Search results for: molecular diagnosis
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 4038

Search results for: molecular diagnosis

438 Assessment of Serum Osteopontin, Osteoprotegerin and Bone-Specific Alp as Markers of Bone Turnover in Patients with Disorders of Thyroid Function in Nigeria, Sub-Saharan Africa

Authors: Oluwabori Emmanuel Olukoyejo, Ogra Victor Ogra, Bosede Amodu, Tewogbade Adeoye Adedeji

Abstract:

Background: Disorders of thyroid function are the second most common endocrine disorders worldwide, with a direct relationship with metabolic bone diseases. These metabolic bone complications are often subtle but manifest as bone pains and an increased risk of fractures. The gold standard for diagnosis, Dual Energy X-ray Absorptiometry (DEXA), is limited in this environment due to unavailability, cumbersomeness and cost. However, bone biomarkers have shown prospects in assessing alterations in bone remodeling, which has not been studied in this environment. Aim: This study evaluates serum levels of bone-specific alkaline phosphatase (bone-specific ALP), osteopontin and osteoprotegerin biomarkers of bone turnover in patients with disorders of thyroid function. Methods: This is a cross-sectional study carried out over a period of one and a half years. Forty patients with thyroid dysfunctions, aged 20 to 50 years, and thirty-eight age and sex-matched healthy euthyroid controls were included in this study. Patients were further stratified into hyperthyroid and hypothyroid groups. Bone-specific ALP, osteopontin, and osteoprotegerin, alongside serum total calcium, ionized calcium and inorganic phosphate, were assayed for all patients and controls. A self-administered questionnaire was used to obtain data on sociodemographic and medical history. Then, 5 ml of blood was collected in a plain bottle and serum was harvested following clotting and centrifugation. Serum samples were assayed for B-ALP, osteopontin, and osteoprotegerin using the ELISA technique. Total calcium and ionized calcium were assayed using an ion-selective electrode, while the inorganic phosphate was assayed with automated photometry. Results: The hyperthyroid and hypothyroid patient groups had significantly increased median serum B-ALP (30.40 and 26.50) ng/ml and significantly lower median OPG (0.80 and 0.80) ng/ml than the controls (10.81 and 1.30) ng/ml respectively, p < 0.05. However, serum osteopontin in the hyperthyroid group was significantly higher and significantly lower in the hypothyroid group when compared with the controls (11.00 and 2.10 vs 3.70) ng/ml, respectively, p < 0.05. Both hyperthyroid and hypothyroid groups had significantly higher mean serum total calcium, ionized calcium and inorganic phosphate than the controls (2.49 ± 0.28, 1.27 ± 0.14 and 1.33 ± 0.33) mmol/l and (2.41 ± 0.04, 1.20 ± 0.04 and 1.15 ± 0.16) mmol/l vs (2.27 ± 0.11, 1.17 ± 0.06 and 1.08 ± 0.16) mmol/l respectively, p < 0.05. Conclusion: Patients with disorders of thyroid function have metabolic imbalances of all the studied bone markers, suggesting a higher bone turnover. The routine bone markers will be an invaluable tool for monitoring bone health in patients with thyroid dysfunctions, while the less readily available markers can be introduced as supplementary tools. Moreover, bone-specific ALP, osteopontin and osteoprotegerin were found to be the strongest independent predictors of metabolic bone markers’ derangements in patients with thyroid dysfunctions.

Keywords: metabolic bone diseases, biomarker, bone turnover, hyperthyroid, hypothyroid, euthyroid

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437 Pond Site Diagnosis: Monoclonal Antibody-Based Farmer Level Tests to Detect the Acute Hepatopancreatic Necrosis Disease in Shrimp

Authors: B. T. Naveen Kumar, Anuj Tyagi, Niraj Kumar Singh, Visanu Boonyawiwat, A. H. Shanthanagouda, Orawan Boodde, K. M. Shankar, Prakash Patil, Shubhkaramjeet Kaur

Abstract:

Early mortality syndrome (EMS)/Acute Hepatopancreatic Necrosis Disease (AHPND) has emerged as a major obstacle for the shrimp farming around the world. It is caused by a strain of Vibrio parahaemolyticus. The possible preventive and control measure is, early and rapid detection of the pathogen in the broodstock, post-larvae and monitoring the shrimp during the culture period. Polymerase chain reaction (PCR) based early detection methods are good, but they are costly, time taking and requires a sophisticated laboratory. The present study was conducted to develop a simple, sensitive and rapid diagnostic farmer level kit for the reliable detection of AHPND in shrimp. A panel of monoclonal antibodies (MAbs) were raised against the recombinant Pir B protein (rPirB). First, an immunodot was developed by using MAbs G3B8 and Mab G3H2 which showed specific reactivity to purified r-PirB protein with no cross-reactivity to other shrimp bacterial pathogens (AHPND free Vibrio parahaemolyticus (Indian strains), V. anguillarum, WSSV, Aeromonas hydrophila, and Aphanomyces invadans). Immunodot developed using Mab G3B8 is more sensitive than that with the Mab G3H2. However, immunodot takes almost 2.5 hours to complete with several hands-on steps. Therefore, the flow-through assay (FTA) was developed by using a plastic cassette containing the nitrocellulose membrane with absorbing pads below. The sample was dotted in the test zone on the nitrocellulose membrane followed by continuos addition of five solutions in the order of i) blocking buffer (BSA) ii) primary antibody (MAb) iii) washing Solution iv) secondary antibody and v) chromogen substrate (TMB) clear purple dots against a white background were considered as positive reactions. The FTA developed using MAbG3B8 is more sensitive than that with MAb G3H2. In FTA the two MAbs showed specific reactivity to purified r-PirB protein and not to other shrimp bacterial pathogens. The FTA is simple to farmer/field level, sensitive and rapid requiring only 8-10 min for completion. Tests can be developed to kits, which will be ideal for use in biosecurity, for the first line of screening (at the port or pond site) and during monitoring and surveillance programmes overall for the good management practices to reduce the risk of the disease.

Keywords: acute hepatopancreatic necrosis disease, AHPND, flow-through assay, FTA, farmer level, immunodot, pond site, shrimp

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436 Antiplatelets and Anticoagulants in Rural Emergency General Surgery

Authors: Jeong-Moh John Yahng, Angelika Na

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Introduction: Increasing numbers of general surgical patients are being prescribed antiplatelet and anticoagulant medications (APAC) for various cardiovascular and cerebrovascular conditions. Surgical patients who are on APAC present a management challenge as bleeding risk needs to be balanced with thromboembolic risk. Although guidelines exist in regards to APAC management in elective surgery, there is a lack of guidelines in the emergency surgery setting. In this study we aim to characterise APAC usage in emergency general surgical patients admitted to a rural hospital. We also assess the impact of APAC usage on clinical management of these patients. Methods: Prospective study of emergency general surgical admissions at Northeast Health Wangaratta (Victoria) from 2 July to 25 Oct 2014. Questionnaire collected demographics data, admission diagnosis, APAC usage, anaesthesia techniques, operation types, transfusion requirement and morbidity / mortality data. Results: During the 4 month study, 118 patients were classified into two groups: non-APAC (n=96, 81%) and APAC (n=22, 19%). Patients in the APAC group were older compared to the non-APAC patients (mean age 72 vs 42 years old). Amongst patients younger than 60 years old, only 1% of them were on APAC. In contrast, 49% of patients older than 60 years old were on APAC (p<0.001). Patients who were admitted with a bleeding problem were more likely to be on APAC (p<0.05). 19% of emergency general surgery patients were on APAC. The majority (91%) of them were on antiplatelet medication, with two patients being on dual antiplatelet agents (aspirin + clopidogrel or ticagrelor). 15% of emergency general surgical patients requiring operations were on APAC. 11% of all laparotomies and 33% of gastroscopy for haematemesis/melaena patients were on APAC. Both of the patients operated for bleeding following surgery at another hospital were in the APAC group. In regards to impact on clinical management, 59% of APAC patients had their medications interrupted or ceased, on average by 3.5 days (range 1-13 days). 2 out of 75 operations were delayed due to APAC usage. There was no difference in the use of central venous or arterial line for increased monitoring (p=0.14) or in the use of warming blanket (Bair Hugger™) (p=0.94). Overall, transfusion rate was higher amongst APAC patients (14% vs 3%) (p 0.04). The recorded morbidity (n=2) and mortality (n=1) in this study were all in the APAC group. Discussion: Nineteen percent of emergency general surgical admissions and fifteen percent of operated patients were on APAC. The prevalence of APAC usage was higher in those aged sixty and above. General surgical patients who were admitted with a bleeding problem were more likely to be on APAC. Two patients who were operated for bleeding following surgery at another hospital were in the APAC group. Note that there was no patient in the non-APAC group who was admitted for post-operative bleeding. We observed two cases in which operation was delayed due to APAC usage. Transfusion, morbidity and mortality rate were higher in the APAC group. Conclusion: In this study, nineteen percent of emergency general surgical admissions were on APAC. The use of APAC is more prevalent in the older age group, particularly those aged sixty and above. Higher proportion of APAC compared to non-APAC patients were admitted and operated for bleeding problems. There is an urgent need for clinical guidelines regarding APAC management in emergency general surgical patients.

Keywords: antiplatelet, anticoagulants, emergency general surgery, rural general surgery, morbidity, mortality

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435 Care Experience of a Female Breast Cancer Patient Undergoing Modified Radical Mastectomy

Authors: Ting-I Lin

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Purpose: This article explores the care experience of a 34-year-old female breast cancer patient who was admitted to the intensive care unit after undergoing a modified radical mastectomy. The patient discovered a lump in her right breast during a self-examination and, after mammography and ultrasound-guided biopsy, was diagnosed with a malignant tumor in the right breast. The tumor measured 1.5 x 1.4 x 2 cm, and the patient underwent a modified radical mastectomy. Postoperatively, she exhibited feelings of inferiority due to changes in her appearance. Method: During the care period, we engaged in conversations, observations, and active listening, using Gordon's Eleven Functional Health Patterns for a comprehensive assessment. In collaboration with the critical care team, a psychologist, and an oncology case manager, we conducted an interdisciplinary discussion and reached a consensus on key nursing issues. These included pain related to postoperative tumor excision and disturbed body image due to changes in appearance after surgery. Result: During the care period, a private space was provided to encourage the patient to express her feelings about her altered body image. Communication was conducted through active listening and a non-judgmental approach. The patient's anxiety level, as measured by the depression and anxiety scale, decreased from moderate to mild, and she was able to sleep for 6-8 hours at night. The oncology case manager was invited to provide education on breast reconstruction using breast models and videos to both the patient and her husband. This helped rebuild the patient's confidence. With the patient's consent, a support group was arranged where a peer with a similar experience shared her journey, offering emotional support and encouragement. This helped alleviate the psychological stress and shock caused by the cancer diagnosis. Additionally, pain management was achieved through adjusting the dosage of analgesics, administering Ultracet 37.5 mg/325 mg 1# Q6H PO, along with distraction techniques and acupressure therapy. These interventions helped the patient relax and alleviate discomfort, maintaining her pain score at a manageable level of 3, indicating mild pain. Conclusion: Disturbance in body image can cause significant psychological stress for patients. Through support group discussions, encouraging patients to express their feelings, and providing appropriate education on breast reconstruction and dressing techniques, the patient's self-concept was positively reinforced, and her emotions were stabilized. This led to renewed self-worth and confidence.

Keywords: breast cancer, modified radical mastectomy, acupressure therapy, Gordon's 11 functional health patterns

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434 A Theragnostic Approach for Alzheimer’s Disease Focused on Phosphorylated Tau

Authors: Tomás Sobrino, Lara García-Varela, Marta Aramburu-Núñez, Mónica Castro, Noemí Gómez-Lado, Mariña Rodríguez-Arrizabalaga, Antía Custodia, Juan Manuel Pías-Peleteiro, José Manuel Aldrey, Daniel Romaus-Sanjurjo, Ángeles Almeida, Pablo Aguiar, Alberto Ouro

Abstract:

Introduction: Alzheimer’s disease (AD) and other tauopathies are primary causes of dementia, causing progressive cognitive deterioration that entails serious repercussions for the patients' performance of daily tasks. Currently, there is no effective approach for the early diagnosis and treatment of AD and tauopathies. This study suggests a theragnostic approach based on the importance of phosphorylated tau protein (p-Tau) in the early pathophysiological processes of AD. We have developed a novel theragnostic monoclonal antibody (mAb) to provide both diagnostic and therapeutic effects. Methods/Results: We have developed a p-Tau mAb, which was doped with deferoxamine for radiolabeling with Zirconium-89 (89Zr) for PET imaging, as well as fluorescence dies for immunofluorescence assays. The p-Tau mAb was evaluated in vitro for toxicity by MTT assay, LDH activity, propidium iodide/Annexin V assay, caspase-3, and mitochondrial membrane potential (MMP) assay in both mouse endothelial cell line (bEnd.3) and cortical primary neurons cell cultures. Importantly, non-toxic effects (up to concentrations of p-Tau mAb greater than 100 ug/mL) were detected. In vivo experiments in the tauopathy model mice (PS19) show that the 89Zr-pTau-mAb and 89Zr-Fragments-pTau-mAb are stable in circulation for up to 10 days without toxic effects. However, only less than 0.2% reached the brain, so further strategies have to be designed for crossing the Brain-Blood-Barrier (BBB). Moreover, an intraparenchymal treatment strategy was carried out. The PS19 mice were operated to implement osmotic pumps (Alzet 1004) at two different times, at 4 and 7 months, to stimulate the controlled release for one month each of the B6 antibody or the IgG1 control antibody. We demonstrated that B6-treated mice maintained their motor and memory abilities significantly compared with IgG1 treatment. In addition, we observed a significant reduction in p-Tau deposits in the brain. Conclusions /Discussion: A theragnostic pTau-mAb was developed. Moreover, we demonstrated that our p-Tau mAb recognizes very-early pathology forms of p-Tau by non-invasive techniques, such as PET. In addition, p-Tau mAb has non-toxic effects, both in vitro and in vivo. Although the p-Tau mAb is stable in circulation, only 0.2% achieve the brain. However, direct intraventricular treatment significantly reduces cognitive impairment in Alzheimer's animal models, as well as the accumulation of toxic p-Tau species.

Keywords: alzheimer's disease, theragnosis, tau, PET, immunotherapy, tauopathies

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433 The Usefulness of Premature Chromosome Condensation Scoring Module in Cell Response to Ionizing Radiation

Authors: K. Rawojć, J. Miszczyk, A. Możdżeń, A. Panek, J. Swakoń, M. Rydygier

Abstract:

Due to the mitotic delay, poor mitotic index and disappearance of lymphocytes from peripheral blood circulation, assessing the DNA damage after high dose exposure is less effective. Conventional chromosome aberration analysis or cytokinesis-blocked micronucleus assay do not provide an accurate dose estimation or radiosensitivity prediction in doses higher than 6.0 Gy. For this reason, there is a need to establish reliable methods allowing analysis of biological effects after exposure in high dose range i.e., during particle radiotherapy. Lately, Premature Chromosome Condensation (PCC) has become an important method in high dose biodosimetry and a promising treatment modality to cancer patients. The aim of the study was to evaluate the usefulness of drug-induced PCC scoring procedure in an experimental mode, where 100 G2/M cells were analyzed in different dose ranges. To test the consistency of obtained results, scoring was performed by 3 independent persons in the same mode and following identical scoring criteria. Whole-body exposure was simulated in an in vitro experiment by irradiating whole blood collected from healthy donors with 60 MeV protons and 250 keV X-rays, in the range of 4.0 – 20.0 Gy. Drug-induced PCC assay was performed on human peripheral blood lymphocytes (HPBL) isolated after in vitro exposure. Cells were cultured for 48 hours with PHA. Then to achieve premature condensation, calyculin A was added. After Giemsa staining, chromosome spreads were photographed and manually analyzed by scorers. The dose-effect curves were derived by counting the excess chromosome fragments. The results indicated adequate dose estimates for the whole-body exposure scenario in the high dose range for both studied types of radiation. Moreover, compared results revealed no significant differences between scores, which has an important meaning in reducing the analysis time. These investigations were conducted as a part of an extended examination of 60 MeV protons from AIC-144 isochronous cyclotron, at the Institute of Nuclear Physics in Kraków, Poland (IFJ PAN) by cytogenetic and molecular methods and were partially supported by grant DEC-2013/09/D/NZ7/00324 from the National Science Centre, Poland.

Keywords: cell response to radiation exposure, drug induced premature chromosome condensation, premature chromosome condensation procedure, proton therapy

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432 Noncovalent Antibody-Nanomaterial Conjugates: A Simple Approach to Produce Targeted Nanomedicines

Authors: Nicholas Fletcher, Zachary Houston, Yongmei Zhao, Christopher Howard, Kristofer Thurecht

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One promising approach to enhance nanomedicine therapeutic efficacy is to include a targeting agent, such as an antibody, to increase accumulation at the tumor site. However, the application of such targeted nanomedicines remains limited, in part due to difficulties involved with biomolecule conjugation to synthetic nanomaterials. One approach recently developed to overcome this has been to engineer bispecific antibodies (BsAbs) with dual specificity, whereby one portion binds to methoxy polyethyleneglycol (mPEG) epitopes present on synthetic nanomedicines, while the other binds to molecular disease markers of interest. In this way, noncovalent complexes of nanomedicine core, comprising a hyperbranched polymer (HBP) of primarily mPEG, decorated with targeting ligands are able to be produced by simple mixing. Further work in this area has now demonstrated such complexes targeting the breast cancer marker epidermal growth factor receptor (EGFR) to show enhanced binding to tumor cells both in vitro and in vivo. Indeed the enhanced accumulation at the tumor site resulted in improved therapeutic outcomes compared to untargeted nanomedicines and free chemotherapeutics. The current work on these BsAb-HBP conjugates focuses on further probing antibody-nanomaterial interactions and demonstrating broad applicability to a range of cancer types. Herein are reported BsAb-HBP materials targeted towards prostate-specific membrane antigen (PSMA) and study of their behavior in vivo using ⁸⁹Zr positron emission tomography (PET) in a dual-tumor prostate cancer xenograft model. In this model mice bearing both PSMA+ and PSMA- tumors allow for PET imaging to discriminate between nonspecific and targeted uptake in tumors, and better quantify the increased accumulation following BsAb conjugation. Also examined is the potential for formation of these targeted complexes in situ following injection of individual components? The aim of this approach being to avoid undesirable clearance of proteinaceous complexes upon injection limiting available therapeutic. Ultimately these results demonstrate BsAb functionalized nanomaterials as a powerful and versatile approach for producing targeted nanomedicines for a variety of cancers.

Keywords: bioengineering, cancer, nanomedicine, polymer chemistry

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431 Modeling Engagement with Multimodal Multisensor Data: The Continuous Performance Test as an Objective Tool to Track Flow

Authors: Mohammad H. Taheri, David J. Brown, Nasser Sherkat

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Engagement is one of the most important factors in determining successful outcomes and deep learning in students. Existing approaches to detect student engagement involve periodic human observations that are subject to inter-rater reliability. Our solution uses real-time multimodal multisensor data labeled by objective performance outcomes to infer the engagement of students. The study involves four students with a combined diagnosis of cerebral palsy and a learning disability who took part in a 3-month trial over 59 sessions. Multimodal multisensor data were collected while they participated in a continuous performance test. Eye gaze, electroencephalogram, body pose, and interaction data were used to create a model of student engagement through objective labeling from the continuous performance test outcomes. In order to achieve this, a type of continuous performance test is introduced, the Seek-X type. Nine features were extracted including high-level handpicked compound features. Using leave-one-out cross-validation, a series of different machine learning approaches were evaluated. Overall, the random forest classification approach achieved the best classification results. Using random forest, 93.3% classification for engagement and 42.9% accuracy for disengagement were achieved. We compared these results to outcomes from different models: AdaBoost, decision tree, k-Nearest Neighbor, naïve Bayes, neural network, and support vector machine. We showed that using a multisensor approach achieved higher accuracy than using features from any reduced set of sensors. We found that using high-level handpicked features can improve the classification accuracy in every sensor mode. Our approach is robust to both sensor fallout and occlusions. The single most important sensor feature to the classification of engagement and distraction was shown to be eye gaze. It has been shown that we can accurately predict the level of engagement of students with learning disabilities in a real-time approach that is not subject to inter-rater reliability, human observation or reliant on a single mode of sensor input. This will help teachers design interventions for a heterogeneous group of students, where teachers cannot possibly attend to each of their individual needs. Our approach can be used to identify those with the greatest learning challenges so that all students are supported to reach their full potential.

Keywords: affective computing in education, affect detection, continuous performance test, engagement, flow, HCI, interaction, learning disabilities, machine learning, multimodal, multisensor, physiological sensors, student engagement

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430 Morphological and Molecular Evaluation of Dengue Virus Serotype 3 Infection in BALB/c Mice Lungs

Authors: Gabriela C. Caldas, Fernanda C. Jacome, Arthur da C. Rasinhas, Ortrud M. Barth, Flavia B. dos Santos, Priscila C. G. Nunes, Yuli R. M. de Souza, Pedro Paulo de A. Manso, Marcelo P. Machado, Debora F. Barreto-Vieira

Abstract:

The establishment of animal models for studies of DENV infections has been challenging, since circulating epidemic viruses do not naturally infect nonhuman species. Such studies are of great relevance to the various areas of dengue research, including immunopathogenesis, drug development and vaccines. In this scenario, the main objective of this study is to verify possible morphological changes, as well as the presence of antigens and viral RNA in lung samples from BALB/c mice experimentally infected with an epidemic and non-neuroadapted DENV-3 strain. Male BALB/c mice, 2 months old, were inoculated with DENV-3 by intravenous route. After 72 hours of infection, the animals were euthanized and the lungs were collected. Part of the samples was processed by standard technique for analysis by light and transmission electronic microscopies and another part was processed for real-time PCR analysis. Morphological analyzes of lungs from uninfected mice showed preserved tissue areas. In mice infected with DENV-3, the analyzes revealed interalveolar septum thickening with presence of inflammatory infiltrate, foci of alveolar atelectasis and hyperventilation, bleeding foci in the interalveolar septum and bronchioles, peripheral capillary congestion, accumulation of fluid in the blood capillary, signs of interstitial cell necrosis presence of platelets and mononuclear inflammatory cells circulating in the capillaries and/or adhered to the endothelium. In addition, activation of endothelial cells, platelets, mononuclear inflammatory cell and neutrophil-type polymorphonuclear inflammatory cell evidenced by the emission of cytoplasmic membrane prolongation was observed. DEN-like particles were seen in the cytoplasm of endothelial cells. The viral genome was recovered from 3 in 12 lung samples. These results demonstrate that the BALB / c mouse represents a suitable model for the study of the histopathological changes induced by DENV infection in the lung, with tissue alterations similar to those observed in human cases of DEN.

Keywords: BALB/c mice, dengue, histopathology, lung, ultrastructure

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429 Neonatal Sepsis in Dogs Attend in Veterinary Hospital of the Sao Paulo State University, Botucatu, Brazil – Incidence, Clinical Aspects and Mortality

Authors: Maria Lucia G. Lourenco, Keylla H. N. P. Pereira, Vivane Y. Hibaru, Fabiana F. Souza, Joao C. P. Ferreira, Simone B. Chiacchio, Luiz H. A. Machado

Abstract:

Neonatal sepsis is a systemic response to the acute generalized infection caused by one or more bacterial agents, representing the main infectious cause of neonatal mortality in dogs during the first three weeks of life. This study aims to describe the incidence of sepsis in neonate dogs, as well as the main clinical signs and mortality rates. The study included 735 neonates admitted to the Sao Paulo State University (UNESP) Veterinary Hospital, Botucatu, Sao Paulo, Brazil, between January 2018 and November 2019. Seven hundred thirty-five neonates, 14% (98/703) presented neonatal sepsis. The main sources of infection for the neonates were intrauterine (72.5%, 71/98), lactogenic (13.2%, 13/98), umbilical (5.1%, 5/98) and unidentified sources (9.2%, 9/98). The main non-specific clinical signs observed in the newborns were weakness, depression, impaired or absent reflexes, hypothermia, hypoglycemia, dehydration, reduced muscle tonus and diarrhea. The newborns also manifested clinical signs of severe infection, such as hyperemia in the abdominal and anal regions, omphalitis, hematuria, abdomen and extremities with purplish-blue coloration necrosing injuries in the pads, bradycardia, dyspnea, epistaxis, hypotension and evolution to septic shock. Infections acquired during intrauterine life led to the onset of the clinical signs at the time of birth, with fast evolution during the first hours of life. On the other hand, infections acquired via milk or umbilical cord presented clinical signs later. The total mortality rate was 5.4% (38/703) and the mortality rate among the neonates with sepsis was 38.7% (38/98). The early mortality rate (0 to 2 days) accounted for 86.9% (33/38) and the late mortality rate (3 to 30 days) for 13.1% (5/38) of the deaths among the newborns with sepsis. The main bacterial agents observed were Staphylococcus spp., Streptococcus spp., Proteus spp. Mannheimia spp. and Escherichia coli. Neonatal sepsis evolves quickly and may lead to high mortality in a litter. The prognosis is usually favorable if the diagnosis is reached early and the antibiotic therapy instituted as soon as possible, even before the results of blood cultures and antibiograms. The therapeutic recommendations should meet the special physiological conditions of a neonate in terms of metabolism and excretion of medication. Therefore, it is of utmost importance that the veterinarian is knowledgeable regarding neonatology to provide effective intervention and improve the survival rates of these patients.

Keywords: Neonatal infection , bacteria, puppies, newborn

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428 De Novo Assembly and Characterization of the Transcriptome from the Fluoroacetate Producing Plant, Dichapetalum Cymosum

Authors: Selisha A. Sooklal, Phelelani Mpangase, Shaun Aron, Karl Rumbold

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Organically bound fluorine (C-F bond) is extremely rare in nature. Despite this, the first fluorinated secondary metabolite, fluoroacetate, was isolated from the plant Dichapetalum cymosum (commonly known as Gifblaar). However, the enzyme responsible for fluorination (fluorinase) in Gifblaar was never isolated and very little progress has been achieved in understanding this process in higher plants. Fluorinated compounds have vast applications in the pharmaceutical, agrochemical and fine chemicals industries. Consequently, an enzyme capable of catalysing a C-F bond has great potential as a biocatalyst in the industry considering that the field of fluorination is virtually synthetic. As with any biocatalyst, a range of these enzymes are required. Therefore, it is imperative to expand the exploration for novel fluorinases. This study aimed to gain molecular insights into secondary metabolite biosynthesis in Gifblaar using a high-throughput sequencing-based approach. Mechanical wounding studies were performed using Gifblaar leaf tissue in order to induce expression of the fluorinase. The transcriptome of the wounded and unwounded plant was then sequenced on the Illumina HiSeq platform. A total of 26.4 million short sequence reads were assembled into 77 845 transcripts using Trinity. Overall, 68.6 % of transcripts were annotated with gene identities using public databases (SwissProt, TrEMBL, GO, COG, Pfam, EC) with an E-value threshold of 1E-05. Sequences exhibited the greatest homology to the model plant, Arabidopsis thaliana (27 %). A total of 244 annotated transcripts were found to be differentially expressed between the wounded and unwounded plant. In addition, secondary metabolic pathways present in Gifblaar were successfully reconstructed using Pathway tools. Due to lack of genetic information for plant fluorinases, a transcript failed to be annotated as a fluorinating enzyme. Thus, a local database containing the 5 existing bacterial fluorinases was created. Fifteen transcripts having homology to partial regions of existing fluorinases were found. In efforts to obtain the full coding sequence of the Gifblaar fluorinase, primers were designed targeting the regions of homology and genome walking will be performed to amplify the unknown regions. This is the first genetic data available for Gifblaar. It has provided novel insights into the mechanisms of metabolite biosynthesis and will allow for the discovery of the first eukaryotic fluorinase.

Keywords: biocatalyst, fluorinase, gifblaar, transcriptome

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427 Anti-Neuroinflammatory and Anti-Apoptotic Efficacy of Equol, against Lipopolysaccharide Activated Microglia and Its Neurotoxicity

Authors: Lalita Subedi, Jae Kyoung Chae, Yong Un Park, Cho Kyo Hee, Lee Jae Hyuk, Kang Min Cheol, Sun Yeou Kim

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Neuroinflammation may mediate the relationship between low levels of estrogens and neurodegenerative disease. Estrogens are neuroprotective and anti-inflammatory in neurodegenerative disease models. Due to the long term side effects of estrogens, researches have been focused on finding an effective phytoestrogens for biological activities. Daidzein present in soybeans and its active metabolite equol (7-hydroxy-3-(4'-hydroxyphenyl)-chroman) bears strong antioxidant and anticancer showed more potent anti-inflammatory and neuroprotective role in neuroinflammatory model confirmed its in vitro activity with molecular mechanism through NF-κB pathway. Three major CNS cells Microglia (BV-2), Astrocyte (C6), Neuron (N2a) were used to find the effect of equol in inducible nitric oxide synthase (iNOS), cyclooxygenase (COX-2), MAPKs signaling proteins, apoptosis related proteins by western blot analysis. Nitric oxide (NO) and prostaglandin E2 (PGE2) was measured by the Gries method and ELISA, respectively. Cytokines like tumor necrosis factor-α (TNF-α) and IL-6 were also measured in the conditioned medium of LPS activated cells with or without equol. Equol inhibited the NO production, PGE-2 production and expression of COX-2 and iNOS in LPS-stimulated microglial cells at a dose dependent without any cellular toxicity. At the same time Equol also showed promising effect in modulation of MAPK’s and nuclear factor kappa B (NF-κB) expression with significant inhibition of the production of proinflammatory cytokine like interleukin -6 (IL-6), and tumor necrosis factor -α (TNF-α). Additionally, it inhibited the LPS activated microglia-induced neuronal cell death by downregulating the apoptotic phenomenon in neuronal cells. Furthermore, equol increases the production of neurotrophins like NGF and increase the neurite outgrowth as well. In conclusion the natural daidzein metabolite equol are more active than daidzein, which showed a promising effectiveness as an anti-neuroinflammatory and neuroprotective agent via downregulating the LPS stimulated microglial activation and neuronal apoptosis. This work was supported by Brain Korea 21 Plus project and High Value-added Food Technology Development Program 114006-4, Ministry of Agriculture, Food and Rural Affairs.

Keywords: apoptosis, equol, neuroinflammation, phytoestrogen

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426 Neighborhood Sustainability Assessment Tools: A Conceptual Framework for Their Use in Building Adaptive Capacity to Climate Change

Authors: Sally Naji, Julie Gwilliam

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Climate change remains a challenging matter for the human and the built environment in the 21st century, where the need to consider adaptation to climate change in the development process is paramount. However, there remains a lack of information regarding how we should prepare responses to this issue, such as through developing organized and sophisticated tools enabling the adaptation process. This study aims to build a systematic framework approach to investigate the potentials that Neighborhood Sustainability Assessment tools (NSA) might offer in enabling both the analysis of the emerging adaptive capacity to climate change. The analysis of the framework presented in this paper aims to discuss this issue in three main phases. The first part attempts to link sustainability and climate change, in the context of adaptive capacity. It is argued that in deciding to promote sustainability in the context of climate change, both the resilience and vulnerability processes become central. However, there is still a gap in the current literature regarding how the sustainable development process can respond to climate change. As well as how the resilience of practical strategies might be evaluated. It is suggested that the integration of the sustainability assessment processes with both the resilience thinking process, and vulnerability might provide important components for addressing the adaptive capacity to climate change. A critical review of existing literature is presented illustrating the current lack of work in this field, integrating these three concepts in the context of addressing the adaptive capacity to climate change. The second part aims to identify the most appropriate scale at which to address the built environment for the climate change adaptation. It is suggested that the neighborhood scale can be considered as more suitable than either the building or urban scales. It then presents the example of NSAs, and discusses the need to explore their potential role in promoting the adaptive capacity to climate change. The third part of the framework presents a comparison among three example NSAs, BREEAM Communities, LEED-ND, and CASBEE-UD. These three tools have been selected as the most developed and comprehensive assessment tools that are currently available for the neighborhood scale. This study concludes that NSAs are likely to present the basis for an organized framework to address the practical process for analyzing and yet promoting Adaptive Capacity to Climate Change. It is further argued that vulnerability (exposure & sensitivity) and resilience (Interdependence & Recovery) form essential aspects to be addressed in the future assessment of NSA’s capability to adapt to both short and long term climate change impacts. Finally, it is acknowledged that further work is now required to understand impact assessment in terms of the range of physical sectors (Water, Energy, Transportation, Building, Land Use and Ecosystems), Actor and stakeholder engagement as well as a detailed evaluation of the NSA indicators, together with a barriers diagnosis process.

Keywords: adaptive capacity, climate change, NSA tools, resilience, sustainability

Procedia PDF Downloads 381
425 Isotope Effects on Inhibitors Binding to HIV Reverse Transcriptase

Authors: Agnieszka Krzemińska, Katarzyna Świderek, Vicente Molinier, Piotr Paneth

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In order to understand in details the interactions between ligands and the enzyme isotope effects were studied between clinically used drugs that bind in the active site of Human Immunodeficiency Virus Reverse Transcriptase, HIV-1 RT, as well as triazole-based inhibitor that binds in the allosteric pocket of this enzyme. The magnitudes and origins of the resulting binding isotope effects were analyzed. Subsequently, binding isotope effect of the same triazole-based inhibitor bound in the active site were analyzed and compared. Together, these results show differences in binding origins in two sites of the enzyme and allow to analyze binding mode and place of newly synthesized inhibitors. Typical protocol is described below on the example of triazole ligand in the allosteric pocket. Triazole was docked into allosteric cavity of HIV-1 RT with Glide using extra-precision mode as implemented in Schroedinger software. The structure of HIV-1 RT was obtained from Protein Data Bank as structure of PDB ID 2RKI. The pKa for titratable amino acids was calculated using PROPKA software, and in order to neutralize the system 15 Cl- were added using tLEaP package implemented in AMBERTools ver.1.5. Also N-terminals and C-terminals were build using tLEaP. The system was placed in 144x160x144Å3 orthorhombic box of water molecules using NAMD program. Missing parameters for triazole were obtained at the AM1 level using Antechamber software implemented in AMBERTools. The energy minimizations were carried out by means of a conjugate gradient algorithm using NAMD. Then system was heated from 0 to 300 K with temperature increment 0.001 K. Subsequently 2 ns Langevin−Verlet (NVT) MM MD simulation with AMBER force field implemented in NAMD was carried out. Periodic Boundary Conditions and cut-offs for the nonbonding interactions, range radius from 14.5 to 16 Å, are used. After 2 ns relaxation 200 ps of QM/MM MD at 300 K were simulated. The triazole was treated quantum mechanically at the AM1 level, protein was described using AMBER and water molecules were described using TIP3P, as implemented in fDynamo library. Molecules 20 Å apart from the triazole were kept frozen, with cut-offs established on range radius from 14.5 to 16 Å. In order to describe interactions between triazole and RT free energy of binding using Free Energy Perturbation method was done. The change in frequencies from ligand in solution to ligand bounded in enzyme was used to calculate binding isotope effects.

Keywords: binding isotope effects, molecular dynamics, HIV, reverse transcriptase

Procedia PDF Downloads 432
424 Selective Separation of Amino Acids by Reactive Extraction with Di-(2-Ethylhexyl) Phosphoric Acid

Authors: Alexandra C. Blaga, Dan Caşcaval, Alexandra Tucaliuc, Madalina Poştaru, Anca I. Galaction

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Amino acids are valuable chemical products used in in human foods, in animal feed additives and in the pharmaceutical field. Recently, there has been a noticeable rise of amino acids utilization throughout the world to include their use as raw materials in the production of various industrial chemicals: oil gelating agents (amino acid-based surfactants) to recover effluent oil in seas and rivers and poly(amino acids), which are attracting attention for biodegradable plastics manufacture. The amino acids can be obtained by biosynthesis or from protein hydrolysis, but their separation from the obtained mixtures can be challenging. In the last decades there has been a continuous interest in developing processes that will improve the selectivity and yield of downstream processing steps. The liquid-liquid extraction of amino acids (dissociated at any pH-value of the aqueous solutions) is possible only by using the reactive extraction technique, mainly with extractants of organophosphoric acid derivatives, high molecular weight amines and crown-ethers. The purpose of this study was to analyse the separation of nine amino acids of acidic character (l-aspartic acid, l-glutamic acid), basic character (l-histidine, l-lysine, l-arginine) and neutral character (l-glycine, l-tryptophan, l-cysteine, l-alanine) by reactive extraction with di-(2-ethylhexyl)phosphoric acid (D2EHPA) dissolved in butyl acetate. The results showed that the separation yield is controlled by the pH value of the aqueous phase: the reactive extraction of amino acids with D2EHPA is possible only if the amino acids exist in aqueous solution in their cationic forms (pH of aqueous phase below the isoeletric point). The studies for individual amino acids indicated the possibility of selectively separate different groups of amino acids with similar acidic properties as a function of aqueous solution pH-value: the maximum yields are reached for a pH domain of 2–3, then strongly decreasing with the pH increase. Thus, for acidic and neutral amino acids, the extraction becomes impossible at the isolelectric point (pHi) and for basic amino acids at a pH value lower than pHi, as a result of the carboxylic group dissociation. From the results obtained for the separation from the mixture of the nine amino acids, at different pH, it can be observed that all amino acids are extracted with different yields, for a pH domain of 1.5–3. Over this interval, the extract contains only the amino acids with neutral and basic character. For pH 5–6, only the neutral amino acids are extracted and for pH > 6 the extraction becomes impossible. Using this technique, the total separation of the following amino acids groups has been performed: neutral amino acids at pH 5–5.5, basic amino acids and l-cysteine at pH 4–4.5, l-histidine at pH 3–3.5 and acidic amino acids at pH 2–2.5.

Keywords: amino acids, di-(2-ethylhexyl) phosphoric acid, reactive extraction, selective extraction

Procedia PDF Downloads 431
423 Determination of the Knowledge Level of Healthcare Professional's Working at the Emergency Services in Turkey about Their Approaches to Common Forensic Cases

Authors: E. Tuğba Topçu, Ebru E. Kazan, Erhan Büken

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Emergency nurses are the first health care professional to generally observe the patients, communicate patients’ family or relatives, touch the properties of patients and contact to laboratory sample of patients. Also, they are the encounter incidents related crime, people who engage in violence or suspicious injuries frequently. So, documentation of patients’ condition came to the hospital and conservation of evidence are important in the inquiry of forensic medicine. The aim of the study was to determine the knowledge level of healthcare professional working at the emergency services regarding their approaches to common forensic cases. The study was comprised of 404 healthcare professional working (nurse, emergency medicine technician, health officer) at the emergency services of 6 state hospitals, 6 training and 6 research hospitals and 3 university hospitals in Ankara. Data was collected using questionnaire form which was developed by researches in the direction of literature. Questionnaire form is comprised of two sections. The first section includes 17 questions related demographic information about health care professional and 4 questions related Turkish laws. The second section includes 43 questions to the determination of knowledge level of health care professional’s working in the emergency department, about approaches to frequently encountered forensic cases. For the data evaluation of the study; Mann Whitney U test, Bonferroni correction Kruskal Wallis H test and Chi Square tests have been used. According to study, it’s said that there is no forensic medicine expert in the foundation by 73.4% of health care professionals. Two third (66%) of participants’ in emergency department reported daily average 7 or above forensic cases applied to the emergency department and 52.1% of participants did not evaluate incidents came to the emergency department as a forensic case. Most of the participants informed 'duty of preservation of evidence' is health care professionals duty related forensic cases. In result, we determinated that knowledge level of health care professional working in the emergency department, about approaches to frequently encountered forensic cases, is not the expected level. Because we found that most of them haven't received education about forensic nursing.Postgraduates participants, educated health professional about forensic nursing, staff who applied to sources about forensic nursing and staff who evaluated emergency department cases as forensic cases have significantly higher level of knowledge. Moreover, it’s found that forensic cases diagnosis score is the highest in health officer and university graduated. Health care professional’s deficiency in knowledge about forensic cases can cause defects in operation of the forensic process because of mistakes in collecting and conserving of evidence. It is obvious that training about the approach to forensic nursing should be arranged.

Keywords: emergency nurses, forensic case, forensic nursing, level of knowledge

Procedia PDF Downloads 294
422 Clinical Validation of C-PDR Methodology for Accurate Non-Invasive Detection of Helicobacter pylori Infection

Authors: Suman Som, Abhijit Maity, Sunil B. Daschakraborty, Sujit Chaudhuri, Manik Pradhan

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Background: Helicobacter pylori is a common and important human pathogen and the primary cause of peptic ulcer disease and gastric cancer. Currently H. pylori infection is detected by both invasive and non-invasive way but the diagnostic accuracy is not up to the mark. Aim: To set up an optimal diagnostic cut-off value of 13C-Urea Breath Test to detect H. pylori infection and evaluate a novel c-PDR methodology to overcome of inconclusive grey zone. Materials and Methods: All 83 subjects first underwent upper-gastrointestinal endoscopy followed by rapid urease test and histopathology and depending on these results; we classified 49 subjects as H. pylori positive and 34 negative. After an overnight, fast patients are taken 4 gm of citric acid in 200 ml water solution and 10 minute after ingestion of the test meal, a baseline exhaled breath sample was collected. Thereafter an oral dose of 75 mg 13C-Urea dissolved in 50 ml water was given and breath samples were collected upto 90 minute for 15 minute intervals and analysed by laser based high precisional cavity enhanced spectroscopy. Results: We studied the excretion kinetics of 13C isotope enrichment (expressed as δDOB13C ‰) of exhaled breath samples and found maximum enrichment around 30 minute of H. pylori positive patients, it is due to the acid mediated stimulated urease enzyme activity and maximum acidification happened within 30 minute but no such significant isotopic enrichment observed for H. pylori negative individuals. Using Receiver Operating Characteristic (ROC) curve an optimal diagnostic cut-off value, δDOB13C ‰ = 3.14 was determined at 30 minute exhibiting 89.16% accuracy. Now to overcome grey zone problem we explore percentage dose of 13C recovered per hour, i.e. 13C-PDR (%/hr) and cumulative percentage dose of 13C recovered, i.e. c-PDR (%) in exhaled breath samples for the present 13C-UBT. We further explored the diagnostic accuracy of 13C-UBT by constructing ROC curve using c-PDR (%) values and an optimal cut-off value was estimated to be c-PDR = 1.47 (%) at 60 minute, exhibiting 100 % diagnostic sensitivity , 100 % specificity and 100 % accuracy of 13C-UBT for detection of H. pylori infection. We also elucidate the gastric emptying process of present 13C-UBT for H. pylori positive patients. The maximal emptying rate found at 36 minute and half empting time of present 13C-UBT was found at 45 minute. Conclusions: The present study exhibiting the importance of c-PDR methodology to overcome of grey zone problem in 13C-UBT for accurate determination of infection without any risk of diagnostic errors and making it sufficiently robust and novel method for an accurate and fast non-invasive diagnosis of H. pylori infection for large scale screening purposes.

Keywords: 13C-Urea breath test, c-PDR methodology, grey zone, Helicobacter pylori

Procedia PDF Downloads 301
421 Enhancement Effect of Superparamagnetic Iron Oxide Nanoparticle-Based MRI Contrast Agent at Different Concentrations and Magnetic Field Strengths

Authors: Bimali Sanjeevani Weerakoon, Toshiaki Osuga, Takehisa Konishi

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Magnetic Resonance Imaging Contrast Agents (MRI-CM) are significant in the clinical and biological imaging as they have the ability to alter the normal tissue contrast, thereby affecting the signal intensity to enhance the visibility and detectability of images. Superparamagnetic Iron Oxide (SPIO) nanoparticles, coated with dextran or carboxydextran are currently available for clinical MR imaging of the liver. Most SPIO contrast agents are T2 shortening agents and Resovist (Ferucarbotran) is one of a clinically tested, organ-specific, SPIO agent which has a low molecular carboxydextran coating. The enhancement effect of Resovist depends on its relaxivity which in turn depends on factors like magnetic field strength, concentrations, nanoparticle properties, pH and temperature. Therefore, this study was conducted to investigate the impact of field strength and different contrast concentrations on enhancement effects of Resovist. The study explored the MRI signal intensity of Resovist in the physiological range of plasma from T2-weighted spin echo sequence at three magnetic field strengths: 0.47 T (r1=15, r2=101), 1.5 T (r1=7.4, r2=95), and 3 T (r1=3.3, r2=160) and the range of contrast concentrations by a mathematical simulation. Relaxivities of r1 and r2 (L mmol-1 Sec-1) were obtained from a previous study and the selected concentrations were 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 2.0, and 3.0 mmol/L. T2-weighted images were simulated using TR/TE ratio as 2000 ms /100 ms. According to the reference literature, with increasing magnetic field strengths, the r1 relaxivity tends to decrease while the r2 did not show any systematic relationship with the selected field strengths. In parallel, this study results revealed that the signal intensity of Resovist at lower concentrations tends to increase than the higher concentrations. The highest reported signal intensity was observed in the low field strength of 0.47 T. The maximum signal intensities for 0.47 T, 1.5 T and 3 T were found at the concentration levels of 0.05, 0.06 and 0.05 mmol/L, respectively. Furthermore, it was revealed that, the concentrations higher than the above, the signal intensity was decreased exponentially. An inverse relationship can be found between the field strength and T2 relaxation time, whereas, the field strength was increased, T2 relaxation time was decreased accordingly. However, resulted T2 relaxation time was not significantly different between 0.47 T and 1.5 T in this study. Moreover, a linear correlation of transverse relaxation rates (1/T2, s–1) with the concentrations of Resovist can be observed. According to these results, it can conclude that the concentration of SPIO nanoparticle contrast agents and the field strengths of MRI are two important parameters which can affect the signal intensity of T2-weighted SE sequence. Therefore, when MR imaging those two parameters should be considered prudently.

Keywords: Concentration, resovist, field strength, relaxivity, signal intensity

Procedia PDF Downloads 352
420 Nano-Immunoassay for Diagnosis of Active Schistosomal Infection

Authors: Manal M. Kame, Hanan G. El-Baz, Zeinab A.Demerdash, Engy M. Abd El-Moneem, Mohamed A. Hendawy, Ibrahim R. Bayoumi

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There is a constant need to improve the performance of current diagnostic assays of schistosomiasis as well as develop innovative testing strategies to meet new testing challenges. This study aims at increasing the diagnostic efficiency of monoclonal antibody (MAb)-based antigen detection assays through gold nanoparticles conjugated with specific anti-Schistosoma mansoni monoclonal antibodies. In this study, several hybidoma cell lines secreting MAbs against adult worm tegumental Schistosoma antigen (AWTA) were produced at Immunology Department of Theodor Bilharz Research Institute and preserved in liquid nitrogen. One MAb (6D/6F) was chosen for this study due to its high reactivity to schistosome antigens with highest optical density (OD) values. Gold nanoparticles (AuNPs) were functionalized and conjugated with MAb (6D/6F). The study was conducted on serum samples of 116 subjects: 71 patients with S. mansoni eggs in their stool samples group (gp 1), 25 with other parasites (gp2) and 20 negative healthy controls (gp3). Patients in gp1 were further subdivided according to egg count in their stool samples into Light infection {≤ 50 egg per gram(epg) (n= 17)}, moderate {51-100 epg (n= 33)} and severe infection {>100 epg(n= 21)}. Sandwich ELISA was performed using (AuNPs -MAb) for detection of circulating schistosomal antigen (CSA) levels in serum samples of all groups and the results were compared with that after using MAb/ sandwich ELISA system. Results Gold- MAb/ ELISA system reached a lower detection limit of 10 ng/ml compared to 85 ng/ml on using MAb/ ELISA and the optimal concentrations of AuNPs -MAb were found to be 12 folds less than that of MAb/ ELISA system for detection of CSA. The sensitivity and specificity of sandwich ELISA for detection of CSA levels using AuNPs -MAb were 100% & 97.8 % respectively compared to 87.3% &93.38% respectively on using MAb/ ELISA system. It was found that CSA was detected in 9 out of 71 S.mansoni infected patients on using AuNPs - MAb/ ELISA system and was not detected by MAb/ ELISA system. All those patients (9) was found to have an egg count below 50 epg feces (patients with light infections). ROC curve analyses revealed that sandwich ELISA using gold-MAb was an excellent diagnostic investigator that could differentiate Schistosoma patients from healthy controls, on the other hand it revealed that sandwich ELISA using MAb was not accurate enough as it could not recognize nine out of 71 patients with light infections. Conclusion Our data demonstrated that: Loading gold nanoparticles with MAb (6D/6F) increases the sensitivity and specificity of sandwich ELISA for detection of CSA, thus active (early) and light infections could be easily detected. Moreover this binding will decrease the amount of MAb consumed in the assay and lower the coast. The significant positive correlation that was detected between ova count (intensity of infection) and OD reading in sandwich ELISA using gold- MAb enables its use to detect the severity of infections and follow up patients after treatment for monitoring of cure.

Keywords: Schistosomiasis, nanoparticles, gold, monoclonal antibodies, ELISA

Procedia PDF Downloads 371
419 Biocontrol Potential of Trichoderma longibrachiatum as an Entomopathogenic Fungi against Bemisia tabaci

Authors: Waheed Anwar, Kiran Nawaz, Muhammad Saleem Haider, Ahmad Ali Shahid, Sehrish Iftikhar

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The whitefly, Bemisia tabaci (Gennadius), is a complex insect species, including many cryptic species or biotypes. Whitefly causes damage to many ornamental and horticultural crops through directly feeding on phloem sap, resulting in sooty mould and critically decreases the rate of photosynthesis of many host plants. Biological control has emerged as one of the most important methods for the management of soil-borne plant pathogens. Among the natural enemies of insects different entomopathogenic fungi are mostly used as biological control of the pest. The purpose of this research was to find indigenous insect-associated fungi and their virulence against Bemisia tabaci. A detailed survey of cotton fields in sample collection was conducted during July and August 2013 from the central mixed zone of Punjab, Pakistan. For the isolation of T. longibrachiatum, sabouraud dextrose peptone yeast extract agar (SDAY) media was used and morphological characterization of isolated T. longibrachiatum was studied using different dichotomous keys. Molecular Identification of the pathogen was confirmed by amplifying the internal transcribed spacer region. Blastn analysis showed 100% homology with already reported sequences on the database. For these bioassays, two conidial concentrations 4 × 108/mL & 4 × 104/mL of T. longibrachiatum was sprayed in clip cages for nymph and adult B. tabaci respectively under controlled environmental conditions. The pathogenicity of T. longibrachiatum was tested on nymph and adult whitefly to check mortality. Mortality of B. tabaci at nymphal and adult stages were observed after 24-hour intervals. Percentage mortality of nymphs treated with 4 x 104/mL conidia of T. longibrachiatum was 20, 24, 36 and 40% after 48, 72, 96, 72, 96, 120 and 144 hours respectively. However, no considerable difference was recorded in percentage mortality of whitefly after 120 and 144 hours. There were great variations after 24, 48, 72 and 96 hours in the rate of mortality. The efficacy of T. longibrachiatum as entomopathogenic fungi was evaluated in adult and nymphal stages of whitefly. Trichoderma longibrachiatum showed maximum activity on nymphal stages of whitefly as compared to adult stages. The percentage of conidial germination was also recorded on the outer surface of adult and nymphal stages of B. tabaci. The present findings indicated that T. longibrachiatum is an entomopathogenic fungus against B. tabaci and many species of Trichoderma were already reported as an antagonistc organism against a wide range of bacterial and fungal pathogens.

Keywords: efficacy, Trichoderma, virulence, bioassay

Procedia PDF Downloads 287
418 The Valuable Triad of Adipokine Indices to Differentiate Pediatric Obesity from Metabolic Syndrome: Chemerin, Progranulin, Vaspin

Authors: Mustafa M. Donma, Orkide Donma

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Obesity is associated with cardiovascular disease risk factors and metabolic syndrome (MetS). In this study, associations between adipokines and adipokine as well as obesity indices were evaluated. Plasma adipokine levels may exhibit variations according to body adipose tissue mass. Besides, upon consideration of obesity as an inflammatory disease, adipokines may play some roles in this process. The ratios of proinflammatory adipokines to adiponectin may act as highly sensitive indicators of body adipokine status. The aim of the study is to present some adipokine indices, which are thought to be helpful for the evaluation of childhood obesity and also to determine the best discriminators in the diagnosis of MetS. 80 prepubertal children (aged between 6-9.5 years) included in the study were divided into three groups; 30 children with normal weight (NW), 25 morbid obese (MO) children and 25 MO children with MetS. Physical examinations were performed. Written informed consent forms were obtained from the parents. The study protocol was approved by Ethics Committee of Namik Kemal University Medical Faculty. Anthropometric measurements, such as weight, height, waist circumference (C), hip C, head C, neck C were recorded. Values for body mass index (BMI), diagnostic obesity notation model assessment Index-II (D2 index) as well as waist-to-hip, head-to-neck ratios were calculated. Adiponectin, resistin, leptin, chemerin, vaspin, progranulin assays were performed by ELISA. Adipokine-to-adiponectin ratios were obtained. SPSS Version 20 was used for the evaluation of data. p values ≤ 0.05 were accepted as statistically significant. Values of BMI and D2 index, waist-to-hip, head-to-neck ratios did not differ between MO and MetS groups (p ≥ 0.05). Except progranulin (p ≤ 0.01), similar patterns were observed for plasma levels of each adipokine. There was not any difference in vaspin as well as resistin levels between NW and MO groups. Significantly increased leptin-to-adiponectin, chemerin-to-adiponectin and vaspin-to-adiponectin values were noted in MO in comparison with those of NW. The most valuable adipokine index was progranulin-to-adiponectin (p ≤ 0.01). This index was strongly correlated with vaspin-to-adiponectin ratio in all groups (p ≤ 0.05). There was no correlation between vaspin-to-adiponectin and chemerin-to--adiponectin in NW group. However, a correlation existed in MO group (r = 0.486; p ≤ 0.05). Much stronger correlation (r = 0.609; p ≤ 0.01) was observed in MetS group between these two adipokine indices. No correlations were detected between vaspin and progranulin as well as vaspin and chemerin levels. Correlation analyses showed a unique profile confined to MetS children. Adiponectin was found to be correlated with waist-to-hip (r = -0.435; p ≤ 0.05) as well as head-to-neck (r = 0.541; p ≤ 0.05) ratios only in MetS children. In this study, it has been investigated if adipokine indices have priority over adipokine levels. In conclusion, vaspin-to-adiponectin, progranulin-to-adiponectin, chemerin-to-adiponectin along with waist-to-hip and head-to-neck ratios were the optimal combinations. Adiponectin, waist-to-hip, head-to-neck, vaspin-to-adiponectin, chemerin-to-adiponectin ratios had appropriate discriminatory capability for MetS children.

Keywords: adipokine indices, metabolic syndrome, obesity indices, pediatric obesity

Procedia PDF Downloads 205
417 Blunt Abdominal Trauma Management in Adult Patients: An Investigation on Safety of Discharging Patients with Normal Initial Findings

Authors: Rahimi-Movaghar Vafa, Mansouri Pejman, Chardoli Mojtaba, Rezvani Samina

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Introduction: Blunt abdominal trauma is one of the leading causes of morbidity and mortality in all age groups, but diagnosis of serious intra-abdominal pathology is difficult and most of the damages are obscure in the initial investigation. There is still controversy about which patients should undergo abdomen/pelvis CT, which patients needs more observation and which patients can be discharged safely The aim of this study was to determine that is it safe to discharge patients with blunt abdominal trauma with normal initial findings. Methods: This non-randomized cross-sectional study was conducted from September 2013 to September 2014 at two levels I trauma centers, Sina hospital and Rasoul-e-Akram hospital (Tehran, Iran). Our inclusion criteria were all patients were admitted for suspicious BAT and our exclusion criteria were patients that have serious head and neck, chest, spine and limb injuries which need surgical intervention, those who have unstable vital signs, pregnant women with a gestational age over 3 months and homeless or without exact home address. 390 patients with blunt trauma abdomen examined and the necessary data, including demographic data, the abdominal examination, FAST result, patients’ lab test results (hematocrit, base deficit, urine analysis) on admission and at 6 and 12 hours after admission were recorded. Patients with normal physical examination, laboratory tests and FAST were discharged from the ED during 12 hours with the explanation of the alarm signs and were followed up after 24 hours and 1 week by a telephone call. Patients with abnormal findings in physical examination, laboratory tests, and FAST underwent abdomino-pelvic CT scan. Results: The study included 390 patients with blunt abdominal trauma between 12 and 80 years of age (mean age, 37.0 ± 13.7 years) and the mean duration of hospitalization in patients was 7.4 ± 4.1 hours. 88.6% of the patients were discharged from hospital before 12 hours. Odds ratio (OR) for having any symptoms for discharge after 6 hours was 0.160 and after 12 hours was 0.117 hours, which is statistically significant. Among the variables age, systolic and diastolic blood pressure, heart rate, respiratory rate, hematocrit and base deficit at admission, 6 hours and 12 hours after admission showed no significant statistical relationship with discharge time. From our 390 patients, 190 patients have normal initial physical examination, lab data and FAST findings that didn’t show any signs or symptoms in their next assessment and in their follow up by the phone call. Conclusion: It is recommended that patients with no symptoms at admission (completely normal physical examination, ultrasound, normal hematocrit and normal base deficit and lack of microscopic hematuria) and good family and social status can be safely discharged from the emergency department.

Keywords: blunt abdominal trauma, patient discharge, emergency department, FAST

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416 The Effects of a Hippotherapy Simulator in Children with Cerebral Palsy: A Pilot Study

Authors: Canan Gunay Yazici, Zubeyir Sarı, Devrim Tarakci

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Background: Hippotherapy considered as global techniques used in rehabilitation of children with cerebral palsy as it improved gait pattern, balance, postural control, balance and gross motor skills development but it encounters some problems (such as the excess of the cost of horses' care, nutrition, housing). Hippotherapy simulator is being developed in recent years to overcome these problems. These devices aim to create the effects of hippotherapy made with a real horse on patients by simulating the movements of a real horse. Objectives: To evaluate the efficacy of hippotherapy simulator on gross motor functions, sitting postural control and dynamic balance of children with cerebral palsy (CP). Methods: Fourteen children with CP, aged 6–15 years, seven with a diagnosis of spastic hemiplegia, five of diplegia, two of triplegia, Gross Motor Function Classification System level I-III. The Horse Riding Simulator (HRS), including four-speed program (warm-up, level 1-2-3), was used for hippotherapy simulator. Firstly, each child received Neurodevelopmental Therapy (NDT; 45min twice weekly eight weeks). Subsequently, the same children completed HRS+NDT (30min and 15min respectively, twice weekly eight weeks). Children were assessed pre-treatment, at the end of 8th and 16th week. Gross motor function, sitting postural control, dynamic sitting and standing balance were evaluated by Gross Motor Function Measure-88 (GMFM-88, Dimension B, D, E and Total Score), Trunk Impairment Scale (TIS), Pedalo® Sensamove Balance Test and Pediatric Balance Scale (PBS) respectively. Unit of Scientific Research Project of Marmara University supported our study. Results: All measured variables were a significant increase compared to baseline values after both intervention (NDT and HRS+NDT), except for dynamic sitting balance evaluated by Pedalo®. Especially HRS+NDT, increase in the measured variables was considerably higher than NDT. After NDT, the Total scores of GMFM-88 (mean baseline 62,2 ± 23,5; mean NDT: 66,6 ± 22,2; p < 0,05), TIS (10,4 ± 3,4; 12,1 ± 3; p < 0,05), PBS (37,4 ± 14,6; 39,6 ± 12,9; p < 0,05), Pedalo® sitting (91,2 ± 6,7; 92,3 ± 5,2; p > 0,05) and Pedalo® standing balance points (80,2 ± 10,8; 82,5 ± 11,5; p < 0,05) increased by 7,1%, 2%, 3,9%, 5,2% and 6 % respectively. After HRS+NDT treatment, the total scores of GMFM-88 (mean baseline: 62,2 ± 23,5; mean HRS+NDT: 71,6 ± 21,4; p < 0,05), TIS (10,4 ± 3,4; 15,6 ± 2,9; p < 0,05), PBS (37,4 ± 14,6; 42,5 ± 12; p < 0,05), Pedalo® sitting (91,2 ± 6,7; 93,8 ± 3,7; p > 0,05) and standing balance points (80,2 ± 10,8; 86,2 ± 5,6; p < 0,05) increased by 15,2%, 6%, 7,3%, 6,4%, and 11,9%, respectively, compared to the initial values. Conclusion: Neurodevelopmental therapy provided significant improvements in gross motor functions, sitting postural control, sitting and standing balance of children with CP. When the hippotherapy simulator added to the treatment program, it was observed that these functions were further developed (especially with gross motor functions and dynamic balance). As a result, this pilot study showed that the hippotherapy simulator could be a useful alternative to neurodevelopmental therapy for the improvement of gross motor function, sitting postural control and dynamic balance of children with CP.

Keywords: balance, cerebral palsy, hippotherapy, rehabilitation

Procedia PDF Downloads 143
415 Structural Investigation of the GAF Domain Protein BPSL2418 from Burkholderia pseudomallei

Authors: Mona G. Alharbi

Abstract:

A new family of methionine-sulfoxide reductase (Msr) was recently discovered and was named free methionine sulfoxide reductase (fRMsr). This family includes enzymes with a reductase activity toward the free R isomer of a methionine sulfoxide substrate. The fRMsrs have a GAF domain topology, a domain, which was previously identified as having in some cases a cyclic nucleotide phosphodiesterase activity. The classification of fRMsrs as GAF domains revealed a new function can be added to the GAF domain family. Interestingly the four members identified in the fRMsr family share the GAF domain structure and the presence of three conserved cysteines in the active site with free R methionine sulfoxide substrate specificity. This thesis presents the crystal structures of reduced, free Met-SO substrate-bound and MES-bound forms of a new fRMsr from Burkholderia pseudomallei (BPSL2418). BPSL2418 was cloned, overexpressed and purified to enable protein crystallization. The crystallization trials for reduced, Met-SO-bound and MES-bound forms of BPSL2418 were prepared and reasonable crystals of each form were produced. The crystal structures of BPSL2418MES, BPSL2418Met-SO and BPSL2418Reduced were solved at 1.18, 1.4 and 2.0Å, respectively by molecular replacement. The BPSL2418MES crystal belongs to space group P 21 21 21 while BPSL2418Met-SO and BPSL2418Reduced crystals belong to space group P 1 21 1. All three forms share the GAF domain structure of six antiparallel β-strands and four α-helices with connecting loops. The antiparallel β-strands (β1, β2, β5 and β6) are located in the center of the BPSL2418 structure flanked on one side by a three α-helices (α1, α2 and α4) and on the other side by a (loop1, β3, loop2, α3, β4 loop4) unit where loop4 forms a capping flap and covers the active site. The structural comparison of the three forms of BPSL2418 indicates that the catalytically important cysteine is CYS109, where the resolving cysteine is CYS75, which forms a disulfide bond with CYS109. They also suggest that the third conserved cysteine in the active site, CYS85, which is located in α3, is a non-essential cysteine for the catalytic function but it may play a role in the binding of the substrate. The structural comparison of the three forms reveals that conformational changes appear in the active site particularly involving loop4 and CYS109 during catalysis. The 3D structure of BPSL2418 shows strong structure similarity to fRMsrs enzymes, which further suggests that BPSL2418 acts as a free Met-R-SO reductase and shares the catalytic mechanism of fRMsr family.

Keywords: Burkholderia pseudomallei, GAF domain protein, methionine sulfoxide reductase, protein crystallization

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414 Deep Convolutional Neural Network for Detection of Microaneurysms in Retinal Fundus Images at Early Stage

Authors: Goutam Kumar Ghorai, Sandip Sadhukhan, Arpita Sarkar, Debprasad Sinha, G. Sarkar, Ashis K. Dhara

Abstract:

Diabetes mellitus is one of the most common chronic diseases in all countries and continues to increase in numbers significantly. Diabetic retinopathy (DR) is damage to the retina that occurs with long-term diabetes. DR is a major cause of blindness in the Indian population. Therefore, its early diagnosis is of utmost importance towards preventing progression towards imminent irreversible loss of vision, particularly in the huge population across rural India. The barriers to eye examination of all diabetic patients are socioeconomic factors, lack of referrals, poor access to the healthcare system, lack of knowledge, insufficient number of ophthalmologists, and lack of networking between physicians, diabetologists and ophthalmologists. A few diabetic patients often visit a healthcare facility for their general checkup, but their eye condition remains largely undetected until the patient is symptomatic. This work aims to focus on the design and development of a fully automated intelligent decision system for screening retinal fundus images towards detection of the pathophysiology caused by microaneurysm in the early stage of the diseases. Automated detection of microaneurysm is a challenging problem due to the variation in color and the variation introduced by the field of view, inhomogeneous illumination, and pathological abnormalities. We have developed aconvolutional neural network for efficient detection of microaneurysm. A loss function is also developed to handle severe class imbalance due to very small size of microaneurysms compared to background. The network is able to locate the salient region containing microaneurysms in case of noisy images captured by non-mydriatic cameras. The ground truth of microaneurysms is created by expert ophthalmologists for MESSIDOR database as well as private database, collected from Indian patients. The network is trained from scratch using the fundus images of MESSIDOR database. The proposed method is evaluated on DIARETDB1 and the private database. The method is successful in detection of microaneurysms for dilated and non-dilated types of fundus images acquired from different medical centres. The proposed algorithm could be used for development of AI based affordable and accessible system, to provide service at grass root-level primary healthcare units spread across the country to cater to the need of the rural people unaware of the severe impact of DR.

Keywords: retinal fundus image, deep convolutional neural network, early detection of microaneurysms, screening of diabetic retinopathy

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413 Combining Bio-Molecular and Isotopic Tools to Determine the Fate of Halogenated Compounds in Polluted Groundwater

Authors: N. Balaban, A. Buernstein, F. Gelman, Z. Ronen

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Brominated flame retardants are widespread pollutants, and are known to be toxic, carcinogenic, endocrinic disrupting as well as recalcitrant. The industrial complex Neot Hovav, in the Northern Negev, Israel, is situated above a fractured chalk aquitard, which is polluted by a wide variety of halogenated organic compounds. Two of the abundant pollutants found in the site are Dibromoneopentyl-glycol (DBNPG) and tribromoneopentyl-alcohol (TBNPA). Due to the elusive nature of the groundwater flow, it is difficult to connect between the spatial changes in contaminant concentrations to degradation. In this study, we attempt to determine whether these compounds are biodegraded in the groundwater, and to gain a better understanding concerning the bacterial community in the groundwater. This was achieved through the application of compound-specific isotope analysis (CSIA) of carbon (13^C/12^C) and bromine (81^Br/79^Br), and new-generation MiSeq pyrosequencing. The sampled boreholes were distributed among three main areas of the industrial complex: around the production plant of TBNPA and DBNPG; along the Hovav Wadi (small ephemeral stream) which crosses and drains the industrial complex; and downstream to the industrial area. TBNPA and DBNPG are found in all three areas, with no clear connection to the proximity of the borehole to the production plant. Initial isotopic data of TBNPA from boreholes in the area surrounding the production plant, reveal no changes in the carbon and bromine isotopic values. When observing the microbial groundwater community, the dominant phylum is Proteobacteria. Known anaerobic dehalogenating bacteria such as Dehalococcoides from the Chloroflexi phylum have also been detected. A statistical comparison of the groundwater microbial diversity using a multi-variant ordination of non-metric multidimensional scaling (NMDS) reveals three main clusters in accordance to spatial location in the industrial complex: all the boreholes sampled adjacent to the production plant cluster together and separately from the Wadi Hovav boreholes cluster and the downstream to the industrial area borehole cluster. This work provides the basis for the development and implication of an isotopic fractionation based tool for assessing the biodegradation of brominated organic compounds in contaminated environments, and a novel attempt to characterize the spatial microbial diversity in the contaminated site.

Keywords: biodegradation, brominated flame retardants, groundwater, isotopic fractionation, microbial diversity

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412 Forensic Nursing in the Emergency Department: The Overlooked Roles

Authors: E. Tugba Topcu

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The emergency services are usually the first places to encounter forensic cases. Hence, it is important to consider forensics from the perspective of the emergency services staff and the physiological and psychological consequences that may arise as a result of behaviour by itself or another person. Accurate and detailed documentation of the situation in which the patient first arrives at the emergency service and preservation of the forensic findings is pivotal for the subsequent forensic investigation. The first step in determining whether or not a forensic case exists is to perform a medical examination of the patient. For each individual suspected to be part of a forensic case, police officers should be informed at the same time as the medical examination is being conducted. Violent events are increasing every year and with an increase in the number of forensic cases, emergency service workers have increasing responsibility and consequently play a key role in protecting, collecting and arranging the forensic evidence. In addition, because the emergency service workers involved in forensic events typically have information about the accused and/or victim, as well as evidence related to the events and the cause of injuries, police officers often require their testimony. However, both nurses and other health care personnel do not typically have adequate expertise in forensic medicine. Emergency nurses should take an active role for determining that whether any patient admitted to the emergency services is a clinical forensic patient the emergency service with injury and requiring possible punishment and knowing of their roles and responsibilities in this area provides legal protection as well as the protection of the judicial affair. Particularly, in emergency services, where rapid patient turnover and high workload exists, patient registration and case reporting may not exist. In such instances, the witnesses, typically the nurses, are often consulted for information. Knowledge of forensic medical matters plays a vital role in achieving justice. According to the Criminal Procedure Law, Article 75, Paragraph 3, ‘an internal body examination or the taking of blood or other biological samples from the body can be performed only by a doctor or other health professional member’. In favour of this item, the clinic nurse and doctor are mainly responsible for evaluating forensic cases in emergency departments, performing the examination, collecting evidence, and storing and reporting data. The courts place considerable importance on determining whether a suspect is the victim or accused and, thus, in terms of illuminating events, it is crucial that any evidence is gathered carefully and appropriately. All the evidence related to the forensic case including the forensic report should be handed over to the police officers. In instances where forensic evidence cannot be collected and the only way to obtain the evidence is the hospital environment, health care personnel in emergency services need to have knowledge about the diagnosis of forensic evidence, the collection of evidence, hiding evidence and provision of the evidence delivery chain.

Keywords: emergency department, emergency nursing, forensic cases, forensic nursing

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411 Serological Evidence of Enzootic Bovine Leukosis in Dairy Cattle Herds in the United Arab Emirates

Authors: Nabeeha Hassan Abdel Jalil, Lulwa Saeed Al Badi, Mouza Ghafan Alkhyeli, Khaja Mohteshamuddin, Ahmad Al Aiyan, Mohamed Elfatih Hamad, Robert Barigye

Abstract:

The present study was done to elucidate the prevalence of enzootic bovine leucosis (EBL) in the UAE, the seroprevalence rates of EBL in dairy herds from the Al Ain area, Abu Dhabi (AD) and indigenous cattle at the Al Ain livestock market (AALM) were assessed. Of the 949 sera tested by ELISA, 657 were from adult Holstein-Friesians from three farms and 292 from indigenous cattle at the AALM. The level of significance between the proportions of seropositive cattle were analyzed by the Marascuilo procedure and questionnaire data on husbandry and biosecurity practices evaluated. Overall, the aggregated farm and AALM data demonstrated a seroprevalence of 25.9%, compared to 37.0% for the study farms, and 1.0% for the indigenous cattle. Additionally, the seroprevalence rates at farms #1, #2 and #3 were 54.7%, 0.0%, and 26.3% respectively. Except for farm #2 and the AALM, statistically significant differences were noted between the proportions of seropositive cattle for farms #1 and #2 (Critical Range or CR=0.0803), farms #1 and #3 (p=0.1069), and farms #2 and #3 (CR=0.0707), farm #1 and the AALM (CR=0.0819), and farm #3 and the AALM (CR=0.0726). Also, the proportions of seropositive animals on farm #1 were 9.8%, 59.8%, 29.3%, and 1.2% in the 12-36, 37-72, 73-108, and 109-144-mo-old age groups respectively compared to 21.5%, 60.8%, 15.2%, and 2.5% in the respective age groups for farm #2. On both farms and the AALM, the 37-72-mo-old age group showed the highest EBL seroprevalence rate while all the 57 cattle on farm #2 were seronegative. Additionally, farms #1 and #3 had 3,130 and 2,828 intensively managed Holstein-Friesian cattle respectively, and all animals were routinely immunized against several diseases except EBL. On both farms #1 and #3, artificial breeding was practiced using semen sourced from the USA, and USA and Canada respectively, all farms routinely quarantined new stock, and farm #1 previously imported dairy cattle from an unspecified country, and farm #3 from the Netherlands, Australia and South Africa. While farm #1 provided no information on animal nutrition, farm #3 cited using hay, concentrates, and ad lib water. To the authors’ best knowledge, this is the first serological evidence of EBL in the UAE and as previously reported, the seroprevalence rates are comparatively higher in the intensively managed dairy herds than in indigenous cattle. As two of the study farms previously sourced cattle and semen from overseas, biosecurity protocols need to be revisited to avoid inadvertent EBL incursion and the possibility of regional transboundary disease spread also needs to be assessed. After the proposed molecular studies have adduced additional data, the relevant UAE animal health authorities may need to develop evidence-based EBL control policies and programs.

Keywords: cattle, enzootic bovine leukosis, seroprevalence, UAE

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410 Estimating the Efficiency of a Meta-Cognitive Intervention Program to Reduce the Risk Factors of Teenage Drivers with Attention Deficit Hyperactivity Disorder While Driving

Authors: Navah Z. Ratzon, Talia Glick, Iris Manor

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Attention Deficit Hyperactivity Disorder (ADHD) is a chronic disorder that affects the sufferer’s functioning throughout life and in various spheres of activity, including driving. Difficulties in cognitive functioning and executive functions are often part and parcel of the ADHD diagnosis, and thus form a risk factor in driving. Studies examining the effectiveness of intervention programs for improving and rehabilitating driving in typical teenagers have been conducted in relatively small numbers; while studies on similar programs for teenagers with ADHD have been especially scarce. The aim of the present study has been to examine the effectiveness of a metacognitive occupational therapy intervention program for reducing risk factors in driving among teenagers with ADHD. The present study included 37 teenagers aged 17 to 19. They included 23 teenagers with ADHD divided into experimental (11) and control (12) groups; as well as 14 non-ADHD teenagers forming a second control group. All teenagers taking part in the study were examined in the Tel Aviv University driving lab, and underwent cognitive diagnoses and a driving simulator test. Every subject in the intervention group took part in 3 assessment meetings, and two metacognitive treatment meetings. The control groups took part in two assessment meetings with a follow-up meeting 3 months later. In all the study’s groups, the treatment’s effectiveness was tested by comparing monitoring results on the driving simulator at the first and second evaluations. In addition, the driving of 5 subjects from the intervention group was monitored continuously from a month prior to the start of the intervention, a month during the phase of the intervention and another month until the end of the intervention. In the ADHD control group, the driving of 4 subjects was monitored from the end of the first evaluation for a period of 3 months. The study’s findings were affected by the fact that the ADHD control group was different from the two other groups, and exhibited ADHD characteristics manifested by impaired executive functions and lower metacognitive abilities relative to their peers. The study found partial, moderate, non-significant correlations between driving skills and cognitive functions, executive functions, and perceptions and attitudes towards driving. According to the driving simulator test results and the limited sampling results of actual driving, it was found that a metacognitive occupational therapy intervention may be effective in reducing risk factors in driving among teenagers with ADHD relative to their peers with and without ADHD. In summary, the results of the present study indicate a positive direction that speaks to the viability of using a metacognitive occupational therapy intervention program for reducing risk factors in driving. A further study is required that will include a bigger number of subjects, add actual driving monitoring hours, and assign subjects randomly to the various groups.

Keywords: ADHD, driving, driving monitoring, metacognitive intervention, occupational therapy, simulator, teenagers

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409 Improving Patient and Clinician Experience of Oral Surgery Telephone Clinics

Authors: Katie Dolaghan, Christina Tran, Kim Hamilton, Amanda Beresford, Vicky Adams, Jamie Toole, John Marley

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During the Covid 19 pandemic routine outpatient appointments were not possible face to face. That resulted in many branches of healthcare starting virtual clinics. These clinics have continued following the return to face to face patient appointments. With these new types of clinic it is important to ensure that a high standard of patient care is maintained. In order to improve patient and clinician experience of the telephone clinics a quality improvement project was carried out to ensure the patient and clinician experience of these clinics was enhanced whilst remaining a safe, effective and an efficient use of resources. The project began by developing a process map for the consultation process and agreed on the design of a driver diagram and tests of change. In plan do study act (PDSA) cycle1 a single consultant completed an online survey after every patient encounter over a 5 week period. Baseline patient responses were collected using a follow-up telephone survey for each patient. Piloting led to several iterations of both survey designs. Salient results of PDSA1 included; patients not receiving appointment letters, patients feeling more anxious about a virtual appointment and many would prefer a face to face appointment. The initial clinician data showed a positive response with a provisional diagnosis being reached in 96.4% of encounters. PDSA cycle 2 included provision of a patient information sheet and information leaflets relevant to the patients’ conditions were developed and sent following new patient telephone clinics with follow-up survey analysis as before to monitor for signals of change. We also introduced the ability for patients to send an images of their lesion prior to the consultation. Following the changes implemented we noted an improvement in patient satisfaction and, in fact, many patients preferring virtual clinics as it lead to less disruption of their working lives. The extra reading material both before and after the appointments eased patients’ anxiety around virtual clinics and helped them to prepare for their appointment. Following the patient feedback virtual clinics are now used for review patients as well, with all four consultants within the department continuing to utilise virtual clinics. During this presentation the progression of these clinics and the reasons that these clinics are still operating following the return to face to face appointments will be explored. The lessons that have been gained using a QI approach have helped to deliver an optimal service that is valid and reliable as well as being safe, effective and efficient for the patient along with helping reduce the pressures from ever increasing waiting lists. In summary our work in improving the quality of virtual clinics has resulted in improved patient satisfaction along with reduced pressures on the facilities of the health trust.

Keywords: clinic, satisfaction, telephone, virtual

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