niloofar taghipour

Many cancer patients suffer from irreparable complications under the influence of oxidative stress. The administration of effective supplements to control and inhibit the action of oxidative radicals has recently attracted the attention of doctors and researchers. In particular, zinc supplementation is one of the minerals most relevant to the human health because of its antioxidant properties. Zinc acts as a cofactor for important enzymes involved in the proper functioning of the antioxidant defense system. In addition, zinc protects cells from oxidative damage. By conducting the present study, the effect of zinc dietary supplements in improving the treatment process of multiple myeloma (MM) patients who underwent autologous bone - marrow transplantation was investigated in terms of oxidative radical changes.

This study was a double - blind, placebo - controlled clinical trial. The study participants were randomly divided into two groups of 20, zinc gluconate (Zn) and placebo. On days 0, +15, and +30 after transplantation, patients received three 30 mg zinc gluconate tablets or placebo daily. The serum levels of zinc and copper of the patients were measured before the intervention and on day 30 after transplantation. The change in NADPH oxidase 2 (Nox2) gene expression was measured by real - time PCR method. The activity of nitrite (Nitric Oxide) and malondialdehyde (Malondialdehyde) metabolites were measured with Thiobarbituric acid and Griess methods, respectively. Analysis of the findings and graphs were done using SPSS Version 27 software. Mann - Whitney test was used to measure the difference in the number of biochemical variables between the intervention group and the placebo group, since thedata was not distributed normally. GraphPad Prism 8 software and one-way ANOVA and unpaired t-test were used to measure the difference in gene expression changes.

During the nutritional questionnaire, it was observed that the placebo group had more zinc, and the estimated difference between the two groups was determined to be 1.236. The expression of NOX2 gene showed a significant decrease (P-value < 0.05) compared to the control group after 30 days. The MDA values and Griess test findings did not change significantly after zinc supplementation, but a decrease was seen in the zinc group compared to the placebo group (P-value > 0.05).

It seems that zinc supplementation is effective for controlling oxidative stress and inflammation.

Immune cells and their secreted cytokines are known as the first barrier against pathogens. Leishmania major as an intracellular protozoan produces anti-inflammatory cytokines that lead to proliferation and survival of the parasite in the macrophages. miRNAs are small non-coding RNA molecules that regulate mRNAs expression. We aimed to investigate the relationship between the expression of TGF-β and a bioinformatically candidate miRNA, in leishmaniasis as a model of TGF-β overexpression.

The miRNAs that target TGF-β -3´UTR were predicted and scored by bioinformatic tools. After cloning of TGF-β-3'UTR in psi-CHECK TM- 2 vector, targeting validation was confirmed using Luciferase assay. After miRNA mimic transfection, the expression of miR-27a, TGF-β, as well as Nitric Oxide concentration was evaluated.

miR-27a received the highest score for targeting TGF-β in bioinformatic predictions. Luciferase assay confirmed that miR-27a is targeting TGF-β-3'UTR, since miR-27a transfection decreased the luciferase activity. After miRNA transfection, TGF-β expression and Nitric Oxide concentration were declined in L. major infected macrophages.

Bioinformatic prediction, luciferase assay, and miRNA transfection results showed that miR-27a targets TGF-β. Since miRNA and cytokine-base therapies are developing in infectious diseases, finding and validating miRNAs targeting regulatory cytokines can be a novel strategy for controlling and treating leishmaniasis.

The great gerbil (Rhombomys opimus), is widely distributed in Asia and is a natural reservoir for zoonotic cutaneous leishmaniasis in many endemic areas, as well as Iran.

In this study, infection to Leishmania species was investigated by two methods, parasitological and molecular survey, in the small number of R. opimus collected from Jovain, a Zoonotic Cutaneous Leishmaniasis (ZCL) focus located in North East of Iran.

Parasitological observation showed infection in only one of five rodents. But, ITS2-Nested-PCR revealed Leishmania infection in three out of 5 gerbils, including the parasitological positive one. Based on the PCR amplified size, two cases of infections were Leishmania major and one Leishmania turanica, their sequences are accessible in GenBank. The results of sequence analysis were consistent with the results obtained based on the size of the PCR.

These findings re-confirm the important role of R. opimus in the natural circulation of Leishmania spp and indicate the need to be concerned about the disease in the study area.

The presence of ambient particulate matter (PM) poses more dangers to human health than that of other common air pollutants such as Carbon dioxide (Co2) and ozone.  Epidemiologic studies show a direct correlation between PM and the risk of respiratory and cardiovascular diseases. The immune system seems to play a critical role in the process of these diseases. The main goal of this study was to investigate the effect of Tehran particulate matter in two aerodynamic diameters (PM2.5 and PM10) on alveolar macrophages (AM) from C57/BL6 mice. To evaluate the inflammatory effects of PMs, cultured alveolar, and peritoneal macrophages were treated with PM2.5 and PM10 (concentrations of 5 µg/mL and 10 µg/mL). Tumor necrosis factor-alpha (TNF-α) and IL-10 (representatives of inflammatory and anti-inflammatory cytokines, respectively) were assessed in the culture supernatant by ELISA. Expression of arginase and inducible nitric oxide synthase (iNOS) genes was carried out by quantitative real-time PCR. Different functional types of cultured alveolar macrophages (M1, M2) were also determined in this study. Our results suggest that PM2.5 induces M1 inflammatory phenotype in comparison with PM10. We found Also, an increase in TNF-α and M1-related gene expression (iNOS), as well as a decrease in both IL-10 and M2 phenotype genes (Arginase). Moreover, a reduction in phagocytic capacity and increased apoptosis function of macrophage cells were detected. PM2.5 as a major component in hydrocarbons has a considerable effect on polarizing the alveolar macrophages to an inflammatory phenotype and eliciting lung inflammation in mice.

Different kinds of substances has been used for wound dressing, however, some disadvantages such as unsatisfactory mechanical stability, poor flexibility, severe shrinkage, low porosity, hard separation from the wound site, and non-antibacterial activity, has been reported. Over the last two decades, much effort has been made to find suitable biopolymer materials for wound healing applications. Chitosan has revealed various biological properties like biodegradable, biocompatible, non-toxic and non-allergenic, antibacterial effects thus can be used for the production of biofilms and nano-scaffolds. The poor solubility and thermal properties of chitosan restrict its widespread uses, but this polysaccharide is highly compatible with other biopolymers, and researchers are using this property to improve the limitations of chitosan and produce various types of chitosan-based hybrids materials. The purpose of this study is to provide an overview of various chitosan-based nanoscaffolds as wound healing dressings.

This narrative review was performed using ISI Web of Science, PubMed, SID, Scholar, Scopus, and Science Direct and articles published up to Jan 2020 were included. The keywords of chitosan, chitosan-based scaffolds, chitosan-based composite, and wound dressing were used.

Many researches have been accomplished to obtain chitosan-based scaffolds, including the construction of chitosan based blends and composite scaffolds and etc. The results of most of these researches showed positive effects of chitosan, and its nanocomposite scaffolds/biofilms in blood clotting, activated platelet activity, facilitated tissue regeneration and wound healing process.

The use of chitosan-based scaffolds is effective in biological dressings and wound healing. Futuristic and innovative approaches in chitosan derivatives and nanocomposites can lead to the preparation of suitable co-polymers and the production of wound dressings with the desired properties. the authors hope that this review will help for researchers.

Cutaneous Leishmaniasis (CL) is an emerging uncontrollable and neglected infectious disease worldwide including Iran. The aim of this study was to investigate the expression profile of apoptosis- related miRNA and its target gene in macrophages.

This study was carried out in the Department of Medical Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran from January 2016 to November 2018. Applying literature reviews, bioinformatics software, and microarray expression analysis, we selected miRNA-24-3p interfering in apoptosis pathway. The expression profile of this miRNA and target gene were investigated in Leishmania major (MRHO/IR/75/ER)-infected primary and RAW 264.7 macrophages (IBRC-C10072) compared with non-infected macrophages (control group) using quantitative Real-time PCR.

Results of bioinformatics analysis showed that miR-24-3p as anti-apoptotic miRNA inhibits pro-apoptotic genes (Caspases 3 and 7). Microarray expression data presented in Gene Expression Omnibus (GEO) revealed a significant difference in the expression level of selected miRNA and its target gene between two groups. QRT-PCR results showed that the expression of miR-24-3p was upregulated in L. major infectioned macrophages that approved the results of bioinformatics and microarray analysis.

Parasite can alter miRNAs expression pattern in the host cells to establish infection and its survival. Alteration in miRNAs levels likely plays an important role in regulating macrophage functions following L. major infection. These results could highlight current understanding and new insights concerning the gene expression in macrophages during leishmaniasis and will help to development of novel strategies for control and treatment of CL.

Fascioliasis is one of the most important food-borne worm disease caused by Fasciola sp. Parasitological diagnosis is more difficult due to the low parasite burden and a few eggs shedding of helminths. Therefore, it will be valuable to development of simple, fast and reliable diagnostic tests for detection of human and animal fascioliasis.

Infected liver collected from abattoir in Tehran, Iran in 2017. F. hepatica eggs were detached from the uterus of worms under a stereo microscope. Various numbers of eggs were spiked to 200 mgr. of negative feces samples. DNA was extracted and then target regions (nuclear IGS) were amplified by LAMP assay using six primers. Fecal specimens without egg and DNA of other helminths were used as negative controls. F. hepatica sample which confirmed by morphologic criteria and PCR- RFLP was used as positive control.

LAMP products by using SYBR Green I could detect even a single egg in fecal samples which was visible by change of color from orange to green. There was no cross amplification by other helminths including; Taenia saginata, Dicrosolium dendriticum and F. gigantica.

LAMP seems a rapid, sensitive, cost-effective technique for detection of human fascioliasis.

The fatal form of leishmaniasis is visceral form (VL), found in some of the countries in the world. Visceral leishmaniasis has been reported sporadically from all provinces in Iran, including Lorestan. This study aimed to characterize parasite species in DAT positive and some of the DAT negative human blood samples of Delfan district, Lorestan Province, central Iran.
Blood samples were collected from different geographical areas of Delfan. Serum was used for DAT test and remained part of molecular study. DNA was extracted by using DNG-plus extracted kit (Cinagen, Iran). Poly­merase chain reaction amplification of Leishmania kDNA and PCR-RFLP of ITS1 was done to identify Leishmania species. Some amplicons were sequenced, submitted to GenBank and analyzed by BLASTn.
Expected band of kDNA for L. infantum (720bp) was amplified in 16 out of 186 (8.6%) samples which showed previously anti-Leishmania antibody at different titers or were negative serologically. Using BLASTn, 93% similarity with L. infantum has been shown. The rDNA-ITS1 was amplified only in 9 samples (4.7%). RFLP pattern was similar to what expected for L. infantum.
A new emerging hypo-endemic focus, caused by L. infantum, is going to be established in Delphan District, Lorestan Province. Further studies on vector and reservoirs are necessary for the region and other parts of Lorestan Province.

Trichinellosis is an important and neglected foodborne zoonotic infectious disease in worldwide. The most human outbreaks in recent years have been related to consumption of wild boar meat. This cross-sectional study determined the prevalence of Trichinella spp. infections in hunted wild boars in northern Iran.
Thirty-five hunted wild boars were subjected in this study in 2015. All samples were examined by conventional artificial digestion method to detect of muscle larvae. Genomic DNA was extracted by phenol-chloroform method from isolated larvae. To identify the Trichinella species, a PCR-based method was applied using the internal transcribed spacer 2 (ITS2) and mitochondrial small-subunit ribosomal RNA (rRNA) gene sequences.
The overall prevalence of Trichinella spp. infection was 5.7% (2/35, 95%CI= 0-13.4). The mean larval burdens in two positive samples were 0.05 and 6 larvae per gr tissue muscle, respectively. The PCR reaction, using specific primers, yielded two 367 bp and 195 bp bands on agarose gel for ITS 2 and rrnS, respectively.
There is a hidden burden of Trichinella spp. infection in wild boar population in Iran. Moreover, T. britovi is the prevalent species circulating in wild boars of Iran. Therefore, education of the hunters and other consumers should be performed about the risk of consumption of raw or undercooked meat and meat products from wild boars.

Cryptosporidium species are recognized as important gastrointestinal pathogens. This study was conducted to identify the prevalence, clinical manifestations and genotyping of Cryptosporidium spp. in patients with gastrointestinal illnesses (GIs) in western Iran.
Overall, 1301 fecal samples were collected from patients with GIs referred to the 12 clinical laboratories in Nahavand County, west of Iran. Modified Ziehl-Neelsen staining method was used to identify the oocysts. DNA was extracted from positive samples and Cryptosporidium spp. were characterized by Nested PCR and sequence analysis of the 60-kDa glycoprotein (gp60) gene. Data analysis was performed using SPSS ver. 16.
Prevalence of cryptosporidiosis was 1.3% (17/1301). Cryptosporidium infection was significantly associated with vomiting and nausea (P=0.001, OR=0.013; CI 95%=0.004- 0.044), abdominal pain (P=0.018, OR=0.073; CI 95%=0.008- 0.633) and diarrhea (P=0.001, OR=0.092; CI 95%=0.023- 0.362). Of the 17 isolates typed, 11 belonged to the C. parvum IId subtype family (subtypes IIdA26G1 and IIdA20G1) and six belonged to the C. parvum IIa subtype family (subtypes IIaA15G2R1 and IIaA16G3R1). There was no significant difference between subtype families IIa and IId in occurrence of clinical symptoms (P= 0.75).
Improved hygiene and avoidance of contact with animals and contaminated soil should be advocated to reduce the occurrence of Cryptosporidium infections, especially in children.

Cutaneous leishmaniasis (CL) is one of the most important neglected tropical diseases and a major public health challenge in Iran caused by Leishmania spp and transmitted by phlebotomine sand flies. The number of CL cases has shown an increasing pattern all over the country, including the district of Varamin, southeast of Tehran, Iran. This study aimed to identify the Leishmania spp isolated from CL patients using molecular methods in Varamin during 2012–2013.
Exudate materials collected from the swollen edge of the skin lesions of 44 parasitological positive CL patients by disposable lancet. They were referred to Varamin Health Center by physician. The samples were sub­jected to molecular method for Leishmania species identification.
The digestion pattern of restriction enzyme revealed that 37 (84.1%) CL patients were infected with L. ma­jor and 7 (15.9%) were infected with L. tropica. They were mostly male than female. More than half of the patients (58%) had multiple lesions, and they were mostly observed on extremities, 34.1% on legs and 29.5% on hands. Le­sions were mostly of wet ulcerative type.
Dominancy of L. major provides more evidence that Varamin District probably could be considered as Zoonotic Cutaneous Leishmaniasis (ZCL) areas. More investigation on other epidemiological aspects of disease is needed.

Leishmaniasis is a zoonotic disease caused by species of protozoa of the genus Leishmania. In recent years, incidence of cutaneous leishmaniasis has increasing trend in Golestan Province, North of Iran. The aim of the present study was to identify the frequency of cutaneous leishmaniasis using PCR-RFLP in patients referred to Kalaleh Health Center, during 2013-14.
This descriptive cross-sectional study was conducted on 70 individuals with suspected cutaneous leishmaniasis that referred to health center of Kalaleh County, Golestan Province, Northern Iran, from Sep 2013 to Nov 2014. Samples of cutaneous lesions were examined microscopically. DNA was extracted from all of the positive smears and PCR was done on ITS-1 gene. RFLP was performed using HaeIII enzyme for species identification.
Totally, 38 out of the 70 (54.3%) suspected individuals including 22 males (57.9%) were found positive by microscopic examination. All of microscopically positive samples were confirmed to be positive for Leishmania DNA (approximately 340 bp bands were detected). RFLP revealed 140 bp and 200 bp bands (approximate size), indicative of L. major.
The detected species of studied region was L. major. Cutaneous leishmaniasis has high prevalence in Kalaleh County, thus more studies on leishmaniasis in the animal reservoirs, comparison of homology of animal and human isolates and a survey regarding natural infection of vectors in this region is highly recommended.

The purpose of this study is to model Cryptosporidiosis in laboratory animals. The parasites were inoculated into animalsand thenmultiplied. The process of proliferation was compared to controlCryptosporidiosis in humans.
Twenty-five laboratory mice (4-7 days of age) and twenty-five laboratory rats (5 days of age) were assigned to the category I while the category II (control group) consisted oftwenty-fiverats and twenty-five mice. The two categories were infected with 5×105 Cryptosporidium parvum oocysts originated from a calf by using a 24-gauge & 20 gauge ball-point feeding needle. On 4-9 days of post inoculation the intestine, colon, and rectum were removed. Cryptosporidium infection was determined by detecting oocysts in intestinal homogenates by Staining and PCR method. Simple extraction and purification method was used by ficoll gradient centrifugation. Also, twenty laboratory rats (4-6 weeks of age) were intramuscularly injected with dexamethasone(Sigma, Chemical Co. UK) two times per week, and the last injection was given with 5×105 Cryptosporidium parvum oocyst on the same day as oral inoculation. The water was supplemented with tetracycline to avoiding secondary infections.
Two to four million purified oocysts with a maximum of 10 million were routinely obtained per mouse and rat. Also the day in which oocyst excretion is the highest was determined. The number of oocyst per neonatal mouse was (11±2)×105 on 9-12 days of post infection while similarly it was (10±1)×105 per neonatal rat.
The evaluation of the cryptosporidiosis in immunocompromised animal models can help us to understand and control the Cryptosporidium infections.

To induce acute colitis progresses to chronicity in C57BL/6 mice by dextran sulfate sodium. Murine models are essential tools to understand IBD pathogenesis. Among different types of chemically induced colitis models, the dextran sulfate sodium (DSS)-induced colitis model is the most common model of IBD, due to its simplicity. Patients and Male C57BL/6 mice 6–8 weeks old, were collected and matched by age with controls. C57BL/6 mice treated with 2 cycles of 3.5% DSS for 4 days and 4 days of pure water between each cycle. After that, mice were sacrificed and the entire colon was removed. Small sections of the colon were fixed in formaldehyde, embedded in paraffin and sectioned with a microtome. Sections were stained with hematoxylin eosin to analyses the degree of inflammation. After the first cycle oral administration of DSS, mice with severe and visible rectal bleeding and diarrhea entered into the acute phase. After day 4-5, bleeding and diarrhea were improved and mice entered into the chronic phase with peak levels of weight loss. Macroscopically, the inflammation was predominantly located in the distal colon. Microscopically, examination of the distal colon sections showed a decrease number of goblet cells, loss of crypts, signs of surface epithelial regeneration and moderate to severe infiltration of inflammatory cells in the mucosa. In order to achieve an experimental colitis model, our protocol is recommended for future therapies in IBD experimental modeling.

Carboxy-terminal 42 kDa region of Plasmodium vivax merozoite surface protein-1 is considered as an important antigen in blood stage. Since, this region has been observed to be polymorphic among isolates of P. vivax, it is signifi­cant to survey on different regions of this antigen in various areas of the world. In the present study, the genetic diversity of cloned PvMSP-142 kDa gene from an Iranian patient is analyzed. Parasite DNA was extracted from a P. vivax - infected patient in Iran. The region of PvMSP-142 kDa was amplified by PCR, cloned into pTZ57R/T vector and then sequenced. Sequencing of cloned PvMSP-142 kDa gene clearly has a high degree of homology (95%) with reference Sal-I sequence and also with the homogeneous sequences from some studied countries (97%). 38 SNPs (single nucleotide polymor­phism) were identified in cloned PvMSP-142 kDa gene which the muta­tions had localized in the 33 kDa fragment (PvMSP-133 kDa), while there was nearly no variation in the 19 kDa fragment (PvMSP-119 kDa). 2 out of 38 muta­tions were found as to be novel haplotypes. High similarity of cloned PvMSP-142 kDa gene in comparison to reference sequence and other sequences could be beneficial as a remarkable molecu­lar marker for serological diagnostic kits of P. vivax in malarious neighbor­ing countries of Iran and around the world.

Entamoeba moshkovskii and E. dispar are impossible to differenti­ate microscopically from the pathogenic species E. histolytica. Multiplex polymer­ase chain reaction (Multiplex PCR) is a widespread molecular biology technique for amplification of multiple targets in a single PCR experiment. For detection and differentiation of the three-microscopy indistinguish­able Entamoeba species in human, multiplex PCR assay using differ­ent DNA extraction methods was studied. A conserved forward primer was derived from the middle of the small-subunit rRNA gene, and reverse primers were designed from signature sequences specific to each of these three Entamoeba species. A 166-bp PCR product with E. histolytica DNA, a 580-bp product with E. moshkovskii DNA and a 752-bp product with E. dispar DNA were gener­ated in a single-round and multiplex PCR reaction. We recommend this PCR assay as an accurate, rapid, and effec­tive diagnostic method for the detection and discrimination of these three Enta­moeba species in both routine diagnosis of amoebiasis and epidemiological surveys.

The aim of the present study was to determine the species and genotypes of Cryptosporidium spp. among children with diarrhea by PCR- RFLP using the TRAP-C2 gene. Cryptosporidium is a globally distributed protozoan parasite and one of the most common causes of infection and diarrhea in humans.Patients and Four hundred and sixty nine stool samples were collected from children less than 12 years with diarrhea who had been referred to Pediatrics Medical Centers in Gazvin provinces. The presence of Cryptosporidium oocysts was determined by Ziehl-Neelsen acid fast staining, then, genomic DNA was extracted from positive samples and nested PCR-RFLP was performed to amplify the TRAP-C2 gene. The overall prevalence of Cryptosporidium infection in children was 2.5 %. Results of nested PCR amplification showed that of 12 positive children samples, 10 (83.3%) were belonged to C. parvum, followed by C. hominis in 1 (8.3%) and mixed infection in 1 isolate (8.3%). This study showed that Cryptosporidium parvum (the zoonotic genotypes) is more prevalent than other Cryptosporidium species in children from this area. This suggests that zoonotic transmission is the main mode of transmission of Cryptosporidium infection in Iran

Backgraound: Entamoeba histolytica antigenic markers such as Serine-Rich E. histolytica protein (SREHP) have recently been used for vaccine preparation, genetic diversity studies of Entamoeba histolytica isolates and for differentiation between E. histolytica and E. dispar species. This study was carried out with the aim of expression of a recombinant Serine Rich E. histolytica protein in the laboratory to use it in the ELISA kit.

In this study which is an exploration method, an Iranian isolate of Serine-Rich E. histolytica gene which had previously been cloned in bluescript plasmid (pBSc), was cut using BamHI restriction enzyme. After extracting and purification from gel, the SREHP gene was sub cloned into pET32a expression vector. The inserted gene was confirmed with Rosconis solution, PCR and sequencing methods. PCR was performed with the SREHP specific primers as well as pET T7 promoter primer. The cloned gene was also digested with HindIII and BamHI restriction enzymes. Recombinant plasmid was conveyed to competent cell BL21 (DE3). A colony of the plasmid including SREHP gene was cultivated and induced with IPTG. The result of expressed protein was observed on the SDS-PAGE gel. The SREHP gene was sub cloned into pET32a expression vector. A recombinant plasmid including an inserted SREHP gene was screened and confirmed with quick check method using Ruscoins solution, as well as PCR by special primers (SREHP and universal pET primer), digested with BamHI and HindIII restriction enzymes. Finally an open reading frame of 666 nucleotides from inserted SREHP gene was obtained with the sequencing method.

The recombinant protein of Serine-Rich E. histolytica in presence of IPTG was expressed in five hours and the result of expressed protein in the length of 44 KDa was observed on SDS-PAGE gel.

SREHP protein was successfully cloned and expressed in this study. However additional studies are recommended for preparation and purification of the SREHP in a large quantity and the using it for the ELISA test.

Cutaneous Leishmaniasis is endemic in plenty of Iranian provinces. This study aimed to determine the epidemiological status of the cutaneous Leishmaniasis outbreak, isolation and identification of the parasite using a PCR method in burden rural areas of Gonbad-e-Qabus County, north Iran. Data was collected on the prevalence of scars and ulcers over a period of three months among 6990 inhabitants of five villages around Gonbad-e-Qabus county, during 2006-2007