Search results for: erythrocytes

Authors: Leah Ghebreberhan, Liya Abraham, John Issac, Atul Kaushik

Abstract:

Medicinal plants are used for various indications in Eritrea according to traditional systems of medicine. Safety concerns, however, are dubious since some medicinal plants have toxic effects indeed. The medicinal plants under study (Commicarpus pedunculosis, Steganotaenia araliaceae, Boscia angustifolia, Solanum incanum, and Calpurnia aurea) are used in the treatment of various diseases. Thus, safety studies must be performed prior to usage since they are rich in phytoconstituents like saponins. Saponns are natural glycosides with several pharmacologic activities including hemolysis. The study was done to assess the level of saponin and toxic effects (hemolysis) of medicinal plants used in folk medicine. The plant extracts were subject to phytochemical analysis, foam test, and hemolytic assay. Regarding the Fh value, Solanam incanum consisted highest Fh value (20mm), whereas Boscia angustifolia showed the lowest Fh value (10mm). The level of hemolysis of all the plant extracts ranged between 9.0 to 20.2 %. All the plant extracts were suitable for treatment with respect to saponin level since they exhibited minimal hemolytic effect against erythrocytes.

Keywords: Boscia angustifolia, Calpurnia aurea, Commicarpus pedunculosis, hemolysis, saponin, Solanum incanum, Steganotaenia araliaceae

Procedia PDF Downloads 237

Authors: H. Bouaziz-Ketata, G. Ben Salah, Z. Aidi, C. Kallel, H. Kammoun, F. Fakhfakh, N. Zeghal

Abstract:

The present study was performed to evaluate the potential protective effect of Hyparrhenia hirta methanolic extract in NaNO3-induced genotoxic and hematotoxic effects. Male Wistar rats were randomly divided into three groups: a control group and two treated groups during 50 days with NaNO3 administered at a dose of 400 mg kg-1 bw either alone in drinking water or co-administered with Hyparrhenia hirta at a dose of 200 mg kg-1 bw. NaNO3 treatment showed a significant increase in the frequencies of total chromosomal aberrations, aberrant metaphases and micronucleus in bone-marrow cells. In parallel, the NaNO3-treated group showed a significant decrease in red blood cell count, hemoglobin and hematocrit and a significant increase in total white blood cell, in neutrophil and eosinophil counts. Platelet count, mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration remained unchanged in treated groups compared to those of controls. Hyparrhenia hirta methanolic extract appeared to be effective against genotoxic and hematotoxic changes induced by nitrate, as evidenced by the improvement of the markers cited above.

Keywords: Hyparrhenia hirta, sodium nitrate, erythrocytes, genotoxicity

Procedia PDF Downloads 249

Authors: Nail Nazarov, Vladimir Zobov, Alexandra Vyshtakalyuk, Vyacheslav Semenov, Irina Galyametdinova, Vladimir Reznik

Abstract:

There have been studied effects of xymedon and six new pyrimidine derivatives, that are close and distant analogs of xymedon, on rats' working capacity in the test 'swimming to failure'. It has been shown that a single administration of the studied compounds did not have a statistically significant effect in the test. In the conditions of multiple intraperitoneal administration of the studied pyrimidine derivatives, the compound L-ascorbate, 1-(2-hydroxyethyl)-4.6-dimethyl-1.2-dihydropyrimidine-2-one had the lowest toxicity and the most pronounced actoprotective effect. Introduction in the dose of 20 mg/kg caused a statistically significant increase 440 % in the duration of swimming of rats on the 14th day of the experiment compared with the control group. Multiple administration of the compound in the conditions of physical load did not affect leucopoiesis but stimulates erythropoiesis resulting in an increase in the number of erythrocytes and a hemoglobin level. The substance introduction under mixed exhausting loads prevented such changes of blood biochemical parameters as reduction of glucose, increased of urea and lactic acid levels, what indicates improvement in the animals' tolerability of loads and an anti-catabolic effect of the compound. Absence of hepato and cardiotoxic effects of the substance has been shown. This work was performed with the financial support of Russian Science Foundation (grant № 14-50-00014).

Keywords: actoprotectors, physical working capacity, pyrimidine derivatives, xymedon

Procedia PDF Downloads 280

Authors: N. Boussoualim, H. Trabsa, I. Krache, S. Aouachria, S. Boumerfeg, L. Arrar, A. Baghiani

Abstract:

The aim of this study consisted in evaluating the antioxidant in vivo properties, anti-hemolytic and XOR inhibitory effect of Globularia alypum L. (GA) extracts. GA was submitted to extraction and fractionation to give crude (CrE), chloroformique (ChE), ethyle acetate (EAE) and aqueos (AqE) extracts. Total polyphenols contents of GA extracts were determined; EAE is the most rich in polyphenols (157,74±5,27 mg GAE/mg of extract). GA Extracts inhibited XO in a concentration-dependent manner, the EAE showed the highest inhibitory properties on the XOR activity (IC50=0,083±0,001 mg/ml), followed by CrE and ChE. The antioxidant activities of the CrE, EAE, and AqE were tested by an in vivo assay in mice, the plasma ability to inhibit DPPH radical was measured, The CrE was found to exhibit the greatest scavenger activity with 48.41±2.763%, followed by AqE and EAE (40.54±7.51% and 41.79±1.654%, respectively). Total antioxidant capacity of red blood cells was measured, from the kinetics of hemolysis obtained. The calculated HT50 reveal an extension of time for half hemolysis in all treated groups compared with the control group. CrE increase significantly HT50 (112,8±2,427). The hemolysis is lagged, indicating that endogenous antioxidants in the erythrocytes can trap radicals to protect them against free-radical-induced hemolysis. Antimicrobial activities of the extracts were determined by the disc diffusion method. Test microorganisms were; 4 Gram positive, 7 gram negative bacteria, most active extracts were EAE and CrE. We deduce a great relationship between the effect on the extracts antibacterial effect and their contents in flavonoid.

Keywords: Globularia alypum, Xanthine oxidoreductase, in vivo-antioxidant activity, hemolysis, polyphenol

Procedia PDF Downloads 317

Authors: Erfan Niazi, Marianne Fenech

Abstract:

Red Blood Cells (RBCs) or erythrocytes tend to form chain-like aggregates under low shear rate called rouleaux. This is a reversible process and rouleaux disaggregate in high shear rates. Therefore, RBCs aggregation occurs in the microcirculation where low shear rates are present but does not occur under normal physiological conditions in large arteries. Numerical modeling of RBCs interactions is fundamental in analytical models of a blood flow in microcirculation. Population Balance Modeling (PBM) is particularly useful for studying problems where particles agglomerate and break in a two phase flow systems to find flow characteristics. In this method, the elementary particles lose their individual identity due to continuous destructions and recreations by break-up and agglomeration. The aim of this study is to find RBCs aggregation in a dynamic situation. Simplified PBM was used previously to find the aggregation rate on a static observation of the RBCs aggregation in a drop of blood under the microscope. To find aggregation rate in a dynamic situation we propose an experimental set up testing RBCs sedimentation. In this test, RBCs interact and aggregate to form rouleaux. In this configuration, disaggregation can be neglected due to low shear stress. A high-speed camera is used to acquire video-microscopic pictures of the process. The sizes of the aggregates and velocity of sedimentation are extracted using an image processing techniques. Based on the data collection from 5 healthy human blood samples, the aggregation rate was estimated as 2.7x10³(±0.3 x10³) 1/s.

Keywords: red blood cell, rouleaux, microfluidics, image processing, population balance modeling

Procedia PDF Downloads 345

Authors: Orkide Donma, M. Metin Donma, Burcin Nalbantoglu, Birol Topcu, Feti Tulubas, Murat Aydin, Tuba Gokkus, Ahmet Gurel

Abstract:

Increasing prevalence of childhood obesity has increased the interest in early and late indicators of gaining weight. Cell blood counts may be indicators of proinflammatory states. The aim was to evaluate associations of hematological parameters, including Hematocrit (HTC), hemoglobin, blood cell counts, and their indices with the degree of obesity in pediatric population. A total of 249; -139 morbidly obese (MO), 82 healthy Normal Weight (NW) and 28 Overweight (OW) children were included into the scope of the study. WHO BMI-for age percentiles were used to form age- and sex-matched groups. Informed consent forms and the Ethics Committee approval were obtained. Anthropometric measurements were performed. Hematological parameters were determined. Statistical analyses were performed using SPSS. The degree for statistical significance was p≤0.05. Significant differences (p=0.000) between waist-to-hip ratios and head-to-neck ratios (hnrs) of MO and NW children were detected. A significant difference between hnrs of OW and MO children (p=0.000) was observed. Red cell Distribution Width (RDW) was higher in OW children than NW group (p=0.030). Such finding couldn’t be detected between MO and NW groups. Increased RDW was prominent in OW children. The decrease in Mean Corpuscular Hemoglobin Concentration (MCHC) values in MO children was sharper than the values in OW children (p=0.006 vs p=0.042) compared to those in NW group. Statistically higher HTC levels were observed between MO-NW (p=0.014), but none between OW-NW. Though the cause-effect relationship between obesity and erythrocyte indices still needs further investigation, alterations in RDW, HTC, MCHC during obesity may be of significance in the early life.

Keywords: anthropometry, children, erythrocytes, obesity

Procedia PDF Downloads 340

Authors: Sanja Manchevska, Vaska Antevska, Lidija Todorovska, Beti Dejanova, Sunchica Petrovska, Ivanka Karagjozova, Elizabeta Sivevska, Jasmina Pluncevic Gligoroska

Abstract:

Introduction: A number of positive effects on the player’s physical status, including the body mass components are attributed to training process. As young soccer players grow up qualitative and quantitative changes appear and contribute to better performance. Player’s anthropometric and physiologic characteristics are recognized as important determinants of performance. Material: A sample of 52 soccer players with an age span from 9 to 14 years were divided in two groups differentiated by age. The younger group consisted of 25 boys under 11 years (mean age 10.2) and second group consisted of 27 boys with mean age 12.64. Method: The set of basic anthropometric parameters was analyzed: height, weight, BMI (Body Mass Index) and body mass components. Maximal oxygen uptake was tested using the treadmill protocol by Brus. Results: The group aged under 11 years showed the following anthropometric and physiological features: average height= 143.39cm, average weight= 44.27 kg; BMI= 18.77; Err = 5.04; Hb= 13.78 g/l; VO2=37.72 mlO2/kg. Average values of analyzed parameters were as follows: height was 163.7 cm; weight= 56.3 kg; BMI = 19.6; VO2= 39.52 ml/kg; Err=5.01; Hb=14.3g/l for the participants aged 12 to14 years. Conclusion: Physiological parameters (maximal oxygen uptake, erythrocytes and Hb) were insignificantly higher in the older group compared to the younger group. There were no statistically significant differences between analyzed anthropometric parameters among the two groups except for the basic measurements (height and weight).

Keywords: body composition, young soccer players, BMI, physical status

Procedia PDF Downloads 391

Authors: Aymen Laadhari

Abstract:

The cardiovascular system is centered on the heart and is characterized by a very complex structure with different physical scales in space (e.g. micrometers for erythrocytes and centimeters for organs) and time (e.g. milliseconds for human brain activity and several years for development of some pathologies). The development and numerical implementation of mathematical models of the cardiovascular system is a tremendously challenging topic at the theoretical and computational levels, inducing consequently a growing interest over the past decade. The accurate computational investigations in both healthy and pathological cases of processes related to the functioning of the human cardiovascular system can be of great potential in tackling several problems of clinical relevance and in improving the diagnosis of specific diseases. In this talk, we focus on the specific task of simulating three particular phenomena related to the cardiovascular system on the macroscopic, mesoscopic and microscopic scales, respectively. Namely, we develop numerical methodologies tailored for the simulation of (i) the haemodynamics (i.e., fluid mechanics of blood) in the aorta and sinus of Valsalva interacting with highly deformable thin leaflets, (ii) the hyperelastic anisotropic behaviour of cardiomyocytes and the influence of calcium concentrations on the contraction of single cells, and (iii) the dynamics of red blood cells in microvasculature. For each problem, we present an appropriate fully Eulerian finite element methodology. We report several numerical examples to address in detail the relevance of the mathematical models in terms of physiological meaning and to illustrate the accuracy and efficiency of the numerical methods.

Keywords: finite element method, cardiovascular system, Eulerian framework, haemodynamics, heart valve, cardiomyocyte, red blood cell

Procedia PDF Downloads 239

Authors: E. A. Alaswad, H. M. Choudhry, F. Z. Filimban

Abstract:

Sickle Cell Anemia (SCA) is one type of blood diseases related to autosomal disorder. The sickle shaped red blood cells are the main cause of many problems in the blood vessels and capillaries. Aerva Javanica (J) and Ficus Palmata (P) are medicinal plants that have many popular uses and have been proved their efficacy. The aim of this study was to assess the antioxidants activity and the antisickling effect of J and P extractions. The period of this study, air-dried leaves of J, and P plants were ground and the active components were extracted by maceration in water (W) and methanol (M) as solvents. The antioxidants activity of JW, PW, JM, and PM were assessed by way of the radical scavenging method using 2,2-diphenyl-1-picrylhydrazyl (DPPH). To determine the antisickling effect of J and P extracts. 20 samples were collected from sickle cell anemia patients. Different concentrations of J and P extracts (200 and 110 μg/mL) were added on the sample and incubated. A drop of each sample was examined with light microscope. Normal and sickled RBCs were calculated and expressed as the percent of sickling. The stabilization effect of the extracts was measured by the osmotic fragility test for erythrocytes. The finding suggests as estimated by DPPH method, all the extracts showed an antioxidant activity with a significant inhibition of the DPPH radicals. PM has the least IC50% with 71.49 μg/ml while JM was the most with 408.49 μg/ml. Sickle cells treated with extracts at different concentrations significantly reduced the percentage of sickling compering to control samples. However, JM 200 μg/mL give the highest anti-sickling affect with 17.4% of sickling compared to control 67.5 of sickling while PM at 200 μg/mL showed the highest membrane cell stability. In a conclusion, the results showed that J and P extracts have antisickling effects. Therefore, the Aerva javanica and Ficus palmata may have a role in SCA management and a good impact on the patient's lives.

Keywords: Aerva javanica, antioxidant, antisickling, Ficus palmata, sickle cell anemia

Procedia PDF Downloads 147

Authors: Ivan Zupan, Tomislav Saric, Suzana Tkalcic

Abstract:

IMUNO-2865® is a commercially available, β–glucan based, natural hemicellulose compound with proven immunostimulative properties in people, domestic and some aquatic animals. During the experimental feeding trial with IMUNO-2865® in juvenile wild-caught chub under laboratory conditions, supplementation resulted in overall higher growth performance for all experimental groups regardless of the concentration of the added compound. The maximum, 5% concentration of the supplement, resulted in highest weight gain and calculated specific growth rate. In sea bream, as economically most important species in the Mediterranean aquaculture, significant increases in numbers of monocytes and heterophils were observed in the group supplemented with 2.5 % of IMUNO-2865® in the feed. An overall increase of erythrocytes was noted by the end of the experiment, although with variable distribution among groups. Blood Ca++ levels, total proteins, and total NH₃ were significantly higher after 60 days of feeding in all treatment groups compared to the control and remained elevated in the treated group following the secession of supplementation. Superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and serum paraoxonase PON1 (U/L) showed similar trends. All these parameters are playing a significant role in either oxygen supplementation of tissues, or anabolic and catabolic processes that on molecular levels contribute to the overall health and immune-building capacity of cells and tissues. The complete lack of mortality in sea bream and presented increases in cellular, biochemical and oxidative stress parameters in the blood suggest that the IMUNO-2865® represents a safe dietary supplement for in aquaculture, with an overall positive and potentially immunostimulative effect on farmed fish.

Keywords: IMUNO-2865®, β–glucans, Mediterranean aquaculture, fish imunnostimulans

Procedia PDF Downloads 132

Authors: Shazia Anwer Bukhari, Madiha Javeed Ghani, Muhammad Ibrahim Rajoka

Abstract:

Objective: Biochemical, environmental, physical and genetic factors have a strong effect on the development of coronary disease (CAD). Plasma homocysteine (Hcy) level and DNA damage play a pivotal role in its development and progression. The aim of this study was to investigate the predictive strength of an oxidative stress, clinical biomarkers and total antioxidant status (TAS) in CAD patients to find the correlation of homocysteine, TOS and oxidative DNA damage with other clinical parameters. Methods: Sixty confirmed patients with CAD and 60 healthy individuals as control were included in this study. Different clinical and laboratory parameters were studied in blood samples obtained from patients and control subjects using commercially available biochemical kits and statistical software Results: As compared to healthy individuals, CAD patients had significantly higher concentrations of indices of oxidative stress: homocysteine (P=0.0001), total oxidative stress (TOS) (P=0.0001), serum cholesterol (P=0.04), low density lipoprotein cholesterol (LDL) (P=0.01), high density lipoprotein-cholesterol (HDL) (P=0.0001), and malondialdehyde (MDA) (P=0.001) than those of healthy individuals. Plasma homocysteine level and oxidative DNA damage were positively correlated with cholesterol, triglycerides, systolic blood pressure, urea, total protein and albumin (P values= 0.05). Both Hcy and oxidative DNA damage were negatively correlated with TAS and proteins. Conclusion: Coronary artery disease patients had a significant increase in homocysteine level and DNA damage due to increased oxidative stress. In conclusion, our study shows a significantly increase in lipid peroxidation, TOS, homocysteine and DNA damage in the erythrocytes of patients with CAD. A significant decrease level of HDL-C and TAS was observed only in CAD patients. Therefore these biomarkers may be useful diagnosis of patients with CAD and play an important role in the pathogenesis of CAD.

Keywords: antioxidants, coronary artery disease, DNA damage, homocysteine, oxidative stress, malondialdehyde, 8-Hydroxy-2’deoxyguanosine

Procedia PDF Downloads 477

Authors: Hadiza Abdullahi

Abstract:

Sialic Acids and sialidases have been implicated in many disease states particularly bacterial and viral infections which are common opportunist infections of HIV disease. Their role in HIV/AIDS is contemplated. A study was carried out to determine Sialic Acid profile and Sialidase Activity in HIV infected and Apparently Healthy individuals, and also determine the relationship between the sialic acid levels and sialidase activity. Blood samples were collected from 200 subjects (150 HIV infected individuals and 50 apparently healthy individuals divided into four groups- HIV ART Naïve, HIV Stable (on ART but have been stable with no clinical episodes), HIV-OI (on ART with opportunistic infections), and Apparently Healthy). Complete Blood Count, Erythrocyte Surface Sialic Acid (ESSA), Free Serum Sialic Acid (FSSA) concentrations and Sialidase activity were determined for all 200 subjects. Analysis of variance (ANOVA) was used to compare the results of the different groups of HIV infected individuals as well as controls. The mean haemoglobin (HGB), Packed Cell Volume (PCV) and Red Blood Cells (RBC) concentrations were significantly lower (P ≤ 0.05) in the HIV groups compared with the apparently healthy groups. Anaemia and neutropaenia were the most common heamatological abnormalities observed in this study with highest prevalence of anaemia found in the ART naive group. The mean FSSA was 0.4±0.4mg/ml. There was a significant difference (p ≤ 0.05) between some groups. The highest levels of FSSA was observed in the HIV ART naïve (0.65±0.5mg/ml). The mean ESSA value for the study population was 0.54±0.35mg/ml with no significant difference (p ≤ 0.05) between groups. The mean sialidase activity values were 0.52±0.1 µmol/min/µl, 0.40±0.1 µmol/min/µl, 0.45±0.1 µmol/min/µl and 0.41±0.1 µmol/min/µl for the HIV ART naïve, HIV stable, HIV/OIs and apparently healthy groups respectively. No significant difference (p ≤ 0.05) was found between groups and also in gender and age. The finding in this study of higher mean sialidase activity and FSSA levels in the ART naïve HIV group compared with other groups indicate that the virus and other opportunistic pathogens may be sialidase producers in vivo which cleave off sialic acids from erythrocytes surface, leading to high levels of FSSA, anaemia and neutropaenia seen in this group. The higher ESSA concentration found in the HIV stable group along with lowest FSSA concentration in the group suggests the presence of sialyltransferases.

Keywords: erythrocyte surface sialic acid, free serum sialic acid, HIV, sialidase

Procedia PDF Downloads 196

Authors: Raghuram Kandimalla, Sanjeeb Kalita, Bhaswati Choudhury, Jibon Kotoky

Abstract:

The quest for developing an ideal suture material prompted our interest to develop a novel suture with advantageous characteristics to market available ones. We developed novel suture biomaterial from muga silk (Antheraea assama) and ramie (Boehmeria nivea) plant fiber. Field emission scanning electron microscopy (FE-SEM), energy-dispersive X-ray spectroscopy (EDX), attenuated total reflection fourier transform infrared spectroscopy (ATR-FTIR) and thermo gravimetric analysis (TGA) results revealed the physicochemical properties of the fibers which supports the suitability of fibers for suture fabrication. Tensile properties of the prepared sutures were comparable with market available sutures and it found to be biocompatible towards human erythrocytes and nontoxic to mammalian cells. The prepared sutures completely healed the superficial deep wound incisions within seven days in adult male wister rats leaving no rash and scar. Histopathology studies supports the wound healing ability of sutures, as rapid synthesis of collagen, connective tissue and other skin adnexal structures were observed within seven days of surgery. Further muga suture surface modified by exposing the suture to oxygen plasma which resulted in formation of nanotopography on suture surface. Broad spectrum antibiotic amoxicillin was functionalized on the suture surface to prepare an advanced antimicrobial muga suture. Surface hydrophilicity induced by oxygen plasma results in an increase in drug-impregnation efficiency of modified muga suture by 16.7%. In vitro drug release profiles showed continuous and prolonged release of amoxicillin from suture up to 336 hours. The advanced muga suture proves to be effective against growth inhibition of Staphylococcus aureus and Escherichia coli, whereas normal muga suture offers no antibacterial activity against both types of bacteria. In vivo histopathology studies and colony-forming unit count data revealed accelerated wound healing activity of advanced suture over normal one through rapid synthesis and proliferation of collagen, hair follicle and connective tissues.

Keywords: sutures, biomaterials, silk, Ramie

Procedia PDF Downloads 299

Authors: Eszter J. Tóth, Alexandra Hoffmann, Csaba Vágvölgyi, Tamás Papp

Abstract:

Members of the genera Curvularia and Bipolaris are closely related melanin producing filamentous fungi; both of them have the teleomorph states in genus Cochliobolus. While Bipolaris species infect only plants and may cause serious agriculture damages, some Curvularia species was recovered from opportunistic human infections. The human pathogenic species typically cause phaeohyphomycoses, i.e. mould infections caused by melanised fungi, which can manifest as invasive mycoses with frequent involvement of the central nervous system in immunocompromised patients or as local infections (e.g. keratitis, sinusitis, and cutaneous lesions) in immunocompetent people. Although their plant-fungal interactions have been intensively studied, there is only little information available about the human pathogenic feature of these fungi. The aim of this study was to investigate the neutrophil granulocytes’ response to hyphal forms of Curvularia and Bipolaris in comparison with the response to Aspergillus. In the present study Curvularia lunata SZMC 23759 and Aspergillus fumigatus SZMC 23245 both isolated from human eye infection, and Bipolaris zeicola BRIP 19582b isolated from plant leaf were examined. Neutrophils were isolated from heparinised venous blood of healthy donors with dextran sedimentation followed by centrifugation over Ficoll and hypotonic lysis of erythrocytes. Viability and purity of the cells were checked with trypan blue and Wright staining, respectively. Infection of neutrophils was carried out with germinated conidia in a ratio of 5:1. Production of hydrogen peroxide, superoxide anion, and nitrogen monoxide was measured both intracellularly and extracellularly in response to the germinated spores with or without the supernatant and after serum treatment. ROS and NOS production of neutrophils in interaction with the three fungi were compared. It is already known that Aspergillus species induce ROS production of neutrophils only after serum treatment. Although, in case of C. lunata, serum opsonisation also induced an intensive production of reactive species, lower level of production was measured in the lack of serum as well. After interaction with the plant pathogenic B. zeicola, amount of reactive species found to be similar with and without serum treatment. The presence of germination supernatant decreased the reactive species production in case of each fungus. Interaction with Curvularia, Bipolaris and Aspergillus species induced different response of neutrophils. It seems that recognition of C. lunata and B. zeicola is independent of serum opsonisation, albeit it increases the level of the produced reactive species in response for C. lunata. The study was supported by the grant LP2016-8/2016.

Keywords: Curvularia, neutrophils, NOS, ROS, serum opsonisation

Procedia PDF Downloads 190

Authors: Santi Maneewatchararangsri, Wanpen Chaicumpa, Patcharin Saengjaruk, Urai Chaisri

Abstract:

Leptospirosis is a globally neglected disease that continues to be a significant public health and veterinary burden, with millions of cases reported each year. Early and accurate differential diagnosis of leptospirosis from other febrile illnesses and the development of a broad spectrum of leptospirosis vaccines are needed. The LipL32 outer membrane lipoprotein is a member of Leptospira adhesive matrices and has been found to exert hemolytic activity to erythrocytes in vitro. Therefore, LipL32 is regarded as a potential target for diagnosis, broad-spectrum leptospirosis vaccines, and for passive immunotherapy. In this study, we established LipL32-specific mouse monoclonal antibodies, mAbLPF1 and mAbLPF2, and their respective mouse- and humanized-engineered single chain variable fragment (ScFv). Their antibodies’ neutralizing activities against Leptospira-mediated hemolysis in vitro, and the therapeutic efficacy of mAbs against heterologous Leptospira infected hamsters were demonstrated. The epitope peptide of mAb LPF1 was mapped to a non-contiguous carboxy-terminal β-turn and amphipathic α-helix of LipL32 structure contributing to phospholipid/host cell adhesion and membrane insertion. We found that the mAbLPF2 epitope was located on the interacting loop of peptide binding groove of the LipL32 molecule responsible for interactions with host constituents. Epitope sequences are highly conserved among Leptospira spp. and are absent from the LipL32 superfamily of other microorganisms. Both epitopes are surface-exposed, readily accessible by mAbs, and immunogenic. However, they are less dominant when revealed by LipL32-specific immunoglobulins from leptospirosis-patient sera and rabbit hyperimmune serum raised by whole Leptospira. Our study also demonstrated an adhesion inhibitory activity of LipL32 protein to host membrane components and cells mediated by mAbs as well as an anti-hemolytic activity of the respective antibodies. The therapeutic antibodies, particularly the humanized-ScFv, have a potential for further development as non-drug therapeutic agent for human leptospirosis, especially in subjects allergic to antibiotics. The epitope peptides recognized by two therapeutic mAbs have potential use as tools for structure-function studies. Finally, protective peptides may be used as a target for epitope-based vaccines for control of leptospirosis.

Keywords: leptospira lipl32-specific antibodies, therapeutic epitopes, epitopes characterization, immunotherapy

Procedia PDF Downloads 289

Authors: Nina Ivanova

Abstract:

More than one third of the population and animals are infected with parasitic helminths. It is especially difficult to cure the larval forms of parasites. The larvae of Hymenolepis nana invade the villi of the intestinal mucosa. Toxocara larvae can live in the liver, heart, lungs, brain, eyes, and pancreas. Commercial antiparasitic drugs cannot guarantee a 100% cure after a single course of treatment, because parasite larvae invade the villi of the intestinal mucosa and the anthelmintics do not reach and kill cestode larvae. The aim of this work was to conduct a study of liposomal antihelminthics on the laboratory animals under the certification received from the Committee on Bioethics and Deontology. It has been checked: 1) anthelmintic activity of the liposomal form of fenasal in experimental hymenolepidosis of white mice (larval stage - Hymenolepis nana). 2) anthelmintic activity of the liposomal form of albendazole in experimental toxocariasis of white mice (in the lungs at the stage of larval migration). Since some helminths cause hemolysis of erythrocytes as we used a mixture of polar lipids developed by us with antihemolytic activity to obtain liposomes Fenasal and albendazole were included in the liposome membrane in the ratio of anthelmintic: lipids 1:10. . The average size of liposomes was 180 nm, and the concentration of lipids in liposomes was 1%. The researches were carried out on white male mice who were infected with Hymenolepis Nana invasional eggs in a peroral way with a doze of 100 eggs per animal. On the 5th day after infection, a liposomal fenasal and commercial fenasal were administered orally for comparison. The animals were observed for 15 days. Before the introduction of liposomes and on the 3rd, 5th, 15th day after the administration of the drug, studies were carried out on the presence of helminths in the organs of animals. The liposomal fenasal, when administered orally, had an anthelmintic effect on Hymenolepis Nana cysticercoids at a dose of 25 mg/kg. The percentage of efficiency was 90.06, 91.36 96.85% on days 3, 5, 15, respectively. For comparison, the commercial activity was at a dose of 200 mg/ml, which is 8 times higher than the dose of liposomal fenasal. To evaluate the anthelmintic effect of the liposomal form of albendazole in experimental toxocariasis of white mice (toxascaris in the lungs at the stage of larval migration), studies were also carried out on white mice. The animals were infected with invasive eggs of Toxocara canis, orally, at a dose of 100 eggs per animal. On the 5th day after infection, the liposomal albendazole was administered orally. The efficacy of the study dosage form was determined by counting Larva mirgans larvae in the lungs. The results obtained showed that the liposomal albendazole had the greatest anthelmintic effect on Toxocara larvae at a dose of 2.0 mg/kg, which was 3.75 times less than the therapeutic one. At the same time, the percentage of efficiency was 93.75% on the 3rd day, and 98.66% on the 5th day.

Keywords: hymenolepis, Toxocara, larvae, liposomes

Procedia PDF Downloads 99

Authors: Akram Abi, Mohammad Hossein Sarrafzadeh

Abstract:

Because of their several advantages over chemical surfactants, biosurfactants have given rise to a growing interest in the past decades. Advantages such as lower toxicity, higher biodegradability, higher selectivity and applicable at extreme temperature and pH which enables them to be used in a variety of applications such as: enhanced oil recovery, environmental and pharmaceutical applications, etc. Bacillus subtilis produces a cyclic lipopeptide, called surfactin, which is one of the most powerful biosurfactants with ability to decrease surface tension of water from 72 mN/m to 27 mN/m. In addition to its biosurfactant character, surfactin exhibits interesting biological activities such as: inhibition of fibrin clot formation, lyses of erythrocytes and several bacterial spheroplasts, antiviral, anti-tumoral and antibacterial properties. Surfactin is an antibiotic substance and has been shown recently to possess anti-HIV activity. However, application of biosurfactants is limited by their high production cost. The cost can be reduced by optimizing biosurfactant production using cheap feed stock. Utilization of inexpensive substrates and unconventional carbon sources like urban or agro-industrial wastes is a promising strategy to decrease the production cost of biosurfactants. With suitable engineering optimization and microbiological modifications, these wastes can be used as substrates for large-scale production of biosurfactants. As an effort to fulfill this purpose, in this work we have tried to utilize olive oil as second carbon source and also yeast extract as second nitrogen source to investigate the effect on both biomass and biosurfactant production improvement in Bacillus subtilis cultures. Since the turbidity of the culture was affected by presence of the oil, optical density was compromised and no longer could be used as an index of growth and biomass concentration. Therefore, cell Dry Weight measurements with applying necessary tactics for removing oil drops to prevent interference with biomass weight were carried out to monitor biomass concentration during the growth of the bacterium. The surface tension and critical micelle dilutions (CMD-1, CMD-2) were considered as an indirect measurement of biosurfactant production. Distinctive and promising results were obtained in the cultures containing olive oil compared to cultures without it: more than two fold increase in biomass production (from 2 g/l to 5 g/l) and considerable reduction in surface tension, down to 40 mN/m at surprisingly early hours of culture time (only 5hr after inoculation). This early onset of biosurfactant production in this culture is specially interesting when compared to the conventional cultures at which this reduction in surface tension is not obtained until 30 hour of culture time. Reducing the production time is a very prominent result to be considered for large scale process development. Furthermore, these results can be used to develop strategies for utilization of agro-industrial wastes (such as olive oil mill residue, molasses, etc.) as cheap and easily accessible feed stocks to decrease the high costs of biosurfactant production.

Keywords: agro-industrial waste, bacillus subtilis, biosurfactant, fermentation, second carbon and nitrogen source, surfactin

Procedia PDF Downloads 287

Authors: Sergey Kozlov, Juliya Kozlova

Abstract:

Introduction: The increased attention of researchers to an explosive trauma around the world is associated with a constant renewal of military weapons and a significant increase in terrorist activities using explosive devices. Explosive wave is a well known damaging factor of explosion. The most sensitive to the action of explosive wave in the human body are the head brain, lungs, intestines, urine bladder. The severity of damage to these organs depends on the distance from the explosion epicenter to the object, the power of the explosion, presence of barriers, parameters of the body position, and the presence of protective clothing. One of the places where a shock wave acts, in human tissues and organs, is the vascular endothelial barrier, which suffers the greatest damage in the head brain and lungs. The objective of the study was to determine the pathomorphological changes of the head brain followed the action of explosive wave. Materials and methods of research: To achieve the purpose of the study, there have been studied 6 male corpses delivered to the morgue of Municipal Institution "Dnipropetrovsk regional forensic bureau" during 2014-2016 years. The cause of death of those killed was a military explosive injury. After a visual external assessment of the head brain, for histological study there was conducted the 1 x 1 x 1 cm/piece sampling from different parts of the head brain, i.e. the frontal, parietal, temporal, occipital sites, and also from the cerebellum, pons, medulla oblongata, thalamus, walls of the lateral ventricles, the bottom of the 4th ventricle. Pieces of the head brain were immersed in 10% formalin solution for 24 hours. After fixing, the paraffin blocks were made from the material using the standard method. Then, using a microtome, there were made sections of 4-6 micron thickness from paraffin blocks which then were stained with hematoxylin and eosin. Microscopic analysis was performed using a light microscope with x4, x10, x40 lenses. Results of the study: According to the results of our study, injuries of the head brain were divided into macroscopic and microscopic. Macroscopic injuries were marked according to the results of visual assessment of haemorrhages under the membranes and into the substance, their nature, and localisation, areas of softening. In the microscopic study, our attention was drawn to both vascular changes and those of neurons and glial cells. Microscopic qualitative analysis of histological sections of different parts of the head brain revealed a number of structural changes both at the cellular and tissue levels. Typical changes in most of the studied areas of the head brain included damages of the vascular system. The most characteristic microscopic sign was the separation of vascular walls from neuroglia with the formation of perivascular space. Along with this sign, wall fragmentation of these vessels, haemolysis of erythrocytes, formation of haemorrhages in the newly formed perivascular spaces were found. In addition to damages of the cerebrovascular system, destruction of the neurons, presence of oedema of the brain tissue were observed in the histological sections of the brain. On some sections, the head brain had a heterogeneous step-like or wave-like nature. Conclusions: The pathomorphological microscopic changes in the brain, identified in the study on the died of explosive traumas, can be used for diagnostic purposes in conjunction with other characteristic signs of explosive trauma in forensic and pathological studies. The complex of microscopic signs in the head brain, i.e. separation of blood vessel walls from neuroglia with the perivascular space formation, fragmentation of walls of these blood vessels, erythrocyte haemolysis, formation of haemorrhages in the newly formed perivascular spaces is the direct indication of explosive wave action.

Keywords: blast wave, neurotrauma, human, brain

Procedia PDF Downloads 179