In Vitro Study of the Leishmanicidal Activity of Perovskia Abrotanoides Terpenoid-Rich Fractions Against Leishmania Major (MRHO/IR/75/ER).

Background: Cutaneous leishmaniasis (CL) is an ulcerative skin disease caused by some species of the genus Leishmania. Evidence shows that Perovskia abrotanoides is an important herbal medicine against Leishmania. This study was conducted to investigate the killing effect of terpenoid-rich fractions on promastigotes of L. major (MRHO/IR/75/ER). Material and Method: The eluates of reverse phased medium pressure liquid chromatography (RP-MPLC) of the extract were subjected to thin-layer chromatography (TLC) and categorized into six final fractions. Primary proton nuclear magnetic resonance (H-NMR) spectroscopy confirmed fractions' nature. Fractions 4, 5, and 6 (F4, F5, F6) were identified as terpenoid-rich content. Two concentrations of 50 and 100 µg/ml were prepared to test leishmanicidal activity. Followed by treating promastigotes of L. major by the fractions in incubation times of 12, 24, and 48 hours, their viability was determined using a cell proliferation MTS ((3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay. Result: F4, F5, and F6 showed significant killing activity on promastigotes of L. major in a concentration-dependent manner. The viability of promastigotes was significantly reduced at a concentration of 100 µg/ml compared to 50 µg/ml (P-value <0.05). Also, over time a significant decreasing trend in the viability of promastigotes confirmed the time-dependent manner of the fractions (P-value <0.01). Furthermore, F5 had the highest leishmanicidal activity at the first incubation time compared with other fractions. Conclusion: Terpenoid-rich fractions of the P. abrotanoides have a leishmanicidal activity that depends on time and concentration. Among them, F5 has the highest potency that may contain potent terpenoid constituents.

Wetting dynamics in two-liquid systems: Effect of the surrounding phase viscosity.

This paper reports the experimental results of a water droplet spreading on a glass substrate submerged in an oil phase. The radius of the wetted area grows exponentially over time forming two distinct regimes. The early time dynamics of wetting is characterized with the time exponent of 1, referred to as the viscous regime, which is ultimately transitioned to the Tanner's regime with the time exponent of 0.1. It is revealed that an increase in the ambient phase viscosity over three decades considerably slows down the rate of three-phase contact line movement. A scaling law is developed where the three-phase contact line velocity is a function of both spreading radius and mean viscosity, close to the geometric mean of the droplet and ambient fluids' viscosities. Using the proposed scaling and mean viscosity, all plots of spreading radius for different viscosity ratios collapse to a master curve. Furthermore, several cases with multiple rupture and spreading points, i.e., wetting in a nonideal system, are considered. The growth of an equivalent wetting radius in a multiple point spreading system is predicted by the developed scaling law.

A wide diversity of zoonotic intestinal parasites in domestic and stray dogs in rural areas of Kermanshah province, Iran.

Dogs can act as reservoirs, carriers, and transmitters of several zoonotic intestinal parasites that can cause serious health problems for humans. The aim of this study was to determine the prevalence of intestinal parasites in dogs in Kermanshah Province, west of Iran. Faecal samples were collected from domestic and stray dogs from 30 rural areas of Kermanshah province from August 2014 to April 2015 and were analyzed by formalin-ether sedimentation, sucrose otation technique and the modied Ziehl-Neelsen method. Out of 301 dogs examined, 230 (76.4%) were infected with at least one parasite. The incidence on the different types of intestinal parasitic species recovered from domestic and stray dogs are as follows: Toxocara leonina (20.8% and 27.6%), T. canis (7.5% and 9.4%), Taenia spp. (9.2% and 9.4%), hookworm spp. (18.3% and 33.7%), Capillaria spp. (0.8 and 1.7%), Dicrocoelium dendriticum (0.8% and 3.3%), Fasciola spp. (0.8% and 2.2%), Acanthocephal spp. (3.3% and 5.5%), Trichuris vulpis (0.8% and 1.7%), Dipylidium caninum (4.2% and 3.3%), Physaloptera spp. (6.7% and 6.6%), Cryptosporidium spp.(21.7% and 25.4%), Eimeria spp. (35.0% and 34.3%), Giardia spp. (6.7% and 12.7%), Cystoisospora spp. (7.5% and 5.5%), Blastocystis spp. (18.3% and 20.4%) and Sarcocystis spp. (6.7% and 7.2%), respectively. Signicant difference in infection rates was observed between domestic and stray dogs (P>0.05). Hookworm and Eimeria spp. were the most common intestinal helminth and protozoa detected with 83 (27.6%) out of 301 and 104 (34.6%), out of 301, respectively. There was no significant difference in prevalence and distribution of intestinal parasites between male and female dogs (P>0.05). The wide range of zoonotic parasites indicated that people residing in rural areas in Kermanshah province are at risk of exposure to these pathogens. In this respect, appropriate implementation of control programs by public health authorities and veterinarians should be taken into account.

Analysis of the cox1 gene in Echinococcus granulosus from sheep in northeast Iran using PCR high-resolution melting (qPCR-HRM) curve analysis.

Echinococcus granulosus, the etiologic agent of echinococcosis, is one of the most important zoonotic helminthes worldwide. Knowledge of E. granulosus species and genotypes has important implications for epidemiology, control, and prevention of diseases as well as future vaccine and drug designs. There are many molecular methods developed to define genotypes of E. granulosus, among them high resolution melting (HRM) analysis, as a new approach, is a single step and closed tube method. It is appropriate for fast screening of large number of isolates. This technique is an accurate, user friendly, cost-effective, fast and simple method, which does not need post-PCR processes. Between March and lst august 2016, of 726 sheep examined in abattoirs in Razavi Khorasan province, Northeast Iran, 109 harboured cystic echincoccosis lesions (liver samples= 65 and lung samples= 44) which were collected for analysis. Total genomic DNA was extracted from each sample and amplified for the presence of polymorphism in the mitochondrial cox1 gene of Echinococcus granulosus using a high resolution melting curve (HRM) method. A total of 109 hydatid cyst samples analyzed by PCR high-resolution melting (qPCR-HRM) curve of the cox1 gene, all isolates were identified as G1 genotype (sheep strain). G1 is the predominant genotype in sheep in northeast of Iran. The high incidence of the G1 genotype (known to be the predominant E. granulosus genotype infecting humans globally) in sheep has considerable implications for hydatid disease control programs in this area.

Geo-material surface modification of microchips using layer-by-layer (LbL) assembly for subsurface energy and environmental applications.

A key constraint in the application of microfluidic technology to subsurface flow and transport processes is the surface discrepancy between microchips and the actual rocks/soils. This research employs a novel layer-by-layer (LbL) assembly technology to produce rock-forming mineral coatings on microchip surfaces. The outcome of the work is a series of 'surface-mimetic micro-reservoirs (SMMR)' that represent multi-scales and multi-types of natural rocks/soils. For demonstration, the clay pores of sandstones and mudrocks are reconstructed by representatively coating montmorillonite and kaolinite in polydimethylsiloxane (PDMS) microchips in a wide range of channel sizes (width of 10-250 µm, depth of 40-100 µm) and on glass substrates. The morphological and structural properties of mineral coatings are characterized using a scanning electron microscope (SEM), optical microscope and profilometer. The coating stability is tested by dynamic flooding experiments. The surface wettability is characterized by measuring mineral oil-water contact angles. The results demonstrate the formation of nano- to micro-scale, fully-covered and stable mineral surfaces with varying wetting properties. There is an opportunity to use this work in the development of microfluidic technology-based applications for subsurface energy and environmental research.

Evaluation of a novel lentiviral vaccine expressing KMP11-HASPB fusion protein against Leishmania infantum in BALB/c mice.

Hydrophilic acylated surface protein B (HASPB) is an immunogenic Leishmania protein against which antibodies are produced in the sera of cutaneous and visceral Leishmaniasis (VL) patients. Kinetoplastid membrane protein 11 (KMP11) is another protein antigen of Leishmania which is reported as a promising candidate for vaccination of VL. It is a highly conserved surface protein present in all members of kinetoplastid family and is expressed in both promastigotes and amastigotes. In this study, the coding sequence of KMP11 and HASPB was cloned into a pCDH-cGFP lentiviral vector as a fusion protein. The gene expression was confirmed using RT-PCR and Western blot methods. After injection of the recombinant KMP11-HASPB-expressing lentiviruses to BALB/c mice, using ELISA technique, a significant increase in IFN-γ and IL-4 as well as IgG1 and IgG2a was observed compared to the control group. Furthermore, the number of parasites in the liver and spleen of vaccinated mice decreased significantly compared with the control group.

PCR-RFLP diagnosis and characterization of Leishmania species causing human cutaneous leishmaniasis and evaluation of treatment times with glucantime in these patients.

Antimony compounds are first line treatments for cutaneous leishmaniasis. The prognosis of the disease varies depending on the type of medicine and species. We aimed to determine the species responsible for cutaneous leishmaniasis in patients referred to Skin Diseases and Leishmaniasis Research Center in Isfahan and Bam Health Center (Kerman) in order to follow and assess the complete healing of the lesions. A total of 40 skin lesions samples were collected from patients with cutaneous leishmaniasis (CL) form January 2014 to 2015. Dermal scrapings were analyzed by examination of Giemsa-stained smears. Parasites were cultured and isolated in NNN and RPMI 1640 medium and DNA was extracted. We used PCR-RFLP assays of ITS1 genes for direct identification of Leishmania species. Treatment process was assessed after a treatment period with glucantime and healing of the studied cases was followed up. All the samples from Isfahan and Bam regions were L. major and L. tropica species respectively. In patients infected with L. major and L. tropica treated with glucantime, the shortest healing period was 40 days in 5(25%) and 60 days in 3(15.8%) patients, respectively and the longest healing period was 100 days in 1 (5%) and 160 days in 1 (5.3%) patient, respectively. The mean complete healing periods in patients with L. tropica and L. major were 100 and 58 days, respectively (P<0.001). Average recovery period for people with dry cutaneous leishmaniasis is longer than average recovery period for people with wet cutaeous leishmaniasis.

TLR2 and TLR4 in cutaneous leishmaniasis caused by Leishmania major.

Cutaneous leishmaniasis (CL) is a self-healing skin disease which rarely for unknown reason(s) the lesion develops to a non-healing form. It seems that the initial contact of Leishmania parasites with the host innate immune system is an important step in the outcome of the disease. Recent studies suggested that toll-like receptors (TLRs) play a role in Leishmania recognition. In this study, the level of TLR2 and TLR4 was checked in patients with healing form of lesion and compared with that of patients with non-healing form of lesion caused by Leishmania major. Gene expression of TLR2 and TLR4 in peripheral blood-derived macrophages, before and after stimulation with live L. major promastigotes, was evaluated using quantitative real-time reverse transcription PCR and flow cytometry. The results showed that the mean relative gene expression and difference membrane expression of TLR2 in macrophages of patients with healing form of lesion were significantly higher than patients with non-healing form of lesion (P < 0.0001 and P = 0.0034), respectively, and the mean relative gene expression and difference in protein expression of TLR4 in macrophages of patients with healing form of lesion were significantly higher than that of patients with non-healing form of lesion (P = 0.021 and P = 0.002), respectively. The data suggested a possible role for TLR2 and TLR4 in the outcome of CL lesion. Further studies are needed to understand more about the detail role of the immune factors in leishmaniasis.

Hydrodynamic instability in the transport of miscible reactive slices through porous media.

Miscible displacement of a slice of a pollutant solution by a carrier solution in homogeneous porous media is examined. The carrier solution reacts with the slice solution to generate a chemical product, and as a result of differences in viscosities of the three species, a hydrodynamic instability known as viscous fingering is observed. The dynamics of the instability and the rate of consumption as well as spread of the pollutant are examined through numerical simulations. The study shows that the rate of consumption of the pollutant is the highest when the chemical product is the most or the least viscous solution in the system. It was also found that displacements in which the pollutant viscosity is the smallest or the largest of all three species lead to the widest spread of the pollutant in the porous media. In addition, the most complex finger structures are observed when the carrier solution has the smallest or largest viscosity in the flow. Furthermore, a mechanism of channeling whereby the carrier is able to break through the slice, therefore bypassing the pollutant, is found in cases where the chemical product is more viscous than the carrier solution. The dynamics of the displacement are analyzed and physical interpretations of their development are presented.

Karyosystematic and morphometric characterization of the rodents as reservoir hosts of zoonotic cutaneous leishmaniasis in an endemic focus of Isfahan Province, Iran.

BACKGROUND & OBJECTIVES: Rodents belonging to Gerbillinae subfamily are the main reservoir hosts of zoonotic cutaneous leishmaniasis (ZCL) in Iran. Regarding the important role of these rodents in the maintenance of Leishmania major in the nature, their identification with morphometric, cytogenetic and molecular methods seems to be essential. The karyotype study of these species, captured from a new focus of zoonotic cutaneous leishmaniasis located in the south of Isfahan Province was carried out in 2007. METHODS: Twenty specimens containing seventeen Meriones persicus and three Nesokia indica were captured from Mobarakeh rural district south of Isfahan. Giemsa-stained karyotypes of these two species were prepared from bone marrow chromosome preparations. Systematic important characters of the body and cranium (incisors, molars, occipitonasal, condylobasal, zygomatic, tympanic bullae, etc.) of these rodents were studied. Cranium size was measured using a Vernier calipers. RESULTS: Specimens of M. persicus and N. indica had 2n = 42. The karyotype study of these species included metacentric, sub-metacentric and acrocentric chromosomes. Morphological studies were completely matched with the reported characters of these species and further confirmed the diagnoses. INTERPRETATION & CONCLUSION: Based on the results of this study, M. persicus and N. indica are two completely differentiated rodents species that were collected from a new focus and they can also be differentiated morphologically.

Distribution frequency of pathogenic bacteria isolated from cutaneus leishmaniasis lesions.

Cutaneous leishmaniasis (CL) is a parasitic disease characterized by single or multiple ulcerations. Secondary bacterial infections are one of the complications that can increase the tissue destruction and the resulting scar. To better determine the incidence of real secondary bacterial infections in CL, we designed the current study. This was a cross-sectional study performed in Skin Diseases and Leishmaniasis Research Centre, Isfahan, Iran. A total of 1,255 patients with confirmed CL enrolled in the study. Sterile swaps were achieved for ulcer exudates and scraping was used for non-ulcerated lesions. All samples were transferred to tryptic soy broth medium. After 24 hr of incubation at 37 degrees C they were transferred to eosin methylene blue agar (EMB) and blood agar. Laboratory tests were used to determine the species of bacteria. Among 1,255 confirmed CL patients, 274 (21.8%) had positive cultures for secondary bacterial infections. The bacteria isolated from the lesions were Staphylococcus aureus in 190 cases (69.3%), coagulase negative Staphylococcus in 63 cases (23.0%), E. coli in 10 cases (3.6%), Proteus sp. in 6 cases (2.2%), and Klebsiella sp. in 5 cases (1.9%). The results show that the overall incidence of secondary bacterial infections in the lesions of CL was 21.8%, considerably high. The incidence of secondary bacterial infections was significantly higher in ulcerated lesions compared with non-ulcerated lesions.