Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 6066

Search results for: Leishmania major

6066 Infection of Phlebotomus Sergenti with Leishmania Tropica in a Classical Focus of Leishmania Major in Tunisia

Authors: Kaouther Jaouadi, Jihene Bettaieb, Amira Bennour, Ghassen Kharroubi, Sadok Salem, Afif Ben Salah

Abstract:

In Tunisia, chronic cutaneous leishmaniasis due to Leishmania (L) tropica is an important health problem. Its spreading has not been fully elucidated. Information on sandfly vectors, as well as their associated Leishmania species, is of paramount importance since vector dispersion is one of the major factors responsible for pathogen dissemination. In total, 650 sandflies were captured between June and August 2015 using sticky paper traps in the governorate of Sidi Bouzid, a classical focus of L. major in the Central-West of Tunisia. Polymerase chain reaction-restriction fragment length polymorphism analysis of the internal transcribed spacer 1 and sequencing were used for Leishmania detection and identification. Ninety-seven unfed females were tested for the presence of Leishmania parasite DNA. Six Phlebotomus sergenti were found positive for L. tropica. This finding enhances the understanding of the cycle extension of L. tropica outside its original focus of Tataouine in the South-East of the country.

Keywords: cutaneous leishmaniasis, Leishmania tropica, sandflies, Tunisia

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6065 Green Synthesis of Nano Liposomes Containing Berberine Chlorideagainst Leishmania major

Authors: Ali Fattahi Bafghi, Abolghasem Siyadatpanah, Farzaneh Mirzaei, Fahimeh Pournasir, Roghayeh Norouzi, Maria De Lourdes Pereira

Abstract:

Leishmaniasis caused by Leishmania major is one of the main infectious diseases that affect populations in developing countries around the world. We assessed the effectiveness of berberine chloride nano-liposome (BcNLs) against L. major promastigotes in vitro. Nano-liposomal berberine chloride was prepared using the thin-film hydration method and characterized based on encapsulation efficiency, size, and zeta potential. Anti-Leishmania effect of different concentrations (0.05-60 µg/ml) of BcNLs as studied in L. major [MRHO/IR/75/ER] at 24, 48, and 72 h using the hemocytometer technique. Berberine chloride was successfully loaded into nano-liposomes with an encapsulation efficiency of 85.54%. The surface charge of nanoparticles is neutral, and the morphology of nano-liposomal berberine chloride is spherical without any agglomeration. Cell viability assay was performed on the HFF cell line to show the biocompatibility of liposome nanoparticles. IC50 of BcNPs at 24, 48, and 72 h against L. major were found to be 7.6, 5.96, and 3.19 µg/ml, respectively. BcNLs showed a significant anti-Leishmania effect and induced a better and more tangible effect on the survival of L. major promastigotes and could be suitable candidates for further investigation. The results showed that the BcNLs agent is effective against L. major promastigotes and may be a promising alternative to current treatments.

Keywords: Leishmania major, berberine chloride, nano-liposomes, cutaneous leishmaniasis

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6064 Detection of Leishmania Mixed Infection from Phlebotomus papatasi in Central Iran

Authors: Nassibeh Hosseini-Vasoukolaei, Amir Ahmad Akhavan, Mahmood Jeddi-Tehrani, Ali Khamesipour, Mohammad Reza Yaghoobi Ershadi, Kamhawi Shaden, Valenzuela Jesus, Hossein Mirhendi, Mohammad Hossein Arandian

Abstract:

Zoonotic cutaneous leishmaniasis (ZCL) is an endemic disease in many rural areas of Iran. Sand flies were collected from rural areas of Esfahan province and were identified using valid identification keys. DNA was extracted from sand flies and Nested PCRs were done using specific primers. In this study, 44 out of 152 (28.9 %) sand flies were infected with L. majoralone. Eight sand flies showed mixed infection: four sand flies (2.6 %) were infected with L. major, L. turanicaand L. gerbili, one sand fly (0.7 %) was infected with L. major and L. turanica and three sand flies (2 %) were infected with L. turanicaand L. gerbili. Our results demonstrate the natural infection of P. papatasi sand fly with three species of L. major, L. turanica and L. gerbili which are circulating among R. opimusreservoir host and P. papatasi sand fly vector in central Iran.

Keywords: Phlebotomus papatasi, Leishmania major, Leishmania turanica, Leishmania gerbili, mixed infection, Iran

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6063 Epidemiology, Prevention and Treatment of Leishmaniasis in Afghanistan

Authors: Mohammad Reza Mohammadi, Layegheh Daliri

Abstract:

Introduction: Leishmaniasis occurs in infectious diseases of Leishmania protozoa in Afghanistan, anthroponotic leishmaniasis and common cutaneous leishmaniasis (ZCL). Anthroponotic skin leishmania tropica may cause urban diseases and transmitted by Phlebotomus Sergenti. In different parts of Afghanistan, different species of Leishmania are observed. We report the epidemiological characteristics of prevention and treatment in this study. Methods: This study examines the epidemiology and prevention of religious diseases in Afghanistan. Knowledge gaps were analyzed and collected with our own data. Results: In Afghanistan, most of the Lishmania Tropic seekers are Four species of Leishmania in northern Afghanistan, including Leishmania Tropica, L. Major and L. Donovani, cause skin lesions, but L. Donovani and L. infantum are visible. Even combined prevention can significantly reduce the amount of infection. Conclusion: Skinny, as well as visceral leishmaniasis, can occur among the returnees from Afghanistan. Unusual and poor skin lesions can be created by L. Donovani. In most pathogenic areas, the transmission of common diseases between humans and animals. Home dogs are the main reservoir, transferring in some areas such as India and Sudan.

Keywords: leishmania donovani, leishmania tropica, treatment, disease, epidemiology

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6062 A Comprehensive Analysis of LACK (Leishmania Homologue of Receptors for Activated C Kinase) in the Context of Visceral Leishmaniasis

Authors: Sukrat Sinha, Abhay Kumar, Shanthy Sundaram

Abstract:

The Leishmania homologue of activated C kinase (LACK) is known T cell epitope from soluble Leishmania antigens (SLA) that confers protection against Leishmania challenge. This antigen has been found to be highly conserved among Leishmania strains. LACK has been shown to be protective against L. donovani challenge. A comprehensive analysis of several LACK sequences was completed. The analysis shows a high level of conservation, lower variability and higher antigenicity in specific portions of the LACK protein. This information provides insights for the potential consideration of LACK as a putative candidate in the context of visceral Leishmaniasis vaccine target.

Keywords: bioinformatics, genome assembly, leishmania activated protein kinase c (lack), next-generation sequencing

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6061 Ribosomal Protein S4 Gene: Exploring the Presence in Syrian Strain of Leishmania Tropica Genome, Sequencing it and Evaluating Immune Response of pCI-S4 DNA Vaccine

Authors: Alyaa Abdlwahab

Abstract:

Cutaneous leishmaniasis represents a serious health problem in Syria; this problem has become noticeably aggravated after the civil war in the country. Leishmania tropica parasite is the main cause of cutaneous leishmaniasis in Syria. In order to control the disease, we need an effective vaccine against leishmania parasite. DNA vaccination remains one of the favorable approaches that have been used to face cutaneous leishmaniasis. Ribosomal protein S4 is responsible for important roles in Leishmania parasite life. DNA vaccine based on S4 gene has been used against infections by many species of Leishmania parasite but leishmania tropica parasite, so this gene represents a good candidate for DNA vaccine construction. After proving the existence of ribosomal protein S4 gene in a Syrian strain of Leishmania tropica (LCED Syrian 01), sequencing it and cloning it into pCI plasmid, BALB/C mice were inoculated with pCI-S4 DNA vaccine. The immune response was determined by monitoring the lesion progression in inoculated BALB/C mice for six weeks after challenging mice with Leishmania tropica (LCED Syrian 01) parasites. IL-12, IFN-γ, and IL-4 were quantified in draining lymph nodes (DLNa) of the immunized BALB/C mice by using the RT-qPCR technique. The parasite burden was calculated in the final week for the footpad lesion and the DLNs of the mice. This study proved the existence and the expression of the ribosomal protein S4 gene in Leishmania tropica (LCED Syrian 01) promastigotes. The sequence of ribosomal protein cDNA S4 gene was determined and published in Genbank; the gene size was 822 bp. Expression was also demonstrated at the level of cDNA. Also, this study revealed that pCI-S4 DNA vaccine induces TH1\TH2 response in immunized mice; this response prevents partially developing a dermal lesion of Leishmania.

Keywords: ribosomal protein S4, DNA vaccine, Leishmania tropica, BALB\c

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6060 Insight into Structure and Functions of of Acyl CoA Binding Protein of Leishmania major

Authors: Rohit Singh Dangi, Ravi Kant Pal, Monica Sundd

Abstract:

Acyl-CoA binding protein (ACBP) is a housekeeping protein which functions as an intracellular carrier of acyl-CoA esters. Given the fact that the amastigote stage (blood stage) of Leishmania depends largely on fatty acids as the energy source, of which a large part is derived from its host, these proteins might have an important role in its survival. In Leishmania major, genome sequencing suggests the presence of six ACBPs, whose function remains largely unknown. For functional and structural characterization, one of the ACBP genes was cloned, and the protein was expressed and purified heterologously. Acyl-CoA ester binding and stoichiometry were analyzed by isothermal titration calorimetry and Dynamic light scattering. Our results shed light on high affinity of ACBP towards longer acyl-CoA esters, such as myristoyl-CoA to arachidonoyl-CoA with single binding site. To understand the binding mechanism & dynamics, Nuclear magnetic resonance assignments of this protein are being done. The protein's crystal structure was determined at 1.5Å resolution and revealed a classical topology for ACBP, containing four alpha-helical bundles. In the binding pocket, the loop between the first and the second helix (16 – 26AA) is four residues longer from other extensively studied ACBPs (PfACBP) and it curls upwards towards the pantothenate moiety of CoA to provide a large tunnel space for long acyl chain insertion.

Keywords: acyl-coa binding protein (ACBP), acyl-coa esters, crystal structure, isothermal titration, calorimetry, Leishmania

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6059 Evaluation of Anti-Leishmanial Activity of Albaha Medicinal Plants against Leishmania amazonensis

Authors: Saeed S. Al-Sokari, Nasser A. Awadh Ali, Lianet Monzote

Abstract:

Leishmaniasis (CL) is endemic in at least 82 countries and considered to be a major public-health problem (1). The annual incidence of CL is 1–1.5 million cases of which 90% occur in only seven countries: Afghanistan, Algeria, Brazil, Iran, Peru, Saudi Arabia and Syria (2). In Saudi Arabia, the disease was first described in 1973 by Moursy and Shoura (3). Currently, CL is common in the human population in different localities, including the Eastern Province of Saudi Arabia and in particular the Al-Hassa Oasis that is a known endemic area for CL (4). Five methanolic extracts obtained from Achillea biebersteinii (flower leaf), Euphorbia antiquorm, Solanum incanum (leaf and fruit extracts), collected from Albaha region and selected from ethno-botanical data, were screened for their anti-leishmanial activity against Leishmania amazonensis (6). The cytotoxic activity against normal peritoneal macrophages from normal BALB/c mice was also determined (6). The five extracts had IC50 values ranging from < 12.5 to 37.8 µg/ml against promastigotes. Achillea biebersteinii flower, Euphorbia antiquorm, Solanum incanum leaf extracts showed anti-leishmanial activities with IC50 between < 12.5 - 26.9µg/mL and acceptable selectivity indices of 8 - 5.

Keywords: plant extracts, Albaha, Leishmania amazonensis, Medicinal

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6058 Brazilian Brown Propolis as a Natural Source against Leishmania amazonensis

Authors: Victor Pena Ribeiro, Caroline Arruda, Jennyfer Andrea Aldana Mejia, Jairo Kenupp Bastos

Abstract:

Leishmaniasis is a serious health problem around the world. The treatment of infected individuals with pentavalent antimonial drugs is the main therapeutic strategy. However, they present high toxicity and persistence side effects. Therefore, the discovery of new and safe natural-derived therapeutic agents against leishmaniasis is important. Propolis is a resin of viscous consistency produced by Apis mellifera bees from parts of plants. The main types of Brazilian propolis are green, red, yellow and brown. Thus, the aim of this work was to investigate the chemical composition and leishmanicidal properties of a brown propolis (BP). For this purpose, the hydroalcoholic crude extract of BP was obtained and was fractionated by liquid-liquid chromatography. The chemical profile of the extract and its fractions were obtained by HPLC-UV-DAD. The fractions were submitted to preparative HPLC chromatography for isolation of the major compounds of each fraction. They were analyzed by NMR for structural determination. The volatile compounds were obtained by hydrodistillation and identified by GC/MS. Promastigote forms of Leishmania amazonensis were cultivated in M199 medium and then 2×106 parasites.mL-1 were incubated in 96-well microtiter plates with the samples. The BP was dissolved in dimethyl sulfoxide (DMSO) and diluted into the medium, to give final concentrations of 1.56, 3.12, 6.25, 12.5, 25 and 50 µg.mL⁻¹. The plates were incubated at 25ºC for 24 h, and the lysis percentage was determined by using a Neubauer chamber. The bioassays were performed in triplicate, using a medium with 0.5% DMSO as a negative control and amphotericin B as a positive control. The leishimnicidal effect against promastigote forms was also evaluated at the same concentrations. Cytotoxicity experiments also were performed in 96-well plates against normal (CHO-k1) and tumor cell lines (AGP01 and HeLa) using XTT colorimetric method. Phenolic compounds, flavonoids, and terpenoids were identified in brown propolis. The major compounds were identified as follows: p-coumaric acid (24.6%) for a methanolic fraction, Artepelin-C (29.2%) for ethyl acetate fraction and the compounds of hexane fraction are in the process of structural elucidation. The major volatile compounds identified were β-caryophyllene (10.9%), germacrene D (9.7%), nerolidol (10.8%) and spathulenol (8.5%). The propolis did not show cytotoxicity against normal cell lines (CHO) with IC₅₀ > 100 μg.mL⁻¹, whereas the IC₅₀ < 10 μg.mL⁻¹ showed a potential against the AGP01 cell line, propolis did not demonstrate cytotoxicity against HeLa cell lines IC₅₀ > 100 μg.mL⁻¹. In the determination of the leishmanicidal activity, the highest (50 μg.mL⁻¹) and lowest (1.56 μg.mL⁻¹) concentrations of the crude extract caused the lysis of 76% and 45% of promastigote forms of L. amazonensis, respectively. To the amastigote form, the highest (50 μg.mL⁻¹) and lowest (1.56 μg.mL⁻¹) concentrations caused the mortality of 89% and 75% of L. amazonensis, respectively. The IC₅₀ was 2.8 μg.mL⁻¹ to amastigote form and 3.9 μg.mL⁻¹ to promastigote form, showing a promising activity against Leishmania amazonensis.

Keywords: amastigote, brown propolis, cytotoxicity, promastigote

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6057 Peptide-Gold Nanocluster as an Optical Biosensor for Glycoconjugate Secreted from Leishmania

Authors: Y. A. Prada, Fanny Guzman, Rafael Cabanzo, John J. Castillo, Enrique Mejia-Ospino

Abstract:

In this work, we show the important results about of synthesis of photoluminiscents gold nanoclusters using a small peptide as template for biosensing applications. Interestingly, we design one peptide (NBC2854) homologue to conservative domain from 215 250 residue of a galactolectin protein which can recognize the proteophosphoglycans (PPG) from Leishmania. Peptide was synthetized by multiple solid phase synthesis using FMoc group methodology in acid medium. Finally, the peptide was purified by High-Performance Liquid Chromatography using a Vydac C-18 preparative column and the detection was at 215 nm using a Photo Diode Array detector. Molecular mass of this peptide was confirmed by MALDI-TOF and to verify the α-helix structure we use Circular Dichroism. By means of the methodology used we obtained a novel fluorescents gold nanoclusters (AuNC) using NBC2854 as a template. In this work, we described an easy and fast microsonic method for the synthesis of AuNC with ≈ 3.0 nm of hydrodynamic size and photoemission at 630 nm. The presence of cysteine residue in the C-terminal of the peptide allows the formation of Au-S bond which confers stability to Peptide-based gold nanoclusters. Interactions between the peptide and gold nanoclusters were confirmed by X-ray Photoemission and Raman Spectroscopy. Notably, from the ultrafine spectra shown in the MALDI-TOF analysis which containing only 3-7 KDa species was assigned to Au₈-₁₈[NBC2854]₂ clusters. Finally, we evaluated the Peptide-gold nanocluster as an optical biosensor based on fluorescence spectroscopy and the fluorescence signal of PPG (0.1 µg-mL⁻¹ to 1000 µg-mL⁻¹) was amplified at the same wavelength emission (≈ 630 nm). This can suggest that there is a strong interaction between PPG and [email protected], therefore, the increase of the fluorescence intensity can be related to the association mechanism that take place when the target molecule is sensing by the [email protected] conjugate. Further spectroscopic studies are necessary to evaluate the fluorescence mechanism involve in the sensing of the PPG by the [email protected] To our best knowledge the fabrication of an optical biosensor based on [email protected] for sensing biomolecules such as Proteophosphoglycans which are secreted in abundance by parasites Leishmania.

Keywords: biosensing, fluorescence, Leishmania, peptide-gold nanoclusters, proteophosphoglycans

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6056 Molecular Detection of Leishmania from the Phlebotomus Genus: Tendency towards Leishmaniasis Regression in Constantine, North-East of Algeria

Authors: K. Frahtia, I. Mihoubi, S. Picot

Abstract:

Leishmaniasis is a group of parasitic disease with a varied clinical expression caused by flagellate protozoa of the Leishmania genus. These diseases are transmitted to humans and animals by the sting of a vector insect, the female sandfly. Among the groups of dipteral disease vectors, Phlebotominae occupy a prime position and play a significant role in human pathology, such as leishmaniasis that affects nearly 350 million people worldwide. The vector control operation launched by health services throughout the country proves to be effective since despite the prevalence of the disease remains high especially in rural areas, leishmaniasis appears to be declining in Algeria. In this context, this study mainly concerns molecular detection of Leishmania from the vector. Furthermore, a molecular diagnosis has also been made on skin samples taken from patients in the region of Constantine, located in the North-East of Algeria. Concerning the vector, 5858 sandflies were captured, including 4360 males and 1498 females. Male specimens were identified based on their morphological. The morphological identification highlighted the presence of the Phlebotomus genus with a prevalence of 93% against 7% represented by the Sergentomyia genus. About the identified species, P. perniciosus is the most abundant with 59.4% of the male identified population followed by P. longicuspis with 24.7% of the workforce. P. perfiliewi is poorly represented by 6.7% of specimens followed by P. papatasi with 2.2% and 1.5% S. dreyfussi. Concerning skin samples, 45/79 (56.96%) collected samples were found positive by real-time PCR. This rate appears to be in sharp decline compared to previous years (alert peak of 30,227 cases in 2005). Concerning the detection of Leishmania from sandflies by RT-PCR, the results show that 3/60 PCR performed genus are positive with melting temperatures corresponding to that of the reference strain (84.1 +/- 0.4 ° C for L. infantum). This proves that the vectors were parasitized. On the other side, identification by RT-PCR species did not give any results. This could be explained by the presence of an insufficient amount of leishmanian DNA in the vector, and therefore support the hypothesis of the regression of leishmaniasis in Constantine.

Keywords: Algeria, molecular diagnostic, phlebotomus, real time PCR

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6055 Invitro Study of Anti-Leishmanial Property of Nigella Sativa Methanalic Black Seed Extract

Authors: Tawqeer Ali Syed, Prakash Chandra

Abstract:

This study aims to evaluate the antileishmanial activity of Nigella sativa black seed extract. This well-known plant extract was taken from the botanical garden of Kashmir. Materials and Methods: The methanolic extracts of these plants were screened for their antileishmanial activity against Leishmania major using 3‑(4.5‑dimethylthiazol‑2yl)‑2.5‑diphenyltetrazolium bromide assay or MTT assay. Results: The methanolic extract of Nigella sativa showed potential antileishmanial activity at an inhibition% value of 80.29% ± 0.65%. IC 50 was calculated after 48 hours to be 964.3 µg/ml. Conclusion: Considering these results, these medicinal plants from Kashmir could serve as potential drug sources for antileishmanial compounds.

Keywords: MTT assay, antileishmanial, cell viability, Nigella sativa

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6054 Retrospective Evaluation of Vector-borne Infections in Cats Living in Germany (2012-2019)

Authors: I. Schäfer, B. Kohn, M. Volkmann, E. Müller

Abstract:

Introduction: Blood-feeding arthropods transmit parasitic, bacterial, or viral pathogens to domestic animals and wildlife. Vector-borne infections are gaining significance due to the increase of travel, import of domestic animals from abroad, and the changing climate in Europe. Aims of the study: The main objective of this retrospective study was to assess the prevalence of vector-borne infections in cats in which a ‘Feline Travel Profile’ had been conducted. Material and Methods: This retrospective study included test results from cats for which a ‘Feline Travel Profile’ established by LABOKLIN had been requested by veterinarians between April 2012 and December 2019. This profile contains direct detection methods via polymerase chain reaction (PCR) for Hepatozoon spp. and Dirofilaria spp. as well as indirect detection methods via immunofluorescence antibody test (IFAT) for Ehrlichia spp. and Leishmania spp. This profile was expanded to include an IFAT for Rickettsia spp. from July 2015 onwards. The prevalence of the different vector-borne infectious agents was calculated. Results: A total of 602 cats were tested using the ‘Feline Travel Profile’. Positive test results were as follows: Rickettsia spp. IFAT 54/442 (12.2%), Ehrlichia spp. IFAT 68/602 (11.3%), Leishmania spp. IFAT 21/602 (3.5%), Hepatozoon spp. PCR 51/595 (8.6%), and Dirofilaria spp. PCR 1/595 cats (0.2%). Co-infections with more than one pathogen could be detected in 22/602 cats. Conclusions: 170/602 cats (28.2%) were tested positive for at least one vector-borne pathogen. Infections with multiple pathogens could be detected in 3.7% of the cats. The data emphasizes the importance of considering vector-borne infections as potential differential diagnoses in cats.

Keywords: arthopod-transmitted infections, feline vector-borne infections, Germany, laboratory diagnostics

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6053 Molecular Characterization of Arginine Sensing Response in Unravelling Host-Pathogen Interactions in Leishmania

Authors: Evanka Madan, Madhu Puri, Dan Zilberstein, Rohini Muthuswami, Rentala Madhubala

Abstract:

The extensive interaction between the host and pathogen metabolic networks decidedly shapes the outcome of infection. Utilization of arginine by the host and pathogen is critical for determining the outcome of pathogenic infection. Infections with L. donovani, an intracellular parasite, will lead to an extensive competition of arginine between the host and the parasite donovani infection. One of the major amino acid (AA) sensing signaling pathways in mammalian cells are the mammalian target of rapamycin complex I (mTORC1) pathway. mTORC1, as a sensor of nutrient, controls numerous metabolic pathways. Arginine is critical for mTORC1 activation. SLC38A9 is the arginine sensor for the mTORC1, being activated during arginine sufficiency. L. donovani transport arginine via a high-affinity transporter (LdAAP3) that is rapidly up-regulated by arginine deficiency response (ADR) in intracellular amastigotes. This study, to author’s best knowledge, investigates the interaction between two arginine sensing systems that act in the same compartment, the lysosome. One is important for macrophage defense, and the other is essential for pathogen virulence. We hypothesize that the latter modulates lysosome arginine to prevent host defense response. The work presented here identifies an upstream regulatory role of LdAAP3 in regulating the expression of SLC38A9-mTORC1 pathway, and consequently, their function in L. donovani infected THP-1 cells cultured in 0.1 mM and 1.5 mM arginine. It was found that in physiological levels of arginine (0.1 mM), infecting THP-1 with Leishmania leads to increased levels of SLC38A9 and mTORC1 via an increase in the expression of RagA. However, the reversal was observed with LdAAP3 mutants, reflecting the positive regulatory role of LdAAP3 on the host SLC38A9. At the molecular level, upon infection, mTORC1 and RagA were found to be activated at the surface of phagolysosomes which was found to form a complex with phagolysosomal localized SLC38A9. To reveal the relevance of SLC38A9 under physiological levels of arginine, endogenous SLC38A9 was depleted and a substantial reduction in the expression of host mTORC1, its downstream active substrate, p-P70S6K1 and parasite LdAAP3, was observed, thereby showing that silencing SLC38A9 suppresses ADR. In brief, to author’s best knowledge, these results reveal an upstream regulatory role of LdAAP3 in manipulating SLC38A9 arginine sensing in host macrophages. Our study indicates that intra-macrophage survival of L. donovani depends on the availability and transport of extracellular arginine. An understanding of the sensing pathway of both parasite and host will open a new perspective on the molecular mechanism of host-parasite interaction and consequently, as a treatment for Leishmaniasis.

Keywords: arginine sensing, LdAAP3, L. donovani, mTORC1, SLC38A9, THP-1

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6052 Combining in vitro Protein Expression with AlphaLISA Technology to Study Protein-Protein Interaction

Authors: Shayli Varasteh Moradi, Wayne A. Johnston, Dejan Gagoski, Kirill Alexandrov

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The demand for a rapid and more efficient technique to identify protein-protein interaction particularly in the areas of therapeutics and diagnostics development is growing. The method described here is a rapid in vitro protein-protein interaction analysis approach based on AlphaLISA technology combined with Leishmania tarentolae cell-free protein production (LTE) system. Cell-free protein synthesis allows the rapid production of recombinant proteins in a multiplexed format. Among available in vitro expression systems, LTE offers several advantages over other eukaryotic cell-free systems. It is based on a fast growing fermentable organism that is inexpensive in cultivation and lysate production. High integrity of proteins produced in this system and the ability to co-express multiple proteins makes it a desirable method for screening protein interactions. Following the translation of protein pairs in LTE system, the physical interaction between proteins of interests is analysed by AlphaLISA assay. The assay is performed using unpurified in vitro translation reaction and therefore can be readily multiplexed. This approach can be used in various research applications such as epitope mapping, antigen-antibody analysis and protein interaction network mapping. The intra-viral protein interaction network of Zika virus was studied using the developed technique. The viral proteins were co-expressed pair-wise in LTE and all possible interactions among viral proteins were tested using AlphaLISA. The assay resulted to the identification of 54 intra-viral protein-protein interactions from which 19 binary interactions were found to be novel. The presented technique provides a powerful tool for rapid analysis of protein-protein interaction with high sensitivity and throughput.

Keywords: AlphaLISA technology, cell-free protein expression, epitope mapping, Leishmania tarentolae, protein-protein interaction

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6051 Exploring the Role of Immune-Modulators in Pathogen Recognition Receptor NOD2 Mediated Protection against Visceral Leishmaniasis

Authors: Junaid Jibran Jawed, Prasanta Saini, Subrata Majumdar

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Background: Leishmania donovani infection causes severe host immune-suppression through the modulation of pathogen recognition receptors. Apart from TLRs (Toll Like Receptor), recent studies focus on the important contribution of NLR (NOD-Like Receptor) family member NOD1 and NOD2 as these receptors are capable of triggering host innate immunity. The aim of this study was to decipher the role of NOD1/NOD2 receptors during experimental visceral leishmaniasis (VL) and the important link between host failure and parasite evasion strategy. Method: The status of NOD1 and NOD2 receptors were analysed in uninfected and infected cells through western blotting and RT-PCR. The active contributions of these receptors in reducing parasite burden were confirmed by siRNA mediated silencing, and over-expression studies and the parasite numbers were calculated through microscopic examination of the Giemsa-stained slides. In-vivo studies were done by using non-toxic dose of Mw (Mycobacterium indicus pranii), Ara-LAM(Arabinoasylated lipoarabinomannan) along with MDP (Muramyl dipeptide) administration. Result: Leishmania donovani infection of the macrophages reduced the expression of NOD2 receptors whereas NOD1 remain unaffected. MDP, a NOD2-ligand, treatment during over-expression of NOD2, reduced the parasite burden effectively which was associated with increased pro-inflammatory cytokine generation and NO production. In experimental mouse model, Ara-LAM treatment increased the expression of NOD2 and in combination with MDP it showed active therapeutic potential against VL and found to be more effective than Mw which was already reported to be involved in NOD2 modulation. Conclusion: This work explores the essential contribution of NOD2 during experimental VL and mechanistic understanding of Ara-LAM + MDP combination therapy to work against this disease and highlighted NOD2 as an essential therapeutic target.

Keywords: Ara-LAM (Arabinoacylated Lipoarabinomannan), NOD2 (nucleotide binding oligomerization receptor 2), MDP (muramyl di peptide), visceral Leishmaniasis

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6050 Artemisia Species from Iran as Valuable Resources for Medicinal Uses

Authors: Mohammad Reza Naghavi, Farzad Alaeimoghadam, Hossein Ghafoori

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Artemisia species, which are medically beneficial, are widespread in temperate regions of both Northern and Southern hemispheres among which Iran is located. About 35 species of Artemisia are indigenous in Iran among them some are widespread in all or most provinces, yet some are restricted to some specific regions. In this review paper, initially, GC-Mass results of some experiments done in different provinces of Iran are mentioned among them some compounds are common among species, some others are mostly restricted to other species; after that, medical advantages based on some researches on species of this genus are reviewed; different qualities such as anti-leishmania, anti-bacteria, antiviral as well as anti-proliferative could be mentioned.

Keywords: artemisia, GC-Mass analysis, medical advantage, antiviral

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6049 Differential Effect of Technique Majors on Isokinetic Strength in Youth Judoka Athletes

Authors: Chungyu Chen, Yi-Cheng Chen, Po-Hsian Hsu, Hsin-Ying Chen, Yen-Po Hsiao

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The purpose of this study was to assess the muscular strength performance of upper and lower extremity in isokinetic system for the youth judo players, and also to compare the strength difference between major techniques. Sixteen male and 20 female judo players (age: 16.7 ± 1.6 years old, training age: 4.5 ± 0.8 years) were served as the volunteers for this study. There were 21 players major hand techniques and 15 players major foot techniques. The Biodex S4 Pro was used to assess the strength performance of extensor and flexor of concentric action under the load condition of 30 degree/sec, 60 degree/sec, and 120 degree/sec for elbow joints and knee joints. The strength parameters were included the maximal torque, the normalized maximal torque, the average power, and the average maximal torque. A t test for independent groups was used to evaluate whether hand major and foot major differ significantly with an alpha level of .05. The result showed the maximal torque of left knee extensor in foot major players (243.5 ± 36.3 Nm) was higher significantly than hand major (210.7 ± 21.0 Nm) under the load of 30 degree/sec (p < .05). There were no differences in upper extremity strength between the hand and foot techniques major in three loads (ps < .05). It indicated that the judo player is required to develop the upper extremity strength overall to secure the execution of major techniques.

Keywords: knee, elbow, power, judo

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6048 Major Incident Tier System in the Emergency Department: An Approach

Authors: Catherine Bernard, Paul Ransom

Abstract:

Recent events have prompted emergency planners to re-evaluate their emergency response to major incidents and mass casualties. At the Royal Sussex County Hospital, we have adopted a tiered system comprised of three levels, anticipating an increasing P1, P2 or P3 load. This will aid planning in the golden period between Major Incident ‘Standby,’ and ‘Declared’. Each tier offers step-by-step instructions on appropriate patient movement within and out of the department, as well as suggestions for overflow areas and additional staffing levels. This system can be adapted to individual hospitals and provides concise instructions to be followed in a potentially overwhelming situation.

Keywords: disaster planning, emergency preparedness, major incident planning, mass casualty event

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6047 Refractory Visceral Leishmaniasis Responding to Second-Line Therapy

Authors: Preet Shah, Om Shrivastav

Abstract:

Introduction : In India, Leishmania donovani is the only parasite causing Leishmaniasis. The parasite infects the reticuloendothelial system and is found in the bone marrow, spleen and liver. Treatment of choice is amphotericin-B with sodium stibogluconate being an alternative. Miltefosine is useful in refractory cases. In our case, Leishmaniasis occurred in a person residing in western India (which is quite rare) and it failed to respond to two different drugs (again an uncommon feature) before it finally responded to a third one. Description: A 50 year old lady, a resident of western India, with no history of recent travel, presented with an ulcer on the left side of the nose since 8 months. She was apparently alright 8 months back, when she noticed a small ulcerated lesion on the left ala of the nose which was immediately biopsied. The biopsy revealed amastigotes of Leishmania for which she was administered intra-lesional sodium stibogluconate for 1 month (4 doses every 8 days).Despite this, there was no regression of the ulcer and hence she presented to us for further management. On examination, her vital parameters were normal. Barring an ulcer on the left side of the nose, rest of the examination findings were unremarkable. Complete blood count was normal. Ultrasound abdomen showed hepatomegaly. PET-CT scan showed increased metabolic activity in left ala of nose, hepatosplenomegaly and increased metabolic activity in spleen and bone marrow. Bone marrow biopsy was done which showed hypercellular marrow with erythroid preponderance. Considering a diagnosis of leishmaniasis which had so far been unresponsive to sodium stibogluconate, she was started on liposomal amphotericin-B. At the time of admission, her creatinine level was normal, but it started rising with the administration of liposomal amphotericin-B, hence the dose was reduced. Despite this, creatinine levels did not improve, and she started developing hypokalemia and hypomagnesemia as side effects of the drug, hence further reductions in the dosage were made. Despite a total of 3 weeks of liposomal amphotericin-B, there was no improvement in the ulcer. As had so far failed to respond to sodium stibogluconate and liposomal amphotericin-B, it was decided to start her on miltefosine. She received the miltefosine for a total of 12 weeks. At the end of this duration, there was a marked regression of the cutaneous lesion. Conclusion: Refractoriness to amphotericin-B in leishmaniasis may be seen in up to 5 % cases. Here, an alternative drug such as miltefosine is useful and hence we decided to use it, to which she responded adequately. Furthermore, although leishmaniasis is common in the eastern part of India, it is a relatively unknown entity in the western part of the country with the occurrence being very rare. Because of these 2 reasons, we consider our case to be a unique one.

Keywords: amphotericin-b, leishmaniasis, miltefosine, tropical diseases

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6046 An Optimal Control Model for the Dynamics of Visceral Leishmaniasis

Authors: Ibrahim M. Elmojtaba, Rayan M. Altayeb

Abstract:

Visceral leishmaniasis (VL) is a vector-borne disease caused by the protozoa parasite of the genus leishmania. The transmission of the parasite to humans and animals occurs via the bite of adult female sandflies previously infected by biting and sucking blood of an infectious humans or animals. In this paper we use a previously proposed model, and then applied two optimal controls, namely treatment and vaccination to that model to investigate optimal strategies for controlling the spread of the disease using treatment and vaccination as the system control variables. The possible impact of using combinations of the two controls, either one at a time or two at a time on the spread of the disease is also examined. Our results provide a framework for vaccination and treatment strategies to reduce susceptible and infection individuals of VL in five years.

Keywords: visceral leishmaniasis, treatment, vaccination, optimal control, numerical simulation

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6045 Influence of Major Axis on the Aerodynamic Characteristics of Elliptical Section

Authors: K. B. Rajasekarababu, J. Karthik, G. Vinayagamurthy

Abstract:

This paper is intended to explain the influence of major axis on aerodynamic characteristics of elliptical section. Many engineering applications such as off shore structures, bridge piers, civil structures and pipelines can be modelled as a circular cylinder but flow over complex bodies like, submarines, Elliptical wing, fuselage, missiles, and rotor blades, in which the parameters such as axis ratio can influence the flow characteristics of the wake and nature of separation. Influence of Major axis in Flow characteristics of elliptical sections are examined both experimentally and computationally in this study. For this research, four elliptical models with varying major axis [*AR=1, 4, 6, 10] are analysed. Experimental works have been conducted in a subsonic wind tunnel. Furthermore, flow characteristics on elliptical model are predicted from k-ε turbulence model using the commercial CFD packages by pressure based transient solver with Standard wall conditions.The analysis can be extended to estimation and comparison of Drag coefficient and Fatigue analysis of elliptical sections.

Keywords: elliptical section, major axis, aerodynamic characteristics, k-ε turbulence model

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6044 Effects of Major and Minor Modes to Emotional Perceptions of 'Happy' and 'Sad' in Piano Music among Students Aged 9-17

Authors: Nurezlin Mohd Azib, Pan Kok Chang

Abstract:

This quantitative study investigates the effects of major and minor modes, and contributing musical parameter of tempo, to the emotional perceptions of ‘happy’ and ‘sad’ in piano music among subjects aged 9-17 years old. The study was conducted in two phases; survey-questionnaire, and listening activity. Subjects (N=31) were sampled from piano music students’ population in Bangi, Selangor. In the survey-questionnaire, subjects answered 20 questions on demographic characteristics, music listening and preference, and understanding of emotional perception in music. In the listening activity, subjects listened to 20 untitled piano music excerpts and rated the emotion perceived for each excerpt, whether ‘happy’ or ‘sad’. Results from survey-questionnaire show that most percentage of subjects are 11 years old, in Grade 1, of 3 years of learning piano, prefer classical music, always listen to music, prefer both major and minor modes’ music, and find it easy to understand emotion in music, as well as major and minor modes. Results from listening activity show that 60 % of major mode music are perceived as ‘major-happy’, while 60 % too, of minor mode music are perceived as ‘minor-sad’. However, Chi-square test of independence statistical analysis indicates that there are no association and significant relationship between modes (major and minor) and ‘happy’, as well as ‘sad’ perceptions (x2 (1, N = 20) = 0.80, p = 0.371), at the significance level of p ≤ 0.05. Contrastingly, there are association and significant relationship between tempo (fast and slow), and ‘happy’, as well as ‘sad’ perceptions (x2 (1, N = 20) = 9.899, p = 0.005). Therefore, it is concluded that tempo plays an important role in effects of major and minor mode to ‘happy’ and ‘sad’ emotional perceptions in piano music among subjects aged 9 to 17 in this study.

Keywords: effects, emotional perceptions, major and minor modes, piano music

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6043 Appropriate Depth of Needle Insertion during Rhomboid Major Trigger Point Block

Authors: Seongho Jang

Abstract:

Objective: To investigate an appropriate depth of needle insertion during trigger point injection into the rhomboid major muscle. Methods: Sixty-two patients who visited our department with shoulder or upper back pain participated in this study. The distance between the skin and the rhomboid major muscle (SM) and the distance between the skin and rib (SB) were measured using ultrasonography. The subjects were divided into 3 groups according to BMI: BMI less than 23 kg/m2 (underweight or normal group); 23 kg/m2 or more to less than 25 kg/m2 (overweight group); and 25 kg/m2 or more (obese group). The mean ±standard deviation (SD) of SM and SB of each group were calculated. A range between mean+1 SD of SM and the mean-1 SD of SB was defined as a safe margin. Results: The underweight or normal group’s SM, SB, and the safe margin were 1.2±0.2, 2.1±0.4, and 1.4 to 1.7 cm, respectively. The overweight group’s SM and SB were 1.4±0.2 and 2.4±0.9 cm, respectively. The safe margin could not be calculated for this group. The obese group’s SM, SB, and the safe margin were 1.8±0.3, 2.7±0.5, and 2.1 to 2.2 cm, respectively. Conclusion: This study will help us to set the standard depth of safe needle insertion into the rhomboid major muscle in an effective manner without causing any complications.

Keywords: pneumothorax, rhomboid major muscle, trigger point injection, ultrasound

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6042 Organization of the Olfactory System and the Mushroom Body of the Weaver Ant, Oecophylla smaragdina

Authors: Rajashekhar K. Patil, Martin J. Babu

Abstract:

Weaver ants-Oecophylla smaragdina live in colonies that have polymorphic castes. The females which include the queen, major and minor workers are haploid. The individuals of castes are dependent on olfactory cues for carrying out caste-specific behaviour. In an effort to understand whether organizational differences exist to support these behavioural differences, we studied the olfactory system at the level of the sensilla on the antennae, olfactory glomeruli and the Kenyon cells in the mushroom bodies (MB). The MB differ in major and minor workers in terms of their size, with the major workers having relatively larger calyces and peduncle. The morphology of different types of Kenyon cells as revealed by Golgi-rapid staining was studied and the major workers had more dendritic arbors than minor workers. This suggests a greater degree of olfactory processing in major workers. Differences in caste-specific arrangement of sensilla, olfactory glomeruli and celluar architecture of MB indicate a developmental programme that forms basis of differential behaviour.

Keywords: ant, oecophylla, caste, mushroom body

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6041 Examining the Level of Career Maturity on Cultural Aspect among Undergraduate Foreign Students in A Public University in Malaysia

Authors: Mustafa Tekke, Nurullah Kurt

Abstract:

This study examined the level of career maturity of undergraduate foreign students in a public university in Malaysia by examining on cultural aspect by using the Career Maturity Inventory. Two hundred and twenty nine (Male = 106, Female = 123) foreign students studying in various majors completed the Career Maturity Inventory and the scores of the foreign students on the CMI suggested that they had slightly higher levels than the mean level of maturity in career. Result was also supported by testing the feeling about major, consideration of changing major and planning after graduation, which indicated that foreign students had their own career decision making. However, this result should be viewed with caution within ethnic difference.

Keywords: career maturity, foreign students, career decision making, feeling about major, knowledge about major

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6040 Time Overrun in Pre-Construction Planning Phase of Construction Projects

Authors: Hafiz Usama Imad, Muhammad Akram Akhund, Tauha Hussain Ali, Ali Raza Khoso, Fida Hussain Siddiqui

Abstract:

Construction industry plays a significant role in fulfilling the major requirements of the human being. It is one of the major constituents of every developed country. Although the construction industry of both the developing and developed countries encompasses a major part of the economy, and millions of rupees are utilized every year on various kinds of construction projects. But, this industry is facing numerous hurdles in terms of its budget and timely completion. Construction projects generally consist of several phases like planning, designing, execution, and finishing. This research study aims to determine the significant factors of time overrun in pre-construction planning (PCP) phase of construction projects in Pakistan. Questionnaires were distributed by various means and responses of respondents were compiled and collected data were then analyzed through a statistical technique using SPSS version 24. Major causes of time overrun in pre-construction planning phase; which is an extremely important phase of construction projects, were revealed. The research conclusion will provide a pathway for stakeholders to pay attention to the mentioned causes to overcome the major issue of time overrun.

Keywords: construction industry, Pakistan, pre-construction planning phase, time overrun

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6039 Synthesis and Structural Characterization of 6-Nitroindazole Derivatives

Authors: Mohamed El Moctar Abeidi

Abstract:

The indazole derivatives exhibit a wide spectrum of biological activities. They are known for their anti-tumor, antiplatelet, anti-viral, anti-microbial, anti-inflammatory, anti-leishmania and even anti-spermatogen. As part of our research on the synthesis of a number of heterocycles capable of exhibiting a biological and pharmacological property, due to our ongoing interest in the development of a simple and low-cost procedure for obtaining heterocyclic compounds that may have an interest for medicinal purposes. We present in this work the synthesis of 6-nitro-indazoles derivatives, using two different methods. the first method is the alkylation of Nitroindazole by two different alkylating agents under the conditions of solid/liquid phase transfer catalysis in N, N-dimethylformamide (DMF) in the presence of potassium carbonate (K₂CO₃) as a base, and tetra-n-butylammonium bromide (BTBA) as a catalyst. While the other method is the 1,3-dipolar cycloaddition, in this case, we have undertaken the preparation of bi-heterocyclic containing the 6-nitroindazole associate with group of isoxazoline, isoxazole or 1,2,3-Triazole under normal conditions and, under the catalytic conditions of the click chemistry we were also able to determine the structures without any ambiguity by the ¹H and ¹³C NMR.

Keywords: indazole, 6-nitroindazole, isoxazole, 1, 2, 3-Triazole

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6038 Self-Inflicted Major Trauma: Inpatient Mental Health Management and Patient Outcomes

Authors: M. Walmsley, S. Elmatarri, S. Mannion

Abstract:

Introduction: Self-inflicted injury is a recognised cause of major trauma in adults and is an independent indicator of a reduced functional outcome compared to non-intentional major trauma. There is little literature available on the inpatient mental health (MH) management of this vulnerable group. A retrospective review was conducted of inpatient MH management of major trauma patients admitted to a UK regional Major Trauma Centre (MTC). Their outcomes were compared to all major trauma patients. This group of patients required multiple MH interventions whilst on the Major Trauma Ward (MTW) and a had worse functional outcome compared to non-intentional trauma. Method: The national TARN (Trauma Audit and Research Network) database was used to identify patients admitted to a regional MTC over a 2-year period from June 2018 to July 2020. Patients with an ISS (Injury Severity Score) of greater than 15 with a mechanism of either self-harm or high-risk behavior were included for further analysis. Inpatient medical notes were reviewed for MH interventions on the MTW. Further outcomes, including mortality, length of stay (LOS) and Glasgow Outcome Score (GOS) were compared with all major trauma patients for the same time period. Results: A total of 60 patients were identified in the time period and of those, 27 spent time on the MTW. A total of 23 (85%) had a prior MH diagnosis, with 11 (41%) under the care of secondary MH services. Adequate inpatient records for review were available for 24 patients. During their inpatient stay, 8 (33%) were reviewed on the ward by the inpatient MH team. There were 10 interventions required for 6 (25%) patients on the MTW including, sections under the Mental Health Act, transfer to specialist MH facility, pharmacological sedation and security being called to the MTW. When compared to all major trauma patients, those admitted due to self-harm or high-risk behavior had a statistically significantly higher ISS (31.43 vs 24.22, p=0.0001) and LOS (23.51d vs 16.06d, p=0.002). Functional outcomes using the GOS were reduced in this group of patients, GOS 5 (low disability) (51.66% vs. 61.01%) and they additionally had a higher level of mortality, GOS 1 (15.00% vs 11.67%). Discussion: Intentional self-harm is a recognised cause of major trauma in adults and this patient group sustains more severe injuries, requiring a longer hospital stay with worse outcomes compared to all major trauma patients. Inpatient MH interventions are required for a significant proportion of these patients and therefore, there needs to be a close relationship with MH services. There is limited available evidence for how this patient group is best managed as an inpatient to aid their recovery and further work is needed on how outcomes in this vulnerable group can be improved.

Keywords: adult major trauma, attempted suicide, self-inflicted major trauma, inpatient management

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6037 Determining Inventory Replenishment Policy for Major Component in Assembly-to-Order of Cooling System Manufacturing

Authors: Tippawan Nasawan

Abstract:

The objective of this study is to find the replenishment policy in Assembly-to-Order manufacturing (ATO) which some of the major components have lead-time longer than customer lead-time. The variety of products, independent component demand, and long component lead-time are the difficulty that has resulted in the overstock problem. In addition, the ordering cost is trivial when compared to the cost of material of the major component. A conceptual design of the Decision Supporting System (DSS) has introduced to assist the replenishment policy. Component replenishment by using the variable which calls Available to Promise (ATP) for making the decision is one of the keys. The Poisson distribution is adopted to realize demand patterns in order to calculate Safety Stock (SS) at the specified Customer Service Level (CSL). When distribution cannot identify, nonparametric will be applied instead. The test result after comparing the ending inventory between the new policy and the old policy, the overstock has significantly reduced by 46.9 percent or about 469,891.51 US-Dollars for the cost of the major component (material cost only). Besides, the number of the major component inventory is also reduced by about 41 percent which helps to mitigate the chance of damage and keeping stock.

Keywords: Assembly-to-Order, Decision Supporting System, Component replenishment , Poisson distribution

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