Development, Characterization, and In Vitro Biological Performance of Amphotericin B and Terbinafine Microemulsions Against Leishmania major.

This study evaluated the effect of microemulsion (ME) containing Amphotericin B (AmB) alone and associated with Terbinafine (Tbf) on Leishmania major (L. major) using in vitro models. The ME formulations of these drugs were formulated and described. After evaluating their cytotoxicity on the J774A1 macrophage (MΦ), their potency against promastigotes and intracellular amastigotes models was evaluated using an in vitro MTT assay and Giemsa stain, respectively. Based on pseudo ternary phase diagrams, unloaded ME, Tbf-loaded ME (ME-Tbf), and, AmB-loaded ME (ME-AmB) with mean droplet sizes 3.4 ± 0.81, 10.05 ± 0.21, and 8.21 ± 0.46 were successfully prepared, respectively. Concerning toxicity, ME-AmB and ME-Tbf indicated lower toxicity on MΦs compared to the free drugs. The ME formulations showed considerably inhibitory effects compared to the free drug forms when the IC50 was examined. The IC50 values of AmB (59.19 ± 1.74 and 36.4 ± 3.2 µg/mL), ME-AmB (7.5 ± 0.9 and 0.8 ± 0.05 µg/mL), Tbf (234.5 ± 9 and 128.8 ± 0.28 µg/mL), ME-Tbf (26.27 ± 0.2 and 11.97 ± 0.6 µg/mL), AmB + Tbf (30.18 and 24.93 µg/mL), and ME-AmB + ME-Tbf (4.79 and 0.37 µg/mL) were estimated after 48 and 72 h, respectively.

Evaluation of the Efficacy of Amphotericin B and Terbinafine Microemulsions and Their Combination on Cutaneous Leishmaniasis and Comparison with the Conventional Drug Form in BALB/c Mice.

Intracellular parasitic protozoa of Leishmania sp. causes leishmaniasis. The restricted access of the drugs to affected cells in the treatment of intracellular infections such as leishmaniasis is frequently hampered. Furthermore, most of today's drugs have limited uses due to some containing toxic compounds, and drug resistance is on the rise. In the present investigation, Amphotericin B (AmB) and Terbinafine (Tbf) were loaded in microemulsion (ME) in combination and alone, and the in vivo efficacy was considered in BALB/c mice infected with Leishmania major (L. major). The wound size at the base of the mouse tail was measured, and real-time PCR was performed to quantify the parasite load after the infection challenge. The study demonstrated that the ME-AmB and ME-Tbf formulations are safe and effective compounds for the treatment of cutaneous leishmaniasis by enhancing the effectiveness of AmB and Tbf in reducing the parasite burden.

In vitro antifungal susceptibility patterns of Trichophyton benhamiae complex isolates from diverse origin.

BACKGROUND: Species from the Trichophyton benhamiae complex are mostly zoophilic dermatophytes which cause inflammatory dermatophytosis in animals and humans worldwide. OBJECTIVES: This study was purposed to (a) to identify 169 reference and clinical dermatophyte strains from the T benhamiae complex species by molecular method and adhering to the newest taxonomy in the complex (b) to evaluate the in vitro antifungal susceptibility profile of these strains against eight common and new antifungal agents that may be used for the treatment of dermatophytosis. METHODS: All isolates, mainly originated from Europe but also from Iran, Japan and USA, were subjected to ITS-rDNA sequencing. The in vitro antifungal susceptibility profiles of eight common and new antifungal drugs against the isolates were determined by CLSI M38-A2 protocol and according to microdilution method. RESULTS: Based on the ITS-rDNA sequencing, T benhamiae was the dominant species (n = 102), followed by T europaeum (n = 29), T erinacei (n = 23), T japonicum (n = 10), Trichophyton sp (n = 4) and T eriotrephon (n = 1). MIC ranges across all isolates were as follows: luliconazole: 0.0002-0.002 µg/ml, terbinafine: 0.008-0.125 µg/ml, efinaconazole: 0.008-0.125 µg/ml, ciclopirox olamine: 0.03-0.5 µg/ml, itraconazole: 0.06-2 µg/ml, griseofulvin: 0.25-4 µg/ml, amorolfine hydrochloride: 0.125-4 µg/ml and tavaborole: 1-16 µg/ml. CONCLUSION: Luliconazole, efinaconazole and terbinafine were the most potent antifungals against T benhamiae complex isolates, regardless of the geographic locations where strains were isolated. These data might help dermatologists to develop effective therapies for successful treatment of infections due to T benhamiae complex species.

Gene expression in human liver fibrosis associated with Echinococcus granulosus sensu lato.

Liver fibrosis is a dynamic process that occurs in response to chronic liver disease resulting from factors such as chronic infections, autoimmune reactions, allergic responses, toxins, radiation, and infectious agents. Among the infectious agents, multicellular parasites cause chronic inflammation and fibrosis. Twenty-five patients with different stages of cystic echinococcosis (CE) were enrolled in the study. The expression of ACTA2, COL3A1, IFN-γ, MMP2, MMP9, TGF-ß1, and TNF-α genes was determined by qRT-PCR in healthy and fibrotic liver tissue of the CE patients. TGF-ß1 expression was evaluated by immunohistochemistry, and histology was conducted to assess the development of liver fibrosis. Expression of MMP9, ACTA2, COL3A1, and MMP2 was found significantly higher in the fibrotic tissue compared to healthy tissue. We observed a significant correlation between TGF-ß1 and TNF-α gene expressions and liver fibrosis. The mRNA level of IFN-γ was lower in the fibrotic than in the healthy hepatic tissue. Immunohistochemistry analysis revealed TGF-ß1 upregulation in the fibrotic tissue. Histology showed inflammation and fibrosis to be significantly higher in the fibrotic tissue. The findings of this study suggest that Echinococcus granulosussensu lato can promotes fibrosis through the overexpression of TGF-ß1, MMP9, ACTA2, COL3A1, and MMP2. The downregulation of IFN-γ mRNA in fibrotic samples is probably due to the increased production of TGF-ß1 and the suppression of potential anti-fibrotic role of IFN-γ during advanced liver injury caused by E. granulosussensu lato.

Trichophyton mentagrophytes and T interdigitale genotypes are associated with particular geographic areas and clinical manifestations.

The fungi Trichophyton mentagrophytes and T interdigitale account for significant amount of dermatophytosis cases worldwide. These two dermatophytes form a species complex and have a number of ribosomal internal transcribed spacer (ITS) region genotypes, allowing simultaneous species identification and strain typing. Our aim was to describe the geographic distribution of T mentagrophytes/T interdigitale ITS region genotypes and find an association between the genotypes and clinical presentations of respective infections. We performed rDNA ITS region sequencing in 397 Iranian T mentagrophytes/T interdigitale isolates and analysed all available in GenBank entries with sequences of this kind. For the study, 515 clinical annotations were available. Statistical analysis was performed by chi-squared test and Spearman rank correlation analysis. A total of 971 sequences belonged to genotypes with at least 10 geographic annotations and were classified on the basis of exclusive occurrence in a particular region or high relative contribution to a regional sample. We discerned Asian and Oceanian ("KU496915" Type V, "KT192500" Type VIII, "KU315316"), European ("FM986750" Type III, "MF926358" Type III*, "KT285210" Type VI) and cosmopolitan ("FM986691" Type I, "JX122216" Type II, "KP132819" Type II* and "AF170453" Type XXIV) genotypes. There was statistically significant difference in the ITS genotype distribution between different affected body sites. Trichophyton mentagrophytes "KT192500" Type VIII correlated with tinea cruris, T mentagrophytes "KU496915" Type V correlated with tinea corporis, T interdigitale "JX122216" Type II correlated with tinea pedis and onychomycosis. Trichophyton mentagrophytes and T interdigitale genotypes can be associated with distinct geographic locations and particular clinical presentations.

Multiple zoonotic helminth infections in domestic dogs in a rural area of Khuzestan Province in Iran.

BACKGROUND: Echinococcosis and toxocarosis caused by the genus of Echinococcus and Toxocara spp. are among important helminthic diseases worldwide. Limited data on the prevalence of these parasites persuaded us to determine the prevalence of E. granulosus, E. multilocularis, and T. canis infections in domestic dogs in rural areas of Ahvaz, southwestern Iran. Fecal samples from 167 domestic dogs were examined using both microscopy and PCR techniques. Multiplex PCR was performed for the presence of Echinococcus, and Taenia spp. and single PCR for detection of T. canis and Toxascaris leonina. RESULTS: The total occurrence of identified parasites was 65 (38.9%). The microscopic examinations showed that 40 (24%), 18 (10.8%), and four (2.4%) of dogs were infected with taeniid-like, ascarid, and both genera eggs, respectively. Echinococcus granulosus was identified in seven (4.2%), Taenia spp. in 29 (17.4%), and mixed infection with both in 11 (6.6%) samples. Sequencing of PCR-positive samples identified E. granulosus s.s. (G1), 18 T. hydatigena (10.8%), five T. multiceps (3%), three T. serialis (1.8%), one T. ovis (0.6%), one Spirometra erinaceieuropaei voucher (0.6%), and two Mesocestoides corti (1.2%). This is the first report of S. erinaceieuropaei voucher and M. corti in dogs in Iran. Nine (5.4%) and 16 (9.6%) dogs showed infection with T. canis and T. leonina, respectively. Two samples showed coinfection with both ascarids. CONCLUSIONS: Several studies have reported echinococcosis and toxocarosis in intermediate hosts from the southwest of Iran; however, this study is the first molecular research on E. granulosus and T. canis in domestic dogs in a rural area of southwestern Iran. Furthermore, issues of soil contamination with dogs' feces and recent dust storms in Khuzestan may have a role in the spreading of these zoonotic infections to other provinces close to it, and neighboring countries such as Iraq.

Molecular Characterization and In Vitro Antifungal Susceptibility of 316 Clinical Isolates of Dermatophytes in Iran.

Dermatophytosis is a common mycotic infection of the skin, nail, and hair, associated with major public health concern worldwide. Various species of dermatophytes show significant differences in susceptibility to antifungals. Here, we present the antifungal susceptibility of a large collection of molecularly identified dermatophyte isolates obtained from tropical region of south of Iran. A total of 9485 patients clinically suspected to have cutaneous fungal infections were examined. Dermatophytosis was confirmed in 1502 cases by direct microscopy and culture. Three hundred and sixteen isolates recovered in culture were identified to species level using PCR sequencing of ITS region and RFLP. Tinea corporis was the most prevalent type of clinical manifestation (35.2 %), followed by tinea cruris (17 %), tinea capitis (12.8 %), tinea pedis (11.3 %), tinea manuum (11 %), tinea unguium (6.9 %), and tinea barbae (5.8 %). Trichophyton interdigitale was the most common isolate (49.36 %), followed by Trichophyton rubrum (18.98 %), Epidermophyton floccosum (13.29 %), Microsporum canis (9.17 %), Arthroderma benhamiae (T. anamorph of A. benhamiae; 5.38 %), and Trichophyton tonsurans (3.79 %). Overall, irrespective of the geographical region, terbinafine was the most potent antifungal against all isolates, with an MIC range of 0.002-0.25 µg/mL, followed by itraconazole (0.004-0.5 µg/mL), griseofulvin (0.125-8 µg/mL), and fluconazole (4-128 µg/mL). Analysis of our data revealed a significant increase in the frequency of A.benhamiae, which definitely warrants further investigation to explore source of this infection in south of Iran. Moreover, terbinafine was the most effective antifungal against all isolates, in vitro.

Epidemiological Aspects of Dermatophytosis in Khuzestan, southwestern Iran, an Update.

Dermatophytosis is among the most common superficial mycoses in Iran. The purpose of this report was to update the clinical and mycological features of human dermatophytosis in the Khuzestan, southwestern Iran. In the framework of a one-year survey, a total of 4120 skin, hair and nail samples obtained from the outpatients with symptoms suggestive of tinea were analyzed by using direct microscopy, culture and molecular identification methods. Strains isolated from cultures were subjected to amplification of the nuclear rDNA ITS regions in a PCR assay followed by an early established RFLP analysis. For confirmation of species identification, 100 isolates as representatives of all presumable species were subjected to ITS sequencing. Infection was confirmed in 1123 individuals (27.25 %) in the age range of 1-89 years by direct microscopy and/or culture including 603 males versus 520 females. Frequencies of infections were the highest and the lowest in age groups of 21-30 and 11-20 years, respectively. Tinea corporis was the most prevalent clinical manifestation followed by tinea cruris, tinea capitis, tinea manuum, tinea pedis, tinea unguium, tinea faciei and tinea barbae. Trichophyton interdigitale (58.7 %) was the most dominant isolate followed by Epidermophyton floccosum (35.4 %), Microsporum canis (3 %), T. rubrum (1.5 %), T. species of Arthroderma benhamiae (0.5 %), T. tonsurans (0.3 %) and T. violaceum (0.3 %). Other species included M. gypseum, M. fulvum and T. verrucosum (each one 0.1 %). Such a high occurrence of infection with T. interdigitale, which has not been reported from Iran, is due to the use of accurate molecular methods based on new species concept in dermatophytes. The prevalence of dermatophytoses caused by zoophilic species remarkably increased and Trichophyton species of A. benhamiae has emerged as a new agent of dermatophytosis in southwestern Iran, while infections due to anthropophilic species, except E. floccosum, took a decreasing trend.

Dermatophytosis due to Microsporum incurvatum: Notification and Identification of a Neglected Pathogenic Species.

A 4-year-old Iranian boy developed erythematous, itchy and annular lesion on his face. Microscopic examination of the scraped samples with 10 % potassium hydroxide (KOH) revealed fungal septate hyphae and arthroconidia. The etiological agent was found to be Microsporum gypseum in mycological examinations. Amplification and restriction digestion of the internal transcribed spacers (ITS) of rDNA was not helpful for identification, but in ITS sequencing the isolate showed 98 % homology to Microsporum incurvatum strain CBS 172.64. Empirical treatment of the patient with griseofulvin for 4 weeks was successful. Other than our isolate, the ITS1 sequences of 38 strains from related species were retrieved from GenBank and phylogenetic tree using maximum likelihood method was constructed. The case isolate clustered apart from other strains of M. incurvatum. Pairwise comparison of ITS1 showed intraspecies variations of 0-13 nucleotides among M. incurvatum strains and an extensive interspecies variation of 33-80 bp and remarkable interspecies size polymorphism between the three sister species in the M. gypseum complex. The high level of ITS1 intraspecific variation is suitable for species identification rather than phylogeographic analysis of M. gypseum complex.

Expression of Matrix Metalloproteinases in Human Cystic Echinococcosis.

BACKGROUND: Cystic echinococcosis (CE) is a zoonotic disease caused by the Echinococcus granulosus senso lato (E. granulosus s.l.) larval stages. Parasitederived products have been shown to regulate host matrix metalloproteinases (MMPs), contributing to CE pathogenesis and progressive liver fibrosis in intermediate hosts. The current study aimed to investigate the potential role of MMP1, 7, 8, and 13 in E. granulosus s.l-induced liver fibrosis. METHODS: Thirty CE patients with active, transitional, or inactive hydatid cysts were enrolled in this study to determine the inductive effects of E. granulosus on the expression of MMP-1, MMP-7, MMP-8, and MMP-13 in healthy liver tissue and fibrotic liver tissue using qRT-PCR. RESULTS: According to the WHO-IWGE classification, patients with functional cysts (CE1 and CE2) had the highest percentage (46.6%). MMP-1, MMP-7, MMP-8, and MMP-13 expression levels were significantly higher in fibrotic liver than in normal liver tissue. MMP-13 and MMP-1 had the highest and lowest expression levels among MMPs. Compared to the normal group, the fold change for MMP-13 in the fibrotic group was greater than 12 and had the highest AUC value (AUC= 0.8283). CONCLUSION: Our findings suggest that E. granulosus-derived products might be involved in regulating host MMPs. Thus, MMPs may be considered potential biomarkers for predicting CE prognosis. Because of the non-normal distribution of our patients' CE types, further research, particularly on circulation MMPs, is needed to confirm the potential role of MMPs in CE pathogenesis and to follow up on CE patients.

Strongyloides stercoralis genotyping in a human population in southwestern Iran.

BACKGROUND: Strongyloidiasis is a neglected tropical disease (NTD) that is caused mainly by Strongyloides stercoralis, with an estimated 600 million people infected worldwide, and in fewer cases by Strongyloides fuelleborni fuelleborni and Strongyloides fuelleborni kellyi. A number of studies have been conducted on the genetic diversity of S. stercoralis in East and Southeast Asia; however, there is very limited corresponding information from West Asian countries, including Iran. METHODS: For Strongyloides worms collected from patients in southwestern Iran, the hypervariable regions I (HVR-I) and IV (HVR-IV) of the nuclear 18S ribosomal DNA (rDNA) locus (SSU) and a fragment of the subunit 1 mitochondrial cytochrome c oxidase gene (cox-1) were sequenced. For a subset of the worms, whole-genome sequencing data were generated. RESULTS: The cox-1 sequences of 136 worms isolated from 23 patients indicated that all isolates were S. stercoralis. Among the cox-1 sequences, 33 polymorphic sites and 13 haplotypes were found. The phylogenetic analysis demonstrated that some sequences clustered fairly closely with sequences from humans and dogs from other parts of the world, while others formed a separate, Iran-specific group. Among 64 S. stercoralis analyzed, we found three of the previously described SSU HVR-I haplotypes, with haplotype II being the most frequent haplotype. In contrast to Southeast Asia, where S. stercoralis heterozygous for different haplotypes at the HVR-I locus are rare, we found 20 worms to be heterozygous for two different HVR-I haplotypes, 18 of which fell into the Iran-specific cox-1 cluster. SSU-heterozygous worms also showed elevated heterozygosity at the whole-genome level. CONCLUSIONS: We conclude that the S. stercoralis population from the Khuzestan province shares much of the genetic diversity with the population in Southeast Asia, but there is an indication of additional genetic input. There appears to be some population structure with different subpopulations, which however do interbreed at least occasionally.

Prevalence of asymptomatic strongyloidiasis co-infection in COVID-19 patients residing in endemic areas.

BACKGROUND: Fatal forms of strongyloidiasis, hyperinfection syndrome (HS) and disseminated strongyloidiasis (DS), are caused by exaggerated autoinfection of the intestinal nematode, Strongyloides stercoralis (S. stercoralis). Corticosteroids, frequently administered to patients with severe COVID-19, can transform chronic asymptomatic strongyloidiasis into the above-mentioned fatal diseases. This study aimed to investigate the prevalence of strongyloidiasis in COVID-19 patients receiving corticosteroids in a hypoendemic region. METHODS: The present cross-sectional study enrolled 308 COVID-19 patients admitted to two hospitals in Ahvaz and Abadan in the southwest of Iran between 2020 and 2022. A real-time reverse transcription polymerase chain reaction (RT-PCR) test and chest computed tomography (CT) scan were employed to detect and monitor the disease's severity in the patients, respectively. All patients were evaluated for IgG/IgM against S. stercoralis using Enzyme-linked immunosorbent assay (ELISA) test. Subsequently, individuals with a positive ELISA test were confirmed using parasitological methods, including direct smear and agar plate culture (APC). RESULTS: The patients were between 15 and 94 years old, with a mean age of 57.99 ± 17.4 years. Of the 308 patients, 12 (3.9%) had a positive ELISA test, while 296 (96.1%) had negative results. Three of the 12 patients with a positive ELISA result died, and three failed to provide a stool sample. To this end, only six cases were examined parasitologically, in which S. stercoralis larvae were observed in five patients. Significant differences were found between S. stercoralis infection with sex (p = 0.037) and age (p = 0.027). Binary regression analysis revealed that strongyloidiasis was positively associated with sex (odds ratio [OR]: 5.137; 95% confidence interval [CI]: 1.107-23.847), age (OR: 5.647; 95% CI 1.216-26.218), and location (OR: 3.254; 95% CI: 0.864-12.257). CONCLUSIONS: Our findings suggest that screening for latent strongyloidiasis in COVID-19 patients in endemic areas using high-sensitivity diagnostic methods, particularly ELISA, before receiving suppressive drugs should be given more consideration.

Serum Level of egr-miR-2a-3p as a Potential Diagnostic Biomarker for Cystic Echinococcosis.

INTRODUCTION: Cystic Echinococcosis (CE) is a chronic parasitic disease caused by the metacestodes of Echinococcus granulosus senso lato (E. granulosus s.l.). The larval stages of this parasite, hydatid cyst, are usually diagnosed using imaging modalities and serological testing; however, several studies have recently suggested using the parasite-derived microRNAs (miRNAs) as novel diagnostic biomarkers. MATERIALS AND METHODS: The present study included 31 CE patients who were older than 5 years and were admitted to the hospitals of Ahvaz, Iran for hydatid cyst removal surgery during 2019-2021. The egr-miR-125-5p and egr-miR-2a-3p levels were evaluated in the sera of the CE patients before and 6 months after the surgery using Quantitative Real-Time PCR (qRT-PCR), and the results were compared with the serum samples from 15 healthy volunteers. Then, the intergroup comparisons were performed using the t test. RESULTS: The patients' age range was 6-72 years, with a mean age of 34.6 years. Moreover, based on the classification by the WHO-IWGE, one patient (3.2%) had CE1, 14 patients (45.2%) had CE2, 5 patients (16.1%) had CE3, 2 patients (6.5%) had CE4, and 9 patients (29%) had CE5. Also, 21 patients (67.74%) had a positive antigen test using the ELISA method, while 10 patients (32.26%) had a negative ELISA. The pre-operative expression level of egr-miR-2a-3p was 10.36 folds higher compared to 6 months after the surgery, with an AUC value of 0.8176. However, the expression levels of egr-miR-125-5p did not change significantly 6 months after the surgery compared to pre-operative levels. CONCLUSIONS: According to the present study results, the serum levels of egr-miR-2a-3p can be a promising non-invasive biomarker for diagnosing CE and monitoring its potential recurrence after cystectomy.

Can Echinococcus granulosus-Derived MicroRNAs be Biomarkers for Diagnosis and Follow-up of Cystic Echinococcosis Patients?

INTRODUCTION: Cystic echinococcosis (CE) is a neglected tropical disease caused by the larval stages of Echinococcus granulosus (E. granulosus). MicroRNAs (miRNAs) are small noncoding RNAs acting as mediators in host-parasite interaction. Recently, numerous studies have been conducted on miRNAs in infectious diseases; however, little data are available about the role of miRNAs in pathogenesis and early diagnosis of CE. METHODS: The current study evaluated the expression of four E. granulosus-derived miRNAs, including egr-miR-125,5p, egr-let-7,5p, egr-miR-2, and egr-miR-71 in fibrotic and healthy liver tissues of 31 CE patients with active and inactive hydatid cysts by qRT-PCR. RESULTS: Of the 31 patients, 48.4% had active cysts (CE1 and CE2), while the remainder had transitional (16.1%) and inactive (35.5%) CE types cysts. The qRT-PCR analysis revealed a significant increase of 11.2, 9.91, 6.2, and 13.1-fold in the fibrotic tissue group for egr-miR-125,5p, egr-let-7,5p, egr-miR-2, and egr-miR-71, respectively. Among these miRNAs, egr-miR-125-5p exhibited the highest area under the curve (AUC) value of 0.8050 for predicting liver fibrosis. CONCLUSIONS: Our findings provide new data about the role of E. granulosus-derived miRNAs in pathogenesis of CE. The high AUC of egr-miR125,5p reflecting the possibility of using egr-miR125,5p as biomarker in CE diagnosis. Further studies on serum of CE patients are needed to confirm the potential role of circulating egr-miR-2a-3p and egr-miR-125-5p in the early diagnosis of CE.

Evaluation of the Protoscolicidal Effectiveness of Hypertonic Saline, Silver Nitrate, Ethanol, Using Sponge Pad Method and Injecting into Fertile Hydatid Cysts.

Background: We aimed to evaluate currently protoscolices agents by using two methods including injection and impregnated sponges with protoscolices agents. Methods: Hypertonic saline 20%, silver nitrate 0.5% and Ethanol 96% were injected into sheep hydatid cysts. Then, after different exposure times (5, 10, and 15 min), the protoscolices were aspirated from the cyst and viability of the protoscolices were evaluated microscopically using vital staining. In the second part, the live protoscolices were sprayed on the soaked sponge with the mentioned protoscolicidal agents. After different exposure times live protoscolices were counted microscopically. One-way ANOVA and Tukey tests were used for showing difference significant between studied groups. Results: Cyst injection method showed that ethanol, Nacl and AgNo3 have 80%, 60% and 6% protoscolocidal efficacy, respectively (P<0.05). In sponge method the most and fastest protoscolocidal efficacy were observed in all used protoscolicidal agents. Conclusion: Ethanol and NaCl and AgNo3 with sponge method is more effective than the injection of protoscolices agents in the cyst.

Nephrotoxicity of Albendazole and Albendazole Loaded Solid Lipid Nanoparticles in Mice with Experimental Hydatidosis.

Purpose: Cystic echinococcosis (CE) is a serious contemporary public health problem. Different CE treatment methods are of considerable importance, with albendazole (ABZ) being one of the most preferred drugs for CE treatment and prophylaxis. In this study, we evaluated the nephrotoxicity caused by ABZ and ABZ-loaded solid lipid nanoparticles (SLNs) in mice with experimental hydatid cyst. Methods: ABZ-loaded SLNs were produced by micro-emulsification and a high shear homogenization technique. Thereafter, we evaluated the physicochemical characterization of the product. Live protoscolices were injected into mice to induce experimental hydatidosis. Mice were then treated with ABZ and ABZ-loaded SLNs. The nephrotoxicity effects were evaluated by biochemical and histopathological surveys. Results: Significantly different blood urea nitrogen (BUN) levels were observed between the two infected groups (ABZ treatment and ABZ-loaded SLN treatment) and the control group. The kidney malondialdehyde (MDA) and glutathione (GSH) levels of the infected groups were not significantly different from those of the control group. The histopathological study revealed nephropathic and pathologic changes in the ABZ and ABZ-loaded SLN groups. Conclusion: ABZ formulated for ABZ-loaded SLNs had a more prominent chemoprophylactic efficacy on CE and fewer side effects than ABZ alone. Neither ABZ nor ABZ-loaded SLNs caused significant biochemical and histopathological defects on the kidney, and all functional biochemical markers stayed within the normal range. Therefore, ABZ-loaded SLNs could be a potential new product for CE treatment.

The Role of Tissue Inhibitor of Metalloproteinase-1 and 2 in Echinococcus granulosus senso lato-Induced Human Hepatic Fibrosis.

INTRODUCTION: The main mechanism underlying hepatic fibrosis is the imbalance between tissue Matrix Metalloproteinases (MMPs) and Tissue Inhibitors of Metalloproteinases (TIMPs). This study aimed to investigate the potential role of TIMP-1 and TIMP-2 in the process of hepatic fibrosis caused by Echinococcus granulosus senso lato (E. granulosus s.l.). METHODS: The expressions levels of TIMP-1 and TIMP-2 mRNAs were evaluated in fibrotic and normal hepatic tissues of 30 patients with Cystic Echinococcus (CE) using qRT-PCR. Moreover, their serum levels of TIMP-1 were assessed before CE cyst removal and 6 months after surgery using ELISA. RESULTS: The qRT-PCR results showed that the expression levels of TIMP-1 and TIMP-2 mRNAs were significantly higher in the fibrotic hepatic tissue compared to the normal liver tissue, in a way that the TIMP-1 and TIMP-2 mRNA expression levels were 19.07 and 6.58 folds higher in the fibrotic tissue compared to the normal liver tissue. Among these TIMPs, TIMP-1 exhibited the higher area under the curve (AUC) value for predicting liver fibrosis. However, we could not find a significant difference in the serum levels of TIMP-1 before and after the cyst removal procedure (p = 0.48). CONCLUSIONS: For the first time, our study showed that the significant overexpression of both TIMP mRNAs in the fibrotic liver tissue of the CE patients may be due to the increased expression of MMPs in the peri-cystic tissue. However, we could not find a significant difference in the pre- and post-operative TIMP-1 levels, which may be due to recurrence or heterogeneity in the cyst type. Therefore, performing further studies with a larger sample size of the CE patients is recommended.

Primary disseminated intraabdominal hydatidosis: a case report.

BACKGROUND: Hydatidosis, a zoonotic disease caused by the larvae of Echinococcus granulosus sensu lato (E. granulosus s.l.), can be primary or secondary. However, primary disseminated intraabdominal hydatidosis is a rare form of the disease, accounting for about 2% of all intraabdominal cysts. CASE PRESENTATION: We report herein a case of primary disseminated intraabdominal hydatidosis with multiple organ involvement in a 51-year-old Iranian man presenting to a healthcare facility with abdominal pain. During the physical examination, two abdominal masses were palpated. Ultrasound and computed tomography revealed six cystic lesions in the patient's liver, subhepatic region, pelvic, and omentum. Afterward, he underwent surgery, during which the cystic lesions were completely removed. The patient received albendazole (400 mg/kg/day) postoperatively and was recommended to continue the treatment for 4 months. CONCLUSIONS: Although primary disseminated intraabdominal hydatidosis is rare, this problem is of great importance due to the fertility of cysts and the high risk of recurrence. Therefore, it is recommended to follow such patients with imaging modalities and enzyme-linked immunosorbent assay for native antigen B (AgB). In addition, patients should undergo albendazole therapy postoperatively for 4 months.

Comparison of survival time of Echinococcus granulosus protoscolices in different culture media and temperatures and evaluation of their ability to generate cysts in mice.

Hydatidosis is a disease caused by the larval stage of Echinococcus granulosus, which has great importance in medicine and veterinary medicine. Protoscolices (PSCs) of fertile hydatid cysts play a critical role in secondary echinococcosis after surgery. Fertile cysts were acquired from infected sheep at the local municipal abattoir in Ahvaz, southwest of Iran. PSCs were obtained aseptically and transferred to 10 different culture media and kept at 4°C and 37°C to determine the duration of PSCs' survival. Then, 2000 live PSCs from each of the culture media were injected into the peritoneal cavity of BALB/c mice. After five months, the mice were evaluated in terms of cyst number, size, and weight. The highest PSCs survival time at 4°C was related to RPMI-1640 medium and cyst fluid for 50 and 45 days, respectively. Also, at 37°C, the longest survival time of PSCs was related to cyst fluid and RPMI-1640 media for 30 and 29 days, respectively. The highest level of infection and median cyst number was observed in mice received PSCs from the RPMI-1640 medium at 4°C, and the highest level of infection in mice at 37°C was related to the DMEM low glucose (L) medium. The current study indicated that 4°C was a more suitable temperature for in vitro storage of live PSCs. The maximum amount of infection was observed in mice received PSCs from the RPMI-1640 medium at 4°C. The present study is the first attempt to compare the ability of PSCs to generate hydatid cysts in mice after being cultured in different media and at various temperatures.

PCR Detection and Sequencing of Trichomonas vaginalis in Women with Suspected Vaginitis in Southwestern Iran.

INTRODUCTION: The human being is considered a natural host for Trichomonas vaginalis (T. vaginalis), which causes trichomoniasis, the most frequent non-viral sexually transmitted infectious disease in the world. The current study aimed to evaluate the prevalence and sequencing of T. vaginalis in women with vaginitis. ; Methods: In the current research, 514 vaginal discharge samples were obtained from women with vaginitis. The specimens were evaluated by the direct wet mount examination, Dorset culture medium, and PCR technique. Primers were designed for the detection of TVK3/TVK7, TVA5/TVA6 genes specific for the identification of T. vaginalis. The PCR-positive samples were sequenced and compared with the sequences registered in the GenBank database. ; Results: Among the collected samples, 30 (5.83%), 45 (8.75%), 90 (17.50%), and 62 (12.06%) cases were positive for T. vaginalis when assayed by the direct wet mount examination, Dorset culture medium, and PCR technique (TVK3/TVK7, TVA5/TVA6 genes), respectively. There was no significant relationship between trichomoniasis and demographic characteristics of women, such as age, occupational status, mode of delivery, number of deliveries, educational level, and contraceptive methods (p˃0.05). The range of vaginal pH was between 5-7 in women with vaginitis, and there was a significant statistical correlation between the pH values and the infection rate (p<0.05). The PCR-positive samples had 100% sequence homology with the reference sequence in the GenBank database (accession number L23861.1). ; Conclusion: This study confirmed a relatively high prevalence of T. vaginalis in the southwestern region of Iran. According to our results, the PCR method, especially detecting TVK3/TVK7 genes, was more sensitive than the direct wet mount examination and Dorset culture medium methods.