Isoenzyme characterization of Leishmania infantum toward checking the antioxidant activity of superoxide dismutase and glutathione peroxidase.

BACKGROUND: Leishmania infantum is the major causative agent of visceral leishmaniasis in Mediterranean regions. Isoenzyme electrophoresis (IE), as a biochemical technique, is applied in the characterization of Leishmania species. The current study attempted to investigate the isoenzyme patterns of logarithmic and stationary promastigotes and axenic amastigotes (amastigote-like) of L. infantum using IE. The antioxidant activity of superoxide dismutase (SOD) and glutathione peroxidase (GPX) was also checked in the aforementioned forms. METHOD: After L. infantum cultivation and obtaining logarithmic and stationary promastigotes, axenic amastigotes were achieved by incubation of stationary promastigotes at 37 °C for 48 h. The lysate samples were prepared and examined for six enzymatic systems including glucose-6-phosphate dehydrogenase (G6PD), nucleoside hydrolase 1 (NH1), malate dehydrogenase (MDH), glucose-phosphate isomerase (GPI), malic enzyme (ME), and phosphoglucomutase (PGM). Additionally, the antioxidant activity of SOD and GPX was measured. RESULTS: GPI, MDH, NH1, and G6PD enzymatic systems represented different patterns in logarithmic and stationary promastigotes and axenic amastigotes of L. infantum. PGM and ME showed similar patterns in the aforementioned forms of parasite. The highest level of SOD activity was determined in the axenic amastigote form and GPX activity was not detected in different forms of L. infantum. CONCLUSION: The characterization of leishmanial-isoenzyme patterns and the measurement of antioxidant activity of crucial antioxidant enzymes, including SOD and GPX, might reveal more information in the biology, pathogenicity, and metabolic pathways of Leishmania parasites and consequently drive to designing novel therapeutic strategies in leishmaniasis treatment.

SARS-CoV-2 mechanisms of cell tropism in various organs considering host factors.

A critical step in the drug design for SARS-CoV-2 is to discover its molecular targets. This study comprehensively reviewed the molecular mechanisms of SARS-CoV-2, exploring host cell tropism and interaction targets crucial for cell entry. The findings revealed that beyond ACE2 as the primary entry receptor, alternative receptors, co-receptors, and several proteases such as TMPRSS2, Furin, Cathepsin L, and ADAM play critical roles in virus entry and subsequent pathogenesis. Additionally, SARS-CoV-2 displays tropism in various human organs due to its diverse receptors. This review delves into the intricate details of receptors, host proteases, and the involvement of each organ. Polymorphisms in the ACE2 receptor and mutations in the spike or its RBD region contribute to the emergence of variants like Alpha, Beta, Gamma, Delta, and Omicron, impacting the pathogenicity of SARS-CoV-2. The challenge posed by mutations raises questions about the effectiveness of existing vaccines and drugs, necessitating consideration for updates in their formulations. In the urgency of these critical situations, repurposed drugs such as Camostat Mesylate and Nafamostat Mesylate emerge as viable pharmaceutical options. Numerous drugs are involved in inhibiting receptors and host factors crucial for SARS-CoV-2 entry, with most discussed in this review. In conclusion, this study may provide valuable insights to inform decisions in therapeutic approaches.

First Molecular Identification and Subtyping of Blastocystis sp. in the Most Consumed Edible Marine Fish of Iran: A Foodborne Concern.

PURPOSE: The presence of Blastocystis sp. is commonly observed in humans and different animals, displaying a wide range of genetic variations with the discovery of multiple subtypes (STs). However, the prevalence and distribution of these STs in edible marine fish and marine mammals remain uncertain. This study marks the first survey conducted in Iran and the second global molecular investigation to examine the occurrence and STs distribution of Blastocystis in various species of edible marine fish. METHODS: This study screened 200 fresh intestinal contents from 10 well-known fish species (Narrow-barred mackerel, Indo-pacific king mackerel, Tigertooth croaker, Silver pomfret, Black pomfret, Longtail tuna, John's snapper, Blackspotted croaker, Four-finger threadfin, and Javelin grunter) in southern Iran, caught in the Persian Gulf. All collected samples were evaluated by microscopy and SSU-PCR methods. RESULTS: Based on both microscopy and PCR, the overall prevalence of Blastocystis sp. in evaluated fish species was 2% (4/200). In brief, Blastocystis sp. was reported from Narrow-barred mackerel [10% (2/20)], Silver pomfret [5% (1/20)], and Tigertooth croaker [5% (1/20)]. Interestingly, among infected fish species three zoonotic STs (ST1, ST2, and ST7) were identified. ST2 was the most predominant ST [50% (2/4)], followed by ST1 and ST7, one sample each [5% (1/20)]. CONCLUSION: Overall, the prevalence and STs distribution of Blastocystis in edible marine fish along with the possibility of its zoonotic transmission are still open to question and require extensive and more detailed studies.

A novel outlook in the delivery of artemisinin: production and efficacy in experimental visceral leishmaniasis.

The visceral form of leishmaniasis (VL), due to infection by Leishmania infantum, is a neglected tropical disease. The accessible therapeutic options are limited. Artemisinin is an efficient antileishmanial product with poor biological availability that requires high repetition of therapeutic doses in VL. Solid lipid nanoparticles (SLNs) provide targeted delivery, increase bioavailability and reduce toxicity of the traditional therapeutic strategy. The spherical shape artemisinin-loaded SLNs were prepared in a particle diameter of 222.0 ± 14.0 nm. The SLNs showed no particular toxic effect on the parasites, whereas the native artemisinin demonstrated a significant toxicity rate of 31% in viability of the promastigotes at the 250 µg/ml concentration. The therapeutic efficacy of the artemisinin-loaded SLNs was demonstrated in the experimental VL, using the L. infantum-infected BALB/c mice, in the present study. The 10 and 20 mg/kg doses of artemisinin-loaded SLNs showed higher level of antileishmanial efficacy compared with the free artemisinin. There was a significant diminishing of the parasite burden in liver (84.7 ± 4.9%) and spleen (85.0 ± 3.1%) and hepatosplenomegaly by the artemisinin-loaded SLNs treated at 20 mg/kg compared to the free artemisinin. Therefore, the present study supports the superior efficacy of artemisinin-loaded SLNs over the free artemisinin and could be considered as a new therapeutic strategy in the treatment of leishmaniasis.

The potential role of protein disulfide isomerases (PDIs) during parasitic infections: a focus on Leishmania spp.

Leishmaniasis is a group of vector-borne diseases caused by intracellular protozoan parasites belonging to the genus Leishmania. Leishmania parasites can employ different and numerous sophisticated strategies, including modulating host proteins, cell signaling, and cell responses by parasite proteins, to change the infected host conditions to favor the parasite persistence and induce pathogenesis. In this sense, protein disulfide isomerases (PDIs) have been described as crucial proteins that can be modulated during leishmaniasis and affect the pathogenesis process. The effect of modulated PDIs can be investigated in both aspects, parasite PDIs and infected host cell PDIs, during infection. The information concerning PDIs is not sufficient in parasitology; however, this study aimed to provide data regarding the biological functions of such crucial proteins in parasites with a focus on Leishmania spp. and their relevant effects on the pathogenesis process. Although there are no clinical trial vaccines and therapeutic approaches, highlighting this information might be fruitful for the development of novel strategies based on PDIs for the management of parasitic diseases, especially leishmaniasis.

A Novel Multiplex LAMP Assay for the Rapid and Accurate Diagnosis of Visceral Leishmaniasis Caused by Leishmania infantum from Iran.

Visceral leishmaniosis (VL) is one of the neglected tropical diseases despite being responsible for serious clinical symptoms, some of which lead to fatal outcomes. Thus, there is a need to apply accurate, rapid, and specific diagnostic measurements in order to control the disease and reduce the mortality rate. We aimed to develop and validate a multiplex LAMP assay for the diagnosis of VL caused by Leishmania infantum (L. infantum). Moreover, a thorough assessment was conducted to determine the effectiveness of multiplex LAMP in identifying various Leishmania species, such as Leishmania tropica (L. tropica) and Leishmania major (L. major) in comparison to Leishmania infantum (L. infantum). The diagnostic performance of the multiplex LAMP method for VL was compared to each LAMP assay, real-time polymerase chain reaction (RT-qPCR), and nested PCR technique. Two separated primers were set and used in a multiplex LAMP assay which was designed based on the ITS2 (internal transcribed spacer II) and were selected on the basis of conserved and high copy number region. Multiplex LAMP primers were designed using an online tool available at https://www.primerexplorer.jp/e. The alignment was performed using MEGA5, and the primers were further adjusted utilizing GENE Runner software. All molecular methods were tested on the serial dilution of cloned plasmid containing ITS region from standard strains of L. infantum, L. tropica, and L. major. Moreover, multiplex LAMP assay was evaluated and compared based on both standard strains and 55 clinical samples from humans as well as dogs. Various approaches were applied to interpret the multiplex LAMP reaction which deciphered a higher sensitivity when compared to the RT-qPCR for L. infantum (one copy number of plasmid, equal to 0.85 femtograms (fg) of plasmid concentration, and 0.004 parasite DNA per µL) detection while these three standard strains of Leishmania were confirmed to contain 40 DNA copies using RT-qPCR. Additionally, the multiplex LAMP detection limit was approximately equivalent to RT-qPCR for L. major and L. tropica, which included 0.342 picograms (pg) and 342 femtograms (fg) of plasmid concentration, 4 × 103 and 4 × 102 copy number of plasmid, and 17.1 and 1.71 parasite DNA per µL for L. major and L. tropica, respectively. Nested PCR exhibited a lower detection limit for L. infantum of 4 × 106 plasmid copy number compared to multiplex LAMP and RT-qPCR. Multiplex LAMP has the potential for accurate and rapid detection of infectious disease, successful treatment, and finding and monitoring asymptomatic cases, especially in low-income countries.

Evaluating the Typing Power of Six Isoenzymatic Systems for Differentiation of Clinical and Standard Isolates of Candida Species.

Background: Due to the increasing prevalence of candidiasis, early detection of the causative agents may pave the way for the management of this infection. The present study aimed to assess the discriminative power of the six isoenzymatic systems for differentiating the Candida species. Materials and Methods: Sixteen standard Candida albicans and Candida dubliniensis strains and 30 fluconazole-sensitive and fluconazole-resistant clinical strains of Candida albicans were analyzed using a Multilocus Enzyme Electrophoresis (MLEE) method, including six enzymatic systems consisting of malate dehydrogenase (MDH), phosphoglucomutase (PGM), glucose-phosphate isomerase (GPI), glucose-6-phosphate dehydrogenase (G6PDH), 6-phosphogluconate dehydrogenase (6PGD), and malic enzyme (ME). Results: Among the six enzymatic systems, ME showed no diagnostic activity, whereas MDH provided the best species-specific pattern for species discrimination. In addition, the MDH and G6PD systems provided a discriminatory pattern for differentiating C. dubliniensis from C. albicans isolates. The same isoenzymatic activity was detected in all 36 standard and clinical isolates. Moreover, the results showed no correlation between the isoenzymatic profiles and drug resistance. Conclusion: Among the investigated MLEE systems, MDH was able to differentiate between Candida albicans and Candida dubliniensis. Although no association was detected between isoenzyme patterns and fluconazole resistance in this investigation, isoenzyme patterns are likely correlated with virulence factors between species and even within species. To answer these questions, additional studies should be done on more strains.

In vitro and in vivo antileishmanial activity of the hanging sedge flavonoids based on bio- guided fractionation assay.

As a major public health issue, cutaneous leishmaniasis (CL) has a number of complications, including drug resistance and poor response to conventional treatments. Over the last decade, research on natural sources for finding new antileishmanial agents has been a critical part of tropical disease research. Natural products also should be regarded as one of the most valuable applications for CL infection drug development. In this study, we assessed the in vitro and in vivo antileishmanial potential of Carex pendula Huds. (hanging sedge) methanolic extract and its fractions against Leishmania major produced cutaneous infection. Although the methanolic extract and its fractions exhibited suitable activity, the ethyl acetate fraction showed the best activity (with the half maximal inhibitory concentration IC50  = 1.627 ± 0.211 mg/mL). The toxicity and selectivity indices (SI) of all samples were determined in murine peritoneal macrophage cells (J774A.1) using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test. The flavonoid components of the ethyl acetate fraction were identified using liquid chromatography electrospray ionization mass spectrometry (LC-ESI MS/MS). Nine chemical compounds were identified in this fraction, including three flavonols, four flavanonols, and two flavan derivatives. L. major-infected mice were used as an in vivo model because the methanolic extract was effective against L. major promastigotes in the mammalian cell line J774A.1 with SI = 2.514 (tail lesion size model). In silico analysis of identified compounds also revealed a favorable interaction between compounds 2-5 and L. major protein targets (3UIB, 4JZX, 4JZB, 5L4N, and 5L42). According to the findings of this study, the ethyl acetate fraction (as flavonoid fraction) exhibited considerable in vitro antileishmanial activity.

The novel treatments based on tissue engineering, cell therapy and nanotechnology for cutaneous leishmaniasis.

Cutaneous leishmaniasis (CL) is a global public health issue. Conventional treatments have substantial costs, side effects, and parasite resistance. Due to easy application and inexpensive cost, topical treatment is the optimal approach for CL. It could be used alone or with systemic treatments. Electrospun fibers as drug release systems in treating skin lesions have various advantages such as adjustable drug release rate, maintaining appropriate humidity and temperature, gas exchange, plasticity at the lesion site, similarity with the skin extracellular matrix (ECM) and drug delivery with high efficiency. Hydrogels are valuable scaffolds in the treatment of skin lesions. The important features of hydrogels include preserving unstable drugs from degradation, absorption of wound secretions, high biocompatibility, improving the re-epithelialization of the wound and preventing the formation of scars. One of the issues in local drug delivery systems for the skin is the low permeability of drugs in the skin. Polymeric scaffolds that are designed as microneedle patches can penetrate the skin and overcome this challenge. Also, drug delivery using nanocarriers increases the effectiveness of drugs in lower and more tolerable doses and reduces the toxicity of drugs. The application of cell therapy in the treatment of parasitic and infectious diseases has been widely investigated. The complexity of leishmaniasis treatment requires identifying new treatment options like cell therapy to overcome the disease. Topics investigated in this study include drug delivery systems based on tissue engineering scaffolds, nanotechnology and cell therapy-based studies to reduce the complications of CL.

Superparamagnetic Iron Oxide-Labeled Leishmania major Can Be Traced in Fibroblasts.

Introduction: Leishmaniasis is still a neglected tropical disease that can endanger more than 350 million people among 98 countries. Leishmania can survive in fibroblasts as latent inactive forms. This study was conducted to evaluate the role of superparamagnetic iron oxide nanoparticles (SPIONs) in cell culture for tracking the labeled Leishmania major in fibroblasts. Methods: Dextran-coated SPIONs were used for labeling L. major in co-culture of fibroblasts with the parasite. To quantify and trace SPION-labeled Leishmania, Prussian blue staining was undertaken. Fibroblast characterization was undertaken by real time polymerase chain reaction. Transmission electron microscope (TEM) was used for confirming the entry of the labeled L. major to the cytoplasm and the nucleus of the fibroblast. Results: Fibroblasts were spindle-shaped and adherent to culture flasks. Promastigotes were with thin elongated lance-like morphology with an anterior kinetoplast and an emergent free flagellum. Prussian blue staining revealed that internalized SPIONs were localized within cytoplasm and nucleus of the fibroblasts after 24 hours of culture. Prussian blue staining successfully showed the presence of iron (stained blue) in labeled L. major within the fibroblasts. This finding was confirmed by TEM, and labeled L. major was detected in the fibroblast cytoplasm and nucleus too. Conclusion: We can conclude that SPIONs are safe, inexpensive, easy to use, and accurate, and a fast method to label Leishmania parasite in cells that the parasite can be latent, such as fibroblasts. These findings can open a new window in diagnosis, pathogenesis, and treatment of cutaneous leishmaniasis and can be added to the literature.

Bioinformatics analysis for the purpose of designing a novel multi-epitope DNA vaccine against Leishmania major.

Leishmaniasis is one of the main infectious diseases worldwide. In the midst of all the different forms of the disease, Cutaneous Leishmania (CL) has the highest incidence in the world. Many trial vaccines have been developed with the purpose of generating long-term cell-mediated immunity to Leishmania(L) major. As there is not any multi-epitope DNA vaccine with high efficacy against L.major, the aim of this study is to design a new multi-epitope DNA vaccine in order to have effective control upon this infectious disease through the immune bioinformatics. The L.major antigens: Gp63, LACK, TSA, LmSTI1and KMP11 were selected to design a multi-epitope DNA vaccine. The initial structure of the DNA vaccine was designed, benefiting from Gen Bank's website information. Epitopes of MHC-I antigens were predicted through the Immune Epitope Database (IEDB), and the selected epitopes were used to make vaccines construct along with linkers. New multi-epitope vaccine including 459 nucleic acids designed, and inserted between BamH1 and HindIII restriction sites of pCDNA3.1 mammalian expression vector. 12 epitopes among the chosen antigens were selected by two servers (IEDB and ANTIGEN). They had high stability and high antigenic power. Physicochemical features of vaccine measured by ProtParam server, and this structure was thermostable and hydrophilic. it's a suitable model to study on the animal and human phases. The designed vaccine is expected to be an effective candidate through development of (CL) vaccines. However, the effectiveness of this vaccine should also evaluate in vivo model.

The Spread of Insertion Sequences Element and Transposons in Carbapenem Resistant Acinetobacter baumannii in a Hospital Setting in Southwestern Iran.

BACKGROUND: Acinetobacter baumannii is one of the most important hospital pathogenic bacteria that cause infectious diseases. The present study aimed to determine the frequency of carbapenem resistance genes in association with transposable elements and molecular typing of carbapenem-resistant A. baumannii bacteria collected from patients in Shiraz, Iran. MATERIALS AND METHODS: A total of 170 carbapenem-resistant A. baumannii isolates were obtained from different clinical specimens in two hospitals. The minimum inhibitory concentrations (MIC) of imipenem were determined and the prevalence of OXA Carbapenemases, Metallo-ß-lactamases genes, insertion sequences (IS) elements, and transposons were evaluated by the polymerase chain reaction (PCR) method. Finally, molecular typing of the isolates was performed by the Enterobacterial Repetitive Intergenic Consensus-PCR method. RESULTS: The MICs ranged from 16 to 1,024 µg/mL for imipenem-resistant A. baumannii isolates. Out of the 170 carbapenem resistant A. baumannii isolates, blaOXA-24-like (94, 55.3%) followed by blaOXA-23-like (71, 41.7%) were predominant. In addition, A. baumannii isolates carried blaVIM (71, 41.7%), blaGES (32, 18.8%), blaSPM (4, 2.3%), and blaKPC (1, 0.6%). Moreover, ISAba1 (94.2%) and Tn2009 (39.2%) were the most frequent transposable elements. Furthermore, (71, 44.0%) and (161, 94.7%) of the ISAba1 of the isolates were associated with blaOXA-23 and blaOXA-51 genes, respectively. Besides (3, 1.7%), (1, 0.6%) and (5, 2.9%) of blaOXA-23 were associated with IS18, ISAba4, and ISAba2, respectively. Considering an 80.0% cut off, clusters and four singletons were detected. CONCLUSION: According to the results, transposable elements played an important role in the development of resistance genes and resistance to carbapenems. The results also indicated carbapenem-resistant A. baumannii bacteria as a public health concern.

Molecular-based detection of Leishmania tropica isolates among sensitive, resistant, and relapsed patients treated with Meglumine Antimoniate.

The appearance of resistance to pentavalent antimony, as the mainline of treatment for Cutaneous Leishmaniasis (CL) has been reported from Iran. According to the patients' laboratory and clinical history, 96 archived slides of patients infected with Leishmania tropica (L. tropica) treated with Meglumine Antimoniate (Glucantime®) were selected. After microscopic examination, Nested Polymerase Chain Reaction (Nested-PCR) and Restriction Fragment Length Polymorphism (RFLP) assays were done for each sample. In Nested-PCR, all positive samples were characterized as L. tropica. Additionally, some positive products of sensitive, resistant, and recidivans cases were selected to check their differentiations by sequencing software. In RFLP, various patterns of schizodemes were detected according to the reference patterns. Most sensitive cases of L. tropica (treated with Glucantime®) were categorized as schizodeme B, and most resistant cases were identified as schizodeme B and D. In recidivans cases, 91% of specimens categorized as schizodeme A and B. However, study on the type of L. tropica isolates that are resistant or sensitive to Glucantime® could be helpful before drug therapy.

Current global status, subtype distribution and zoonotic significance of Blastocystis in dogs and cats: a systematic review and meta-analysis.

BACKGROUND: Blastocystis is a common intestinal protozoa found in animal and human fecal samples, with over 1 billion individuals infected worldwide. Since domestication, dogs and cats have had a close bond with humans. However, their close proximity poses a potential health risk since they may harbor several zoonotic agents. A global estimate of Blastocystis infection and subtype (ST) distribution in dogs and cats would therefore be of great health importance to humans. METHODS: We performed a comprehensive systematic search of four English-language databases (PubMed, Scopus, Google Scholar, Web of Science) for relevant articles up to 8 November 2021. The random-effects model was used to make pooled estimates with confidence intervals (CIs). RESULTS: In total, we identified 49 publications that met our inclusion criteria and subsequently analyzed the 65 datasets in these articles, of which 23 and 42 datasets were on cats and dogs, respectively. Among the 2934 cats included in the 23 datasets, which involved 16 countries, the prevalence rate of Blastocystis infection was 9.3% (95% CI 5.3-15.9%). The prevalence of Blastocystis infection was slightly lower [7%, 95% CI 4.7-10.4%) among the 7946 dogs included in the 42 datasets, involving 23 countries. The sensitivity analysis showed that no remarkable variation in the estimates upon the stepwise removal of each dataset. Higher ST diversity was found among the examined dogs (ST1-8, ST10, ST23, ST24) than among cats (ST1-4, ST10, ST14). Among dogs, ST3 was the most frequent ST (41.3%), followed by ST2 (39.3%), ST1 (30.9%), ST4 (13.4%), ST8 (12.7%), ST10 (11%) and ST5 (8.1%). Also among dogs, each of ST6, ST7, ST23 and ST24 was observed in only one study. Of the ST found in the cats examined, ST4 (29.5%), followed by ST10 (22.5%), ST1 (19.8%) and ST3 (17.6%) were the most common. A single study also reported the presence of both ST2 and ST14 in cats. With respect to zoonotic Blastocystis STs (ST1-ST9 and ST12), eight were reported from dogs (ST1-ST8) and four were isolated from cats (ST1-ST4), showing the implication of dog and cats in zoonotic transmission. CONCLUSIONS: Taken together, our results show that elucidation of the true epidemiology and ST distribution of Blastocystis in dogs and cats demands more comprehensive studies, particularly in the negelected regions of the world.

Spatio-temporal variability of malaria infection in Chahbahar County, Iran: association with the ENSO and rainfall variability.

Malaria is one of the most widespread communicable diseases in the southeast regions of Iran, particularly the Chabahar County. Although the outbreak of this disease is a climate-related phenomenon, a comprehensive analysis of the malaria-climate relationship has not yet been investigated in Iran. The aims of this study are as follows: a) analyzing the seasonal characteristics of the various species of the infection; b) differentiating between number of patients during El Niño and La Niña and also during the wet and dry years. The monthly malaria statistics collected from twelve health centers were firstly averaged into seasonal scale and then composited with the corresponding data of the ground-based meteorological records, Southern Oscillation Index (SOI), and the satellite-based rainfall data. The proper statistical tests were used to detect differences in the number of patients between El Niño and La Niña and also between the adopted wet and dry episodes. Infection rate from the highest to the lowest was associated with summer, autumn, spring, and winter, respectively. Plasmodium falciparum, P. vivax, and the other species were responsible for 22%, 75%, and 3% of the sickness, respectively. The outbreak of P. falciparum/P. vivax occurs during autumn/summer. Due to the malaria eradication programs in urban areas, infection statistics collected from the rural areas were found to be more climate-related than that of urban regions. For rural/urban areas, the infection statistics exhibited a significant decline/increase during El Niño episodes. In autumn, spring, and winter, the patient number has significantly increased/decreased during the dry/wet years, respectively. These relationships were, however, reversed in summer.

Microtubule Disruption Without Learning Impairment in the Unicellular Organism, Paramecium: Implications for Information Processing in Microtubules.

Introduction: Information processing in microtubules is an open question that has not been adequately addressed. It was suggested that microtubules could store and process information in the nervous system or even support consciousness. The unicellular organism, Paramecium caudatum, has a microtubular structure but lacks a neuron or neural network. However, it shows intelligent behaviors such as associative learning. This property may suggest that the microtubules are involved in intelligent behavior, information storage, or information processing in this organism. Methods: To test this hypothesis and study the role of microtubules in P. caudatum learning, we utilized a learning task in which the organism associates brightness in its swimming medium with attractive cathodal shocks. To see if microtubules are an integral part of information storage and processing in P. caudatum, we disrupted the microtubular dynamics in the organism using an antimicrotubular agent (parbendazole). Results: We observed that while a partial allosteric modulator of GABA (midazolam) could disrupt the learning process in P. caudatum, the antimicrotubular agent could not interfere with the learning. Conclusion: Microtubules are probably not vital for the learning behavior in P. caudatum. Consequently, our results call for further investigation of the microtubular information processing hypothesis. Highlights: Importance of Information processing in microtubules;Microtubules could store and process information in the nervous system;Unicellular organism, Paramecium caudatum, has a microtubular structure but lacks a neuron or neural network. Plain Language Summary: Information processing in microtubules is an open question that has not been adequately addressed. It was suggested that microtubules could store and process information in the nervous system or even support consciousness. The unicellular organism, Paramecium caudatum, has a microtubular structure but lacks a neuron or neural network. However, it shows intelligent behaviors such as associative learning. This property may suggest that the microtubules are involved in intelligent behavior, information storage, or information processing in this organism. To test this hypothesis and study the role of microtubules in P. caudatum learning, we utilized a learning task in which the organism associates brightness in its swimming medium with attractive cathodal shocks. To see if microtubules are an integral part of information storage and processing in P. caudatum, we disrupted the microtubular dynamics in the organism using an antimicrotubular agent (parbendazole). We observed that while a partial allosteric modulator of GABA (midazolam) could disrupt the learning process in P. caudatum, the antimicrotubular agent could not interfere with the learning. Microtubules are probably not vital for the learning behavior in P. caudatum. Consequently, our results call for further investigation of the microtubular information processing hypothesis.

Geographical distribution and molecular epidemiology of cutaneous leishmaniasis in Fars Province, southern Iran.

Cutaneous leishmaniasis (CL) is one of the important zoonotic diseases in Iran, particularly in Fars Province, southern Iran. The purpose of this descriptive cross-sectional study was to investigate the molecular identification and geographical distribution of anthroponotic and zoonotic CL in southern Iran, during 2018-2019. Overall, 161 patients with CL referred to the Leishmaniasis Diagnostic Laboratory, Valfajr Health Center, Shiraz, Iran, were included in this study. The smears were prepared from the lesion borders of patients and stained with Giemsa and diagnosed microscopically. For molecular identification, the genomic DNA of each sample was extracted, and PCR method was used. The geographical distribution map was prepared using ArcMap software. Finally, the possible correlation between the frequencies of CL in various subgroups was statistically analyzed by Chi-square test, using SPSS software. Of 161 positive samples confirmed by both microscopy and PCR, 126 (78.3%) and 35 (21.7%) samples were shown to be L. major and L. tropica, respectively. Also, 87 (54%) patients were male, and 74 (46%) were female. The study showed that anthroponotic cutaneous leishmaniasis (ACL) was detected only in Shiraz city, while zoonotic cutaneous leishmaniasis (ZCL) was observed both in Shiraz and most counties of Fars Province. Furthermore, the most CL infections occurred in district 8 among the different districts of Shiraz municipality, which requires serious attention.

The effect of integration of basic and clinical aspects of a specific topic in a parasitology course on medical students learning: A randomized controlled trial.

BACKGROUND AND AIM: Parasitology course is one of the basic science courses in the educational curriculum for medical students. Since the integration of basic and clinical sciences has helped students better understand the basic science course content, the aim of the present study was to determine the effect of integration of basic and clinical aspects of a specific topic in a parasitology course on medical students learning. MATERIALS AND METHODS: A randomized controlled trial was conducted on 110 undergraduate fifth-semester medical students from April to July 2018. The students were randomly divided into two groups, based on student identification number: Intervention and control groups. The topic selected for this study from the parasitology course was "cutaneous leishmaniasis." At the beginning of the program, a dermatologist presented the clinical aspects of the topic to the intervention group. Then, a parasitologist delivered a traditional lecture about the basic aspects of the topic to both groups. The students' scores on questions related to cutaneous leishmaniasis in the final exam were used as a measure of learning and teaching outcomes. A questionnaire that consisted of seven items and three open-ended questions was used based on the objectives of the randomized controlled trial. Statistical analysis was performed by SPSS software. RESULTS: Based on the result of the final examination, there was no significant difference in the learning rate of students between the intervention and control groups (P ≥ 0.05). According to students' comments, the teaching of clinical science alongside basic science increased the importance of the topic and the students' interest in basic science. Most students agreed that this method prepares them for their clinical years. CONCLUSION: Many medical students believe that the content of many basic science courses are forgotten in the future. Therefore, basic science education alongside clinical science presentations are suggested.

In vitro effect of artemether-loaded nanostructured lipid carrier (NLC) on Leishmania infantum.

Visceral leishmaniasis (VL) is an acute and deadly form of leishmaniasis, caused by Leishmania infantum parasite. Due to the toxicity and side effects of conventional treatment options, such as glucantime and other pentavalent drugs, finding novel drugs with fewer adverse effects is required. Artemether (ART), is one of the derivatives of artemisinin, which was shown to be effective in treating malaria and more recently, leishmaniasis. In this fundamental-applied research, we compared the effect of ART and nanostructure loaded with artemether (NLC-ART) on Leishmania infantum promastigotes and amastigotes, at different concentrations (2.5-5-10-25-50-100 µg/ml) using the MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay method after 24 and 48 h of treatment. Inhibitory concentration (IC50) values (µg/ml) of promastigote and amastigote of L. infantum to ART/ NLC-ART, after 48 h of treatment, were found to be 37.12 / 32.1 and 16.43 / 15.42, respectively. Moreover, we found that (NLC-ART), had the lowest cytotoxicity against the J774 macrophage cell line. Conclusion: The NLC-ART can be a good candidate for the treatment of visceral leishmaniasis.

Association between Blastocystis sp. infection and immunocompromised patients: a systematic review and meta-analysis.

The significance of opportunistic infections in immunocompromised patients and the enigmatic pathogenicity of Blastocystis directed us to conduct the first global systematic review and meta-analysis on Blastocystis prevalence, odds ratios (ORs), and subtypes distribution in various immunocompromised patients (HIV/AIDS, cancer and hemodialysis patients, as well as transplant recipients). The systematic searching procedure was done in Web of Science, PubMed, Scopus, and Google Scholar databases for relevant published literature until November 11, 2020. Random-effects model was utilized to calculate the weighted estimates and 95% confidence intervals (95% CIs). The computed pooled prevalence of Blastocystis inferred from 118 papers (128 datasets) on immunocompromised patients was 10.3% (95% CI: 8.7-12.2%), with 16.1% (95% CI: 11.3-22.2%), 12.5% (95% CI: 8.5-18%), 8.4% (95 % CI: 6.6-10.6%), and 6% (95% CI: 2.6-13.3%) for hemodialysis patients, cancer patients, HIV/AIDS patients, and transplant recipients, respectively. Based on 50 case-control studies (54 datasets), the highest ORs were associated with cancer [2.81 (95% CI: 1.24-6.38, P = 0.013)] and hemodialysis patients [2.78 (95% CI: 1.19-6.48, P = 0.018)]. The most frequent subtype being found in immunocompromised patients was ST3 [41.7% (95% CI: 31.4-52.7%)], followed by ST1 [31.7% (95% CI: 23.2-41.8%)] and ST2 [23.1% (95% CI: 14.8-34.1%)]. Also, the weighted frequency of Blastocystis in various subgroups (publication year, WHO regions, geographical distribution, continents, and country income) was analyzed separately. In total, the results of the present meta-analysis highlighted that one's immunodeficiency status is probably associated with an increased Blastocystis infection, underpinning strict preventive measures to be taken.