the stool sample of an individual without clinical symptoms. Microsporidia, especially E. cuniculi, should be considered as a cause of respiratory tract infection as well as intestinal infection in cancer patients and should be screened in respiratory samples of these patients when they have pulmonary symptoms.

The Spectrum of Infections in Patients with Lung Cancer.

Although diagnostic and therapeutic advances in lung cancer (LC) have increased the survival of patients, infection and its complications are still among the most important causes of mortality. The disruption of tissue caused by tumor mass, management of cancer therapy and alteration in the humoral/cellular immune systems due to both cancer itself and therapy considerably increase susceptibility to infection in cancer patients. Particularly, opportunistic microorganisms should be considered, then applying rapid and sensitive diagnostic methods for them. Thus, cancer patients who are already exposed to difficult, long-term and expensive treatments can be prevented from dying from complications related to infections.

Retrospective Evaluation of Toxoplasma Serology in Patients Admitted to a Training and Research Hospital Between 2017-2021

Objective: In this study, it was aimed to retrospectively evaluate the anti-Toxoplasma IgG, IgM and avidity index results of patients who were requested for Toxoplasma serology in our hospital between 01.01.2017 and 31.12.2021. Methods: Anti-Toxoplasma antibodies are studied with Abbott Architect I2000 SR device that using the chemiluminescent microparticle immunoassay method (CMIA), according to the company's recommendations. The age, gender, nationality, sending clinic/polyclinic, and pregnancy status information of patients were scanned from the hospital system. Results: In the five-year period between 2017 and 2021, 29.58% of anti-Toxoplasma IgG tests requested from 12694 patients and 0.94% of anti-Toxoplasma IgM tests sent from 12546 patients were found positive. It is striking that the number of test requests is higher in women. IgG positivity is highest in women in the age group of 30-39 (9.97%), and in men in the age group of 60-69 (6.97%). IgM positivity is higher in both women and men in the 20-29 age group (0.48% and 0.38%, respectively). Anti-Toxoplasma IgG was positive in 27.78% and IgM in 0.64% of the pregnant women. IgG positivity in Turkish and Syrian pregnant women were determined as 25.88%; 47.10% and IgM positivity as 0.49% and 1.83%, respectively, and the difference was statistically significant (p<0.001). Conclusion: Our anti-Toxoplasma antibody positivity was found to be compatible with studies conducted in different centers in our country. The fact that IgM positivity in women is high in the 20-29 age group, which is the childbearing age, emphasizes the importance of screening before and during pregnancy. Consistent with other studies in the literature, the rate of seropositivity in Syrian pregnant women was found to be higher than Turkish. This is important in terms of showing the effect of socio-cultural behaviors on prevalence.

[miRNA Expression Profile in Ileocecal Adenocarcinoma Cells Infected with Cryptosporidium]. / Cryptosporidium ile Enfekte Edilmis Ileoçekal Adenokarsinom Hücrelerindeki miRNA Ekspresyon Profili.

Cryptosporidium spp. is an opportunistic protozoan transmitted by fecal-oral route via oocysts. The agent may cause severe infection especially in individuals with suppressed immune system, due to its intracellular location and ability to cause auto-infection. MicroRNAs (miRNAs) are non-translated endogenous RNA molecules with an average of 22 nucleotides in length that regulate the expression of genes involved in important biological functions such as proliferation, differentiation, apoptosis and immune response. Recent studies have focused on the role of miRNAs in pathogenesis of infectious diseases and their potential to be used as biomarkers. The aim of this study was to determine the miRNA profile of human ileocecal adenocarcinoma (HCT-8) cells at 24 hours of infection with Cryptosporidium spp. In the study, the HCT-8 cell line was infected with Cryptosporidium spp. that were isolated from infected human stool samples and RNA was isolated from the cells 24 hours after infection. After this process, cDNA synthesis was performed and the expression of 95 human miRNA profiles were investigated by polymerase chain reaction (PCR) method. Fold changes of expression were determined by comparison with Cryptosporidium spp. uninfected cell lines. Sequence information of miRNAs and their target genes were performed via TargetScanHuman7.1 and miRDB websites, while gene ontology (GO) pathways of target genes were analyzed with the mirPath v.3 program. It was detected that the expression of 10 miRNAs were upregulated and 11 of them were downregulated compared with the control group. It was observed that, this 21 differentially expressed miRNAs were mainly associated with apoptosis, mitotic cell cycle, and immune response. Hsa-miR-612, hsa-miR-6763-5p, hsa-miR-188-5p, hsa-miR-664b-3p, hsa-miR-210-3p, hsa-let-7e-5p hsa-let-7b-3p, hsa-miR-4787-3p, hsa-miR-548ab, hsa-miR-3714 and hsamiR-4803 were found to be associated with apoptosis; and hsa-miR-612, hsa-miR-664b-3p, hsa-miR210-3p, hsa-let-7e-5p, hsa-let-7b-3p, hsa-miR-548ab, and hsa-miR4803 were found to be associated with mitotic cell cycle. The balance of proliferation and apoptosis is very significant in the development of infection and cancer. It is thought that determination of the effect of miRNAs on proliferation-apoptosis balance could provide information related to the etiopathogenesis and prognosis of infections, and on the role of microorganisms in carcinogenesis. In this study, 12 differentially expressed miRNAs were found to be associated with immune response. This may emphasize the role of miRNAs in the prevention and treatment of infections. It was concluded that, miRNAs could be used in the diagnosis, treatment and prevention of infections with the determination of miRNA's role in the infection mechanism as a result of the increasing number of studies.

A Pilot Study on the Evaluation of Cryptosporidium Infection in Patients with Lung Cancer: Respiratory Cryptosporidiosis.

Lung carcinoma is one of the most common cancers and the leading cause of cancer-related death worldwide. Increasing evidence has shown that Cryptosporidium spp., an opportunistic parasite, is associated with cancers, causing life-threatening infections. The most common clinical form of Cryptosporidium is intestinal infections. However, respiratory cryptosporidiosis has rarely been documented, although the parasite infects respiratory epithelial cells and gastrointestinal (GIS) epithelial cells. To evaluate respiratory cryptosporidiosis in patients with lung cancer, we investigated Cryptosporidium spp. in patients with lung cancer (n = 69) in comparison with healthy groups (n = 40). Sputum and stool samples were examined microscopically and by polymerase chain reaction (PCR). Two cancer patients were diagnosed with respiratory cryptosporidiosis (2.9%), on PCR examination of the sputum samples. Cryptosporidium spp. was detected in the stool samples of one patient (1.5%) and 2 healthy individuals (5.4%) by PCR and microscopy. First, respiratory cryptosporidiosis was documented in 2 patients with lung cancer. Cryptosporidium is an important agent of the respiratory tract and GIS infections in cancer patients. These new findings highlight the molecular prevalence of Cryptosporidium spp., an opportunistic infection, in patients with lung cancer. Respiratory cryptosporidiosis should also be considered when patients have respiratory symptoms.

The interaction of gut microbiota with parasitic protozoa.

The human intestinal microbiota is composed of a complex combination of microorganisms including bacteria, virus, and eukaryotes. The microbiota plays a critical role in homeostasis through creating a mucosal barrier, providing protective responses to pathogens, and affecting the immune system and metabolism of the host. Molecules secreted by parasites can alter composition of microbiota both by acting directly on the microbial community and indirectly by affecting the host physiology. On the other hand, the microbiota composition can affect the survival, physiology, and virulence of many parasitic protozoa. Explanation of possible interactions between the microbiota, immune response, and protozoa may further clarify the underlying mechanisms of infectivity, clinical variations, and life-cycle of protozoa.

Impact of the COVID-19 Duration on Neglected Parasitic Diseases

The Coronavirus disease-2019 (COVID-19) pandemic, which started in Wuhan, China in December 2019, has affected the whole world and caused approximately four million deaths. Consequently, scientists have done a great deal of research in such a short time about the disease. Meanwhile, parasites, whose evolutionary process is as old as human history, are often underestimated despite their high prevalence and lethality. Recent studies; however, have shown that immunity changes caused by parasitic infections affect the course of viral diseases. For example, because severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and Plasmodium use a common CD147 receptor to enter the cell and have similarities in their MHC-presented antigenic determinants, scientists suggest that immunity against parasitic infections protects the body against SARS-CoV-2 infections. This could explain the low COVID-19 incidence in malaria-endemic countries. Additionally, the cytokine storm, which is responsible for mortality in COVID-19 infections, is caused by the activation of the immune system to Th1 way. On the other hand, helminth infections, which activate the immune system to Th2 way, can reduce mortality by preventing the cytokine storm. The relationship between COVID-19 and parasites is not limited to changes in the immune system changes. Studies have shown that the pause in the fight against parasitic infections due to the diversion of all attention toward COVID-19 since the beginning of the pandemic will lead to an increase in incidences of malaria, leishmaniasis, schistosomiasis, and soil-transmitted helminths. For this reason, efforts to mitigate this increase should be resumed as soon as possible by taking additional measures globally.

Comparative Gene Expression Profiles of Leishmania major and Leishmania infantum Promastigotesa

Objective: This study aimed to determine the differences between the gene expression profiles of Leismania major and Leishmania infantum promastigotes through comparative analysis of gene expressions. Methods: Cell culture of L. major (MHOM/IL/80) and L. infantum (MHOM/MA/67/ITMAP/263) cell lines was performed. Afterwards, total RNA isolation and cDNA synthesis were performed and fold changes in the expression levels of 30 genes that play a role in metabolic pathways and nucleic acid synthesis and co-expressed in two species were evaluated by reverse transcriptase polymerase chain reaction. Functions of genes were determined using LeishDB and KEGG databases. Results: In this study, profiles of protein-coding 30 genes expressed in L. major and L. infantum promastigotes were evaluated and significant differences were found between the two species (p<0.001). There was a significant fold change in the expression levels of 29% of genes common in the two species. The expression levels of nine genes in L. major were found to be markedly higher than those of L. infantum (fold change >1). These genes include phosphoglycan beta 1.3 galactosyltransferase-like, lathosterol oxidase-like, fatty acid elongase, 3-oxo-5 alpha-steroid 4-dehydrogenase, calpain-like cysteine peptidase, acetyl-coA synthetase, 3'-nucleotidase/nuclease, 3'-nucleotidase/nuclease precursor and 3-ketoacyl-coA thiolase-like. When the functions of the proteins that correspond to the genes common in the two species were examined in detail using the databases, it was determined that these genes play role in lipid, protein, carbohydrate and nucleic acid metabolic functions of the parasite. Conclusion: Alterations in the expression profiles of genes common to L. major and L. infantum species may cause differences in the virulence, pathogenesis, clinical features and treatment modality between these parasite species. In addition, evaluation of gene profiles is important in the selection of species-specific or common targets for vaccine and drug studies.

[Defining the Molecular Signal Pathways and Upstream Regulators in Cutaneous Leishmaniasis with Transcriptomic Data Approach]. / Transkriptomik Veri Yaklasimi ile Kutanöz Leysmanyazisteki Moleküler Sinyal Yolaklarin ve Regülatör Moleküllerin Tanimlanmasi.

Leishmaniasis is a disease caused by the genus Leishmania spp., which are intracellular parasites. Depending on parasite species and host immune response, there are three basic clinical forms of the disease: cutaneous, mucocutaneous, and visceral leishmaniasis. Cutaneous leishmaniasis is a chronic disease and characterized by the presence of ulcerated skin lesions. The type of skin pathology seen during disease is determined in part by the infecting Leishmania spp., but also by a combination of inflammatory and antiinflammatory host immune response factors resulting in diverse clinical outcomes. In this study, it was aimed to determine the genes, molecular signaling mechanisms and biological functions of the molecules that play a role in the pathogenesis of the disease and immune response and determine host-parasite interactions in mice that are naturally resistant and susceptible to Leishmania major and Leishmania braziliensis. For this, transcriptomic series GSE56029 was downloaded from "Gene Expression Omnibus"(GEO) data base, including expression profiling of twenty-four tissue samples that were recovered from both naive mice and mice (BALB/c, C57BL/6) infected with L.major and L.braziliensis. Then, "Differentially Expressed Genes" (DEGs) were identified by limma package in R script. FDR q<0.05 and absolute log2FC> 2 as threshold values were accepted in the analysis. Subsequently, functional and pathway enrichment analyses were performed for the DEGs by "Ingenuity Pathway Analysis" (IPA). For each of DEGs, p<0.01, FDR q<0.01, and absolute log2FC> 1 were used and analyzed with the software program IPA 8.0. Ingenuity Pathway Analysis revealed the most enrichment pathways to be the inflammation, dendritic cell maturation and "Triggering Receptor Expressed on Myeloid Cells 1" (TREM-1) signal mechanisms and that the DEGs related to the regulation of immune system process were closely associated with the progress of cutaneous leishmaniasis. The upstream regulator analysis predicted that TNF-α, IFNy, IL-1 ß, IL-10RA and "Signal Transducer and Activator of Transcription-1" (STAT-1) are the regulators that explained gene expression changes causing biological activities in the tissues. Chemical compounds that may have anti-leishmanial effects were also identified in the study. In this study, the mechanisms belonging to the parasite species and host that determine the resistance/susceptibility phenotype were attempted to elucidate. Assessment of gene expression patterns, cytokine/chemokines, and signaling pathways in BALB/c and C57BL/6 mice infected with L.major and L.braziliensis will provide a better understanding of the potential mechanisms underlying infection from a genetic perspective. These results may guide for the future studies in terms of developing potential biomarkers for the diagnosis and prognosis prediction of cutaneous leishmaniasis and providing information about new treatment targets.

The Role of MicroRNAs in Parasitology.

MicroRNAs (miRNAs), as epigenetic regulators, are small non-coding RNAs regulating gene expression in eukaryotes at the post-transcriptional level to control biological functions. MicroRNAs play a role in development, physiology, infection, immunity and the complex life cycles of parasites. Also, parasite infection can alter host miRNA expression that might result in either parasite clearance or infection. Over the past 20 years, thousands of miRNAs have been identified in the nematode Caenorhabditis elegans and other parasites. Thus, miRNA pathways are potential targets for the diagnostic and therapeutic control of parasitic diseases. Here, we review the current status and potential functions of miRNAs related to protozoans, helminths, and arthropods.