Search results for: immune organs
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1138

Search results for: immune organs

238 Protecting Physicochemical Properties of Black Cumin Seed (Nigella sativa) Oil and Developing Value Added Products

Authors: Zeliha Ustun, Mustafa Ersoz

Abstract:

In the study, a traditional herbal supplement black cumin seed (Nigella sativa) oil properties has been studied to protect the main quality parameters by a new supplement application. Black cumin seed and its oil is used as a dietary supplement and preferred traditional remedy in Africa, Asia and Middle East for centuries. Now it has been consuming by millions of people in America and Europe as natural supplements and/or phytotherapeutic agents to support immune system, asthma, allergic rinnitis etc. by the scientists’ advices. With the study, it is aimed to prove that soft gelatin capsules are a new and more practical way of usage for Nigella sativa oil that has a longer stability. With the study soft gelatin capsules formulation has been developed to protect cold pressed black cumin seed oil physicochemical properties for a longer period. The product design has been developed in laboratory and implemented in pilot scale soft gelatin capsule manufacturing. Physicochemical properties (peroxide value, free fatty acids, fatty acid composition, refractive index, iodine value, saponification value, unsaponifiable matters) of Nigella sativa oil soft gelatin capsules and Nigella sativa oil in liquid form in amber glass bottles have been compared and followed for 8 months. The main parameters for capsules and liquid form found that for free fatty acids 2.29±0.03, 3.92±0.11 % oleic acid, peroxide 23.11±1.18, 27.85±2.50 meqO2/kg, refractive index at 20 0C 1.4738±0.00, 1.4737±0.00, soap 0 ppm, moisture and volatility 0.32±0.01, 0.36±0.01 %, iodine value 123.00±0.00, 122.00±0.00 wijs, saponification value 196.25±0.46, 194.13±0.35 mg KOH/g and unsaponifiable matter 7.72±0.13, 6.88±0.36 g/kg respectively. The main fatty acids are found that linoleic acid 56.17%, oleic acid 24.64%, palmitic acid 11,94 %. As a result, it is found that cold pressed Nigella sativa oil soft gelatin capsules physicochemical properties are more stable than the Nigella sativa oil stored in glass bottles.

Keywords: black cumin seed (Nigella sativa) oil, cold press, nutritional supplements, soft gelatin capsule

Procedia PDF Downloads 335
237 Bioinformatic Design of a Non-toxic Modified Adjuvant from the Native A1 Structure of Cholera Toxin with Membrane Synthetic Peptide of Naegleria fowleri

Authors: Frida Carrillo Morales, Maria Maricela Carrasco Yépez, Saúl Rojas Hernández

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Naegleria fowleri is the causative agent of primary amebic meningoencephalitis, this disease is acute and fulminant that affects humans. It has been reported that despite the existence of therapeutic options against this disease, its mortality rate is 97%. Therefore, the need arises to have vaccines that confer protection against this disease and, in addition to developing adjuvants to enhance the immune response. In this regard, in our work group, we obtained a peptide designed from the membrane protein MP2CL5 of Naegleria fowleri called Smp145 that was shown to be immunogenic; however, it would be of great importance to enhance its immunological response, being able to co-administer it with a non-toxic adjuvant. Therefore, the objective of this work was to carry out the bioinformatic design of a peptide of the Naegleria fowleri membrane protein MP2CL5 conjugated with a non-toxic modified adjuvant from the native A1 structure of Cholera Toxin. For which different bioinformatics tools were used to obtain a model with a modification in amino acid 61 of the A1 subunit of the CT (CTA1), to which the Smp145 peptide was added and both molecules were joined with a 13-glycine linker. As for the results obtained, the modification in CTA1 bound to the peptide produces a reduction in the toxicity of the molecule in in silico experiments, likewise, the prediction in the binding of Smp145 to the receptor of B cells suggests that the molecule is directed in specifically to the BCR receptor, decreasing its native enzymatic activity. The stereochemical evaluation showed that the generated model has a high number of adequately predicted residues. In the ERRAT test, the confidence with which it is possible to reject regions that exceed the error values was evaluated, in the generated model, a high score was obtained, which determines that the model has a good structural resolution. Therefore, the design of the conjugated peptide in this work will allow us to proceed with its chemical synthesis and subsequently be able to use it in the mouse meningitis protection model caused by N. fowleri.

Keywords: immunology, vaccines, pathogens, infectious disease

Procedia PDF Downloads 59
236 Enhanced Cytotoxic Effect of Expanded NK Cells with IL12 and IL15 from Leukoreduction Filter on K562 Cell Line Exhibits Comparable Cytotoxicity to Whole Blood

Authors: Abdulbaset Mazarzaei

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Natural killer (NK) cells are innate immune effectors that play a pivotal role in combating tumors and infected cells. In recent years, the therapeutic potential of NK cells has gained significant attention due to their remarkable cytotoxic ability. This study focuses on investigating the cytotoxic effect of expanded NK cells enriched with interleukin 12 (IL12) and interleukin 15 (IL15), derived from the leukoreduction filter, on the K562 cell line. Firstly, NK cells were isolated from whole blood samples obtained from healthy volunteers. These cells were subsequently expanded ex vivo using a combination of feeder cells, IL12, and IL15. The expanded NK cells were then harvested and assessed for their cytotoxicity against K562, a well-established human chronic myelogenous leukemia cell line. The cytotoxicity was evaluated using flow cytometry assay. Results demonstrate that the expanded NK cells significantly exhibited enhanced cytotoxicity against K562 cells compared to non-expanded NK cells. Interestingly, the expanded NK cells derived specifically from IL12 and IL15-enriched leukoreduction filters showed a robust cytotoxic effect similar to the whole blood-derived NK cells. These findings suggest that IL12 and IL15 in the leukoreduction filter are crucial in promoting NK cell cytotoxicity. Furthermore, the expanded NK cells displayed relatively similar cytotoxicity profiles to whole blood-derived NK cells, indicating their comparable capability in targeting and eliminating tumor cells. This observation is of significant relevance as expanded NK cells from the leukoreduction filter could potentially serve as a readily accessible and efficient source for adoptive immunotherapy. In conclusion, this study highlights the significant cytotoxic effect of expanded NK cells enriched with IL12 and IL15 obtained from the leukoreduction filter on the K562 cell line. Moreover, it emphasizes that these expanded NK cells exhibit comparable cytotoxicity to whole blood-derived NK cells. These findings reinforce the potential clinical utility of using expanded NK cells from the leukoreduction filter as an effective strategy in adoptive immunotherapy for the treatment of cancer. Further studies are warranted to explore the broader implications of this approach in clinical settings.

Keywords: natural killer (NK) cells, Cytotoxicity, Leukoreduction filter, IL-12 and IL-15 Cytokines

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235 Hematological Changes in the Hydatidosed Male Sheep after Experimental Inoculation of Echinococcus granulosus Eggs

Authors: M. Younus, Muhammad Shafique, M. Athar Khan, Tanveer Akhtar , M. Moeen Athar

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A total of 48 apparently healthy weaned sheep lambs (Ovis aries) of 8-10 weeks old weighing 7-10 Kg were purchased from the contractors, maintained in the experimental station of University of the Punjab, Quaid-e-Azam Campus at Lahore, Pakistan. They were dewormed against nematodes with levamisole (ICI) at recommended dose rates. The feces were tested against the parasitic eggs, no helminths ova were seen. All the 48 sheep lambs were divided into two groups i.e. group A & group B. Group 'A' comprising of 40 sheep, kept as infected groups whereas group 'B' comprising of 08 sheep & kept as a new infected control group. Each sheep lamb of group A was given 3-4 fresh gravid segments contains 2-3 thousand eggs of Echinococcus granulosus. These were collected from experimentally infected dogs by feeding fresh hydrated cysts collected from liver & lungs of sheep after slaughtering. Each lamb was fed with fresh gravid segments for a total period of 5 days or each alternate day. Coagulated blood was collected before the start of infected diet and after every month by jugular phlebotomy of each sheep lamb from the infected & new infected control group. One lamb each from group A & group B was slaughtered at the end of each month for the presence of macroscopic hydatid cyst in viscera & abdominal cavity. After 180 days of the experiment, hydatid cysts were confirmed in the abdominal cavity. Hematological parameters of zero days & then at the end of every month revealed that there was a gradual increase (PL 0.05) in the White Blood Cell (WBC), Mean Corpuscular Volume (MCV), Mean Corpuscular Hemoglobin Concentration (MCHC) and Erythrocyte Sedimentation Rates (ESR). The increasing trend was probably due to inflammatory response and lytic effect of the newly developing E. granulosus hydatid cysts. The red blood cell (RBC), Hemoglobin (HB), Packed Cell Volume (PCV) and Mean Corpuscular Hemoglobin (MCH) infected groups were decreased significantly as compared to the control group (PL 0.05). The experiment was terminated at the end of the 7th month. It can be concluded that Echinococcus granulosus can damage livestock and other intermediate hosts such as horses, the development of hydatid cysts affect the organs due to the growing cysts pressuring the organ tissues. Parts of the tissue die, which impairs the functioning of the affected organ. The clinical signs depend on the affected organ. The major damage for livestock is organ condemnation at slaughter.

Keywords: echinococcus granulosus, hydatidosis, sheep, hematology

Procedia PDF Downloads 388
234 Application of Bundle Care to Reduce Invasive Catheter-Associated Infection in High Risk Units at a Medical Center

Authors: Hsin-Hsin Chang, Jann-Tay Wang, Wang-Huei Sheng

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Background: Hospital-associated infections (HAIs) have significant medical and social resource consumption. In view of medical technology change rapidly and the prolonged average life expectancy, the patients' chances of receiving invasive medical devices have also increased. As well as the potential disease of the patients, the aging, and immune dysfunction makes the disease more serious, raising the risk of HAIs. In our adult intensive care units, catheter-associated urinary tract infections (CAUTIs) have an average of 4.6% in 2014, which is much higher than that of the National Healthcare Safety Network (NHSN). Therefore, we started the intervention of CAUTI bundle care. Methods: This 3-year intervention was conducted in adults’ intensive care units (ICUs) during January 2015 to December 2017. The implementation of CAUTI bundle care in order to reduce invasive catheter-associated infections were built on evidence-based infection control measures. Prospective surveillance was performed on all patients admitted to hospital. The four major directions are 'Leader Engagement', 'Educate Personnel', 'Executive Multidisciplinary Teamwork', 'Innovation and Improvement of Tools'. Results: During the intervention period, there were 167,024 patient-days with a total of 508 episodes of CAUTIs in the entire adult ICUs identified. The incidence of CAUTIs in adult ICU was significantly decreased in the intervention period (from 2015 to 2017), from 4.6 to 3.6 per 1000 catheter days (p=0.05). Conclusion: The necessity for the implementation of CAUTI bundle care in the health care system plays an important role in the quality and policy of infection control. Multidisciplinary teamwork, education, a comprehensive checklist and from time to time audit feedback to improve healthcare workers’ compliance are the keys to success.

Keywords: bundle care, hospital-associated infections, leader engagement, multidisciplinary team work

Procedia PDF Downloads 129
233 Factors Contributing to Adverse Maternal and Fetal Outcome in Patients with Eclampsia

Authors: T. Pradhan, P. Rijal, M. C. Regmi

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Background: Eclampsia is a multisystem disorder that involves vital organs and failure of these may lead to deterioration of maternal condition and hypoxia and acidosis of fetus resulting in high maternal and perinatal mortality and morbidity. Thus, evaluation of the contributing factors for this condition and its complications leading to maternal deaths should be the priority. Formulating the plan and protocol to decrease these losses should be our goal. Aims and Objectives: To evaluate the risk factors associated with adverse maternal and fetal outcome in patients with eclampsia and to correlate the risk factors associated with maternal and fetal morbidity and mortality. Methods: All patients with eclampsia admitted in Department of Obstetrics and Gynecology, B. P. Koirala Institute of Health Sciences were enrolled after informed consent from February 2013 to February 2014. Questions as per per-forma were asked to patients, and attendants like Antenatal clinic visits, parity, number of episodes of seizures, duration from onset of seizure to magnesium sulfate and the patients were followed as per the hospital protocol, the mode of delivery, outcome of baby, post partum maternal condition like maternal Intensive Care Unit admission, neurological impairment and mortality were noted before discharge. Statistical analysis was done using Statistical Package for the Social Sciences (SPSS 11). Mean and percentage were calculated for demographic variables. Pearson’s correlation test and chi-square test were applied to find the relation between the risk factors and the outcomes. P value less than 0.05 was considered significant. Results: There were 10,000 antenatal deliveries during the study period. Fifty-two patients with eclampsia were admitted. All of the patients were unbooked for our institute. Thirty-nine patients were antepartum eclampsia. Thirty-one patients required mechanical ventilator support. Twenty-four patients were delivered by emergency c-section and 21 babies were Low Birth Weight and there were 9 stillbirths. There was one maternal mortality and 45 patients were discharged with improvement but 3 patients had neurological impairment. Mortality was significantly related with number of seizure episodes and time interval between seizure onset and administration of magnesium sulphate. Conclusion: Early detection and management of hypertensive complicating pregnancy during antenatal clinic check up. Early hospitalization and management with magnesium sulphate for eclampsia can help to minimize the maternal and fetal adverse outcomes.

Keywords: eclampsia, maternal mortality, perinatal mortality, risk factors

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232 Challenges to Ensure Food Safety through Sanitation and Hygiene Coverage in Bangladesh

Authors: Moshiur Rahman, Tahmida Jakia

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Bangladesh, a densely populated South Asian country is home to more than 160 million people. In two decades ago, the people of this developing nation drank heavily contaminated surface water. Over the past thirty years, the country, and its development partners, has undertaken extensive efforts to provide microbiologically safe groundwater based drinking water through the use of tube-wells. About 85% of the people now drink tube-well water from about 11 million tube-wells/hand pumps. However, diarrhoeal and other water-related diseases are still reported among the major causes of morbidity and mortality among Bangladeshi children. This implies that the mode of transmission of pathogens through water and/or other modes continue. In addition, massive scale arsenic contamination has been recently reported in the ground water. Thirty five million people may be at risk of consuming arsenic contaminated water exceeding 0.05 mg/l in Bangladesh. Drinking of arsenic contaminated water has been linked with skin problems, cancer, cardiovascular diseases, neurological diseases, eye problems, cancer of the internal organs, and other diseases. In the study area, Narail district, recent investigations about existing water quality situations indicated presence of low to high levels of arsenic, salinity, iron, manganese and bacteriological contamination risks. As challenges for safe water exist; it is likely that sanitation and food hygiene practices are poor which lead threat to ensure food security.The main attempt of this study is to find out the challenges to ensure food security andprovide probable solutions to ensure food safety towards 0.7 million of people in study area. A survey has been conducted at Lohagara and Kalia sub district of Narail district with a pretested questionnaire. Primary data are collected through a questionnaire, while secondary data are collected from pertinent offices as well as academic journals. FGD has also been done to know the knowledge regarding water, sanitation as well as food preparation and consumption practice of community people in study area. The major focus of this study is to assess the state of sanitation and food hygiene condition of rural people. It is found that most of the villagers have lack of knowledge about food safety. Open defecation rate is high which lead threat to ensure food security.

Keywords: food safety, challenges, hygiene, Bangladesh

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231 Data Analysis for Taxonomy Prediction and Annotation of 16S rRNA Gene Sequences from Metagenome Data

Authors: Suchithra V., Shreedhanya, Kavya Menon, Vidya Niranjan

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Skin metagenomics has a wide range of applications with direct relevance to the health of the organism. It gives us insight to the diverse community of microorganisms (the microbiome) harbored on the skin. In the recent years, it has become increasingly apparent that the interaction between skin microbiome and the human body plays a prominent role in immune system development, cancer development, disease pathology, and many other biological implications. Next Generation Sequencing has led to faster and better understanding of environmental organisms and their mutual interactions. This project is studying the human skin microbiome of different individuals having varied skin conditions. Bacterial 16S rRNA data of skin microbiome is downloaded from SRA toolkit provided by NCBI to perform metagenomics analysis. Twelve samples are selected with two controls, and 3 different categories, i.e., sex (male/female), skin type (moist/intermittently moist/sebaceous) and occlusion (occluded/intermittently occluded/exposed). Quality of the data is increased using Cutadapt, and its analysis is done using FastQC. USearch, a tool used to analyze an NGS data, provides a suitable platform to obtain taxonomy classification and abundance of bacteria from the metagenome data. The statistical tool used for analyzing the USearch result is METAGENassist. The results revealed that the top three abundant organisms found were: Prevotella, Corynebacterium, and Anaerococcus. Prevotella is known to be an infectious bacterium found on wound, tooth cavity, etc. Corynebacterium and Anaerococcus are opportunist bacteria responsible for skin odor. This result infers that Prevotella thrives easily in sebaceous skin conditions. Therefore it is better to undergo intermittently occluded treatment such as applying ointments, creams, etc. to treat wound for sebaceous skin type. Exposing the wound should be avoided as it leads to an increase in Prevotella abundance. Moist skin type individuals can opt for occluded or intermittently occluded treatment as they have shown to decrease the abundance of bacteria during treatment.

Keywords: bacterial 16S rRNA , next generation sequencing, skin metagenomics, skin microbiome, taxonomy

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230 Antigenic Diversity of Theileria parva Isolates from Cattle and Buffalo at the Wildlife-Livestock Interface in Southern and Eastern Africa

Authors: Mukolwe D. Lubembe, Odongo O. David, Githaka Naftali, Kanduma Esther, Marinda Oosthuizen, Kgomotso P. Sibeko

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Theileriosis is a tick-borne disease of cattle caused by an apicomplexan protozoan parasite of the genus Theileria. In eastern and southern Africa, Theileria infections in cattle are caused by the species Theileria parva whose natural reservoir is the African buffalo (Syncerus caffer). Currently, East Coast Fever (ECF) caused by the cattle-derived Theileria parva is still a major problem in eastern Africa and some parts of southern Africa but not in South Africa following its eradication in the 1950s. However, Corridor disease (CD) caused by the buffalo-derived Theileria parva still remains a concern in South Africa. The diversity of Theileria parva in South Africa in comparison to other affected countries is poorly defined yet its known to be the survival strategy of this parasite. We assessed the antigenic diversity of Theileria parva isolates from Buffalo and cattle at the wildlife-livestock interface comparing samples from South Africa, Zimbabwe, Kenya, Tanzania, and Uganda. Antigenic epitopes of eight schizont antigen genes (Tp1, Tp3, Tp4, Tp5, Tp6, Tp7, Tp8 and Tp10) were amplified by PCR from genomic DNA extracted from blood samples collected from cattle and buffalo at the wildlife-livestock interface. Amplicons were purified and then sequenced on NGS platform. Full length open reading frames (ORFs) of two schizont antigen genes (Tp2 and Tp9) and one sporozoite antigen gene, p67 were also amplified from genomic DNA. Amplicons were then purified and cloned for sequencing. Analysis was based on sequence differences in the genes. Preliminary results show an extensively diverse population of Theileria parva circulating in buffalo and cattle populations at the wildlife-livestock interface. Diversity of the antigen genes contributes to the evasion of the immune system of the host by Theileria parva. This possess a concern in that, some of the Theileria parva populations may re-assort and become adapted to cattle to cause a form of theileriosis that is as fatal as ECF in areas where ECF was eradicated or is absent

Keywords: Theileria parva, east coast fever, corridor diseases, antigen genes, diversity

Procedia PDF Downloads 195
229 Cost Effectiveness Analysis of a Community Intervention for Anti-Retroviral Therapy Delivery in Cambodia

Authors: Esabelle Lo Yan Yam, Pheak Chhoun, Sovannary Tuot, Emily Lancsar, Siyan Yi

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Persons living with HIV (PLHIV) need lifelong antiretroviral treatment (ART) to keep their viral load suppressed to an undetectable level, maintain a healthy immune system, and reduce the risk of transmitting HIV to others. However, many factors affect PLHIV's adherence to ART, including access to antiretrovirals (ARV), stigma, lack of social support, and the burden of seeking lifelong care. Community-based care has been shown to be instrumental in the experience of PLHIV in many countries, including Cambodia. In this study based in Cambodia, a community-based ART delivery (CAD) intervention involving community action workers (CAWs) who are PLHIVs was introduced. These workers collect pre-packaged ARVs from the ART clinics and dispense them to PLHIVs in the communities. The quasi-experimental study involved approximately 2000 stable PLHIV in the intervention arm and another 2000 PLHIV in the control arm (receiving usual care). A cost-effectiveness analysis is currently conducted to complement the clinical effectiveness of the CAD intervention on the care continuum and treatment outcomes for stable PLHIV, as well as the operational effectiveness in increasing the efficiency of the ART clinics and the health system. The analysis will consider health system and societal perspectives based on primary outcomes, including retention in care, viral load suppression, and adherence to ART. Additionally, a consultation with the National Centre for HIV/AIDS, Dermatology, and STD under the Cambodia Ministry of Health will be done to discuss the conduct of a budget impact analysis that can quantify the financial impact on the government's budget when adopting the CAD intervention at the provincial and national levels. The budget impact analysis will take into consideration various scaling-up scenarios for the interventions in the country. The research will assess the cost-effectiveness of the CAD intervention to support national stakeholders in Cambodia to make an informed decision on the adoption and scaling up of the intervention in Cambodia. The results are currently being analyzed and will be available at the time of the conference.

Keywords: Cambodia, community intervention, economic evaluation, global health, HIV/AIDs, implementation research

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228 Automatic Segmentation of 3D Tomographic Images Contours at Radiotherapy Planning in Low Cost Solution

Authors: D. F. Carvalho, A. O. Uscamayta, J. C. Guerrero, H. F. Oliveira, P. M. Azevedo-Marques

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The creation of vector contours slices (ROIs) on body silhouettes in oncologic patients is an important step during the radiotherapy planning in clinic and hospitals to ensure the accuracy of oncologic treatment. The radiotherapy planning of patients is performed by complex softwares focused on analysis of tumor regions, protection of organs at risk (OARs) and calculation of radiation doses for anomalies (tumors). These softwares are supplied for a few manufacturers and run over sophisticated workstations with vector processing presenting a cost of approximately twenty thousand dollars. The Brazilian project SIPRAD (Radiotherapy Planning System) presents a proposal adapted to the emerging countries reality that generally does not have the monetary conditions to acquire some radiotherapy planning workstations, resulting in waiting queues for new patients treatment. The SIPRAD project is composed by a set of integrated and interoperabilities softwares that are able to execute all stages of radiotherapy planning on simple personal computers (PCs) in replace to the workstations. The goal of this work is to present an image processing technique, computationally feasible, that is able to perform an automatic contour delineation in patient body silhouettes (SIPRAD-Body). The SIPRAD-Body technique is performed in tomography slices under grayscale images, extending their use with a greedy algorithm in three dimensions. SIPRAD-Body creates an irregular polyhedron with the Canny Edge adapted algorithm without the use of preprocessing filters, as contrast and brightness. In addition, comparing the technique SIPRAD-Body with existing current solutions is reached a contours similarity at least 78%. For this comparison is used four criteria: contour area, contour length, difference between the mass centers and Jaccard index technique. SIPRAD-Body was tested in a set of oncologic exams provided by the Clinical Hospital of the University of Sao Paulo (HCRP-USP). The exams were applied in patients with different conditions of ethnology, ages, tumor severities and body regions. Even in case of services that have already workstations, it is possible to have SIPRAD working together PCs because of the interoperability of communication between both systems through the DICOM protocol that provides an increase of workflow. Therefore, the conclusion is that SIPRAD-Body technique is feasible because of its degree of similarity in both new radiotherapy planning services and existing services.

Keywords: radiotherapy, image processing, DICOM RT, Treatment Planning System (TPS)

Procedia PDF Downloads 268
227 Building Exoskeletons for Seismic Retrofitting

Authors: Giuliana Scuderi, Patrick Teuffel

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The proven vulnerability of the existing social housing building heritage to natural or induced earthquakes requires the development of new design concepts and conceptual method to preserve materials and object, at the same time providing new performances. An integrate intervention between civil engineering, building physics and architecture can convert the social housing districts from a critical part of the city to a strategic resource of revitalization. Referring to bio-mimicry principles the present research proposes a taxonomy with the exoskeleton of the insect, an external, light and resistant armour whose role is to protect the internal organs from external potentially dangerous inputs. In the same way, a “building exoskeleton”, acting from the outside of the building as an enclosing cage, can restore, protect and support the existing building, assuming a complex set of roles, from the structural to the thermal, from the aesthetical to the functional. This study evaluates the structural efficiency of shape memory alloys devices (SMADs) connecting the “building exoskeleton” with the existing structure to rehabilitate, in order to prevent the out-of-plane collapse of walls and for the passive dissipation of the seismic energy, with a calibrated operability in relation to the intensity of the horizontal loads. The two case studies of a masonry structure and of a masonry structure with concrete frame are considered, and for each case, a theoretical social housing building is exposed to earthquake forces, to evaluate its structural response with or without SMADs. The two typologies are modelled with the finite element program SAP2000, and they are respectively defined through a “frame model” and a “diagonal strut model”. In the same software two types of SMADs, called the 00-10 SMAD and the 05-10 SMAD are defined, and non-linear static and dynamic analyses, namely push over analysis and time history analysis, are performed to evaluate the seismic response of the building. The effectiveness of the devices in limiting the control joint displacements resulted higher in one direction, leading to the consideration of a possible calibrated use of the devices in the different walls of the building. The results show also a higher efficiency of the 00-10 SMADs in controlling the interstory drift, but at the same time the necessity to improve the hysteretic behaviour, to maximise the passive dissipation of the seismic energy.

Keywords: adaptive structure, biomimetic design, building exoskeleton, social housing, structural envelope, structural retrofitting

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226 Molecular Characterisation and Expression of Glutathione S-Transferase of Fasciola Gigantica

Authors: J. Adeppa, S. Samanta, O. K. Raina

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Fasciolosis is a widespread economically important parasitic infection throughout the world caused by Fasciola hepatica and F. gigantica. In order to identify novel immunogen conferring significant protection against fasciolosis, currently, research has been focused on the defined antigens viz. glutathione S-transferase, fatty acid binding protein, cathepsin-L, fluke hemoglobin, paramyosin, myosin and F. hepatica- Kunitz Type Molecule. Among various antigens, GST which plays a crucial role in detoxification processes, i.e. phase II defense mechanism of this parasite, has a unique position as a novel vaccine candidate and a drug target in the control of this disease. For producing the antigens in large quantities and their purification to complete homogeneity, the recombinant DNA technology has become an important tool to achieve this milestone. RT- PCR was carried out using F. gigantica total RNA as template, and an amplicon of 657 bp GST gene was obtained. TA cloning vector was used for cloning of this gene, and the presence of insert was confirmed by blue-white selection for recombinant colonies. Sequence analysis of the present isolate showed 99.1% sequence homology with the published sequence of the F. gigantica GST gene of cattle origin (accession no. AF112657), with six nucleotide changes at 72, 74, 423, 513, 549 and 627th bp found in the present isolate, causing an overall change of 4 amino acids. The 657 bp GST gene was cloned at BamH1 and HindIII restriction sites of the prokaryotic expression vector pPROEXHTb in frame with six histidine residues and expressed in E. coli DH5α. Recombinant protein was purified from the bacterial lysate under non-denaturing conditions by the process of sonication after lysozyme treatment and subjecting the soluble fraction of the bacterial lysate to Ni-NTA affinity chromatography. Western blotting with rabbit hyper-immune serum showed immuno-reactivity with 25 kDa recombinant GST. Recombinant protein detected F. gigantica experimental as well as field infection in buffaloes by dot-ELISA. However, cross-reactivity studies on Fasciola gigantica GST antigen are needed to evaluate the utility of this protein in the serodiagnosis of fasciolosis.

Keywords: fasciola gigantic, fasciola hepatica, GST, RT- PCR

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225 From the Perspective of a Veterinarian: The Future of Plant Raw Materials Used in the Feeding of Farm Animals

Authors: Ertuğrul Yılmaz

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One of the most important occupational groups in the food chain from farm to fork is a veterinary medicine. This occupational group, which has important duties in the prevention of many zoonotic diseases and in public health, takes place in many critical control points from soil to our kitchen. It has important duties from mycotoxins transmitted from the soil to slaughterhouses or milk processing facilities. Starting from the soil, which constitutes 70% of mycotoxin contamination, up to the TMR made from raw materials obtained from the soil, there are all critical control points from feeding to slaughterhouses and milk production enterprises. We can take the precaution of mycotoxins such as Aflatoxin B1, Ochratoxin, Zearalenone, and Fumonisin, which we encounter on farms while in the field. It has been reported that aflatoxin B1 is a casenerogen and passes into milk in studies. It is likely that many mycotoxins pose significant threats to public health and will turn out to be even more dangerous over time. Even raw material storage and TMR preparation are very important for public health. The danger of fumonisin accumulating in the liver will be understood over time. Zoonotic diseases are also explained with examples. In this study, how important veterinarians are in terms of public health is explained with examples. In the two-year mycotoxin screenings, fumonisin mycotoxin was found to be very high in corn and corn by-products, and it was determined that it accumulated in the liver for a long time and remained cornic in animals. It has been determined that mycotoxins are present in all livestock feeds, poultry feeds, and raw materials, not alone, but in double-triple form. Starting from the end, mycotoxin scans should be carried out from feed to raw materials and from raw materials to soil. In this way, we prevent the transmission of mycotoxins to animals and from animals to humans. Liver protectors such as toxin binders, beta-glucan, mannan oligosaccharides, activated carbon, prebiotics, and silymarin were used in certain proportions in the total mixed ratio, and positive results were obtained. Humidity and temperature controls of raw material silos were made at certain intervals. Necropsy was performed on animals that died as a result of mycotoxicosis, and macroscopic photographs were taken of the organs. We have determined that the mycotoxin screening in experimental animals and the feeds made without detecting the presence and amount of bacterial factors affect the results of the project to be made. For this, a series of precautionary plans have been created, starting from the production processes.

Keywords: mycotoxins, feed safety, processes, public health

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224 Rare Diagnosis in Emergency Room: Moyamoya Disease

Authors: Ecem Deniz Kırkpantur, Ozge Ecmel Onur, Tuba Cimilli Ozturk, Ebru Unal Akoglu

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Moyamoya disease is a unique chronic progressive cerebrovascular disease characterized by bilateral stenosis or occlusion of the arteries around the circle of Willis with prominent arterial collateral circulation. The occurrence of Moyamoya disease is related to immune, genetic and other factors. There is no curative treatment for Moyamoya disease. Secondary prevention for patients with symptomatic Moyamoya disease is largely centered on surgical revascularization techniques. We present here a 62-year old male presented with headache and vision loss for 2 days. He was previously diagnosed with hypertension and glaucoma. On physical examination, left eye movements were restricted medially, both eyes were hyperemic and their movements were painful. Other neurological and physical examination were normal. His vital signs and laboratory results were within normal limits. Computed tomography (CT) showed dilated vascular structures around both lateral ventricles and atherosclerotic changes inside the walls of internal carotid artery (ICA). Magnetic resonance imaging (MRI) and angiography (MRA) revealed dilated venous vascular structures around lateral ventricles and hyper-intense gliosis in periventricular white matter. Ischemic gliosis around the lateral ventricles were present in the Digital Subtracted Angiography (DSA). After the neurology, ophthalmology and neurosurgery consultation, the patient was diagnosed with Moyamoya disease, pulse steroid therapy was started for vision loss, and super-selective DSA was planned for further investigation. Moyamoya disease is a rare condition, but it can be an important cause of stroke in both children and adults. It generally affects anterior circulation, but posterior cerebral circulation may also be affected, as well. In the differential diagnosis of acute vision loss, occipital stroke related to Moyamoya disease should be considered. Direct and indirect surgical revascularization surgeries may be used to effectively revascularize affected brain areas, and have been shown to reduce risk of stroke.

Keywords: headache, Moyamoya disease, stroke, visual loss

Procedia PDF Downloads 241
223 Influence of Dietary Inclusion of Butyric Acids, Calcium Formate, Organic Acids and Its Salts on Rabbits Productive Performance, Carcass Traits and Meat Quality

Authors: V. Viliene, A. Raceviciute-Stupeliene, V. Sasyte, V. Slausgalvis, R. Gruzauskas, J. Al-Saifi

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Animal nutritionists and scientists have searched for alternative measures to improve the production. One of such alternative is use of organic acids as feed additive in animal nutrition. The study was conducted to investigate the impact of butyric acids, calcium formate, organic acids, and its salts (BCOS) additives on rabbit’s productive performance, carcass traits and meat quality. The study was conducted with 14 Californian breed rabbits. The rabbits were assigned to two treatment groups (seven rabbits per each treatment group). The dietary treatments were 1) control diet, 2) diet supplemented with a mixture BCOS - 2 kg/t of feed. Growth performance characteristics (body weight, daily weight gain, daily feed intake, feed conversion ratio, mortality) were evaluated. Rabbits were slaughtered; carcass characteristics and meat quality were evaluated. Samples loin and hind leg meat were analysed to determine carcass characteristics, pH and colour measurements, cholesterol, and malonyldialdehyde (MDA) content in loin and hind leg meat. Differences between treatments were significant for body weight (1.30 vs. 1.36 kg; P<0.05), daily weight gain (16.60 vs. 17.85 g; P<0.05), and daily feed intake (78.25 vs. 80.58 g; P<0.05) for control and experimental group respectively for the entire experimental period (from 28–77 days old). No significant differences were found in feed conversion ratio and mortality. The feed additives insertion in the diets did not significantly influence the carcass yield or the proportions of the various carcass parts and organs. Differences between treatments were significant for pH value after 48h in loin (5.86 vs. 5.74; P<0.05), hind leg meat (6.62 vs. 6.65; P<0.05), more intense colour b* of loin (5.57 vs. 6.06; P<0.05), less intense colour a* (14.99 vs. 13.15; P<0.05) in hind leg meat. Cholesterol content in hind leg meat decreased by 17.67 mg/100g compared to control group (P<0.05). After storage for three months, MDA concentration decreased in loin and hind leg meat by 0.3 μmol/kg and 0.26 μmol/kg respectively compared to that of the control group (P<0.05). The results of this study suggest that BCOS could potentially be used in rabbit nutrition with consequent benefits on the rabbits’ productivity and nutritional quality of rabbit meat for consumers.

Keywords: butyric acids, Ca formate, meat quality, organic acids salts, rabbits, productivity

Procedia PDF Downloads 188
222 Adequate Dietary Intake to Improve Outcome of Urine: Urea Nitrogen with Balance Nitrogen and Total Lymphocyte Count

Authors: Mardiana Madjid, Nurpudji Astuti Taslim, Suryani As'ad, Haerani Rasyid, Agussalim Bukhari

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The high level of Urine Urea Nitrogen (UUN) indicates hypercatabolism occurs in hospitalized patients. High levels of Total Lymphocyte Count (TLC) indicates the immune system condition, adequate wound healing, and limit complication. Adequate dietary intake affects to decrease of hypercatabolism status in treated patient’s hospitals. Nitrogen Balance (NB) is simply the difference between nitrogen (N₂) intake and output. If more N₂ intake than output, then positive NB or anabolic will occur. This study aims to evaluate the effect of dietary intake in influencing balance nitrogen and total lymphocyte count. Method: A total of 43 patients admitted to a Wahidin Sudirohusodo Hospital between 2018 and 2019 for 10 days' treats are included. The inclusion criteria were patients who were treated for 10 days and receives food from the hospital orally. Patients did not experience gastrointestinal disorders such as vomiting and diarrhea and experience impair kidney function and liver function and expressed approval to participate in this study. During hospitalization, food intake, UUN, albumin serum, balance nitrogen, and TLC was assessed twice on day 1 and day 10. There is no Physician Clinical Nutritional intervention to correct food intake. UUN is 24 hours of urine collected on the second day after admission and the tenth day. Statistical analysis uses SPSS 24 with observational cohort methods. Result: The Forty-three participants completed the follow-up (27 men and 18 women). The age of fewer than 4 years is 22 people, 45 to 60 years is 16 people, and over 60 years is 4 people. The result of the study on day 1 obtained SGA score A, SGA score B, SGA score C are 8, 32, 3 until day 10 are 8, 31, 4, respectively. According to 24h dietary recalls, the energy intake during observation was from 522.5 ± 400.4 to 1011.9 ± 545.1 kcal/day P < 0.05, protein intake from 20.07 ± 17.2 to 40.3 ± 27.3 g/day P < 0.05, carbohydrates from 92.5 ± 71.6 to 184.8 ± 87.4 g/day, and fat from 5.5 ± 3.86 to 13.9 ± 13.9 g/day. The UUN during the observation was from 6.6 ± 7.3 to 5.5 ± 3.9 g/day, TLC decreased from 1622.9 ± 897.2 to 1319.9 ± 636.3/mm³ value target 1800/mm³, albumin serum from 3.07 ± 0.76 to 2.9 ± 0.57 g/day, and BN from -7.5 ± 7.2 to -3.1 ± 4.86. Conclusion: The high level of UUN needs to correct adequate dietary intake to improve NB and TLC status on hospitalized patients.

Keywords: adequate dietary intake, balance nitrogen, total lymphocyte count, urine urea nitrogen

Procedia PDF Downloads 90
221 Studies on Effect of Nano Size and Surface Coating on Enhancement of Bioavailability and Toxicity of Berberine Chloride; A p-gp Substrate

Authors: Sanjay Singh, Parameswara Rao Vuddanda

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The aim of the present study is study the factual benefit of nano size and surface coating of p-gp efflux inhibitor on enhancement of bioavailability of Berberine chloride (BBR); a p-gp substrate. In addition, 28 days sub acute oral toxicity study was also conducted to assess the toxicity of the formulation on chronic administration. BBR loaded polymeric nanoparticles (BBR-NP) were prepared by nanoprecipitation method. BBR NP were surface coated (BBR-SCNP) with the 1 % w/v of vitamin E TPGS. For bioavailability study, total five groups (n=6) of rat were treated as follows first; pure BBR, second; physical mixture of BBR, carrier and vitamin E TPGS, third; BBR-NP, fourth; BBR-SCNP and fifth; BBR and verapamil (widely used p-gp inhibitor). Blood was withdrawn at pre-set timing points in 24 hrs study and drug was quantified by HPLC method. In oral chronic toxicity study, total four groups (n=6) were treated as follows first (control); water, second; pure BBR, third; BBR surface coated nanoparticles and fourth; placebo BBR surface coated nanoparticles. Biochemical levels of liver (AST, ALP and ALT) and kidney (serum urea and creatinine) along with their histopathological studies were also examined (0-28 days). The AUC of BBR-SCNP was significantly 3.5 folds higher compared to all other groups. The AUC of BBR-NP was 3.23 and 1.52 folds higher compared to BBR solution and BBR with verapamil group, respectively. The physical mixture treated group showed slightly higher AUC than BBR solution treated group but significantly low compared to other groups. It indicates that encapsulation of BBR in nanosize form can circumvent P-gp efflux effect. BBR-NP showed pharmacokinetic parameters (Cmax and AUC) which are near to BBR-SCNP. However, the difference in values of T1/2 and clearance indicate that surface coating with vitamin E TPGS not only avoids the P-gp efflux at its absorption site (intestine) but also at organs which are responsible for metabolism and excretion (kidney and liver). It may be the reason for observed decrease in clearance of BBR-SCNP. No toxicity signs were observed either in biochemical or histopathological examination of liver and kidney during toxicity studies. The results indicate that administration of BBR in surface coated nanoformulation would be beneficial for enhancement of its bioavailability and longer retention in systemic circulation. Further, sub acute oral dose toxicity studies for 28 days such as evaluation of intestine, liver and kidney histopathology and biochemical estimations indicated that BBR-SCNP developed were safe for long use.

Keywords: bioavailability, berberine nanoparticles, p-gp efflux inhibitor, nanoprecipitation method

Procedia PDF Downloads 360
220 Insulin-Producing Cells from Adult Human Bone Marrow Mesenchymal Stem Cells Control Chemically-Induced Diabetes in Dogs

Authors: Maha Azzam, Mahmoud Gabr, Mahmoud Zakaria, Ayman Refaie, Amani Ismail, Sherry Khater, Sylvia Ashamallah, Mohamed Ghoniem

Abstract:

Evidence was provided that human bone marrow-derived mesenhymal stem cells (HBM-MSCs) could be differentiated to form insulin-producing cells (IPCs). Transplantation of these cells was able to cure chemically-induced diabetes in nude mice. The efficacy of these cells to control diabetes in large animals was carried out to evaluate the sufficient number of cells needed/Kg body weight and to determine the functional longevity in vivo. Materials/Methods: Ten male mongrel dogs weighing 15-20 Kg were used in this study. Diabetes was chemically-induced in 7 dogs by a mixture of alloxan and streptozotocin. Three non-diabetic served as normal controls. Differentiated HBM-MSCs (5 million/Kg) were encapsulated in theracyte capsules and transplanted beneath the rectus sheath. Each dog received 2 capsules. One dog died 4 days postoperative from inhalation pneumonia. The remaining 6 dogs were followed up for 6-18 months. Results: Four dogs became normoglycemic within 6-8 weeks with normal glucose tolerance curves providing evidence that the transplanted cells were glucose-sensitive and insulin-responsive. In the remaining 2 dogs, fasting blood glucose was reduced but did not reach euglycemic levels. The sera of all transplanted dogs contained human insulin and c-peptide but negligible levels of canine insulin. When the HBM-MSCs loaded capsules were removed, rapid return of diabetic state was noted. The harvested capsules were examined by immunofluorescence. IPCs were seen and co-expression of with c-peptide was confirmed. Furthermore, all the pancreatic endocrine genes were expressed by the transplanted cells. Conclusions: This study provided evidence that theracyte capsules could protect the xenogenic HBM-MSCs from the host immune response. This is an important issue when clinical stem cell therapy is considered for definitive treatment for T1DM.

Keywords: diabetes, mesenchymal stem cells, dogs, Insulin-producing cells

Procedia PDF Downloads 173
219 Lamellodiscus spp. (Monogenoidea: Diplectanidae) Infecting the Gill Lamellae of Porgies (Spariformes: Sparidae) in Dakar Coast

Authors: Sikhou Drame, Arfang Diamanka

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In Senegal, the fishing sector plays an important role in socio-economic development. However, he is going through enormous difficulties, caused by the scarcity of fish on the Senegalese coast, the overexploitation of fishery resources. Based on this observation, the authorities are betting on the development of aquaculture. It is in this context that the exploration of fish from the highly consumed Sparidae family remains a good solution. Indeed, the Sparidae family has good characteristics for farming at sea. However, parasites can proliferate and destroy the efforts made to cultivate fish in confined areas. the knowledge of these parasites in particular the monogeneans, very specific to the sparidae fishes will allow to better know the bio-ecology of the fishes. Better know the main parasitic monogeneans of the genus Lamellodiscus of sparidae fish of the genus Pagrus harvested in Senegal. It will first be a question of identifying from the observation of the morpho-anatomical characters, Monogeneans of the genus Lamellodiscus, branchial parasites collected from three species of host: Pagrus caeruleostictus , Pagrus auriga and Pagrus africanus. Then to evaluate the spatial and temporary distribution of parasitic indices on two Dakar landing sites (Soumbédioune and Yarakh) and finally to determine their specificity. The fish examined were purchased directly from the landing sites in Dakar and then transported to the laboratory where they were identified, then dissected. The gills were examined under a magnifying glass and the monogeneans were harvested, fixed in 70% ethanol and then mounted between slide and coverslip. The identification of the parasites is based on the observation of the morpho-anatomical characters and on the measurements of the sclerified organs of the haptor and the male copulatory organ. In total out of the 90 individuals examined: Pagrus auriga (30), Pagrus africanus (30) and Pagrus caeruleostictus (30), 6 species of monogeneans of the genus Lamellodiscus (Monogenea, Diplectanidae) are obtained: L. sarculus, L. sigillatus, L.vicinus, L. rastellus, L. africanus n.sp and L. yarakhensis n.sp. Our results show that specimens of small sizes [15-20[cm are the most infested. The values of infestation intensity and abundance are higher in fish from Yarakh and also during the cold season. it is the species Pagrus caeruleostictus which records the highest parasitic loads in the two localities. the majority of the parasites identified have a strict or oioxene specificity. It appears from this study that fish of the genus Pagrus are highly parasitized by monogeneans of the genus Lamellodiscus with a general prevalence of 87.78%. Each infested fish has an average of 30 monogeneans of the genus Lamellodiscus.

Keywords: monogeneans, Lamellodiscus, Dakar coast, genus Pagrus

Procedia PDF Downloads 41
218 Identification and Characterization of Polysaccharide Biosynthesis Protein (CAPD) of Enterococcus faecium

Authors: Liaqat Ali, Hubert E. Blum, Türkân Sakinc

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Enterococcus faecium is an emerging multidrug-resistant nosocomial pathogen increased dramatically worldwide and causing bacteremia, endocarditis, urinary tract and surgical site infections in immunocomprised patients. The capsular polysaccharides that contribute to pathogenesis through evasion of the host innate immune system are also involved in hindering leukocyte killing of enterococci. The gene cluster (enterococcal polysaccharide antigen) of E. faecalis encoding homologues of many genes involved in polysaccharide biosynthesis. We identified two putative loci with 22 kb and 19 kb which contained 11 genes encoding for glycosyltransferases (GTFs); this was confirmed by using genome comparison of already sequenced strains that has no homology to known capsule genes and the epa-locus. The polysaccharide-conjugate vaccines have rapidly emerged as a suitable strategy to combat different pathogenic bacteria, therefore, we investigated a polysaccharide biosynthesis CapD protein in E. faecium contains 336 amino acids and had putative function for N-linked glycosylation. The deletion/knock-out capD mutant was constructed and complemented by homologues recombination method and confirmed by using PCR and sequencing. For further characterization and functional analysis, in-vitro cell culture and in-vivo a mouse infection models were used. Our ΔcapD mutant shows a strong hydrophobicity and all strains exhibited biofilm production. Subsequently, the opsonic activity was tested in an opsonophagocytic assay which shows increased in mutant compared complemented and wild type strains but more than two fold decreased in colonization and adherence was seen on surface of uroepithelial cells. However, a significant higher bacterial colonialization was observed in capD mutant during animal bacteremia infection. Unlike other polysaccharides biosynthesis proteins, CapD does not seems to be a major virulence factor in enterococci but further experiments and attention is needed to clarify its function, exact mechanism and involvement in pathogenesis of enteroccocal nosocomial infections eventually to develop a vaccine/ or targeted therapy.

Keywords: E. faecium, pathogenesis, polysaccharides, biofilm formation

Procedia PDF Downloads 294
217 Assessment of Utilization of Provider Initiated HIV Testing and Counseling and Associated Factors among Adult out Patient Department Patients in Wonchi Woreda, South West Shoa Zone, Central Ethiopia

Authors: Dinka Fikadu, Mulugeta Shegaze

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Background: Currently in health facility, provider-initiated human immunodeficiency virus testing is the key entry point to prevention, care, treatment and support services, but most people remains unaware of their HIV status due to various reasons. In many high-prevalence countries, fewer than one in ten people with HIV are aware of their HIV status. HIV, the virus that causes AIDS, “acquired immunodeficiency syndrome, "has become one of the world’s most serious health and development challenges. Reaching individuals with HIV who do not know their serostatus is a global public health priority. Objective: To assess utilization of provider initiated HIV testing and counseling and associated factors among adult outpatient department patients. Methods: Health facility based cross sectional study was conducted among 392 adult outpatient department patients in Wonchi woreda from February 24 to March 24 /2013. The study participant was recruited patients from all adult outpatient department patients of all four public health facilities of wonchi woreda using systematic sampling. A structured interviewer administered questionnaire was used to elicit all important variables from the study participants and multiple logistic regression analysis was used. Result: A total of 371 adult outpatient department patients aged between 15 to 64 years were actively participated in the study and 291(78.4%) of them utilized provider initiated HIV testing and counseling and 80(21.6%) of them refused. Knowledge on HIV is low in the study population; majority of the participants didn’t have comprehensive knowledge (64.7%) and (35.3%) fail to reject misconception about means of HIV transmission and prevention. Utilization of provider-initiated HIV testing and counseling were associated with divorced/widowed marital status[AOR (95%CI) = 0.32(0.15, 0.69)], being male sex [AOR (95%CI) =1.81(1.01, 3.24)], having comprehensive knowledge on HIV [AOR (95%CI) =0.408(0.220,0.759)],having awareness about provider initiated HIV testing and counseling [AOR(95%CI) =2.89(1.48,5.66)] and receiving test on HIV before[AOR (95%CI)=4.15(2.30, 7.47)]. Conclusion: Utilization of provider initiated HIV testing and counseling among adult outpatient departments in wonchi woreda public health facility was [(78.4%)].Strengthening health information through mass media and peer education on HIV to address barrier to testing in the community such as low awareness on PITC, to increase up take of PITC among adult OPD patients.

Keywords: utilization, human immune deficiency, testing, provider, initiate

Procedia PDF Downloads 263
216 Ethanol Precipitation and Characterization of L-Asparaginase from Aspergillus oryzae

Authors: L. L. Tundisi, A. Pessoa Jr., E. B. Tambourgi, E. Silveira, P. G. Mazzola

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L-asparaginase (L-ASNase) is the gold standard treatment for acute lymphoblastic leukemia that mainly affects pediatric patients; treatment increases survival from 20% to 90%. The characterization of other L-Asparaginases, apart from the most used from Escherichia coli and Erwinia chrysanthemi, has been reported, but the choice of the most appropriate is still under debate. This choice should be based on its pharmacokinetics, immune hypersensitivity, doses, prices, pharmacodynamics. The main factors influencing the antileukemic activity of ASNase are enzymatic activity, Km, glutaminase activity, clearance of the enzyme and development of resistance. However, most of the commercialized enzyme present an intrinsic glutaminase activity, which is responsible for some side effects. In this study, glutaminase free asparaginase produced from Aspergillus oryzae was precipitated in different percentages of ethanol (0–80%), until optimum ethanol concentration of 60% (w/w) was found. Following, precipitation of crude L-ASNase was performed in a single step, using 60% (w/w) ethanol, under constant agitation and temperature. It presented activity of 135.45 U/mg and after gel filtration chromatography with Sephadex G-the enzymatic activity was 322.02 U/mg. The apparent molecular mass of the purified L-ASNase fraction was estimated by 10% SDS-PAGE. Proteins were stained with Coomassie Brilliant Blue R-250. The molar mass range was from 10 kDa to 250 kDa. L-ASNase from Aspergillus oryzae was characterized aiming possible therapeutic use. Four different buffers (phosphate-citrate buffer pH 2.6 to 5.8; phosphate buffer pH 5.8 to 7.4; Tris - HCl pH 7.4 to 9.0; and carbonate buffer pH 9.8 to 10.6) were used to measure the optimum pH for L-ASNase activity. The optimum temperature for enzyme activity was measured at optimal pH conditions (Tris-HCl and phosphate buffer, pH 7.4) at different temperatures ranging from 5 to 55°C. All activities were calculated by quantifying the free ammonia, using the Nessler reagent. The kinetic parameters calculation, e.g. Michaelis-Menten constant (Km), maximum velocity (Vmax) and Hills coefficient (n), were performed by incubating the enzyme in different concentrations of the substrate at optimum conditions of pH and fitted on Hill’s equation. This glutaminase free asparaginase showed a low Km (3.39 mM and 3.81 mM) and enzymatic activity of 135.45 U/mg after precipitation with ethanol. After gel filtration chromatography it rose to 322.02 U/mg. Optimum activity was found between pH 5.8 - 9.0, best activity results with phosphate buffer pH 7.4 and Tris-HCl pH 7.4 and showed activity from 5°C to 55°C. These results indicate that L-ASNase from A. oryzae has the potential for human use.

Keywords: biopharmaceuticals, bioprocessing, bioproducts, biotechnology, enzyme activity, ethanol precipitation

Procedia PDF Downloads 256
215 Evaluation of Existence of Antithyroid Antibodies, Anti-Thyroid Peroxidase and Anti-Thyroglobulin in Patients with Hepatitis C Viral Infections

Authors: Junaid Mahmood Alam, Sana Anwar, Sarah Sughra Asghar

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Chronic hepatitis or Hepatitis C viral (HCV) infection has been identified as one of the factors that could elicit autoimmune disease resulting in the development of auto-antibodies. Furthermore, HCV is implicated in contravening of forbearance to antigens, therefore, inciting auto-reactivity. In this regard, several near and past studies noted the prevalence of thyroid dysfunction and production of anti-thyroid antibodies (ATAb) such as anti-thyroid peroxidase (AntiTPO) and anti-thyroglobulin (AntiTG) in patients with HCV. Likewise, one of the etiologies of augmentation of thyroid disease is basically interferon therapy for HCV infections, for which a number of autoimmune diseases have been noted including Grave’s disease, Hishimoto thyroiditis. A prospectively case-control study was therefore carried out at department of clinical biochemistry lab services and chemical pathology in collaboration with department of clinical microbiology, at Liaquat National Hospital and Medical College, Karachi Pakistan for the period January 2015 to December 2017. Two control groups were inducted for comparison purpose, control group 1 = without HCV infection and with thyroid disorders (n = 20), control group 2 = with HCV infection and without thyroid disorders (n = 20), whereas HCV infected were n = 40 where more than half were noted to be positive for either of HCV IgG and Ag. In HCV group, patients with existing sub-clinical hypothyroidism and clinical hyperthyroidism were less than 5%. Analysis showed the presence of AntiTG in 12 HCV patients (30%), AntiTPO in 15 (37.5%) and both AntiTG and antiTPO in 10 patients (25%). Only 3 patients were found with the history of anti-thyroid auto-antibodies (7.5%) and one with parents and relatives with auto-immune disorders (2.5%). Patients that remained untreated were 12 (30%), under treatment 18 (45%) and with complete-course of treatment 10 (25%). As per review of the literature, meta-analysis of evident data and cross-sectional studies of selective cohorts (as studied in presented research), thyroid connection is designated as one of the most recurrent endocrine ailment associated with chronic HCV infection. Moreover, it also represents an extrahepatic disease in the continuum of HCV syndrome. In conclusion, HCV patients were more likely to encompass thyroid disorders especially related to development of either of ATAb or both antiTG and AntiTPO.

Keywords: Hepatitis C viral (HCV) infection, anti-thyroid antibodies, anti-thyroid peroxidase antibodies, anti-thyroglobulin antibodies

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214 Hierarchical Zeolites as Potential Carriers of Curcumin

Authors: Ewelina Musielak, Agnieszka Feliczak-Guzik, Izabela Nowak

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Based on the latest data, it is expected that the substances of therapeutic interest used will be as natural as possible. Therefore, active substances with the highest possible efficacy and low toxicity are sought. Among natural substances with therapeutic effects, those of plant origin stand out. Curcumin isolated from the Curcuma longa plant has proven to be particularly important from a medical point of view. Due to its ability to regulate many important transcription factors, cytokines, and protein kinases, curcumin has found use as an anti-inflammatory, antioxidant, antiproliferative, antiangiogenic, and anticancer agent. The unfavorable properties of curcumin, such as low solubility, poor bioavailability, and rapid degradation under neutral or alkaline pH conditions, limit its clinical application. These problems can be solved by combining curcumin with suitable carriers such as hierarchical zeolites. This is a new class of materials that exhibit several advantages. Hierarchical zeolites used as drug carriers enable delayed release of the active ingredient and promote drug transport to the desired tissues and organs. In addition, hierarchical zeolites play an important role in regulating micronutrient levels in the body and have been used successfully in cancer diagnosis and therapy. To apply curcumin to hierarchical zeolites synthesized from commercial FAU zeolite, solutions containing curcumin, carrier and acetone were prepared. The prepared mixtures were then stirred on a magnetic stirrer for 24 h at room temperature. The curcumin-filled hierarchical zeolites were drained into a glass funnel, where they were washed three times with acetone and distilled water, after which the obtained material was air-dried until completely dry. In addition, the effect of piperine addition to zeolite carrier containing a sufficient amount of curcumin was studied. The resulting products were weighed and the percentage of pure curcumin in the hierarchical zeolite was calculated. All the synthesized materials were characterized by several techniques: elemental analysis, transmission electron microscopy (TEM), Fourier transform infrared spectroscopy, Fourier transform infrared (FT-IR), N2 adsorption, and X-ray diffraction (XRD) and thermogravimetric analysis (TGA). The aim of the presented study was to improve the biological activity of curcumin by applying it to hierarchical zeolites based on FAU zeolite. The results showed that the loading efficiency of curcumin into hierarchical zeolites based on commercial FAU-type zeolite is enhanced by modifying the zeolite carrier itself. The hierarchical zeolites proved to be very good and efficient carriers of plant-derived active ingredients such as curcumin.

Keywords: carriers of active substances, curcumin, hierarchical zeolites, incorporation

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213 Anti-Colitic and Anti-Inflammatory Effects of Lactobacillus sakei K040706 in Mice with Ulcerative Colitis

Authors: Seunghwan Seo, Woo-Seok Lee, Ji-Sun Shin, Young Kyoung Rhee, Chang-Won Cho, Hee-Do Hong, Kyung-Tae Lee

Abstract:

Doenjang, known as traditional Korean food, is product of a natural mixed fermentation process carried out by lactic acid bacteria (LAB). Lactobacillus sakei K040706 (K040706) has been accepted as the most populous LAB in over ripened doenjang. Recently, we reported the immunostimulatory effects of K040706 in RAW 264.7 macrophages and in a cyclophosphamide-induced mouse model. In this study, we investigated the ameliorative effects of K040706 in a dextran sulfate sodium (DSS)-induced colitis mouse model. We induced colitis using DSS in 5-week-ICR mice over 14 days with or without 0.1, 1 g/kg/day K040706 orally. The body weight, stool consistency, and gross bleeding were recorded for determination of the disease activity index (DAI). At the end of treatment, animals were sacrificed and colonic tissues were collected and subjected to histological experiments and myeloperoxidase (MPO) accumulation, cytokine determination, qRT-PCR and Western blot analysis. Results showed that K040706 significantly attenuated DSS-induced DAI score, shortening of colon length, enlargement of spleen and immune cell infiltrations into colonic tissues. Histological examinations indicated that K040706 suppressed edema, mucosal damage, and the loss of crypts induced by DSS. These results were correlated with the restoration of tight junction protein expression, such as, ZO-1 and occludin in K040706-treated mice. Moreover, K040706 reduced the abnormal secretions and mRNA expressions of pro-inflammatory mediators, such as nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6). DSS-induced mRNA expression of intercellular adhesion molecule (ICAM) and vascular cell adhesion molecule (VCAM) in colonic tissues was also downregulated by K040706 treatment. Furthermore, K040706 suppressed the protein and mRNA expression of toll-like receptor 4 (TLR4) and phosphorylation of NF-κB and signal transducer and activator of transcription 3 (STAT3). These results suggest that K040706 has an anti-colitic effect by inhibition of intestinal inflammatory responses in DSS-induced colitic mice.

Keywords: Lactobacillus sakei, NF-κB, STAT3, ulcerative colitis

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212 Comparative Study between Mesenchymal Stem Cells and Regulatory T-Cells in Macrophage Polarization for Organ Transplant Tolerance: In Vitro Study

Authors: Vijaya Madhuri Devraj, Swarnalatha Guditi, Kiran Kumar Bokara, Gangadhar Taduri

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Cell-based strategies may open therapeutic approaches that promote tolerance through manipulation of macrophages to increase long-term transplant survival rates and minimize side effects of the current immune suppressive regimens. The aim of the present study was, therefore, to test and compare the therapeutic potential of MSC and Tregs on macrophage polarization to develop an alternate cell-based treatment option in kidney transplantation. In the current protocol, macrophages from kidney transplant recipients with graft dysfunction were co-cultured with MSCs and Treg cells with and without cell-cell contact on transwell plates, further to quantitatively assess macrophage polarization in response to MSC and Treg treatment over time, M1 and M2 cell surface markers were used. Additionally, multiple soluble analytes were analyzed in cell supernatant by using bead-based immunoassays. Furthermore, to confirm our findings, gene expression analysis was done. MSCs induced the formation of M2 macrophages more than Tregs when macrophages M0 were cultured in transwell without cell contact. From this, we deduced the mechanism that soluble factors present in the MSCs condition media are involved in skewing of macrophages towards type 2 macrophages; similarly, in co-culture with cell-cell contact, MSCs resulted in more M2 type macrophages than Tregs. And an important finding of this study is the combination of both MSC-Treg showed significantly effective and consistent results in both with and without cell contact setups. Hence, it is suggestive to prefer MSCs over Tregs for secretome-based therapy and a combination of both for either therapy for effective transplantation outcomes. Our findings underline a key role of Tregs and MSCs in promoting macrophage polarization towards anti-inflammatory type. The study has great importance in prolongation of allograft and patient survival without any rejection by cell-based therapy, which induce self-tolerance and controlling infection.

Keywords: graft rejection, graft tolerance, macrophage polarization, mesenchymal stem cells, regulatory T cells, transplant immunology

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211 Ethical Framework in Organ Transplantation and the Priority Line between Law and Life

Authors: Abel Sichinava

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The need for organ transplantation is vigorously increasing worldwide. The numbers on the waiting lists grow, but the number of donors is not keeping up with the demand even though there is a legal possibility of decreasing the gap between the demand and supply. Most countries around the globe are facing an organ donation problem (living or deceased); however, the extent of the problem differs based on how well developed a country is. The determining issues seem to be centered on how aware the society is about the concept of organ donation, as well as cultural and religious factors. Even if people are aware of the benefits of organ donation, they may still have fears that keep them from being in complete agreement with the idea. Some believe that in the case of deceased organ donation: “the brain dead human body may recover from its injuries” or “the sick might get less appropriate treatment if doctors know they are potential donors.” In the case of living organ donations, people sometimes fear that after the donation, “it might reduce work efficiency, cause health deterioration or even death.” Another major obstacle in the organ shortage is a lack of a well developed ethical framework. In reality, there are truly an immense number of people on the waiting list, and they have only two options in order to receive a suitable organ. First is the legal way, which is to wait until their turn. Sadly, numerous patients die while on the waiting list before an appropriate organ becomes available for transplant. The second option is an illegal way: seeking an organ in a country where they can possibly get. To tell the truth, in people’s desire to live, they may choose the second option if their resources are sufficient. This process automatically involves “organ brokers.” These are people who get organs from vulnerable poor people by force or betrayal. As mentioned earlier, the high demand and low supply leads to human trafficking. The subject of the study was the large number of society from different backgrounds of their belief, culture, nationality, level of education, socio-economic status. The great majority of them interviewed online used “Google Drive Survey” and others in person. All statistics and information gathered from trusted sources annotated in the reference list and above mentioned considerable testimonies shared by the respondents are the fundamental evidence of a lack of the well developed ethical framework. In conclusion, the continuously increasing number of people on the waiting list and an irrelevant ethical framework, lead people to commit to atrocious, dehumanizing crimes. Therefore, world society should be equally obligated to think carefully and make vital decisions together for the advancement of an organ donations and its ethical framework.

Keywords: donation, ethical framwork, organ, transplant

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210 Combining the Production of Radiopharmaceuticals with the Department of Radionuclide Diagnostics

Authors: Umedov Mekhroz, Griaznova Svetlana

Abstract:

In connection with the growth of oncological diseases, the design of centers for diagnostics and the production of radiopharmaceuticals is the most relevant area of healthcare facilities. The design of new nuclear medicine centers should be carried out from the standpoint of solving the following tasks: the availability of medical care, functionality, environmental friendliness, sustainable development, improving the safety of drugs, the use of which requires special care, reducing the rate of environmental pollution, ensuring comfortable conditions for the internal microclimate, adaptability. The purpose of this article is to substantiate architectural and planning solutions, formulate recommendations and principles for the design of nuclear medicine centers and determine the connections between the production and medical functions of a building. The advantages of combining the production of radiopharmaceuticals and the department of medical care: less radiation activity is accumulated, the cost of the final product is lower, and there is no need to hire a transport company with a special license for transportation. A medical imaging department is a structural unit of a medical institution in which diagnostic procedures are carried out in order to gain an idea of the internal structure of various organs of the body for clinical analysis. Depending on the needs of a particular institution, the department may include various rooms that provide medical imaging using radiography, ultrasound diagnostics, and the phenomenon of nuclear magnetic resonance. The production of radiopharmaceuticals is an object intended for the production of a pharmaceutical substance containing a radionuclide and intended for introduction into the human body or laboratory animal for the purpose of diagnosis, evaluation of the effectiveness of treatment, or for biomedical research. The research methodology includes the following subjects: study and generalization of international experience in scientific research, literature, standards, teaching aids, and design materials on the topic of research; An integrated approach to the study of existing international experience of PET / CT scan centers and the production of radiopharmaceuticals; Elaboration of graphical analysis and diagrams based on the system analysis of the processed information; Identification of methods and principles of functional zoning of nuclear medicine centers. The result of the research is the identification of the design principles of nuclear medicine centers with the functions of the production of radiopharmaceuticals and the department of medical imaging. This research will be applied to the design and construction of healthcare facilities in the field of nuclear medicine.

Keywords: architectural planning solutions, functional zoning, nuclear medicine, PET/CT scan, production of radiopharmaceuticals, radiotherapy

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209 Taraxacum Officinale (Dandelion) and Its Phytochemical Approach to Malignant Diseases

Authors: Angel Champion

Abstract:

Chemotherapy and radiation use an acidified approach to induce apoptosis, which only kills mature cancer cells while resulting in gene and cell damage with significant levels of toxicity in tumor-affected tissues and organs. The acid approach, where the cells exterminated are not differentiated, induces the disappearance of white blood cells from the blood. This increases susceptibility to infection in severe forms of cancer spread. However, chemotherapy and radiation cannot kill cancer stem cells that metastasize, being the leading cause of 98% of cancer fatalities. With over 12 million new cancer cases symptomatic each year, including common malignancies such as Hepatocellular Carcinoma (HCC), this study aims to assess the bioactive constituents and phytochemical composition of Taraxacum Officinale (Dandelion). This analysis enables pharmaceutical quality and potency to be applied to studies on cancer cell proliferation and apoptosis. A phytochemical screening is carried out to identify the antioxidant components of Dandelion root, stem, and flower extract. The constituents tested for are phlorotannins, carbohydrates, glycosides, saponins, flavonoids, alkaloids, sterols, triterpenes, and anthraquinone glycosides. To conserve the existing phenolic compounds, a portion of the constituent tests will be examined with an acid, alcohol, or aqueous solvent. As a result, the qualitative and quantitative variations within the Dandelion extract that measure uniform effective potency are vital to the conformity for producing medicinal products. These medicines will be constructed with a consistent, uniform composition that physicians can use to control and effectively eradicate malignant diseases safely. Taraxacum Officinale's phytochemical composition comprises a highly-graded potency due to present bioactive contents that will essentially drive out malignant disease within the human body. Its high potency rate is powerful enough to eliminate both mature cancer cells and cancer stem cells without the cell and gene damage induced by chemotherapy and radiation. Correspondingly, the high margins of cancer mortality on a global scale are mitigated. This remarkable contribution to modern therapeutics will essentially optimize the margins of natural products and their derivatives, which account for 50% of pharmaceuticals in modern therapeutics, while preventing the adverse effects of radiation and chemotherapy drugs.

Keywords: antioxidant, apoptosis, metastasize, phytochemical, proliferation, potency

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