Search results for: dried blood spots

Authors: Fahimeh Palizban, Farshad Noravesh, Amir Hossein Saeidian, Mahya Mehrmohamadi

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In the recent decade, the emergence of liquid biopsy has significantly improved cancer monitoring and detection. Dying cells, including those originating from tumors, shed their DNA into the blood and contribute to a pool of circulating fragments called cell-free DNA. Accordingly, identifying the tissue origin of these DNA fragments from the plasma can result in more accurate and fast disease diagnosis and precise treatment protocols. Open chromatin regions are important epigenetic features of DNA that reflect cell types of origin. Profiling these features by DNase-seq, ATAC-seq, and histone ChIP-seq provides insights into tissue-specific and disease-specific regulatory mechanisms. There have been several studies in the area of cancer liquid biopsy that integrate distinct genomic and epigenomic features for early cancer detection along with tissue of origin detection. However, multimodal analysis requires several types of experiments to cover the genomic and epigenomic aspects of a single sample, which will lead to a huge amount of cost and time. To overcome these limitations, the idea of predicting OCRs from WGS is of particular importance. In this regard, we proposed a computational approach to target the prediction of open chromatin regions as an important epigenetic feature from cell-free DNA whole genome sequence data. To fulfill this objective, local sequencing depth will be fed to our proposed algorithm and the prediction of the most probable open chromatin regions from whole genome sequencing data can be carried out. Our method integrates the signal processing method with sequencing depth data and includes count normalization, Discrete Fourie Transform conversion, graph construction, graph cut optimization by linear programming, and clustering. To validate the proposed method, we compared the output of the clustering (open chromatin region+, open chromatin region-) with previously validated open chromatin regions related to human blood samples of the ATAC-DB database. The percentage of overlap between predicted open chromatin regions and the experimentally validated regions obtained by ATAC-seq in ATAC-DB is greater than 67%, which indicates meaningful prediction. As it is evident, OCRs are mostly located in the transcription start sites (TSS) of the genes. In this regard, we compared the concordance between the predicted OCRs and the human genes TSS regions obtained from refTSS and it showed proper accordance around 52.04% and ~78% with all and the housekeeping genes, respectively. Accurately detecting open chromatin regions from plasma cell-free DNA-seq data is a very challenging computational problem due to the existence of several confounding factors, such as technical and biological variations. Although this approach is in its infancy, there has already been an attempt to apply it, which leads to a tool named OCRDetector with some restrictions like the need for highly depth cfDNA WGS data, prior information about OCRs distribution, and considering multiple features. However, we implemented a graph signal clustering based on a single depth feature in an unsupervised learning manner that resulted in faster performance and decent accuracy. Overall, we tried to investigate the epigenomic pattern of a cell-free DNA sample from a new computational perspective that can be used along with other tools to investigate genetic and epigenetic aspects of a single whole genome sequencing data for efficient liquid biopsy-related analysis.

Keywords: open chromatin regions, cancer, cell-free DNA, epigenomics, graph signal processing, correlation clustering

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Authors: Mingqian Zhang

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Tip-enhanced Raman spectroscopy (TERS) is a scanning probe technique for individual objects and structured surfaces investigation that provides a wealth of enhanced spectral information with nanoscale spatial resolution and high detection sensitivity. It has become a powerful and promising chemical and physical information detection method in the nanometer scale. The TERS technique uses a sharp metallic tip regulated in the near-field of a sample surface, which is illuminated with a certain incident beam meeting the excitation conditions of the wave-vector matching. The local electric field, and, consequently, the Raman scattering, from the sample in the vicinity of the tip apex are both greatly tip-enhanced owning to the excitation of localized surface plasmons and the lightning-rod effect. Typically, a TERS setup is composed of a scanning probe microscope, excitation and collection optical configurations, and a Raman spectroscope. In the illumination configuration, an objective lens or a parabolic mirror is always used as the most important component, in order to focus the incident beam on the tip apex for excitation. In this research, a novel TERS setup was built up by introducing a plasmonic lens to the excitation optics as a focusing device. A plasmonic lens with symmetry breaking semi-annular slits corrugated on gold film was designed for the purpose of generating concentrated sub-wavelength light spots with strong longitudinal electric field. Compared to conventional far-field optical components, the designed plasmonic lens not only focuses an incident beam to a sub-wavelength light spot, but also realizes a strong z-component that dominants the electric field illumination, which is ideal for the excitation of tip-enhancement. Therefore, using a PL in the illumination configuration of TERS contributes to improve the detection sensitivity by both reducing the far-field background and effectively exciting the localized electric field enhancement. The FDTD method was employed to investigate the optical near-field distribution resulting from the light-nanostructure interaction. And the optical field distribution was characterized using an scattering-type scanning near-field optical microscope to demonstrate the focusing performance of the lens. The experimental result is in agreement with the theoretically calculated one. It verifies the focusing performance of the plasmonic lens. The optical field distribution shows a bright elliptic spot in the lens center and several arc-like side-lobes on both sides. After the focusing performance was experimentally verified, the designed plasmonic lens was used as a focusing component in the excitation configuration of TERS setup to concentrate incident energy and generate a longitudinal optical field. A collimated linearly polarized laser beam, with along x-axis polarization, was incident from the bottom glass side on the plasmonic lens. The incident light focused by the plasmonic lens interacted with the silver-coated tip apex and enhanced the Raman signal of the sample locally. The scattered Raman signal was gathered by a parabolic mirror and detected with a Raman spectroscopy. Then, the plasmonic lens based setup was employed to investigate carbon nanotubes and TERS experiment was performed. Experimental results indicate that the Raman signal is considerably enhanced which proves that the novel TERS configuration is feasible and promising.

Keywords: longitudinal electric field, plasmonics, raman spectroscopy, tip-enhancement

Procedia PDF Downloads 373

Authors: Aiche Mohamed Amine, Mallem Leila, Yahia El Khansa, Boulakoud Mohamed Salah

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Until recently, toxicological studies focused on the effects of individual chemicals. However, humans and wildlife are exposed to a complex milieu of chemicals from different sources including food and water, personal care products and the environment. The aim of this study is to detect the toxicity of two fungicides and their mixtures in the fertility and oxidative damge induced in the rat. The male of rats (28) were used, they were divided in four groups (7 rats of each group) and one group was used as control. Rats were dosed orally with Propiconazole 60mg/Kg/day, Propinebe 100mg/Kg/day and their mixture 30mg Propiconazole/kg/day + 50mg Propineb/kg/day for 4 weeks. Animals were observed for clinical toxicity. At the end of treatment period, animals of all groups were scarified, blood was collected for hematological and biochemical’s analysis and desired organs were removed and weighted. The results indicated that the fungicide and their mixture were toxic in the treated animals. The semen study showed a decrease in the count and mobility of spermatozoa in all treated group, it was also a decrease in the weight of the testis and epidydimis in the treated group as compared with control. Reduced glutathione (GSH), Glutathione peroxidase (GPx) level was decreased in all treated groups.

Keywords: fungicides, mixtures, fertility, oxidative stress

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Authors: Alice Nwanyioma Ohuoba, Rose Erdoo Kukwa, Ukpabi Joseph Ukpabi

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Cassava root (Manihot esculenta) is one of the important carbohydrates containing crops in Nigeria. It is a staple food, mostly in the southern part of the country, and a source of income to farmers and processors. Cassava gari processing methods result to residue fiber (solid waste) from the sieving operation, these residue fibers ( solid wastes) can be dried and milled into flour and used to prepare cakes, cookies, crackers and chin-chin instead of being thrown away mostly on farmland or near the residential area. Flour for baking or frying may contain carbohydrates and protein (wheat flour) or rich in only carbohydrates (cassava flour). Cake, cookies, crackers, and chin-chin were prepared using the residue flour obtained from the residue fiber of cassava variety NR87184 roots, processed into gari. This study is aimed at evaluating the proximate composition, mineral content and sensory attributes of these selected snacks produced. The proximate composition results obtained showed that crackers had the lowest value in moisture (2.3390%) and fat (1.7130%), but highest in carbohydrates (85.2310%). Amongst the food products, cakes recorded the highest value in protein (8.0910%). Crude fibre values ranges from 2.5265% (cookies) to 3.4165% (crackers). The result of the mineral contents showed cookies ranking the highest in Phosphorus (65.8535 ppm) and Iron (0.1150 mg/L), Calcium (1.3800mg/L) and Potassium (7.2850 mg/L) contents, while chin-chin and crackers were lowest in Sodium ( 2.7000 mg/L). The food products were also subjected to sensory attributes evaluation by thirty member panelists using 9-hedonic scale which ranged from 1 ( dislike extremely) to 9 (like extremely). The means score obtained shows all the food products having above 7.00 (above “like moderately”). This study has shown that food products that may be functional or nutraceuticals could be prepared from the residue flour. There is a call for the use of gluten-free flour in baking due to ciliac disease and other allergic causes by gluten. Therefore local carbohydrates food crops like cassava residue flour that are gluten-free, could be the solution. In addition, this could aid cassava gari processing waste management thereby reducing post-harvest losses of cassava root.

Keywords: allergy, flour, food-products, gluten-free

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Authors: Adele Broodryk, Cindy Pienaar, Martinique Sparks, Ben Coetzee

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Background: Cold water immersion (CWI) is a popular recovery modality utilised. However, discrepancies exist regarding the results over a 48 hour recovery period. Aim: To evaluate the effects of CWI and passive recovery (PAR) on a range of haematological and physical parameters over a 48-hour using a cross-sectional, pre-post-test design. Subjects and Methods: Both the and physical parameters were evaluated at baseline, after a 15-min fitness session, and at 0, 24 and 48 hours post-recovery in 23 male university rugby players. The CWI group sat in a cold water pool (8°C) for 20 min whereas the PAR group remained seated. Results: At 0 hours post-CWI, three (blood lactate (BLa-), Sodium (Na+) and haemoglobin) returned to baseline values, however Vertical Jump Test (VJT) height results decreased whereas after PAR it improved. From 0 to 24 and/or 48 h, four (Partial Oxygen (PO2) VJT-height, plasma glucose, and Na+) significantly increased (p ≤ 0.05) in either and/or both groups. Significant intergroup differences (p ≤ 0.05) were noticed in the physical tests. Conclusions: PAR is superior as an acute modality (0 hours) due to CWI cooling the body down. However, CWI demonstrates advantageous over a 24-hour period in a wide range of haematological variables.

Keywords: cryotherapy, recuperation, haematological, rugby

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Authors: Malarvizhi Selvaraj, Paul Fergus, Andy Shaw

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Uterine contractions produced during labour have the potential to damage the foetus by diminishing the maternal blood flow to the placenta. In order to observe this phenomenon labour and delivery are routinely monitored using cardiotocography monitors. An obstetrician usually makes the diagnosis of foetus hypoxia by interpreting cardiotocography recordings. However, cardiotocography capture and interpretation is time-consuming and subjective, often lead to misclassification that causes damage to the foetus and unnecessary caesarean section. Both of these have a high impact on the foetus and the cost to the national healthcare services. Automatic detection of foetal heart rate may be an objective solution to help to reduce unnecessary medical interventions, as reported in several studies. This paper aim is to provide a system for better identification and interpretation of abnormalities of the fetal heart rate using RStudio. An open dataset of 552 Intrapartum recordings has been filtered with 0.034 Hz filters in an attempt to remove noise while keeping as much of the discriminative data as possible. Features were chosen following an extensive literature review, which concluded with FIGO features such as acceleration, deceleration, mean, variance and standard derivation. The five features were extracted from 552 recordings. Using these features, recordings will be classified either normal or abnormal. If the recording is abnormal, it has got more chances of hypoxia.

Keywords: cardiotocography, foetus, intrapartum, hypoxia

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Authors: Pramote Thangkratok

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The aims of this experimental study were to investigate the effect of Reverse Trendelenburg Position on the Back Pain after Cardiovascular Angiography and Interventions. In addition, to compare bleeding and hematoma occurrences at the Access site between experimental and control groups. The randomized controlled trial (RCT) was conducted in 70 patients who underwent Cardiovascular Angiography and Interventions via the femoral artery and received post procedural care at the intermediate cardiac care unit, Bangkok Heart Hospital. From December 2015 to February 2016. The control group (35 patients) was to get standard care after the intervention, whereas the experimental group (35 patients) was Reverse Trendelenburg Position 30-45 degrees. The groups were not significantly different in terms of demographic characteristics, Age, Gender, BMI, blood pressure, heart rate. While not significantly different from each other, the intensity of back pain control group had a significantly higher pain score than experimental group. Vascular complications in terms of bleeding and hematoma were not significantly different between the control and experimental groups. The findings show that Reverse Trendelenburg Position after Cardiovascular Angiography and Interventions would reduce or prevent the back pain without increasing the chance of bleeding and hematoma.

Keywords: reverse trendelenburg position, back pain, cardiovascular angiography, cardiovascular interventions

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Authors: Ankita Srivastava, Bhupendra S. Butola, Abhijit Majumdar

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Application of shear thickening fluid (STF) has been proved to increase the impact resistance performance of the textile structures to further use it as a body armor material. In the present research, STF was applied on Kevlar woven fabric to make the structure lightweight and flexible while improving its impact resistance performance. It was observed that getting a fair amount of add-on of STF on Kevlar fabric is difficult as Kevlar fabric comes with a pre-coating of PTFE which hinders its absorbency. Hence, a method termed as sequential padding is developed in the present study to improve the add-on of STF on Kevlar fabric. Contrary to the conventional process, where Kevlar fabric is treated with STF once using any one pressure, in sequential padding method, the Kevlar fabrics were treated twice in a sequential manner using combination of two pressures together in a sample. 200 GSM Kevlar fabrics were used in the present study. STF was prepared by adding PEG with 70% (w/w) nano-silica concentration. Ethanol was added with the STF at a fixed ratio to reduce viscosity. A high-speed homogenizer was used to make the dispersion. Total nine STF treated Kevlar fabric samples were prepared by using varying combinations and sequences of three levels of padding pressure {0.5, 1.0 and 2.0 bar). The fabrics were dried at 80°C for 40 minutes in a hot air oven to evaporate ethanol. Untreated and STF treated fabrics were tested for add-on%. Impact resistance performance of samples was also tested on dynamic impact tester at a fixed velocity of 6 m/s. Further, to observe the impact resistance performance in actual condition, low velocity ballistic test with 165 m/s velocity was also performed to confirm the results of impact resistance test. It was observed that both add-on% and impact energy absorption of Kevlar fabrics increases significantly with sequential padding process as compared to untreated as well as single stage padding process. It was also determined that impact energy absorption is significantly better in STF treated Kevlar fabrics when 1st padding pressure is higher, and 2nd padding pressure is lower. It is also observed that impact energy absorption of sequentially padded Kevlar fabric shows almost 125% increase in ballistic impact energy absorption (40.62 J) as compared to untreated fabric (18.07 J).The results are owing to the fact that the treatment of fabrics at high pressure during the first padding is responsible for uniform distribution of STF within the fabric structures. While padding with second lower pressure ensures the high add-on of STF for over-all improvement in the impact resistance performance of the fabric. Therefore, it is concluded that sequential padding process may help to improve the impact performance of body armor materials based on STF treated Kevlar fabrics.

Keywords: body armor, impact resistance, Kevlar, shear thickening fluid

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Authors: Sarwan Dhir, Suma Basak, Dipika Parajulee

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Alfalfa is renowned for its nutritional and biopharmaceutical value as a perennial forage legume. However, establishing a rapid plant regeneration protocol using somatic embryogenesis and efficient transformation frequency are the crucial prerequisites for gene editing in alfalfa. This study was undertaken to establish and improve the protocol for somatic embryogenesis and subsequent plant regeneration. The experiments were conducted in response to natural sensitivity using various antibiotics such as cefotaxime, carbenicillin, gentamycin, hygromycin, and kanamycin. Using 3-week-old leaf tissue, somatic embryogenesis was initiated on Gamborg’s B5 basal (B5H) medium supplemented with 3% maltose, 0.9µM Kinetin, and 4.5µM 2,4-D. Embryogenic callus (EC) obtained from the B5H medium exhibited a high rate of somatic embryo formation (97.9%) after 3 weeks when the cultures were placed in the dark. Different developmental stages of somatic embryos and cotyledonary stages were then transferred to Murashige and Skoog’s (MS) basal medium under light, resulting in a 94% regeneration rate of plantlets. Our results indicate that leaf segments can grow (tolerate) up to 450 mg/L of cefotaxime and 400 mg/L of carbenicillin in the culture medium. However, the survival threshold for hygromycin at 12.5 mg/L, kanamycin at 250 mg/L, gentamycin at 50 mg/L, and timentin (300 mg/L). The experiment to improve the protocol for achieving efficient transient gene expression in alfalfa through genetic transformation with the Agrobacterium tumefaciens pCAMBIA1304 vector was also conducted. The vector contains two reporter genes such as β-glucuronidase (GUS) and green fluorescent protein (GFP), along with a selectable hygromycin B phosphotransferase gene (HPT), all driven under the CaMV 35s promoter. Various transformation parameters were optimized using 3-week-old in vitro-grown plantlets. The different parameters such as types of explant, leaf ages, preculture days, segment sizes, wounding types, bacterial concentrations, infection periods, co-cultivation periods, different concentrations of acetosyringone, silver nitrate, and calcium chloride were optimized for transient gene expression. The transient gene expression was confirmed via histochemical GUS and GFP visualization under fluorescent microscopy. The data were analyzed based on the semi-quantitative observation of the percentage and number of blue GUS spots on different days of agro-infection. The highest percentage of GUS positivity (76.2%) was observed in 3-week-old leaf segments wounded using a scalpel blade of 11 size- after 3 days of post-incubation at a bacterial concentration of 0.6, with 2 days of preculture, 30 min of bacterial-leaf segment co-cultivation, with the addition of 150 µM acetosyringone, 4 mM calcium chloride, and 75 µM silver nitrate. Our results suggest that various factors influence T-DNA delivery in the Agrobacterium-mediated transformation of alfalfa. The stable gene expression in the putative transgenic tissue was confirmed using PCR amplification of both marker genes, indicating that gene expression in explants was not solely due to Agrobacterium, but also from transformed cells. The improved protocol could be used for generating transgenic alfalfa plants using genome editing techniques such as CRISPR/Cas9.

Keywords: Medicago sativa l. (Alfalfa), agrobacterium tumefaciens, β-glucuronidase, green fluorescent protein, transient gene

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Authors: J. H. Bang, H. J. Baek, K. I. Kim, B. J. Lee, H. J. Jung, H. J. Jang, S. K. Jung

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Asthma is a chronic inflammatory lung disease characterized by airway hyper responsiveness (AHR), airway obstruction and airway wall remodeling responsible for significant morbidity and mortality worldwide. Genetic and environment factors may result in asthma, but there are no the exact causes of asthma. Chungpye-tang (CPT) has been prescribed as a representative aerosol agent for patients with dyspnea, cough and phlegm in the respiratory clinic at Kyung Hee Korean Medicine Hospital. This Korean herbal medicines have the effect of dispelling external pathogen and dampness pattern. CPT is composed of 4 species of herbal medicines. The 4 species of herbal medicines are Ephedrae herba, Pogostemonis(Agatachis) herba, Caryophylli flos and Zingiberis rhizoma crudus. CPT suppresses neutrophil infiltration and the production of pro-inflammatory cytokines in lipopolysaccharide (LPS)-induced acute lung injury (ALI) mouse model. Moreover, the anti-inflammatory effects of CPT on a mouse model of Chronic Obstructive Pulmonary Disease (COPD) was proved. Activation of the NF-κB has been proven that it plays an important role in inflammation via inducing transcription of pro-inflammatory genes. Over-expression of NF-κB has been believed be related to many inflammatory diseases such as arthritis, gastritis, asthma and COPD. So we firstly hypothesize whether CPT has an anti-inflammatory effect on asthmatic human airway epithelial tissue via inhibiting NF-κB pathway. In this study, CPT was extracted with distilled water for 3 hours at 100°C. After process of filtration and evaporation, it was freeze dried. And asthmatic human lung tissues were provided by MatTek Corp. We investigated the precise mechanism of the anti-inflammatory effect of CPT by western blotting analysis. We observed whether the decoction extracts could reduce NF-κB activation, COX-2 protein expression and NF-κB-mediated pro-inflammatory cytokines such as TNF-α, eotaxin, IL-4, IL-9 and IL-13 in asthmatic human lung tissue. As results of this study, there was a trend toward decreased NF-κB expression in asthmatic human airway epithelial tissue. We found that the inhibition effects of CPT on COX-2 expression was not determined. IL-9 and IL-13 secretion was significantly reduced in the asthmatic human lung tissue treated with CPT. Overall, our results indicate that CPT has an anti-inflammatory effect through blocking the signaling pathway of NF-κB, thereby CPT may be a potential remedial agent for allergic asthma.

Keywords: Chungpye-tang, allergic asthma, asthmatic human airway epithelial tissue, nuclear factor kappa B (NF-κB) pathway, COX-2

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Authors: Soyeon Kang, Youn-Jee Chung, Jung Namkung

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Objective: This retrospective study investigated the association between uric acid level and bone mineral density (BMD) in the postmenopausal period. Methods: The study included 328 menopausal women (mean age, 57.3 ± 6.5 years; mean serum uric acid level, 4.6 ± 1.0 mg/dL). Patients were divided into three groups by tertile of serum uric acid level. Patients who used hormone treatment (HT), bisphosphonates, or lipid-lowering agents were included. Results: Blood urea nitrogen, serum creatinine, and serum triglyceride levels were significantly higher in the upper uric acid tertiles. No significant difference was found in the mean uric acid levels between medication users and non-users. Distinct HT regimens showed different mean serum uric acid levels. In a cross-sectional analysis, higher serum uric acid levels showed a tendency toward increased BMD in the spine and femoral neck. Longitudinal analysis of 186 women who underwent follow-up examination at a mean interval of 14.6 months revealed a trend toward a smaller reduction in femoral neck BMD in women in the upper serum uric acid tertiles. Conclusion: A positive correlation exists between serum uric acid levels and BMD in menopausal women.

Keywords: menopause, antioxidant, uric acid, bone mineral density

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Authors: Sergey Kozlov, Juliya Kozlova

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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

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Authors: A. M. Almujalli, G. M. Al-Ghamdi

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Mycobacterium avium subspecies paratuberculosis causes paratuberculosis, a chronic debilitating granulomatous enteritis, in camels as well as domestic and wild ruminants. The clinical manifestation of the disease in camel is not well characterized, therefore this study was aimed to investigate the clinical and pathological pictures of camels that are suffering from partuberculosis. Twelve young camels that were presented to the Veterinary Teaching Hospital, King Faisal University were investigated. Clinical and pathological examination were performed. The results revealed highly significant increase in creatinine, blood urea nitrogen, magnesium, AST and ALT in diseased camels, while glucose, total protein and albumin were highly significantly decreased in diseased camels when compared to healthy ones. Post-mortem testing indicated thickening, corrugation of the intestinal wall, folded mucosa, enlarged and oedemated ileocaecal and mesenteric lymph nodes. The microscopic findings detected short, blunt and distorted intestinal villi with hyperactive goblet cells of the villi and the crypts of lieberkuhn contained mucin droplets. The lamina propria was heavily infiltrated with mononuclear cells mostly macrophages. This clinical picture of paratuberculosis may be used to initiate control strategy to limit the spread of the disease in camel herds.

Keywords: camel, partuberclosis, control, Saudi Arabia

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Authors: Masaki Yamaguchi, Daimei Sasayama, Shinsuke Washizuka

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The goal of this study was to examine the possibility of salivary cytokines for the screening of attention deficit hyperactivity disorder (ADHD) in children. We carried out a case-control study, including 19 children with ADHD and 17 healthy children (controls). A multiplex bead array immunoassay was used to conduct a multi-analysis of 27 different salivary cytokines. Six salivary cytokines (interleukin (IL)-1β, IL-8, IL12p70, granulocyte colony-stimulating factor (G-CSF), interferon gamma (IFN-γ), and vascular endothelial growth factor (VEGF)) were significantly associated with the presence of ADHD (p < 0.05). An informative salivary cytokine panel was developed using VEGF by logistic regression analysis (odds ratio: 0.251). Receiver operating characteristic analysis revealed that assessment of a panel using VEGF showed “good” capability for discriminating between ADHD patients and controls (area under the curve: 0.778). ADHD has been hypothesized to be associated with reduced cerebral blood flow in the frontal cortex, due to reduced VEGF levels. Our study highlights the possibility of utilizing differential salivary cytokine levels for point-of-care testing (POCT) of biomarkers in children with ADHD.

Keywords: cytokine, saliva, attention deficit hyperactivity disorder, child

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Authors: Reenu Yadav, Abhay Asthana, S. K. Yadav

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Purpose: The aim of the present work was to develop an oral gel for brushing with an antimicrobial activity which will cure/protect from various periodontal diseases such as periodontitis, gingivitis, and pyorrhea. Methods: Plant materials procured from local suppliers, extracted and standardized. Screening of antimicrobial activity was carried out with the help of disk diffusion method. The gel was formulated by dried extracts of Beautea monosperma and Cordia obliquus. Gels were evaluated on various parameters and standardization of the formulation was performed. The release of drugs was studied in pH 6.8 using a mastication device.Total phenolic and flavonoid contents were estimated by folin-Ciocalteu and aluminium chloride method, and stability studies were performed (40°C and RH 75% ± 5% for 90 days) to assess the effect of temperature and humidity on the concentration of phenolic and flavonoid contents. The results of accelerated stability conditions were compared with that of samples kept at controlled conditions (RT). The control samples were kept at room temperature (25°C, 35% RH for 180 days). Results: Results are encouraging; extracts possess significant antimicrobial activity at very low concentration (15µg/disc, 20µg/disc and 15 µg/ disc) on oral pathogenic bacteria. The formulation has optimal characteristics, as well as has a pleasant appearance, fragrance, texture, and taste, is highly acceptable by the volunteers. The diffusion coefficient values ranged from 0.6655 to 0.9164. Since the R values of korsmayer papas were close to 1, Drug release from formulation follows matrix diffusion kinetics. Hence, diffusion was the mechanism of the drug release. Formulation follows non-Fickian transport mechanism. Most Formulations released 50 % of their contents within 25-30 minutes. Results obtained from the accelerated stability studies are indicative of a slight reduction in flavonoids and phenolic contents with time on long time storage. When measured degradation under ambient conditions, degradation was significantly lower than in accelerated stability study. Conclusion: Plant extracts possess compounds with antimicrobial properties can be used as. Developed formulation will cure/protect from various periodontal diseases. Further development and evaluations oral gel including the isolated compounds on the commercial scale and their clinical and toxicological studies are the future challenges.

Keywords: herbal gel, dental care, ambient conditions, commercial scale

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Authors: Dzatur Rizqi Fathienah Syarifuddin, Isharyah Sunarno, Eddy Hartono, Siti Maisuri T. Chalid

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Introduction: The potential impact of coronavirus disease 2019 (COVID-19) on the health of expectant mothers and fetuses has strained attention. Pregnant women are considered a vulnerable category to respiratory infections. Moreover, several inflammatory cytokines are 2-100 times more abundant in COVID-19 with cytokine storms than in normal individuals; interleukin 6 (IL-6) exhibits much higher elevations. Investigating potential relationships between IL-6 serum levels and the severity of COVID-19 symptoms in pregnant women is the aim of this study. Material and Methods: Sixty-two eligible pregnant women were divided into a positive COVID-19 group (n=31) and a negative COVID-19 group (n=31) in this cross-sectional study. The research subjects were selected using consecutive sampling. The IL-6 was measured from a vein blood specimen using ELISA methods. Results: The COVID-19 positive group had a higher median IL-6 serum level (45.35 (35.15- 153.99) vs. 38.86 ± 11.43 (15.02-59.52), p=0.03) than the negative group. On the other hand, the IL-6 serum level had comparable value according to the COVID-19 symptoms severity (88.35 ± 36.14 ng/mL vs. 51.09 ± 25.48 ng/mL vs. 56.02 ± 33.20 ng/mL in moderate symptoms, mild symptoms, and asymptomatic, respectively; p=0.152). Conclusion: Although the IL-6 serum levels are not related to COVID-19 symptoms severity, an elevated of this biomarker was found in pregnant women with affected diagnoses.

Keywords: interleukin-6, pregnancy, COVID-19, several inflammatory

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Authors: Sipu Guo, Silin Huang

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Background: Most studies investigate the link between executive function and allostatic load (AL) among adults aged 18 years and older. Studies differed regarding the specific biological indicators studied and executive functions accounted for. Specific executive functions may be differentially related to allostatic load. We investigated the comorbidities of executive functions and allostatic load via network analysis. Methods: We included 603 adolescents (49.84% girls; Mean age = 12.38, SD age = 1.79) from junior high school in rural China. Eight biological markers at T1 and four executive function tasks at T2 were used to evaluate networks. Network analysis was used to determine the network structure, core symptoms, and bridge symptoms in the AL-executive function network among rural adolescents. Results: The executive functions were related to 6 AL biological markers, not to cortisol and epinephrine. The most influential symptoms were inhibition control, cognitive flexibility, processing speed, and systolic blood pressure (SBP). SBP, dehydroepiandrosterone, and processing speed were the bridges through which AL was related to executive functions. dehydroepiandrosterone strongly predicted processing speed. The SBP was the biggest influencer in the entire network. Conclusions: We found evidence for differential relations between markers and executive functions. SBP was a driver in the network; dehydroepiandrosterone showed strong relations with executive function.

Keywords: allostatic load, executive function, network analysis, rural adolescent

Procedia PDF Downloads 52

Authors: Illyas Md Isa, Maryam Musfirah Che Sobry, Mohamad Syahrizal Ahmad, Nurashikin Abd Azis

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A single-walled carbon nanotube (SWCNT) that has been modified with magnesium layered hydroxide 3-(4-methoxyphenyl)propionic acid nanocomposite was proposed for the determination of uric acid and bisphenol A by square wave voltammetry. The results obtained denote that MLH-MPP nanocomposites enhance the sensitivity of the voltammetry detection responses. The best performance is shown by the modified carbon nanotube paste electrode (CNTPE) with the composition of single-walled carbon nanotube: magnesium layered hydroxide 3-(4-methoxyphenyl)propionic acid nanocomposite at 100:15 (% w/w). The linear range where the sensor works well is within the concentration 1.0 10-7 – 1.0 10-4 and 3.0 10-7 – 1.0 10-4 for uric acid and bisphenol A respectively with the limit of detection of 1.0 10-7 M for both organics. The interferences of uric acid and bisphenol A with other organic were studied and most of them did not interfere. The results shown for each experimental parameter on the proposed CNTPE showed that it has high sensitivity, good selectivity, repeatability and reproducibility. Therefore, the modified CNTPE can be used for the determination of uric acid and bisphenol A in real samples such as blood, plastic bottles and foods.

Keywords: bisphenol A, magnesium layered hydroxide 3-(4-methoxyphenyl)propionic acid nanocomposite, Nanocomposite, uric acid

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Authors: Murat Mert, Yusuf Ozer, Fatih Kolay

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Behaviour of giving harm to the body in literature has been named as “self-injury”, “self-mutilation” ve “self-harm”. “Self-injury”, or “self-mutilation” is generally used for the same meaning and mentioned as an action which is committed to the body itself directly. As is seen that alcohol and drug users have injured their bodies because of deprivation, whereas behaviour of self-injury in some societies is accepted as religious and cultural, it has nevertheless been diagnosed in people who have a borderline personality disorder, histrionic personality disorder, psychotic personality disorder and mood disorder. There has not been any direct self-murder tendency in people having self-harmed. However, death cases can be seen together with loss of consciousness depending on loss of blood by exceeding the limit in the course of injury action. 34- year old – male person who was alcohol addicted, having had a psycological treatment beforehand, had mutilated his small intestine together with fatty tissue by cutting his body with a razor-blade at the thought of insects strolling around the body (delirium tremens) due to deprivation attack and had died in the result of various cuts. In this study, crime scene investigation and death mechanism of the person having had self-harmed in a result of abstinence syndrome will be explained. Relevant criteria which differentiate this case from homicide will be examined.

Keywords: self-injury, autopsy, abstinence syndrome, CSI

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Authors: I. L. Ibrahim, A. Mann, A. Ndanaimi

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The research involved phytochemical screening, antibacterial activities and mineral determination by flame photometry of the crude extract of Brysocarpus coccineus schum indeed were carried out. The result of Phytochemical screening reveal tha saponins, alkaloids, cardiac glycosides, and anthraquinones were present. This suggests that the plant extract could be used as anti-inflammatory and anti-bleeding agents. Estimation of mineral content shows that the crude extract of B. coccineus contains 0.73 (Na+), 1.06 (K+) and 1.98 (Ca+) which justifies its use to be safe for hypertensive patients and could be used to lower blood pressure. The antibacterial properties of aqueous and ethanol extract were studied against some bacteria; pseudomonas aeruginosa, Escherichia coli, Bacilus subtilis, Klebsilla penmuoniae by disc diffusion method. The aqueous extract showed significant activity against the organisms while the ethanol at concentrations 5-10mg/ml ethanol extract showed significant zone of inhibition against the organisms, E. coli, (19 mm), B. cereus (12 mm), P. aeruginosa (11 mm), K. pnemuoniae (11 mm). Minimum inhibitory concentration (MIC) was carried with considerable effect of inhibition on the organisms. The MIC values observed were 1, 24, 16 and 19 mm against E. coli, B. cereus, P. aeruginosa and K. pnemuoniae respectively. Therefore, the plant could be a potential source of antibacterial agent although more pharmacological and clinical study may be recommended.

Keywords: phytochemicals, microorganisms, screenings, mineral ions

Procedia PDF Downloads 413

Authors: Nadine khadraoui, Rym Essid, Olfa Tabbene, Imen Chniba, Safa Boujemaa, Selim Jallouli, Nadia Fares, Behija Mlik, Boutheina Ben Abdelmoumen Mardassi

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Background and aim: Mycoplasma hominis is an opportunistic pathogen that can cause various gynecological infections such cervicitis, infertility, and, less frequently, extra-genital infections. Previous studies on the antimicrobial susceptibility of Mycoplasma hominis Tunisian strains have highlighted a significant resistance, even multi-resistance, to the most used antibiotic in the therapy of consequential infections. To address this concern, the present study aimed for the alternative of phytotherapy. Peganum harmala seed extract was tested as an antibacterial agent against multidrug-resistant M.hominis clinical strains. Material and Methods: Peganum harmala plant was collected from Ain Sebaa, Tabarka, North West region of Tunisia in April 2018, air-dried, grounded and extracted by different solvents.The crude methanolic extract was further partitioned with n-HEX, DCM, EtOAC and n-BuOl. Antibacterial activity was evaluated against M. hominis ATCC 23114 and 20 M. hominis clinical strains.The antimycoplasmal activity was tested by the microdilution method, and MIC values were determined. Phytochemical analysis and hemolytic activity on human erythrocytes were also performed. The active fraction was then subjected to purification, and the chemical identification of the active compound was investigated. Results: Among the tested fractions, the n-BuOH extract was the most active fraction since it exhibited an inhibitory effect against M. hominis ATCC 23114 and 80% of the tested clinical strains with MIC between 125 and 1000 µg/ml. The phytochemical analysis of the n-BuOH revealed its metabolic abundance in polyphenols, flavonoids and condensed tannin with levels of 257.37 mg AGE/g, 172.27 mg EC/g and 58.27 mg EC/g, respectively. In addition, P. harmala n-BuOH extract exhibited potent bactericidal activity against all M. hominis isolates with CMB values ranging between 125 and 4000 µg/ml. Further, the active fraction exhibited weak cytotoxicity effect at active concentrations when tested on human erythrocytes. The active compound was identified by gas chromatography–mass spectrometry as an indole alkaloid harmaline. Conclusion: In summary, Peganum harmala extract demonstrated an interesting anti-mycoplasmal activity against M. hominis Tunisian strains. Therefore, it could be considered as a potential candidate for the treatment of consequential infections. However, further studies are necessary to evaluate its mechanism of action in mycoplasmas.

Keywords: mycoplasma hominis, peganum harmala, antibioresistance, phytotherapy, phytochemical analysis

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Authors: Amal Yamani, Jaber Elgtou, Aziz Mohammed, Lazaar Jamila, Elachouri Mostafa

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Peganum harmala used traditionally as an emenagogue and abortifacient agent in Morocco phytotherapy. Even thought its benefits effects, Peganum harmala remained severely toxic for the organism especially in strong doses. The present study was initiated to evaluate the nephrotoxic effects of aqueous extract of Peganum harmala seeds (PHS). The solution containing aqueous extract of PHS was administered orally by gavage at the dose of 2g/kg body weight during twenty days. Rats were used in this study, two groups were considered, a treated group received an extract of PHS at dose 2g/kg bodyweight and control group received an amount of tap water equivalent to the volume of the vehicle used for the dose of PHS extract. The data we collected showed that aqueous extracts of PHS administered during twenty days induced a significant changes in renal function expressed in decreases of diuresis (from 10 ± 0,58 to 5,33 ± 0,33 ml/24 hours) and the same profile for mean arterial blood pressure (from 125 ± 2,89 to 96,67 ± 6,01 mmHg). The histopathological study showed an alteration of kidney cells in treated group with regard the control group which is not affected. In conclusion: our results indicate that the aqueous extract of PHS induces toxicity may affect severely kidney function and causes renal histopathology.

Keywords: peganum harmala seeds, nephrotoxic, diuresi, histpathology, kidney

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Authors: Muhammad Abdullah, Asma Rashid Butt, Nauman Raza

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Biomagnetic fluids like blood play key role in different applications of medical science and bioengineering. In this paper, the magnetohydrodynamic flow of a viscous fluid with magnetic particles through a cylindrical tube is investigated. The fluid is electrically charged in the presence of a uniform external magnetic field. The movement in the fluid is produced due to the cylindrical tube. Initially, the fluid and tube are at rest and at time t=0⁺, the tube starts to move along its axis. To obtain the mathematical model of flow with fractional derivatives fractional calculus approach is used. The solution of the flow model is obtained by using Laplace transformation. The Simon's numerical algorithm is employed to obtain inverse Laplace transform. The hybrid technique, we are employing has less computational effort as compared to other methods. The numerical calculations have been performed with Mathcad software. As the special cases of our problem, the solution of flow model with ordinary derivatives and flow without magnetic particles has been procured. Finally, the impact of non-integer fractional parameter alpha, Hartmann number Ha, and Reynolds number Re on flow and magnetic particles velocity is analyzed and depicted by graphs.

Keywords: viscous fluid, magnetic particles, fractional calculus, laplace transformation

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Authors: D. Madhushanka, Y. Liu, H. C. Fernando

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Fires are one of the foremost factors of land surface disturbance in diverse ecosystems, causing soil erosion and land-cover changes and atmospheric effects affecting people's lives and properties. Generally, the severity of the fire is calculated as the Normalized Burn Ratio (NBR) index. This is performed manually by comparing two images obtained afterward. Then by using the bitemporal difference of the preprocessed satellite images, the dNBR is calculated. The burnt area is then classified as either unburnt (dNBR<0.1) or burnt (dNBR>= 0.1). Furthermore, Wildfire Severity Assessment (WSA) classifies burnt areas and unburnt areas using classification levels proposed by USGS and comprises seven classes. This procedure generates a burn severity report for the area chosen by the user manually. This study is carried out with the objective of producing an automated tool for the above-mentioned process, namely the World Wildfire Severity Assessment Tool (WWSAT). It is implemented in Google Earth Engine (GEE), which is a free cloud-computing platform for satellite data processing, with several data catalogs at different resolutions (notably Landsat, Sentinel-2, and MODIS) and planetary-scale analysis capabilities. Sentinel-2 MSI is chosen to obtain regular processes related to burnt area severity mapping using a medium spatial resolution sensor (15m). This tool uses machine learning classification techniques to identify burnt areas using NBR and to classify their severity over the user-selected extent and period automatically. Cloud coverage is one of the biggest concerns when fire severity mapping is performed. In WWSAT based on GEE, we present a fully automatic workflow to aggregate cloud-free Sentinel-2 images for both pre-fire and post-fire image compositing. The parallel processing capabilities and preloaded geospatial datasets of GEE facilitated the production of this tool. This tool consists of a Graphical User Interface (GUI) to make it user-friendly. The advantage of this tool is the ability to obtain burn area severity over a large extent and more extended temporal periods. Two case studies were carried out to demonstrate the performance of this tool. The Blue Mountain national park forest affected by the Australian fire season between 2019 and 2020 is used to describe the workflow of the WWSAT. This site detected more than 7809 km2, using Sentinel-2 data, giving an error below 6.5% when compared with the area detected on the field. Furthermore, 86.77% of the detected area was recognized as fully burnt out, of which high severity (17.29%), moderate-high severity (19.63%), moderate-low severity (22.35%), and low severity (27.51%). The Arapaho and Roosevelt National Forest Park, California, the USA, which is affected by the Cameron peak fire in 2020, is chosen for the second case study. It was found that around 983 km2 had burned out, of which high severity (2.73%), moderate-high severity (1.57%), moderate-low severity (1.18%), and low severity (5.45%). These spots also can be detected through the visual inspection made possible by cloud-free images generated by WWSAT. This tool is cost-effective in calculating the burnt area since satellite images are free and the cost of field surveys is avoided.

Keywords: burnt area, burnt severity, fires, google earth engine (GEE), sentinel-2

Procedia PDF Downloads 235

Authors: Nasib Zaman, Maneesha Kour, Muhammad Rizwan, Fazal Akbar

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Abstract: Chikungunya fever is a re-emerging rapidly spreading mosquito-borne disease cause by Aedes albopictus and Aedes aegypti mosquito vectors. Currently, it is affecting millions of people globally. Objective: This study's main objective was to find the incidence of chikungunya fever in the Swat region and the factors associated with the spread of this infection. Method: This study was carried out in different areas of Swat. Blood samples and data were collected from selected patients, and a questionnaire was filled for each patient. 3-5ml of the specimen was taken from the patient's vein and serum, or plasma was separated by centrifugation. Chikungunya tests were performed for IgG and IgM antibodies. The data was analyzed by SPSS and Graph Paid Prism 5. Results: A total of 169 patients were included in this study, out of which 103 (60.9%) having age less than 30 years were positive for chikungunya infection and 66 (39.1%) having more than 30 years were negative for this infection. Only 1 (0.6%) were positive for both IgG and IgM antibody. About 15 (8.9%) patients have diagnosed with positive IgG antibodies, and 25 (26.6%) patients were positive for IgM positive antibodies. The infection rate was significantly higher in males compared to females 71 (59.6%) vs. 14 (38%) P value=0.088, OR=1.7. Conclusion: This study concludes clinical knowledge and awareness that are necessary for a diagnosis of chikungunya infection properly. Therefore it is important to educate people for the eradication of this infection. Recommendation: This study also recommends investigating the other risk factors associated with this infection.

Keywords: Chikungunya, risk factor, Incidence, antibodies, mosquito

Procedia PDF Downloads 128

Authors: Jma Hannan, Prawej Ansari, Yasser H. A. Abdel-Wahab, Peter R. Flatt

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Spirulina platensis (SP, Blue-green algae) have been accepted as a supplement for the treatment of pre and post-diabetes. The present study investigated the effects of butanol fraction from ethanol extract of S. platensis on insulin release from BRIN BD11 cells, isolated mouse islets, and perfused rat pancreas, as well as glucose homeostasis in type 2 diabetic rats and their molecular pathways. In a dose-dependent manner, S. platensis increased insulin release from mouse islets and pancreatic β-cells. The extract also elevated insulin release in perfused rat pancreas. Glucose, isobutylmethylxanthine, tolbutamide, and a depolarizing concentration of KCl significantly potentiated insulin release from BRIN BD11 cells. The effect of diazoxide and verapamil, as well as the absence of extracellular Ca2+ showed a reduction in insulin secretion. When administered orally together with glucose (2.5g/kg bw), S. platensis extract improved fasting and postprandial blood glucose in type 2 diabetes. These data suggest that the anti-diabetic activity of S. platensis is partly mediated by insulin secretion via the KATP channel-dependent pathway/the intracellular cAMP pathway.

Keywords: Insulin, glucose, S. platensis, type 2 diabetes, medicinal plants

Procedia PDF Downloads 112

Authors: Fateme Afkhami, Mohsen Ehsanpour, Majid Afkhami, Maryam Ehsanpour

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Normal reference ranges of biochemical parameters are considered important for assessing and monitoring the health status of sea turtles. For this means, serum biochemistry determinations were analyzed in normal adult nesting and foraging hawksbill turtles (Eretmochelys imbricata). Blood samples were collected in March–April during nesting season and December-November in the foraging season. Plasma biochemistry values, except for creatinine and lipase were significant between the two periods. FBS, cholesterol, triglycerides, ALP (alkaline phosphatase), AST (aspartate aminotransferase), bilirubin, total protein, LDH (lactate dehydrogenase), CK (creatine kinase) and amylase were significantly higher in nesting season than foraging season (P<0.05). On the other hand urea, ALT (alanine aminotransferase) and albumin in the nesting season were significantly lower than foraging season (P<0.05). It was concluded that the nesting E. imbricata showed significant variation in their biochemical profile due to reproductive output. This study has produced working reference intervals useful for hawksbill turtles for future conservation and rehabilitation projects in the Persian Gulf and may be of assistance in similar programs worldwide.

Keywords: plasma biochemistry, nesting, foraging, hawksbill turtles, Persian Gulf

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Authors: Sami El Khatib, Agnès Leroux, Jean-Louis Merlin, François Guillemin, Marie-Ange D’Hallewin

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Photodynamic therapy (PDT) is a treatment modality based on the cytotoxic effect occurring on the target tissues by interaction of a photosensitizer with light in the presence of oxygen. One of the major advances in PDT can be attributed to the use of topical aminolevulinic (ALA) to induce Protoporphyrin IX (PpIX) for the treatment of early stage cancers as well as diagnosis. ALA is a precursor of the heme synthesis pathway. Locally delivered to the target tissue ALA overcomes the negative feedback exerted by heme and promotes the transient formation of PpIX in situ to reach critical effective levels in cells and tissue. Whereas early steps of the heme pathway occur in the cytosol, PpIX synthesis is shown to be held in the mitochondrial membranes and PpIX fluorescence is expected to accumulate in close vicinity of the initial building site and to progressively diffuse to the neighboring cytoplasmic compartment or other lipophylic organelles. PpIX is known to be highly reactive and will be degraded when irradiated with light. PpIX photobleaching is believed to be governed by a singlet oxygen mediated mechanism in the presence of oxidized amino acids and proteins. PpIX photobleaching and subsequent spectral phototransformation were described widely in tumor cells incubated in vitro with ALA solution, or ex vivo in human and porcine mucosa superfused with hexylaminolevulinate (hALA). PpIX photobleaching was also studied in vivo, using animal models such as normal or tumor mice skin and orthotopic rat bladder model. Hexyl aminolevulinate a more potent lipophilic derivative of ALA was proposed as an adjunct to standard cystoscopy in the fluorescence diagnosis of bladder cancer and other malignancies. We have previously reported the effectiveness of hALA mediated PDT of rat bladder cancer. Although normal and tumor bladder epithelium exhibit similar fluorescence intensities after intravesical instillation of two hALA concentrations (8 and 16 mM), the therapeutic response at 8mM and 20J/cm2 was completely different from the one observed at 16mM irradiated with the same light dose. Where the tumor is destroyed, leaving the underlying submucosa and muscle intact after an 8 mM instillation, 16mM sensitization and subsequent illumination results in the complete destruction of the underlying bladder wall but leaves the tumor undamaged. The object of the current study is to try to unravel the underlying mechanism for this apparent contradiction. PpIX extraction showed identical amounts of photosensitizer in tumor bearing bladders at both concentrations. Photobleaching experiments revealed mono-exponential decay curves in both situations but with a two times faster decay constant in case of 16mM bladders. Fluorescence microscopy shows an identical fluorescence pattern for normal bladders at both concentrations and tumor bladders at 8mM with bright spots. Tumor bladders at 16 mM exhibit a more diffuse cytoplasmic fluorescence distribution. The different response to PDT with regard to the initial pro-drug concentration can thus be attributed to the different cellular localization.

Keywords: bladder cancer, hexyl-aminolevulinate, photobleaching, confocal fluorescence microscopy

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Authors: Evgenia Iakovleva, Juha Koivisto, Pasi Karppinen, J. Inkinen, Mikko Alava

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Preventing the spread of infectious diseases throughout the worldwide is one of the most important tasks of modern health care. Infectious diseases not only account for one fifth of the deaths in the world, but also cause many pathological complications for the human health. Touch surfaces pose an important vector for the spread of infections by varying microorganisms, including antimicrobial resistant organisms. Further, antimicrobial resistance is reply of bacteria to the overused or inappropriate used of antibiotics everywhere. The biggest challenges in bacterial detection by existing methods are non-direct determination, long time of analysis, the sample preparation, use of chemicals and expensive equipment, and availability of qualified specialists. Therefore, a high-performance, rapid, real-time detection is demanded in rapid practical bacterial detection and to control the epidemiological hazard. Among the known methods for determining bacteria on the surfaces, Hyperspectral methods can be used as direct and rapid methods for microorganism detection on different kind of surfaces based on fluorescence without sampling, sample preparation and chemicals. The aim of this study was to assess the relevance of such systems to remote sensing of surfaces for microorganisms detection to prevent a global spread of infectious diseases. Bacillus subtilis and Escherichia coli with different concentrations (from 0 to 10x8 cell/100µL) were detected with hyperspectral camera using different filters as visible visualization of bacteria and background spots on the steel plate. A method of internal standards was applied for monitoring the correctness of the analysis results. Distances from sample to hyperspectral camera and light source are 25 cm and 40 cm, respectively. Each sample is optically imaged from the surface by hyperspectral imaging system, utilizing a JAI CM-140GE-UV camera. Light source is BeamZ FLATPAR DMX Tri-light, 3W tri-colour LEDs (red, blue and green). Light colors are changed through DMX USB Pro interface. The developed system was calibrated following a standard procedure of setting exposure and focused for light with λ=525 nm. The filter is ThorLabs KuriousTM hyperspectral filter controller with wavelengths from 420 to 720 nm. All data collection, pro-processing and multivariate analysis was performed using LabVIEW and Python software. The studied human eye visible and invisible bacterial stains clustered apart from a reference steel material by clustering analysis using different light sources and filter wavelengths. The calculation of random and systematic errors of the analysis results proved the applicability of the method in real conditions. Validation experiments have been carried out with photometry and ATP swab-test. The lower detection limit of developed method is several orders of magnitude lower than for both validation methods. All parameters of the experiments were the same, except for the light. Hyperspectral imaging method allows to separate not only bacteria and surfaces, but also different types of bacteria, such as Gram-negative Escherichia coli and Gram-positive Bacillus subtilis. Developed method allows skipping the sample preparation and the use of chemicals, unlike all other microbiological methods. The time of analysis with novel hyperspectral system is a few seconds, which is innovative in the field of microbiological tests.

Keywords: Escherichia coli, Bacillus subtilis, hyperspectral imaging, microorganisms detection

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Authors: Kye Gilder, Kevin Shan, Amy Halseth, Steve Smith

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This study assessed the effect of prolonged-release naltrexone 32 mg/bupropion 360 mg (NB) on cardiovascular (CV) events in overweight/obese participants at elevated CV risk. Participants must lose ≥2% body weight at 16 wks, without a sustained increase in blood pressure, to continue drug. Only serious adverse events (SAE) and adverse events leading to discontinuation of study drug (AELDSD) were collected. The study was terminated early after second interim analysis with 50% of all CV events. Data on CV endpoints has been published. Current analyses focused on AEs in participants on antidepressants at baseline, as these individuals were excluded from Phase 3 trials. Intent-to-treat (ITT) population (placebo [PBO] N=4450, NB N=4455) was 54.5% female, 83.5% white, mean age of 61 yrs, mean BMI 37.3 kg/m2, 22.8% with a history of depression, 23.1% on antidepressants, including 15.4% on an SSRI. SAEs in participants receiving antidepressants was similar between NB (10.7%) and PBO (9.9%) and also similar to overall population (9.5% NB, 8.1% PBO). SAEs in those on SSRIs were similar, 10.1% NB and PBO 9.4%. For those on SSRIs or other antidepressants, AELDSDs were similar to overall population and were primarily GI disorders. Obesity increases the risk of developing depression. For participants taking NB and antidepressants, including SSRIs, there is a similar AE profile as the overall population and data revealed no evidence of an additional health risk with combined use.

Keywords: antidepressant, Contrave, Mysimba, obesity, pharmacotherapy

Procedia PDF Downloads 259