Background: The possible risk factor of Toxoplasma infection and its possible correlation with Interleukin-12 (IL-12) and Interleukin-23 (IL-23) in breast cancer patients was investigated.
Methods: Overall, 190 female patients referred to the Oncology Teaching Hospital in the Medical City Hospital, Baghdad, Iraq were enrolled from 2017-2018. All serum samples were tested for T. gondii immunoglobulins (IgG and IgM (antibodies and IL-12, IL-23 levels.
Results: In patients with breast cancer, the results revealed a high positivity percentage for anti- Toxoplasma IgG. In breast cancer patients infected with T. gondii, the IL-12 level was lower than the controls while the mean level of IL-23 was higher than the controls. According to the cancer grade in breast cancer patients infected with T. gondii, the higher mean titer of IgG and IL-23 was in grade 3 in contrast the highest mean titer of IL-12 was in grade 1. Concerning the tumor stages in breast cancer patients infected with T. gondii, the higher mean titer of IgG, IL-12 and IL-23 was in stage (III). According to the tumor size in breast cancer patients infected with T. gondii, the highest mean titer of IgG, IL-12 and IL-23 was in size >3cm.
Conclusion: The levels of IL-23 could be a candidate as a non-invasive primitive marker for earlier prediction of breast cancer stage.

Background: We aimed to investigate the effect of miR-15a mimic and inhibitor of miR-155 expression on apoptosis induction in macrophages infected with Iranian strain of Leishmania major in-vitro and in-vivo.
Methods: RAW 264.7 cells were infected with L. major promastigotes (MRHO/IR/75/ER), and then were treated with miRNAs. For in-vivo experiment, BALB/c mice were inoculated with L. major promastigotes, and then they were treated with miRNAs. For evaluation of miRNA therapeutic effect, in-vitro and in-vivo studies were performed using quantitative Real-time PCR, Flow cytometry, lesion size measurement, and Limiting Dilution Assay (LDA). This study was performed in Shahid Beheshti University of Medical Sciences in 2019.
Results: In-vitro results of flow cytometry showed that using miR-15a mimic, miR-155 inhibitor or both of them increased apoptosis of macrophages. In in-vivo, size of lesion increased during experiment in control groups (P<0.05) while application of both miR-155 inhibitor and miR-15a mimic inhibited the increase in the size of lesions within 6 wk of experiment (P=0.85). LDA results showed that microRNA therapy could significantly decrease parasite load in mimic or inhibitor receiving groups compared to the control group (P<0.05).
Conclusion: miR-155 inhibitor and miR-15a mimic in L. major infected macrophages can induce apoptosis and reduce parasite burden. Therefore, miRNA-based therapy can be proposed as new treatment for cutaneous leishmaniasis.

Background: The outcome and complications of liver hydatid cyst surgeries with new and old techniques are not well determined. We intended to present the results of operations done for patients with hepatic hydatid cyst in an endemic area.
Methods: Data of 112 patients referred and operated for liver hydatid cyst, in Imam Khomeini Hospital complex, Tehran, Iran, from 2015 to 2018, were collected including demographic characteristics, operation parameters and complication related statistics. The variables were presented for different surgical methods including operations with and without omentoplasty.
Results: Patients aged 39. 3 ± 13. 9 yr (70 females; 63. 5%). Most frequent clinical complaint was vague abdominal pain (n=45; 40. 2%). The most prevalent comorbidity was hypertension (18; 16.0%). Conservative methods were chosen more frequently including omentoplasty (44; 39. 3%), cyst drainage (27; 24. 1%), cyst resection (19; 17%) and marsupialization (3; 2. 7%). Overall, 56 patients (50%) were operated with omentoplasty as the single method or in combination with segmentectomy. Complications occurred less in patients operated with omentoplasty (41. 1 vs. 23. 2%; P=0. 043); particularly, biloma was more frequent in surgeries without omentoplasty (7. 1 vs 0. 0%; P=0. 042). Persistence and recurrence rates were 12. 5% and 3. 6% with relative predilection in, respectively, segmentectomy and lobectomy surgical methods compared to Omentoplasty. No mortality was recorded.
Conclusion: In our case series of hepatic hydatid cyst patients, omentoplasty was safe with less complication and similar long-term recurrence rate.

Background: Over the last decade, a few cases of visceral leishmaniasis (VL) have been reported in some provinces of northeastern Iran. We aimed to investigate clinical and laboratory findings of VL among children who admitted to the pediatric ward in a referral hospital in Mashhad, northeastern Iran.
Methods: A retrospective study, between 1997 and 2017, was performed on the data sheet registered for children with confirmed VL at the referral Emam Reza Hospital in Mashhad. Hematological and biochemical profiles of the patients were analyzed.
Results: Thirty-five children with VL, confirmed by the presence of amastigotes of Leishmania in Giemsa stained smears of the bone marrow, had been recorded through 20 yr. The mean age of patients was 3.7±4 yr. The majority of the patients suffered from hepatosplenomegaly (100%, n=35/35), followed by prolonged fever and pallor (91%, n=32/35), weight loss (85%, n=30/35). The main laboratory findings were anemia (94.1%), leukopenia (52.9%) and thrombocytopenia (70.5%). Almost one-third (37.1%; 13/35) of VL patients inhabited in rural areas of the Bojnoord district as a known VL endemic focus in northeastern Iran.
Conclusion: Our preliminary data showed that the origin of VL is still in some districts other than Mashhad, where VL just will be diagnosed.

Background: The immunomodulatory role of many parasites is well-documented. The current study designed to assess the immunoregulatory effects of the somatic extract (SE) of Toxocara canis on murine model of airway inflammations.
Methods: The experiment was performed in department of parasitology of Tarbiat Modares University, Tehran, Iran from November 2018 to May 2019. Totally 30 female BALB/c mice divided into one control group and two experimental groups (10 mice in each group). The ovalbumin (OVA) group was sensitized with OVA in alum, while the SE group was administered with SE and OVA in alum intraperitoneally. The control group was injected with PBS in alum. Then, SE and OVA groups were intranasally challenged with OVA for three consecutive days and the control group encountered with PBS at the same time. One day after the last challenge, real-time PCR and histopathology survey were conducted on isolated lung tissues.
Results: The gene expression of IL-25, IL-33, TNF-α and TLR-4 in SE group was significantly lower than OVA group (P<0.05). The level of IL-10, TGF-β and IFN-γ were considerably higher than the OVA group (P<0.05). The inflammation was reduced in SE group, as the total cell number of bronchoalveolar lavage fluid was less than OVA group. Based on the histopathology findings the inflammation was decreased in SE group compared to the OVA group.
Conclusion: Although, an inhibitory effect of SE of T. canis on airway inflammations was detected, there is still a long way ahead regarding the indication of the precise mechanisms.

Background: In this study, mutations in the tripanothion reductase of Leishmania tropica isolated from Iran was investigated using sequencing and simulation of the enzyme by the molecular dynamic method.
Methods: Fifteen susceptible and 15 clinical resistant L. tropica specimens were collected from skin lesions from different regions of Iran in 2017. After DNA extraction, trypanothione reductase (TRYR or TPR), gene fragment was amplified using PCR and sequencing methods. In the case of structural mutations, the components were simulated by molecular dynamics using the GROMACS software.
Results: Some structural mutations were observed in 9 amino acids surrounding the active site of the TRYR gene of L. tropica with three-dimensional trypanothione reductase alteration.
Conclusion: Change in the active site of TRYR of L. tropica, could probably contribute to the development of resistant L. tropica to glucantime. Because of the likely occurrence of mutations in glucantime as well as the ease of development of L. tropica resistant populations, more samples are needed to demonstrate the relationship between mutations in this enzyme and clinical resistance to glucantime. On the other hand, it is recommended that enzymatic studies be performed to confirm the role of mutation in the function and expression of trypanothione reductase in glucantime resistant and susceptible populations.

Background: Toxoplasmosis is suspected to have adverse effects on the male reproductive system. We aimed to determine the possible role of Toxoplasma gondii in oxidative stress in reproductive system of male rats.
Methods: This study was performed from 2018 until 2019 at the Parasitology Re-search Laboratory of Hamadan University of Medical Sciences, Hamadan, Iran. Eighty male Wistar rats were randomly divided to control and test groups. The ani-mals in the test group were inoculated by 107 T. gondii RH strain tachyzoites and the control group were injected by 0.2 ml of phosphate-buffered saline. The both groups were following every 10 days until day 80 post inoculation. Oxidative stress markers (OSMs) including antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and non-enzymatic markers including malondialdehyde (MDA), reduced glutathione (GSH) and total antioxidant capacity (TAC) were as-sessed in testis and serum of T. gondii infected rats.
Results: After post inoculation, the variations of the OSMs in the testis tissue of infected rats were as follows: a significant decrease of SOD on day 80 (P=0.03), and CAT activity were detected on day 60 and 80 (P=0.04 and P=0.01) respectively. In addition, GSH (P =0.01) and TAC (P =0.03) concentration were significantly re-duced on day 80. On the contrary, the concentration of MDA (P =0.01) was in-creased 70 days after infection. In addition, consistent changes with the tissue testis were observed in the serum OSMs of infected rats.
Conclusion: T. gondii infection caused oxidative stress in testis tissue. Thus, the adverse effects of oxidative stress may affect the male rat reproductive system.

Background: This study was designed to detect, if there are asymptomatic malaria infections amongst native and immigrant population from Afghanistan and Pakistan countries in Sistan & Baluchistan Province of Iran, where is under the national malaria elimination program.
Methods: This cross-sectional study was performed among native individuals and resident immigrants in the southeastern province of Sistan & Baluchistan from May 2016 to Jul 2017. A total of 271 individuals were considered in this cross- sectional study based on microscopical method, Rapid Diagnostic Tests (RDTs) and PCR techniques. Out of 271 native and immigrant participants 140 (52%) and 131 (48%) were male and female, respectively.
Results: None of the prepared samples was diagnosed as malaria positive case when was considered via above mentioned three techniques.
Conclusion: Neither native nor immigrant individuals had asymptomatic malaria, hinting that national malaria elimination program is performed according to planned schedule in the studied areas

Background: In the present study, we examined the effects of Fe3O4@bio-MOF nanoparticle (Nano-FO) plus artemisinin (Art) and glucantime (Glu) or shark cartilage extract (ShCE) on Leishmania major in vitro and in vivo.
Methods: This experimental study was conducted at the laboratory of Department of Parasitology, Tarbiat Modares University, Tehran, Iran during 2016-2017. The promastigote and amastigote assays were performed were conducted at the presence of 3.12- 400 µg/mL of the drug combinations. According to in vitro IC50 results, the combinations of 12.5µg/mL Nano-FO with 25 µg/mL Art as well as 200 µg/mL Glu and 0.5 mL of 20 mg/kg of ShCE were used to treat BALB/c mice. During and at the end of the treatment, the lesion sizes were measured. Parasite loads, cytokine levels were evaluated at the end of the treatment.
Results: The IC₅₀ of Fe₃O₄@bio-MOF-Artemisinin (Nano-FO/Art), Fe₃O₄@bio-MOF-Glucantime (Nano-FO/Glu), and Fe₃O₄@bio-MOF-Shark cartilage extract (Nano-FO/ShCE) on promasitigotes were 12.58±0.12, 235±0.17, and 18.54±0.15, respectively. These results on amastigotes were 10.32±0.01, 187±0.03, and 338±0.07 µg/mL, respectively. The apoptosis percentage of these combinations were 32.54%, 20.59%, and 15.68% in promastigotes and 15.68%, 12.84%, and 3.51% in infected macrophages, respectively with no toxicity on uninfected macrophages. In vivo results showed that the size of lesions significantly decreased against all drugs combinations, but Nano-FO/Art combination with Selectivity Index of 23.62 value was safe, and more effective on healing of lesions than other drugs combinations (P=0.003).
Conclusion: This study suggested that Nano-FO/Art combination can be considered as an anti-leishmania combination therapy in CL induced by L. major.

Background: Fasciola hepatica as an important parasite affects health of humans and animals in some tropical and subtropical areas of the world, including Iran. Little is known about the molecular diversity of Fasciola in Equidae. Therefore, this study aimed to characterize the genetic polymorphisms among parasites.
Methods: Eight adult Fasciola spp. isolates were collected from a working donkey after necropsy in Shiraz, southwestern Iran, in 2018. Primarily, various parameters were measured morphologically. Subsequently, DNA was extracted from each fluke and molecular markers of cytochrome C oxidase (cox1) and NADH dehydrogenase 1(nad1) from individual Fasciola isolates were amplified using PCR assay and sequence data were employed for molecular and phylogenetic analysis. Genetic diversity between isolates was evaluated by comparing the sequences of these two mitochondrial regions.
Results: Based on the morphological and analyzed mitochondrial sequences, all of eight donkey isolates (100%) were identified as F. hepatica. Moreover, nine and five nucleotide polymorphisms were identified in the cox1and nad1 region sequences, respectively.
Conclusion: Accordingly, phylogenetic data revealed five and four haplotypes among donkey isolates based on the cox1and nad1 markers. Similarly, some of these haplotypes have been previously reported from different host species in Iran as well as all around the world.

Background: Toxocariasis is one of the most neglected zoonotic diseases, predominantly caused by Toxocara canis. We aimed to evaluate the expression of microRNAs 21 and 103a in seropositive individuals for human toxocariasis as diagnostic biomarkers.

Methods: This study was conducted on 324 individuals for ELISA test on toxocariasis in Tehran and Karaj, Iran 2019. Then positive samples for anti-Toxocara IgG were obtained to quantitative Real-time PCR (qRT-PCR) assays to investigate the transcriptional profiles of miRNAs predicted to be involved in developmental and reproductive processes. qPCR was employed to assess levels of transcription for miRNAs of 103a and 21 in plasma samples.

Results: After the experiments, the results were evaluated by REST software, Livak formula and quantitative t-test. The analyzes performed on human samples showed that in the case group compared to the control group, only in Tc-miR-21 gene, a 0.3-fold increase in expression was obtained with REST software (Fold change ≤ 1.5, P>0.05), which was statistically significant by t-test (P<0.05).

Conclusion: To our knowledge, this is the first study to evaluate miR-21 and miR-103a in toxocariasis, which shed light on the fundamental role of it as a biomarker and diagnostic tool. However, due to the changes in expression of these miRNAs were not vast to be used as biomarkers in diagnosis. Despite of that the changes in the expression of these miRNAs were not vast but they could serve as novel promising biomarkers for diagnosis of toxocariasis.

Background: Toxoplasmosis is a zoonotic disease caused by the obligate intracellular parasite, Toxoplasma gondii. This global infectious disease has been associated with behavioral changes in rodents and can result in humans' neuropsychiatric symptoms. Since the neurotransmitters alteration can cause a behavioral change, in this study, tyrosine level, as a precursor of dopamine, was evaluated in acute murine toxoplasmosis during 2015 and 2016 in Shiraz, Iran.
Methods: At the first, 10⁵ tachyzoites of T. gondii were subcutaneously inoculated to 50 BALB/c mice as experimental groups and 10 mice inoculated by PBS considered as the control group. After that, daily, one group of mice was bled, and sera were collected. Then, their serum tyrosine level was evaluated by HPLC method.
Results: After data analysis, the maximum mean serum tyrosine level was seen at 2^th day of post parasite inoculation (0.0194 mg/ ml), with a significant difference compared to the control group (0.0117 mg/ ml, P=0.025). Moreover, the least quantity of serum tyrosine (0.076 mg/ml) was seen on the 5^th day, after parasite inoculation, however, no significant difference was seen.
Conclusion: Serum tyrosine level increased in 2 d after inoculation of Toxoplasma, but the level regularly decreased in successive days. Tyrosine level increased by phenylalanine hydroxylase 2 days after inoculation, then tyrosine decreased by tyrosine hydroxylase in the next days. Toxoplasma tyrosine hydroxylase enzymes, at primary days of toxoplasmosis, effect on tyrosine production, and after that, the most effect on tyrosine consumption.

Background: We aimed to compare parasite burden in BALB/c mice, using three methods including the direct fluorescent microscopic using recombinant Leishmania major expressing an enhanced green fluorescent protein, limiting dilution assay, and real-time PCR technique.
Methods: The current study was carried out in 2018, to induce stable enhanced green fluorescent protein (EGFP) production. Initially, the linearized DNA expression cassette (pLEXSY-egfp-sat2) was integrated into the ssu locus of L. major. The expression of EGFP in recombinant parasite was analyzed using direct fluorescent microscopy. Afterward, BALB/c mice were infected with the L. major^EGFP, and the infection was evaluated in the foot-pads and inguinal lymph-nodes using an in vivo imaging system. Subsequently, eight BALB/c mice were infected with L. major^EGFP, and the results of evaluating parasite burden by a SYBR-Green based real-time PCR analysis and the limiting dilution assays were compared to the results obtained from the direct fluorescent microscopy.
Results: The results of the direct fluorescent microscopy showed that EGFP gene stably was expressed in parasites. Moreover, the in vivo imaging analysis of foot-pad lesions revealed that the infection caused by L. major^EGFP was progressing over time. Additionally, significant correlations were observed between the results of parasite burden assay using the direct fluorescent microscopy and either limiting dilution assay (r=0.976, P<0.0001) or quantitative real-time PCR assay (r=0.857, P<0.001).
Conclusion: Ultimately, the utilization of the direct fluorescent microscopy by employing a stable EGFP-expressing L. major is a suitable substitution for the existing methods to quantify parasite burden.

Background: When Angiostrongylus cantonensis develops from the third and fourth stage, it needs to change its host from the middle host, snail to the final host, rat. However, the mechanism involved in this change remains unclear.
Methods: The transcriptome differences of the third and fourth stages of A. cantonensis were explored by next-generation Illumina Hiseq/Miseq sequencing in China, in 2018.
Results: Overall, 137 956 488 clean reads and 20 406 213 373 clean bases of the two stages larvae were produced. Based on the queries against the Gene Ontology (GO), NCBI non-redundant protein sequences (Nr), Swissprot, and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases, 14 204 differentially expressed genes (DEGs) were predicted. GO enrichment analysis revealed 5660 DEGs with the top s categories as followings: biological process (GO:0008150, related to 5345 DEGs), cellular component (GO:0005575, related to 5297 DEGs), molecular function (GO:0003674, related to 5290 DEGs). In KEGG enrichment analysis, 116 genes were related to oxidative phosphorylation and 49 genes involved in the glycolytic process.
Conclusion: Metabolism changes, especially oxidative phosphorylation and glycolysis, might play a key role in A. cantonensis infection of its final rat host. Many other pathways might also contribute to the transcriptome changes between these two life stages. Overall, additional studies are needed for further details.

Background: Adenosine deaminase (ADA) is an aminohydrolase involved in the catabolism of purine nucleotides and irreversibly deaminizes adenosine and deoxyadenosine to inosine and deoxyinosine. ADA enzyme deficiency results in the loss of functional properties of B and T lymphocytes. Demodex species have been reported to be transmitted between humans through close contact and to play a role in the pathogenesis of rosacea, acne vulgaris, perioral dermatitis, seborrhoeic dermatitis, micropapillary-pruritic dermatitis and blepharitis. The present study aimed to compare serum ADA levels in D. folliculorum positive patients with the healthy control individuals.
Methods: Serum ADA levels were examined for 30 patients diagnosed with erythematotelangiectatic rosacea and 40 healthy individuals in Malatya Inonu University in 2017. Standardized skin surface biopsy (SSSB) method was used to diagnose D. folliculorum. A significant decrease was found in the ADA levels of Demodex-positive rosacea patients when compared to the control group.
Results: ADA levels were decreased in the Demodex-positive group. The mean ADA level in patient group was significantly lower than the mean in the control group (P<0.001). There was no significant difference between the patient and control groups in terms of age and gender.
Conclusion: During and after treatment of Demodex-positive rosacea patients, determination of ADA levels may give more detailed information on the immune mechanisms.

Background: We aimed to evaluate a potential link between colonization of gingival crevices by the Entamoeba gingivalis as oral parasite microbiome and Hepatitis B infection among gum disease Sudanese patients.
Methods: This study was conducted on 448 gum disease patients attending to Dental Clinic of Kosti Teaching Hospital, Kosti, Sudan in 2017-2018. Oral symptoms were registered in 336 patients at different stages of the HBV infection and in 112 HBV seronegative gum disease control. All participants were screened for HBV using ELISA test. Unstimulated whole saliva and gingival scraping were obtained and analyzed for the presence of the amoeba using a parasitological technique.
Results: Statistically highly significant correlation was found between the detection of the E. gingivalis in Saliva/gingival scraping and gum illness disease with HBV-infected patients than healthy control group (P<0.05). There was high association between the occurrence of the amoeba among the two groups and smoking, snuffing habitats (P<0.05), inversely, no association with the oral personal hygiene.
Conclusion: The presence of the amoeba was not related to the degree of gum diseases only, but to the HBV infection diagnosis. To our knowledge, this is the first study of E. gingivalis in association with HBV infection among gum disease Sudanese patients; maybe predict the role of oral parasitic microbiome in the status of gum disease in HBV infection.

Cutaneous leishmaniasis (CL) is an emergent public health concern, particularly in tropical and subtropical regions. Reports of pregnancy complications are scarce; however, as the endemic range of CL expands in Iran, there is concern of possible detrimental effects on fetal development amongst infected mothers through placental transmission of the parasite or enhanced maternal immune responses. We herein describe the first known case of persistent anthroponotic CL, plausibly responsible for pregnancy complications, preterm birth, and neonatal death in a healthy Iranian primigravida woman. Diagnosis was based on physical examinations of the lesions on the back of both calves of the patient and laboratory analyses including direct smear, culture, and PCR. During active CL infection, the patient gave birth to a premature female neonate who passed three days post-delivery due to immature lung development and subsequent respiratory distress syndrome. This report highlights the challenges associated with CL infection during pregnancy, exacerbation of lesions, and subsequent complications.

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