Search results for: diagnostic toolkit

Authors: Senait Belay Adugna, Mirutse Giday, Tsegahun Manyazewal

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Background: The emergence and resurgence of zoonotic diseases have continued to be a major threat to global health and the economy. Developing countries are particularly vulnerable due to agricultural expansions and the domestication of animals by humans. Scientifically sound clinical trials are important to find better ways to prevent, diagnose, and treat zoonotic diseases, while there is a lack of evidence to inform the clinical trials’ capacity and practice in countries highly affected by the diseases. This study aimed to investigate researchers’ perceptions and experiences in conducting clinical trials on zoonotic diseases in Ethiopia. Methods: This study employed a descriptive, qualitative study design. It included major academic and research institutions in Ethiopia that had active engagements in veterinary and public health research. It included the National Veterinary Institute, the National Animal Health Diagnostic and Investigation Center, the College of Veterinary Medicine at Addis Ababa University, the Ethiopian Public Health Institute, the Armauer Hansen Research Institute, and the College of Health Sciences at Addis Ababa University. In-depth interviews were conducted with 14 senior researcher investigators in the institutions who hold a proven exhibit primarily leading research activities or research units. Data were collected from October 2019 to April 2020. Data analysis was undertaken using open code 4.03 for qualitative data analysis. Results: Five major themes, with 18 sub-themes, emerged from the in-depth interview in connection. These were: challenges in the prevention, control, and treatment of zoonotic diseases; One Health approach to mitigate zoonotic diseases; personal and institutional experiences in conducting clinical trials on zoonotic diseases; barriers in conducting clinical trials towards zoonotic diseases; and strategies that promote conducting clinical trials on zoonotic diseases. Conducting clinical trials on zoonotic diseases in Ethiopia is hampered by a lack of clearly articulated ethics and regulatory frameworks, trial experts, financial resources, and good governance. Conclusions: In Ethiopia, conducting clinical trials on zoonotic diseases deserves due attention. Strengthening institutional and human resources capacity is a precondition to harnessing effective implementation of clinical trials on zoonotic diseases in the country. In Ethiopia, where skilled human resource is scarce, the One Health approach has the potential to form multidisciplinary teams to systematically improve clinical trials capacity and outcomes in the country.

Keywords: Ethiopia, clinical triak, zoonoses, disease

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Authors: Liaqat Ali, Ehsan, Muhammad Shahzad, Nasir Orakzai

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Introduction: Internal optical urethrotomy is the main stay treatment modality in management of urethral stricture. Being minimal invasive with less morbidity, it is commonly performed and favored procedure by urologists across the globe. Although short-term success rate of optical urethrotomy is promising but long-term efficacy of IOU is questionable with high recurrence rate in different studies. Numerous techniques had been adopted to reduce the recurrence after IOU like prolong catheterization and self-clean intermittent catheterization with varying success. Mitomycin C has anti-fibroblast and anti-collagen properties and has been used in trabeculectomy, myringotomy and after keloid scar excision in contemporary surgical practice. Present study according to the best of our knowledge is a pioneer pilot study in Pakistan to determine the efficacy of Mitomycin C in preventing recurrence of urethral stricture after internal optical urethrotomy. Objective: To determine the efficacy of Mitomycin C in reducing the recurrence of anterior urethral stricture after internal optical urethrotomy. Methods: It is a randomized control trial conducted in department of urology, Institute of Kidney Diseases Hayatabad Medical Complex Peshawar from March 2011 till December 2013. After approval of hospital ethical committee, we included maximum of 2 cm anterior urethral stricture irrespective of etiology. Total of 140 patients were equally divided into two groups by lottery method. Group A (Case) comprising of 70 patients in whom Mitomycin C 0.1% was injected sub mucosal in stricture area at 1,11,6 and 12 O clock position using straight working channel paediatric cystoscope after conventional optical urethrotomy. Group B (Control) 70 patients in whom only optical urethrotomy was performed. SCIC was not offered in both the groups. All the patients were regularly followed on a monthly basis for 3 months then three monthly for remaining 9 months. Recurrence was diagnosed by using diagnostic tools of retrograde urethrogram and flexible urethroscopy in selected cased. Data was collected on structured Proforma and was analyzed on SPSS. Result: The mean age in Group A was 33 ±1.5 years and Group B was 35 years. External trauma was leading cause of urethral stricture in both groups 46 (65%) Group A and 50 (71.4%) Group B. In Group A. Iatrogenic urethral trauma was 2nd etiological factor in both groups. 18(25%) Group A while 15( 21.4%) in Group B. At the end of 1 year, At the end of one year, recurrence of urethral stricture was recorded in 11 (15.71%) patient in Mitomycin C Group A and it was recorded in 27 (38.5 %) patients in group B. Significant difference p=0.001 was found in favour of group A Mitomycin group. Conclusion: Recurrence of urethral stricture is high after optical urethrotomy. Mitomycin C is found highly effective in preventing recurrence of urethral stricture after IOU.

Keywords: urethral stricture, mitomycine, internal optical urethrotomy, medical and health sciences

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

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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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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

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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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Authors: Mustafa M. Donma, Orkide Donma

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Metabolic syndrome (MetS) is a severe health problem which is common among obese individuals. The components of MetS are rather stable in adults compared to the components discussed for children. Due to the ambiguity in this group of the population, how to diagnose MetS in morbid obese (MO) children still constitutes a matter of discussion. For this purpose, a formula, which facilitates the diagnosis of MetS in MO children, was investigated. The aim of this study was to develop a formula which was capable of discriminating MO children with and without MetS findings. Study population comprised MO children. Age and sex-dependent body mass index (BMI) percentiles of the children were above 99. Metabolic syndrome components were also determined. Elevated systolic and diastolic blood pressures (SBP and DBP), elevated fasting blood glucose (FBG), elevated triglycerides (TRG), and/or depressed high density lipoprotein cholesterol (HDL-C) in addition to central obesity were listed as MetS components for each child. Presence of at least two of these components confirmed that the case was MetS. Two groups were constituted. In the first group, there were forty-two MO children without MetS components. Second group was composed of forty-four MO children with at least two MetS components. Anthropometric measurements, including weight, height, waist, and hip circumferences, were performed following physical examination. Body mass index and homeostatic model assessment of insulin resistance values were calculated. Informed consent forms were obtained from the parents of the children. Institutional Non-Interventional Ethics Committee approved the study design. Blood pressure values were recorded. Routine biochemical analysis, including FBG, insulin (INS), TRG, HDL-C were performed. The performance and the clinical utility of the Diagnostic Obesity Notation Model Assessment Metabolic Syndrome Index (DONMA MetS index) [(INS/FBG)/(HDL-C/TRG)*100] was tested. Appropriate statistical tests were applied to the study data. p value smaller than 0.05 was defined as significant. Metabolic syndrome index values were 41.6±5.1 in MO group and 104.4±12.8 in MetS group. Corresponding values for HDL-C values were 54.5±13.2 mg/dl and 44.2±11.5 mg/dl. There were statistically significant differences between the groups (p<0.001). Upon evaluation of the correlations between MetS index and HDL-C values, a much stronger negative correlation was found in MetS group (r=-0.515; p=0.001) in comparison with the correlation detected in MO group (r=-0.371; p=0.016). From these findings, it was concluded that the statistical significance degree of the difference between MO and MetS groups was highly acceptable for this recently introduced MetS index as expected. This was due to the involvement of all of the biochemically defined MetS components into the index. This is particularly important because each of these four parameters used in the formula is cardiac risk factor. Aside from discriminating MO children with and without MetS findings, MetS index introduced in this study is important from the cardiovascular risk point of view in MetS group of children.

Keywords: children, fasting blood glucose, high density lipoprotein cholesterol, index, insulin, metabolic syndrome, morbid obesity, triglycerides.

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Authors: Halimat M. Ajose-Adeogun, Zaynab A. Bello

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Artificial Intelligence (AI) and Natural Language Processing (NLP) have transformed computer language comprehension, allowing computers to comprehend spoken and written language with human-like cognition. NLP, a multidisciplinary area that combines rule-based linguistics, machine learning, and deep learning, enables computers to analyze and comprehend human language. NLP applications in medicine range from tackling issues in electronic health records (EHR) and psychiatry to improving diagnostic precision in orthopedic surgery and optimizing clinical procedures with novel technologies like chatbots. The technology shows promise in a variety of medical sectors, including quicker access to medical records, faster decision-making for healthcare personnel, diagnosing dysplasia in Barrett's esophagus, boosting radiology report quality, and so on. However, successful adoption requires training for healthcare workers, fostering a deep understanding of NLP components, and highlighting the significance of validation before actual application. Despite prevailing challenges, continuous multidisciplinary research and collaboration are critical for overcoming restrictions and paving the way for the revolutionary integration of NLP into medical practice. This integration has the potential to improve patient care, research outcomes, and administrative efficiency. The research methodology includes using NLP techniques for Sentiment Analysis and Emotion Recognition, such as evaluating text or audio data to determine the sentiment and emotional nuances communicated by users, which is essential for designing a responsive and sympathetic chatbot. Furthermore, the project includes the adoption of a Personalized Intervention strategy, in which chatbots are designed to personalize responses by merging NLP algorithms with specific user profiles, treatment history, and emotional states. The synergy between NLP and personalized medicine principles is critical for tailoring chatbot interactions to each user's demands and conditions, hence increasing the efficacy of mental health care. A detailed survey corroborated this synergy, revealing a remarkable 20% increase in patient satisfaction levels and a 30% reduction in workloads for healthcare practitioners. The poll, which focused on health outcomes and was administered to both patients and healthcare professionals, highlights the improved efficiency and favorable influence on the broader healthcare ecosystem.

Keywords: natural language processing, artificial intelligence, healthcare communication, electronic health records, patient care

Procedia PDF Downloads 47

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

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Authors: Magdalena B. Kaziuk, Waldemar Kosiba, Marek J. Kuzniewski

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Introduction: Adipose tissue is a typical place for storage water-insoluble toxins in the body. It's connective tissue, where the intercellular substance consist of fat, which level in people with low physical activity should be 18-25% for women and 13-18% for men. Due to the fat distribution in the body we distinquish two types of obesity: android (visceral, abdominal) and gynoidal (gluteal-femoral, peripheral). Abdominal obesity increases the risk of complications of the cardiovascular system diseases, and impaired renal and liver function. Through the influence on disorders of metabolism, lipid metabolism, diabetes and hypertension, leading to emergence of the metabolic syndrome. So thus, obesity will especially overload kidney function in patients after transplantation. Aim: An attempt was made to estimate the impact of amount fat tissue on transplanted kidney function and excretory function of the liver in patients after Ktx. Material and Methods: The study included 108 patients (50 females, 58 male, age 46.5 +/- 12.9 years) with active kidney transplant after more than 3 months from the transplantation. An analysis of body composition was done by using electrical bioimpedance (BIA) and anthropometric measurements. Estimated basal metabolic rate (BMR), muscle mass, total body water content and the amount of body fat. Information about physical activity were obtained during clinical examination. Nutritional status, and type of obesity were determined by using indicators: Waist to Height Ratio (WHR) and Waist to Hip Ratio (WHR). Excretory functions of the transplanted kidney was rated by calculating the estimated renal glomerular filtration rate (eGFR) using the MDRD formula. Liver function was rated by total bilirubin and alanine aminotransferase levels ALT concentration in serum. In our patients haemolitic uremic syndrome (HUS) was excluded. Results: In 19.44% of patients had underweight, 22.37% of the respondents were with normal weight, 11.11% had overweight, and the rest were with obese (49.08%). People with android stature have a lower eGFR compared with those with the gynoidal stature (p = 0.004). All patients with obesity had higher amount of body fat from a few to several percent. The higher amount of body fat percentage, the lower eGFR had patients (p <0.001). Elevated ALT levels significantly correlated with a high fat content (p <0.02). Conclusion: Increased amount of body fat, particularly in the case of android obesity can be a predictor of kidney and liver damage. Due to that obese patients should have more frequent control of diagnostic functions of these organs and the intensive dietary proceedings, pharmacological and regular physical activity adapted to the current physical condition of patients after transplantation.

Keywords: obesity, body fat, kidney transplantation, glomerular filtration rate, liver function

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Authors: Soowon Park, Min-Ji Kim, Jun-Young Lee

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Motivation: The stigma of mental illness has been studied in many disciplines, including social psychology, counseling psychology, sociology, psychiatry, public health care, and related areas, because individuals labeled as ‘mentally ill’ are often deprived of their rights and their life opportunities. To understand the factors that deepen the stigma of mental illness, it is important to understand the influencing factors of the stigma. Problem statement: Depression is a common disorder in adults, but the incidence of help-seeking is low. Researchers have believed that this poor help-seeking behavior is related to the stigma of mental illness, which results from low mental health literacy. However, it is uncertain that increasing mental health literacy decreases mental health stigmatization. Furthermore, even though decreasing stigmatization is important, the stigma of mental illness is still a stable and long-lasting phenomenon. Thus, factors other than knowledge about mental disorders have the power to maintain the stigma. Investigating the influencing factors that facilitate the stigma of psychiatric disease could help lower the social stigmatization. Approach: Face-to-face interviews were conducted with a multi-clustering sample. A total of 700 Korean participants (38% male), ranging in age from 18 to 78 (M(SD)age= 48.5(15.7)) answered demographical questions, Korean version of Link’s Perceived Devaluation and Discrimination (PDD) scale for the assessment of social stigmatization against depression, and the Korean version of the WHO-Composite International Diagnostic Interview for the assessment of mental disorders. Multiple-regression was conducted to find the predicting factors of social stigmatization against depression. Ages, sex, years of education, income, living location, and experience of mental illness were used as the predictors. Results: Predictors accounted for 14% of the variance in the stigma of depressive disorders (F(6, 693) = 20.27, p < .001). Among those, only age, years of education, and experience of mental illness significantly predicted social stigmatization against depression. The standardized regression coefficient of age had a negative association with stigmatization (β = -.20, p < .001), but years of education (β = .20, p < .001) and experience of mental illness (β = .08, p < .05) positively predicted depression stigmatization. Conclusions: The present study clearly demonstrates the association between personal factors and depressive disorder stigmatization. Younger age, more education, and self-stigma appeared to increase the stigmatization. Young, highly educated, and mentally ill people tend to reject patients with depressive disorder as friends, teachers, or babysitters; they also tend to think that those patients have lower intelligence and abilities. These results suggest the possibility that people from a high social class, or highly educated people, who have the power to make decisions, help maintain the social stigma against mental illness patients. To increase the awareness that people from high social classes have more stigmatization against depressive disorders will help decrease the biased attitudes against mentally ill patients.

Keywords: depressive disorder stigmatization, age, education, self-stigma

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Authors: Benyam Zenebe, Tesfaye Sisay, Gurja Belay, Workabeba Abebe

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Background: The prevalence and antibiogram of pathogenic E. coli strains, which cause diarrhea vary from region to region, and even within countries in the same geographical area. In Ethiopia, diagnostic approaches to E. coli induced diarrhea in children less than five years of age are not standardized. The aim of this study was to determine the involvement of pathogenic E. coli strains in child diarrhea and determine the antibiograms of the isolates in children less than 5 years of age with diarrhea at Addis Ababa University College of Health Sciences TikurAnbessa Specialized Hospital, Addis Ababa, Ethiopia. Methods: A purposive study that included 98 diarrheic children less than five years of age was conducted at Addis Ababa University College of Health Sciences, TikurAnbessa Specialized Hospital, Addis Ababa, Ethiopia to detect pathogenic E. coli biotypes. Stool culture was used to identify presumptive E. coliisolates. Presumptive isolates were confirmed by biochemical tests, and antimicrobial susceptibility tests were performed on confirmed E. coli isolates by the disk diffusion method. DNA was extracted from confirmed isolates by a heating method and subjected to Polymerase Chain Reaction or the presence of virulence genes. Amplified PCR products were analyzed by agarose gel electrophoresis. Data were collected on child demographics and clinical conditions using administered questionnaires. The prevalence of E. coli strains from the total diarrheic children, and the prevalence of pathogenic strains from total E. coli isolates along with their susceptibility profiles; the distribution of pathogenic E.coli biotypes among different age groups and between the sexes were determined by using descriptive statistics. Result: Out of 98 stool specimens collected from diarrheic children less than 5 years of age, 75 presumptive E. coli isolates were identified by culture; further confirmation by biochemical tests showed that only 56 of the isolates were E. coli; 29 of the isolates were found in male children and 27 of them in female children. Out of the 58 isolates of E. coli, 25 pathotypes belonging to different classes of pathogenic strains: STEC, EPEC, EHEC, EAEC were detected by using the PCR technique. Pathogenic E. coli exhibited high rates of antibiotic resistance to many of the antibiotics tested. Moreover, they exhibited multiple drug resistance. Conclusion: This study found that the isolation rate of E. coli and the involvement of antibiotic-resistant pathogenic E. coli in diarrheic children is prominent, and hence focus should be given on the diagnosis and antimicrobial sensitivity testing of pathogenic E. coli at Addis Ababa University College of Health Sciences TikurAnbessa Specialized Hospital. Among antibiotics tested, Cefotitan could be a drug of choice to treat E. coli.

Keywords: antibiotic susceptibility profile, children, diarrhea, E. coli, pathogenic

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Authors: Masih Nikgou

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ICU nurses or intensive care nurses are highly specialized and trained healthcare personnel. These nurses provide nursing care for patients with life-threatening illnesses or conditions. They provide the experience, knowledge and specialized skills that patients need to survive and recover. Intensive care nurses (ICU) are trained to make momentary decisions and act quickly when the patient's condition changes. Their primary work environment is in the hospital in intensive care units. Typically, ICU patients require a high level of care. ICU nurses work in challenging and complex fields in their nursing profession. They have the primary duty of caring for and saving patients who are fighting for their lives. Intensive care (ICU) nurses are highly trained to provide exceptional care to patients who depend on 24/7 nursing care. A patient in the ICU is often equipped with a ventilator, intubated and connected to several life support machines and medical equipment. Intensive Care Nurses (ICU) have full expertise in considering all aspects of bringing back their patients. Some of the specific responsibilities of ICU nurses include (a) Assessing and monitoring the patient's progress and identifying any sudden changes in the patient's medical condition. (b) Administration of drugs intravenously by injection or through gastric tubes. (c) Provide regular updates on patient progress to physicians, patients, and their families. (d) According to the clinical condition of the patient, perform the approved diagnostic or treatment methods. (e) In case of a health emergency, informing the relevant doctors. (f) To determine the need for emergency interventions, evaluate laboratory data and vital signs of patients. (g) Caring for patient needs during recovery in the ICU. (h) ICU nurses often provide emotional support to patients and their families. (i) Regulating and monitoring medical equipment and devices such as medical ventilators, oxygen delivery devices, transducers, and pressure lines. (j) Assessment of pain level and sedation needs of patients. (k) Maintaining patient reports and records. As the name suggests, critical care nurses work primarily in ICU health care units. ICUs are completely healthy and have proper lighting with strict adherence to health and safety from medical centers. ICU nurses usually move between the intensive care unit, the emergency department, the operating room, and other special departments of the hospital. ICU nurses usually follow a standard shift schedule that includes morning, afternoon, and night schedules. There are also other relocation programs depending on the hospital and region. Nurses who are passionate about data and managing a patient's condition and outcomes typically do well as ICU nurses. An inquisitive mind and attention to processes are equally important. ICU nurses are completely compassionate and are not afraid to advocate for their patients and family members. who are distressed.

Keywords: nursing, intensive care unit, pediatric intensive care unit, mobile intensive care unit, surgical intensive care unite

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Authors: Ragaa T. Darwish, Maha A. Demellawy, Haidy M. Megahed, Doreen N. Younan, Wael S. Kholeif

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The testing of drug abuse is authentic in order to affirm the misuse of drugs. Several analytical approaches have been developed for the detection of drugs of abuse in pharmaceutical and common biological samples, but few methodologies have been created to identify them from fingerprints. Liquid Chromatography-Mass Spectrometry (LC-MS) plays a major role in this field. The current study aimed at assessing the possibility of detection of some drugs of abuse (tramadol, clonazepam, and phenobarbital) from fingerprints using LC-MS in drug abusers. The aim was extended in order to assess the possibility of detection of the above-mentioned drugs in fingerprints of drug handlers till three days of handling the drugs. The study was conducted on randomly selected adult individuals who were either drug abusers seeking treatment at centers of drug dependence in Alexandria, Egypt or normal volunteers who were asked to handle the different studied drugs (drug handlers). An informed consent was obtained from all individuals. Participants were classified into 3 groups; control group that consisted of 50 normal individuals (neither abusing nor handling drugs), drug abuser group that consisted of 30 individuals who abused tramadol, clonazepam or phenobarbital (10 individuals for each drug) and drug handler group that consisted of 50 individuals who were touching either the powder of drugs of abuse: tramadol, clonazepam or phenobarbital (10 individuals for each drug) or the powder of the control substances which were of similar appearance (white powder) and that might be used in the adulteration of drugs of abuse: acetyl salicylic acid and acetaminophen (10 individuals for each drug). Samples were taken from the handler individuals for three consecutive days for the same individual. The diagnosis of drug abusers was based on the current Diagnostic and Statistical Manual of Mental disorders (DSM-V) and urine screening tests using immunoassay technique. Preliminary drug screening tests of urine samples were also done for drug handlers and the control groups to indicate the presence or absence of the studied drugs of abuse. Fingerprints of all participants were then taken on a filter paper previously soaked with methanol to be analyzed by LC-MS using SCIEX Triple Quad or QTRAP 5500 System. The concentration of drugs in each sample was calculated using the regression equations between concentration in ng/ml and peak area of each reference standard. All fingerprint samples from drug abusers showed positive results with LC-MS for the tested drugs, while all samples from the control individuals showed negative results. A significant difference was noted between the concentration of the drugs and the duration of abuse. Tramadol, clonazepam, and phenobarbital were also successfully detected from fingerprints of drug handlers till 3 days of handling the drugs. The mean concentration of the chosen drugs of abuse among the handlers group decreased when the days of samples intake increased.

Keywords: drugs of abuse, fingerprints, liquid chromatography–mass spectrometry, tramadol

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Authors: Evgeny A. Ermakov, Lyudmila P. Smirnova, Valentina N. Buneva

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Schizophrenia associated with dysregulation of neurotransmitter processes in the central nervous system and disturbances in the humoral immune system resulting in the formation of antibodies (Abs) to the various components of the nervous tissue. Abs to different neuronal receptors and DNA were detected in the blood of patients with schizophrenia. Abs hydrolyzing DNA were detected in pool of polyclonal autoantibodies in autoimmune and infectious diseases, such catalytic Abs were named abzymes. It is believed that DNA-hydrolyzing abzymes are cytotoxic, cause nuclear DNA fragmentation and induce cell death by apoptosis. Abzymes with DNAase activity are interesting because of the mechanism of formation and the possibility of use as diagnostic markers. Therefore, in our work we have set following goals: to determine the level anti-DNA Abs in the serum of patients with schizophrenia and to study DNA-hydrolyzing activity of IgG of patients with schizophrenia. Materials and methods: In our study there were included 41 patients with a verified diagnosis of paranoid or simple schizophrenia and 24 healthy donors. Electrophoretically and immunologically homogeneous IgGs were obtained by sequential affinity chromatography of the serum proteins on protein G-Sepharose and gel filtration. The levels of anti-DNA Abs were determined using ELISA. DNA-hydrolyzing activity was detected as the level of supercoiled pBluescript DNA transition in circular and linear forms, the hydrolysis products were analyzed by agarose electrophoresis followed by ethidium bromide stain. To correspond the registered catalytic activity directly to the antibodies we carried out a number of strict criteria: electrophoretic homogeneity of the antibodies, gel filtration (acid shock analysis) and in situ activity. Statistical analysis was performed in ‘Statistica 9.0’ using the non-parametric Mann-Whitney test. Results: The sera of approximately 30% of schizophrenia patients displayed a higher level of Abs interacting with single-stranded (ssDNA) and double-stranded DNA (dsDNA) compared with healthy donors. The average level of Abs interacting with ssDNA was only 1.1-fold lower than that for interacting with dsDNA. IgG of patient with schizophrenia were shown to possess DNA hydrolyzing activity. Using affinity chromatography, electrophoretic analysis of isolated IgG homogeneity, gel filtration in acid shock conditions and in situ DNAse activity analysis we proved that the observed activity is intrinsic property of studied antibodies. We have shown that the relative DNAase activity of IgG in patients with schizophrenia averaged 55.4±32.5%, IgG of healthy donors showed much lower activity (average of 9.1±6.5%). It should be noted that DNAase activity of IgG in patients with schizophrenia with a negative symptoms was significantly higher (73.3±23.8%), than in patients with positive symptoms (43.3±33.1%). Conclusion: Anti-DNA Abs of patients with schizophrenia not only bind DNA, but quite efficiently hydrolyze the substrate. The data show a correlation with the level of DNase activity and leading symptoms of patients with schizophrenia.

Keywords: anti-DNA antibodies, abzymes, DNA hydrolysis, schizophrenia

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Authors: Natalia Buderatskaya, Igor Ilyin, Julia Gontar, Sergey Lavrynenko, Olga Parnitskaya, Ekaterina Ilyina, Eduard Kapustin, Yana Lakhno

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Introduction: It is known that cryopreservation of oocytes has peculiar features due to the complex structure of the oocyte. One of the most important features is that mature oocytes contain meiotic division spindle which is very sensitive even to the slightest variation in temperature. Thus, the main objective of this study is to analyse the resulting euploid embryos obtained from thawed oocytes in comparison with the data of preimplantation genetic screening (PGS) in fresh embryo cycles. Material and Methods: The study was conducted at 'Medical Centre IGR' from January to July 2016. Data were analysed for 908 donor oocytes obtained in 67 cycles of assisted reproductive technologies (ART), of which 693 oocytes were used in the 51 'fresh' cycles (group A), and 215 oocytes - 16 ART programs with vitrification female gametes (group B). The average age of donors in the groups match 27.3±2.9 and 27.8±6.6 years. Stimulation of superovulation was conducted the standard way. Vitrification was performed in 1-2 hours after transvaginal puncture and thawing of oocytes were carried out in accordance with the standard protocol of Cryotech (Japan). Manipulation ICSI was performed 4-5 hours after transvaginal follicle puncture for fresh oocytes, or after defrosting - for vitrified female gametes. For the PGS, an embryonic biopsy was done on the third or on the fifth day after fertilization. Diagnostic procedures were performed using fluorescence in situ hybridization with the study of such chromosomes as 13, 16, 18, 21, 22, X, Y. Only morphologically quality blastocysts were used for the transfer, the estimation of which corresponded to the Gardner criteria. The statistical hypotheses were done using the criteria t, x^2 at a significance levels p<0.05, p<0.01, p<0.001. Results: The mean number of mature oocytes per cycle in group A was 13.58±6.65 and in group B - 13.44±6.68 oocytes for patient. The survival of oocytes after thawing totaled 95.3% (n=205), which indicates a highly effective quality of performed vitrification. The proportion of zygotes in the group A corresponded to 91.1%(n=631), in the group B – 80.5%(n=165), which shows statistically significant difference between the groups (p<0.001) and explained by non-viable oocytes elimination after vitrification. This is confirmed by the fact that on the fifth day of embryos development a statistically significant difference in the number of blastocysts was absent (p>0.05), and constituted respectively 61.6%(n=389) and 63.0%(n=104) in the groups. For the PGS performing 250 embryos analyzed in the group A and 72 embryos - in the group B. The results showed that euploidy in the studied chromosomes were 40.0%(n=100) embryos in the group A and 41.7% (n=30) - in the group B, which shows no statistical significant difference (p>0.05). The indicators of clinical pregnancies in the groups amounted to 64.7% (22 pregnancies per 34 embryo transfers) and 61.5% (8 pregnancies per 13 embryo transfers) respectively, and also had no significant difference between the groups (p>0.05). Conclusions: The results showed that the vitrification does not affect the resulting euploid embryos in assisted reproductive technologies and are not reflected in their morphological characteristics in ART programs.

Keywords: euploid embryos, preimplantation genetic screening, thawing oocytes, vitrification

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Authors: S. J. Yune, S. Y. Lee, S. J. Im, B. S. Kam, S. Y. Baek

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This study aimed to determine the reliability of clinical performance assessments, having been emphasized by ability-based education, and professors overall assessment methods. We addressed the following problems: First, we try to find out whether there is a difference in what we consider to be the main variables affecting the clinical performance test according to the evaluator’s working period and the number of evaluation experience. Second, we examined the relationship among the global rating score (G), analytic global rating score (Gc), and the sum of the analytical checklists (C). What are the main factors affecting clinical performance assessments in relation to the numbers of times the evaluator had administered evaluations and the length of their working period service? What is the relationship between overall assessment score and analytic checklist score? How does analytic global rating with 6 components in OSCE and 4 components in sub-domains (Gc) CPX: aseptic practice, precision, systemic approach, proficiency, successfulness, and attitude overall assessment score and task-specific analytic checklist score sum (C) affect the professor’s overall global rating assessment score (G)? We studied 75 professors who attended a 2016 Bugyeoung Consortium clinical skills performances test evaluating third and fourth year medical students at the Pusan National University Medical school in South Korea (39 prof. in OSCE, 36 prof. in CPX; all consented to participate in our study). Each evaluator used 3 forms; a task-specific analytic checklist, subsequent analytic global rating scale with sub-6 domains, and overall global scale. After the evaluation, the professors responded to the questionnaire on the important factors of clinical performance assessment. The data were analyzed by frequency analysis, correlation analysis, and hierarchical regression analysis using SPSS 21.0. Their understanding of overall assessment was analyzed by dividing the subjects into groups based on experiences. As a result, they considered ‘precision’ most important in overall OSCE assessment, and ‘precise accuracy physical examination’, ‘systemic approaches to taking patient history’, and ‘diagnostic skill capability’ in overall CPX assessment. For OSCE, there was no clear difference of opinion about the main factors, but there was for CPX. Analytic global rating scale score, overall rating scale score, and analytic checklist score had meaningful mutual correlations. According to the regression analysis results, task-specific checklist score sum had the greatest effect on overall global rating. professors regarded task-specific analytic checklist total score sum as best reflecting overall OSCE test score, followed by aseptic practice, precision, systemic approach, proficiency, successfulness, and attitude on a subsequent analytic global rating scale. For CPX, subsequent analytic global rating scale score, overall global rating scale score, and task-specific checklist score had meaningful mutual correlations. These findings support explanations for validity of professors’ global rating in clinical performance assessment.

Keywords: global rating, clinical performance assessment, medical education, analytic checklist

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Authors: Sadeem Aljamaan, Reem Hariri, Rahaf Alghamdi, Batool Alotaibi, Batool Alsenan, Lama Althunayyan, Areej Alnemer

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The aim of this study is to correlate between radiological findings and histopathological results in regard to the breast imaging-reporting and data system scores, size of breast masses, molecular subtypes and suspicious radiological features, as well as to assess the concordance rate in histological grade between core biopsy and surgical excision among breast cancer patients, followed by analyzing the change of concordance rate in relation to neoadjuvant chemotherapy in a Saudi population. A retrospective review was conducted over 6-year period (2017-2022) on all breast core biopsies of women preceded by radiological investigation. Chi-squared test (χ2) was performed on qualitative data, the Mann-Whitney test for quantitative non-parametric variables, and the Kappa test for grade agreement. A total of 641 cases were included. Ultrasound, mammography, and magnetic resonance imaging demonstrated diagnostic accuracies of 85%, 77.9% and 86.9%; respectively. magnetic resonance imaging manifested the highest sensitivity (72.2%), and the lowest was for ultrasound (61%). Concordance in tumor size with final excisions was best in magnetic resonance imaging, while mammography demonstrated a higher tendency of overestimation (41.9%), and ultrasound showed the highest underestimation (67.7%). The association between basal-like molecular subtypes and the breast imaging-reporting and data system score 5 classifications was statistically significant only for magnetic resonance imaging (p=0.04). Luminal subtypes demonstrated a significantly higher percentage of speculation in mammography. Breast imaging-reporting and data system score 4 manifested a substantial number of benign pathologies in all the 3 modalities. A fair concordance rate (k= 0.212 & 0.379) was demonstrated between excision and the preceding core biopsy grading with and without neoadjuvant therapy, respectively. The results demonstrated a down-grading in cases post-neoadjuvant therapy. In cases who did not receive neoadjuvant therapy, underestimation of tumor grade in biopsy was evident. In summary, magnetic resonance imaging had the highest sensitivity, specificity, positive predictive value and accuracy of both diagnosis and estimation of tumor size. Mammography demonstrated better sensitivity than ultrasound and had the highest negative predictive value, but ultrasound had better specificity, positive predictive value and accuracy. Therefore, the combination of different modalities is advantageous. The concordance rate of core biopsy grading with excision was not impacted by neoadjuvant therapy.

Keywords: breast cancer, mammography, MRI, neoadjuvant, pathology, US

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Authors: Mustafa Metin Donma, Orkide Donma

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Vitamin deficiencies are common in obese individuals. Particularly, the status of vitamin B12 and its association with vitamin B9 (folate) and vitamin D is under investigation in recent time. Vitamin B12 is closely related to many vital processes in the body. In clinical studies, its involvement in fat metabolism draws attention from the obesity point of view. Obesity, in its advanced stages and in combination with metabolic syndrome (MetS) findings, may be a life-threatening health problem. Pediatric obesity is particularly important because it may be a predictor of severe chronic diseases during the adulthood period of the child. Due to its role in fat metabolism, vitamin B12 deficiency may disrupt metabolic pathways of the lipid and energy metabolisms in the body. The association of low B12 levels with obesity degree may be an interesting topic to be investigated. Obesity indices may be helpful at this point. Weight- and fat-based indices are available. Of them, body mass index (BMI) is in the first group. Fat mass index (FMI), fat-free mass index (FFMI) and diagnostic obesity notation model assessment-II (D2I) index lie in the latter group. The aim of this study is to clarify possible associations between vitamin B12 status and obesity indices in the pediatric population. The study comprises a total of one hundred and twenty-two children. Thirty-two children were included in the normal body mass index (N-BMI) group. Forty-six and forty-four children constitute groups with morbid obese children without MetS and with MetS, respectively. Informed consent forms and the approval of the institutional ethics committee were obtained. Tables prepared for obesity classification by World Health Organization were used. Metabolic syndrome criteria were defined. Anthropometric and blood pressure measurements were taken. Body mass index, FMI, FFMI, D2I were calculated. Routine laboratory tests were performed. Vitamin B9, B12, D concentrations were determined. Statistical evaluation of the study data was performed. Vitamin B9 and vitamin D levels were reduced in MetS group compared to children with N-BMI (p>0.05). Significantly lower values were observed in vitamin B12 concentrations of MetS group (p<0.01). Upon evaluation of blood pressure as well as triglyceride levels, there exist significant increases in morbid obese children. Significantly decreased concentrations of high density lipoprotein cholesterol were observed. All of the obesity indices and insulin resistance index exhibit increasing tendency with the severity of obesity. Inverse correlations were calculated between vitamin D and insulin resistance index as well as vitamin B12 and D2I in morbid obese groups. In conclusion, a fat-based index, D2I, was the most prominent body index, which shows a strong correlation with vitamin B12 concentrations in the late stage of obesity in children. A negative correlation between these two parameters was a confirmative finding related to the association between vitamin B12 and obesity degree.

Keywords: body mass index, children, D2I index, fat mass index, obesity

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Authors: G. C. Santini, C. Potrich, L. Lunelli, L. Vanzetti, S. Marasso, M. Cocuzza, C. Pederzolli

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The relevance of circulating miRNAs as non-invasive biomarkers for several pathologies is nowadays undoubtedly clear, as they have been found to have both diagnostic and prognostic value able to add fundamental information to patients’ clinical picture. The availability of these data, however, relies on a time-consuming process spanning from the sample collection and processing to the data analysis. In light of this, strategies which are able to ease this procedure are in high demand and considerable effort have been made in developing Lab-on-a-chip (LOC) devices able to speed up and standardise the bench work. In this context, a very promising polydimethylsiloxane (PDMS)-based microdevice which integrates the processing of the biological sample, i.e. purification of extracellular miRNAs, and reverse transcription was previously developed in our lab. In this study, we aimed at the improvement of the miRNA extraction performances of this micro device by increasing the ability of its surface to absorb extracellular miRNAs from biological samples. For this purpose, we focused on the modulation of two properties of the material: roughness and charge. PDMS surface roughness was modulated by casting with several templates (terminated with silicon oxide coated by a thin anti-adhesion aluminum layer), followed by a panel of curing conditions. Atomic force microscopy (AFM) was employed to estimate changes at the nanometric scale. To introduce modifications in surface charge we functionalized PDMS with different mixes of positively charged 3-aminopropyltrimethoxysilanes (APTMS) and neutral poly(ethylene glycol) silane (PEG). The surface chemical composition was characterized by X-ray photoelectron spectroscopy (XPS) and the number of exposed primary amines was quantified with the reagent sulfosuccinimidyl-4-o-(4,4-dimethoxytrityl) butyrate (s-SDTB). As our final end point, the adsorption rate of all these different conditions was assessed by fluorescence microscopy by incubating a synthetic fluorescently-labeled miRNA. Our preliminary analysis identified casting on thermally grown silicon oxide, followed by a curing step at 85°C for 1 hour, as the most efficient technique to obtain a PDMS surface roughness in the nanometric scaleable to trap miRNA. In addition, functionalisation with 0.1% APTMS and 0.9% PEG was found to be a necessary step to significantly increase the amount of microRNA adsorbed on the surface, therefore, available for further steps as on-chip reverse transcription. These findings show a substantial improvement in the extraction efficiency of our PDMS microdevice, ultimately leading to an important step forward in the development of an innovative, easy-to-use and integrated system for the direct purification of less abundant circulating microRNAs.

Keywords: circulating miRNAs, diagnostics, Lab-on-a-chip, polydimethylsiloxane (PDMS)

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Authors: Cheh-Shyh Ting, Jiin-Liang Lin

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This study was conducted at the Wanglung Farm in Pingtung County to test the groundwater seepage influences on the implemented project for artificial groundwater recharge. The study was divided into three phases. The first phase, conducted on natural groundwater that was recharged through the local climate and growing conditions, observed the natural form of vegetation species. The original plants were flooded, and after 60 days it was observed that of the original plants only Goosegrass (Eleusine indica) and Black heart (Polygonum lapathifolium Linn.) remained. Direct infiltration tests were carried out, and calculations for the effect of vegetation on infiltration velocity of the recharge pool were noted. The second phase was an indoor test. Bahia grass and wild amaranth were selected as vegetation roots. After growth, the distribution of different grassroots was observed in order to facilitate a comparison permeability coefficient calculated by the amount of penetration and to explore the relationship between density and the efficiency to groundwater recharge. The third phase was the root tomography analysis, further observation of the development of plant roots using computed tomography technology. Computed Tomography, also known as (CT), is a diagnostic imaging examination, normally used in the medical field. In the first phase of the feasibility study, most non-aquatic plants wilted and died within seven days. In seven days, the remaining plants were used for experimental infiltration analysis. Results showed that in eight hours of infiltration test, Eleusine indica stems averaged 0.466 m/day and wild amaranth averaged 0.014 m/day. The second phase of the experiment was conducted on the remains of the plant a week in it had died and rotted, and the infiltration experiment was performed under these conditions. The results showed eight hours in end of the infiltration test, Eleusine indica stems averaged 0.033 m/day, and wild amaranth averaged 0.098 m/day. Non-aquatic plants died within two weeks, and their rotted remains clogged the pores of bottom soil particles, causing obstruction of recharge pool infiltration. Experiment results showed that eight hours in the test the average infiltration velocity for Eleusine indica stems was 0.0229 m/day and wild amaranth averaged 0.0117 m/day. Since the rotted roots of the plants blocked the pores of the soil in the recharge pool, which resulted in the obstruction of the artificial infiltration pond and showed an immediate impact on recharge efficiency. In order to observe the development of plant roots, the third phase used computed tomography imaging. Iodine developer was injected into the Black heart, allowing its cross-sectional images to be shown on CT and to be used to observe root development.

Keywords: artificial recharge of groundwater, computed tomography, infiltration velocity, vegetation root system

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Authors: Shamim Mushtaq, Moazzam Shahid

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Breast cancer (BC) claims the lives of half a million women every year and is the most common cause of death in the developing world. In 2019, it was estimated that BC alone accounts for 15% of all cancer deaths in younger women (aged < 45 years old) with advanced-stage lung metastasis. According to the World Health Organization & International Union against Cancer, in Asia, a high number of cancer-related deaths will be observed in 2020, whereas the burden will be reduced in Western countries due to awareness about the disease, better health facilities and advanced treatments. In the last 15 years, it has been reported that the incidence of BC has increased by 1.1% among Asian compared to the US population from 2003 to 2012. To date, several BC biological subtypes have been reported so far, which are associated with different treatment responses. The heterogeneity and diversity of BC reflected these different subtypes, including Luminal A (23.7% prevalence) and B (38.8% prevalence) that have pathological estrogen receptor (ER+)-positive tumors, the human epidermal growth factor receptor 2 (HER2) (11.2% prevalence) and triple-negative breast cancer (TNBC) (25% prevalence). According to Shaukat Khanum Memorial Cancer Hospital and Research Centre – Pakistan, ten years of data showed that among 636 BC patients, 30.5% had TNBC who were <40 years of age, which is an extremely alarming situation. Therefore, there is a dire need to explore and develop therapeutic targets for the treatment of early TNBC. Since the last decade, unfortunately, there has been little success in understanding the complexity of TNBC and in discovering new biological therapeutic targets. However, conventional chemotherapy is the only choice of treatment for TNBC patients. Many investigators revealed advances in multi-omics (multiple "omes", e.g., genome, proteome, transcriptome, epigenome, and microbiome) which were later identified as actionable targets and increased prevalence in TNBC patients. However, various drugs have been identified so far which are related to a particular diagnostic and prognostic biomarker. For example, Epidermal growth factor receptor ( EGFR or ErbB-1), HER-2/neu (ErbB-2), HER-3 (ErbB-3), and HER-4 (ErbB-4). Protein Transglin-2 (TAGLN 2 ) and Profilins-1 (Pfn-1 ) are the ubiquitously expressed large family of proteins present in all eukaryotes, enabling actin cytoskeletal reorganization. It is known that the oncogenic transformation of cells is accompanied by alteration in the actin cytoskeleton. There are causal connections between altered expression of actin cytoskeletal regulators and cancer progression. Our case-control study identified TAGLN-2 and Pfn-1 proteins in TNBC blood by mass spectrometry. Both TAGLN-2 and Pfn-1 proteins are differentially expressed in early and advanced stages of TNBS patients, which could be potential predictors or therapeutic targets for TNBC.

Keywords: TNBC, blood biomarkers, mass spectrometry, qPCR, ELISA

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Authors: Ioannis Panagiotopoulos, Aristidis Anastasakis, Konstantinos Toutouzas, Ioannis Iakovou, Charalampos Vlachopoulos, Vasilis Voudris, Georgios Tziomalos, Konstantinos Tsioufis, Efstathios Kastritis, Alexandros Briassoulis, Kimon Stamatelopoulos, Alexios Antonopoulos, Paraskevi Exadaktylou, Evanthia Giannoula, Anastasia Katinioti, Maria Kalantzi, Evangelos Leontiadis, Eftychia Smparouni, Ioannis Malakos, Nikolaos Aravanis, Argyrios Doumas, Maria Koutelou

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Introduction: Cardiac amyloidosis from Transthyretin (ATTR-CA) is an infiltrative disease characterized by the deposition of pathological transthyretin complexes in the myocardium. This study describes the characteristics of patients diagnosed with ATTR-CA from 2019 until present at the Nuclear Medicine Department of Onassis Cardiac Surgery Center and AHEPA Hospital. These centers have extensive experience in amyloidosis and modern technological equipment for its diagnosis. Materials and Methods: Records of consecutive patients (N=73) diagnosed with any type of amyloidosis were collected, analyzed, and prospectively followed. The diagnosis of amyloidosis was made using specific myocardial scintigraphy with Tc-99m DPD. Demographic characteristics, including age, gender, marital status, height, and weight, were collected in a database. Clinical characteristics, such as amyloidosis type (ATTR and AL), serum biomarkers (BNP, troponin), electrocardiographic findings, ultrasound findings, NYHA class, aortic valve replacement, device implants, and medication history, were also collected. Some of the most significant results are presented. Results: A total of 73 cases (86% male) were diagnosed with amyloidosis over four years. The mean age at diagnosis was 82 years, and the main symptom was dyspnea. Most patients suffered from ATTR-CA (65 vs. 8 with AL). Out of all the ATTR-CA patients, 61 were diagnosed with wild-type and 2 with two rare mutations. Twenty-eight patients had systemic amyloidosis with extracardiac involvement, and 32 patients had a history of bilateral carpal tunnel syndrome. Four patients had already developed polyneuropathy, and the diagnosis was confirmed by DPD scintigraphy, which is known for its high sensitivity. Among patients with isolated cardiac involvement, only 6 had left ventricular ejection fraction below 40%. The majority of ATTR patients underwent tafamidis treatment immediately after diagnosis. Conclusion: In conclusion, the experiences shared by the two centers and the continuous exchange of information provide valuable insights into the diagnosis and management of cardiac amyloidosis. Clinical suspicion of amyloidosis and early diagnostic approach are crucial, given the availability of non-invasive techniques. Cardiac scintigraphy with DPD can confirm the presence of the disease without the need for a biopsy. The ultimate goal still remains continuous education and awareness of clinical cardiologists so that this systemic and treatable disease can be diagnosed and certified promptly and treatment can begin as soon as possible.

Keywords: amyloidosis, diagnosis, myocardial scintigraphy, Tc-99m DPD, transthyretin

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Authors: Mustafa M. Donma, Orkide Donma, Murat Aydin, Muhammet Demirkol, Burcin Nalbantoglu, Aysin Nalbantoglu, Birol Topcu

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Childhood obesity is an ever increasing global health problem, affecting both developed and developing countries. Accurate evaluation of obesity in children requires difficult and detailed investigation. In our study, obesity in children was evaluated using new body fat ratios and indices. Assessment of anthropometric measurements, as well as some ratios, is important because of the evaluation of gender differences particularly during the late periods of obesity. A total of 239 children; 168 morbid obese (MO) (81 girls and 87 boys) and 71 normal weight (NW) (40 girls and 31 boys) children, participated in the study. Informed consent forms signed by the parents were obtained. Ethics Committee approved the study protocol. Mean ages (years)±SD calculated for MO group were 10.8±2.9 years in girls and 10.1±2.4 years in boys. The corresponding values for NW group were 9.0±2.0 years in girls and 9.2±2.1 years in boys. Mean body mass index (BMI)±SD values for MO group were 29.1±5.4 kg/m² and 27.2±3.9 kg/m² in girls and boys, respectively. These values for NW group were calculated as 15.5±1.0 kg/m² in girls and 15.9±1.1 kg/m² in boys. Groups were constituted based upon BMI percentiles for age-and-sex values recommended by WHO. Children with percentiles >99 were grouped as MO and children with percentiles between 85 and 15 were considered NW. The anthropometric measurements were recorded and evaluated along with the new ratios such as trunk-to-appendicular fat ratio, as well as indices such as Index-I and Index-II. The body fat percent values were obtained by bio-electrical impedance analysis. Data were entered into a database for analysis using SPSS/PASW 18 Statistics for Windows statistical software. Increased waist-to-hip circumference (C) ratios, decreased head-to-neck C, height ‘to’ ‘two’-‘to’-waist C and height ‘to’ ‘two’-‘to’-hip C ratios were observed in parallel with the development of obesity (p≤0.001). Reference value for height ‘to’ ‘two’-‘to’-hip ratio was detected as approximately 1.0. Index-II, based upon total body fat mass, showed much more significant differences between the groups than Index-I based upon weight. There was not any difference between trunk-to-appendicular fat ratios of NW girls and NW boys (p≥0.05). However, significantly increased values for MO girls in comparison with MO boys were observed (p≤0.05). This parameter showed no difference between NW and MO states in boys (p≥0.05). However, statistically significant increase was noted in MO girls compared to their NW states (p≤0.001). Trunk-to-appendicular fat ratio was the only fat-based parameter, which showed gender difference between NW and MO groups. This study has revealed that body ratios and formula based upon body fat tissue are more valuable parameters than those based on weight and height values for the evaluation of morbid obesity in children.

Keywords: anthropometry, childhood obesity, gender, morbid obesity

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Authors: Aarti M. Shetty, Estela Davoodi, Subrata Gangooly, Anita Rao-Coppisetty

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Background: Testing Patency of Fallopian Tubes is among one of the several protocols for investigating Subfertile Couples. Both, Hysterosalpingogram (HSG) and Laparoscopy and dye test have been used as Tubal patency test for several years, with well-known limitation. Hysterosalpingo Contrast Sonography (HyCoSy) can be used as an alternative tool to HSG, to screen patency of Fallopian tubes, with an advantage of being non-ionising, and also, use of transvaginal scan to diagnose pelvic pathology. Aim: To determine the indication and analyse the performance of transvaginal scan and HyCoSy in Broomfield Hospital. Methods: We retrospectively analysed fertility workup of 282 women, who attended HyCoSy clinic at our institution from January 2015 to June 2016. An Audit proforma was designed, to aid data collection. Data was collected from patient notes and electronic records, which included patient demographics; age, parity, type of subfertility (primary or secondary), duration of subfertility, past medical history and base line investigation (hormone profile and semen analysis). Findings of the transvaginal scan, HyCoSy and Laparoscopy were also noted. Results: The most common indication for referral were as a part of primary fertility workup on couples who had failure to conceive despite intercourse for a year, other indication for referral were recurrent miscarriage, history of ectopic pregnancy, post reversal of sterilization(vasectomy and tuboplasty), Post Gynaecology surgery(Loop excision, cone biopsy) and amenorrhea. Basic Fertility workup showed 34% men had abnormal semen analysis. HyCoSy was successfully completed in 270 (95%) women using ExEm foam and Transvaginal Scan. Of the 270 patients, 535 tubes were examined in total. 495/535 (93%) tubes were reported as patent, 40/535 (7.5%) tubes were reported as blocked. A total of 17 (6.3%) patients required laparoscopy and dye test after HyCoSy. In these 17 patients, 32 tubes were examined under laparoscopy, and 21 tubes had findings similar to HyCoSy, with a concordance rate of 65%. In addition to this, 41 patients had some form of pelvic pathology (endometrial polyp, fibroid, cervical polyp, fibroid, bicornuate uterus) detected during transvaginal scan, who referred to corrective surgeries after attending HyCoSy Clinic. Conclusion: Our audit shows that HyCoSy and Transvaginal scan can be a reliable screening test for low risk women. Furthermore, it has competitive diagnostic accuracy to HSG in identifying tubal patency, with an additional advantage of screening for pelvic pathology. With addition of 3D Scan, pulse Doppler and other non-invasive imaging modality, HyCoSy may potentially replace Laparoscopy and chromopertubation in near future.

Keywords: hysterosalpingo contrast sonography (HyCoSy), transvaginal scan, tubal infertility, tubal patency test

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Authors: Halima Sadia Qureshi, Urooj Sadiq, Noshi Eram Zaman

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The goal of this study was to see how cognitive behavior hypnotherapy affected nonsuicidal self-injury. DSM 5 invites the researchers to explore the newly added condition under the chapter of conditions under further study named Nonsuicidal self-injury disorder. To date, no empirical sound intervention has been proven effective for NSSI as given in DSM 5. Nonsuicidal self-injury is defined by DSM 5 as harming one's self physically, without suicidal intention. Around 7.6% of teenagers are expected to fulfill the NSSI disorder criteria. 3 Adolescents, particularly university students, account for around 87 percent of self-harm studies. Furthermore, one of the risks associated with NSSI is an increased chance of suicide attempts, and in most cases, the cycle repeats again. 6 The emotional and psychological components of the illness might lead to suicide, either intentionally or unintentionally. 7 According to a research done at a Pakistani military hospital, over 80% of participants had no intention of committing suicide. Furthermore, it has been determined that improvements in NSSI prevention and intervention are necessary as a stand-alone strategy. The quasi-experimental study took place in Islamabad and Rawalpindi, Pakistan, from May 2019 to April 2020 and included students aged 18 to 25 years old from several institutions and colleges in the twin cities. According to the Diagnostic and Statistical Manual of Mental Disorders 5th edition, the individuals were assessed for >2 episodes without suicidal intent using the intentional self-harm questionnaire. The Clinician Administered Nonsuicidal Self-Injury Disorder Index (CANDI) was used to assess the individual for NSSI condition. Symptom checklist-90 (SCL-90) was used to screen the participants for differential diagnosis. Mclean Screening Instrument for Borderline Personality Disorder (MSI-BPD) was used to rule out the BPD cases. The selected participants, n=106 from the screening sample of 600, were selected. They were further screened to meet the inclusion and exclusion criteria, and the total of n=71 were split into two groups: intervention and control. The intervention group received cognitive behavior hypnotherapy for the next three months, whereas the control group received no treatment. After the period of three months, both the groups went through the post assessment, and after the three months’ period, follow-up assessment was conducted. The groups were evaluated, and SPSS 25 was used to analyse the data. The results showed that each of the two groups had 30 (50 percent) of the 60 participants. There were 41 males (68 percent) and 19 girls (32 percent) in all. The bulk of the participants were between the ages of 21 and 23. (48 percent). Self-harm events were reported by 48 (80 percent) of the pupils, and suicide ideation was found in 6 (ten percent). In terms of pre- and post-intervention values (d=4.90), post-intervention and follow-up assessment values (d=0.32), and pre-intervention and follow-up values (d=5.42), the study's effect size was good. The comparison of treatment and no-treatment groups revealed that treatment was more successful than no-treatment, F (1, 58) = 53.16, p.001. The results reveal that the treatment manual of CBH is effective for Nonsuicidal self-injury disorder.

Keywords: NSSI, nonsuicidal self injury disorder, self-harm, self-injury, Cognitive behaviour hypnotherapy, CBH

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Authors: Massimiliano Panella, Sara Bortoluzzi, Sophia Russotto, Daniele Nicolini, Carmela Rinaldi

Abstract:

Osteoporosis affects more than 200 million people worldwide. About 75% of osteoporosis cases are undiagnosed or diagnosed only when a bone fracture occurs. Since osteoporosis related fractures are significant determinants of the burden of disease and health and social costs of aging populations, we believe that this is the early identification and treatment of high-risk patients should be a priority in actual healthcare systems. Screening for osteoporosis by dual energy x-ray absorptiometry (DEXA) is not cost-effective for general population. An alternative is pulse-echo ultrasound (PEUS) because of the minor costs. To this end, we developed an early detection program for osteoporosis with PEUS, and we evaluated is possible impact and sustainability. We conducted a cross-sectional study including 1,050 people in Italy. Subjects with >1 major or >2 minor risk factors for osteoporosis were invited to PEUS bone mass density (BMD) measurement at the proximal tibia. Based on BMD values, subjects were classified as healthy subjects (BMD>0.783 g/cm²) and pathological including subjects with suspected osteopenia (0.783≤BMD>0.719 g/cm²) or osteoporosis (BMD ≤ 0.719 g/cm²). The responder rate was 60.4% (634/1050). According to the risk, PEUS scan was recommended to 436 people, of whom 300 (mean age 45.2, 81% women) accepted to participate. We identified 240 (80%) healthy and 60 (20%) pathological subjects (47 osteopenic and 13 osteoporotic). We observed a significant association between high risk people and reduced bone density (p=0.043) with increased risks for female gender, older ages, and menopause (p<0.01). The yearly cost of the screening program was 8,242 euros. With actual Italian fracture incidence rates in osteoporotic patients, we can reasonably expect in 20 years that at least 6 fractures will occur in our sample. If we consider that the mean costs per fracture in Italy is today 16,785 euros, we can estimate a theoretical cost of 100,710 euros. According to literature, we can assume that the early treatment of osteoporosis could avoid 24,170 euros of such costs. If we add the actual yearly cost of the treatments to the cost of our program and we compare this final amount of 11,682 euros to the avoidable costs of fractures (24,170 euros) we can measure a possible positive benefits/costs ratio of 2.07. As a major outcome, our study let us to early identify 60 people with a significant bone loss that were not aware of their condition. This diagnostic anticipation constitutes an important element of value for the project, both for the patients, for the preventable negative outcomes caused by the fractures, and for the society in general, because of the related avoidable costs. Therefore, based on our finding, we believe that the PEUS based screening performed could be a cost-effective approach to early identify osteoporosis. However, our study has some major limitations. In fact, in our study the economic analysis is based on theoretical scenarios, thus specific studies are needed for a better estimation of the possible benefits and costs of our program.

Keywords: osteoporosis, prevention, public health, screening

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Authors: E. Akdur Öztürk, F. İrvasa Bilgiç, A. Ludovisi , O. Gülbahar, D. Dirim Erdoğan, M. Korkmaz, M. Á. Gómez Morales

Abstract:

Anisakidosis is a zoonotic human fish-borne parasitic disease caused by accidental ingestion of anisakid third-stage larvae (L3) of members of the Anisakidae family present in infected marine fish or cephalopods. Infection with anisakid larvae can lead to gastric, intestinal, extra-gastrointestinal and gastroallergic forms of the disease. Anisakid parasites have been reported in almost all seas, particularly in the Mediterranean Sea. There is a remarkably high level of risk exposure to these zoonotic parasites as they are present in economically and ecologically important fish of Europe. Anisakid L3 larvae have been also detected in several fish species from the Aegean Sea. Turkey is a peninsular country surrounded by Black, Aegean and the Mediterranean Sea. In this country, fishing habit and fishery product consumption are highly common. In recent years, there was also an increase in the consumption of raw fish due to the increasing interest in the cuisine of the Far East countries. In different regions of Turkey, A. simplex (inMerluccius Merluccius Scomber japonicus, Trachurus mediterraneus, Sardina pilchardus, Engraulis encrasicolus, etc.), Anisakis spp., Contraceucum spp., Pseudoterronova spp. and, C. aduncum were identified as well. Although it is accepted both the presence of anisakid parasites in fish and fishery products in Turkey and the presence of Turkish people with allergic manifestations after fish consumption, there are no reports of human anisakiasis in this country. Given the high prevalence of anisakid parasites in the country, the absence of reports is likely not due to the absence of clinical cases rather to the unavailability of diagnostic tools and the low awareness of the presence of this infection. The aim of the study was to set up an IgE-Western Blot (WB) based test to detect the anisakidosis sensitization among Turkish people with a history of allergic manifestation related to fish consumption. To this end, crude worm antigens (CWA) and allergen enriched fraction (50-66% ) were prepared from L3 of A. simplex (s.l.) collected from Lepidopus caudatus fished in the Mediterranean Sea. These proteins were electrophoretically separated and transferred into the nitrocellulose membranes. By WB, specific proteins recognized by positive control serum samples from sensitized patients were visualized on nitrocellulose membranes by a colorimetric reaction. The CWA and 50–66% fraction showed specific bands, mainly due to Ani s 1 (20-22 kD) and Ani s 4 (9-10 kD). So far, a total of 7 serum samples from people with allergic manifestation and positive skin prick test (SPT) after fish consumption, have been tested and all of them resulted negative by WB, indicating the lack of sensitization to anisakids. This preliminary study allowed to set up a specific test and evidence the lack of correlation between both tests, SPT and WB. However, the sample size should be increased to estimate the anisakidosis burden in Turkish people.

Keywords: anisakidosis, fish parasite, serodiagnosis, Turkey

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Authors: Maria C. Almario, Pam Remer, Jeff Resse, Kathy Moran, Linda Theander Adam

Abstract:

The identification of victims of human trafficking and consequential service provision is characterized by a significant disconnection between the estimated prevalence of this issue and the number of cases identified. This poses as tremendous problem for human rights advocates as it prevents data collection, information sharing, allocation of resources and opportunities for international dialogues. The current paper introduces the Inclusive Human Trafficking Checklist (IHTC) as a measurement methodology with theoretical underpinnings derived from dialectic theory. The presence of human trafficking in a person’s life is conceptualized as a dynamic and dialectic interaction between vulnerability and exploitation. The current papers explores the operationalization of exploitation and vulnerability, evaluates the metric qualities of the instrument, evaluates whether there are differences in assessment based on the participant’s profession, level of knowledge, and training, and assesses if users of the instrument perceive it as useful. A total of 201 participants were asked to rate three vignettes predetermined by experts to qualify as a either human trafficking case or not. The participants were placed in three conditions: business as usual, utilization of the IHTC with and without training. The results revealed a statistically significant level of agreement between the expert’s diagnostic and the application of the IHTC with an improvement of 40% on identification when compared with the business as usual condition While there was an improvement in identification in the group with training, the difference was found to have a small effect size. Participants who utilized the IHTC showed an increased ability to identify elements of identity-based vulnerabilities as well as elements of fraud, which according to the results, are distinctive variables in cases of human trafficking. In terms of the perceived utility, the results revealed higher mean scores for the groups utilizing the IHTC when compared to the business as usual condition. These findings suggest that the IHTC improves appropriate identification of cases and that it is perceived as a useful instrument. The application of the IHTC as a multidisciplinary instrumentation that can be utilized in legal and human services settings is discussed as a pivotal piece of helping victims restore their sense of dignity, and advocate for legal, physical and psychological reparations. It is noteworthy that this study was conducted with a sample in the United States and later re-tested in Colombia. The implications of the instrument for treatment conceptualization and intervention in human trafficking cases are discussed as opportunities for enhancement of victim well-being, restoration engagement and activism. With the idea that what is personal is also political, we believe that the careful observation and data collection in specific cases can inform new areas of human rights activism.

Keywords: exploitation, human trafficking, measurement, vulnerability, screening

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Authors: Nyathi Mpumelelo; Moeng Thabiso Maria

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BACKGROUND AND OBJECTIVES: Radiopharmaceuticals are used for diagnosis, treatment, staging and follow up of various diseases. However, there is concern that the ionizing radiation (gamma rays, α and ß particles) emitted by radiopharmaceuticals may result in exposure of radiographers and nurses with limited knowledge of the principles of radiation protection and safety, raising the risk of cancer induction. This study aimed at investigation radiation safety awareness levels among radiographers and nurses at a small tertiary hospital in South Africa. METHODS: An analytical cross-sectional study. A validated two-part questionnaire was implemented to consenting radiographers and nurses working in a Nuclear Medicine Department. Part 1 gathered demographic information (age, gender, work experience, attendance to/or passing ionizing radiation protection courses). Part 2 covered questions related to knowledge and awareness of radiation protection principles. RESULTS: Six radiographers and five nurses participated (27% males and 73% females). The mean age was 45 years (age range 20-60 years). The study revealed that neither professional development courses nor radiation protection courses are offered at the Nuclear Medicine Department understudy. However, 6/6 (100%) radiographers exhibited a high level of awareness of radiation safety principles on handling and working with radiopharmaceuticals which correlated to their years of experience. As for nurses, 4/5 (80%) showed limited knowledge and awareness of radiation protection principles irrespective of the number of years in the profession. CONCLUSION: Despite their major role of caring for patients undergoing diagnostic and therapeutic treatments, the nurses showed limited knowledge of ionizing radiation and associated side effects. This was not surprising since they never received any formal basic radiation safety course. These findings were not unique to this Centre. A study conducted in a Kuwaiti Radiology Department also established that the vast majority of nurses did not understand the risks of working with ionizing radiation. Similarly, nurses in an Australian hospital exhibited knowledge limitations. However, nursing managers did provide the necessary radiation safety training when requested. In Guatemala and Saudi Arabia, where there was shortage of professional radiographers, nurses underwent radiography training, a course that equipped them with basic radiation safety principles. The radiographers in the Centre understudy unlike others in various parts of the world demonstrated substantial knowledge and awareness on radiation protection. Radiations safety courses attended when an opportunity arose played a critical role in their awareness. The knowledge and awareness levels of these radiographers were comparable to their counterparts in Sudan. However, it was much more above that of their counterparts in Jordan, Nigeria, Nepal and Iran who were found to have limited awareness and inadequate knowledge on radiation dose. Formal radiation safety and awareness courses and workshops can play a crucial role in raising the awareness of nurses and radiographers on radiation safety for their personal benefit and that of their patients.

Keywords: radiation safety, radiation awareness, training, nuclear medicine

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Authors: Einat Waizbard, Emilio Ferrer, Meghan Miller, Brianna Heath, Derek S. Andrews, Sally J. Rogers, Christine Wu Nordahl, Marjorie Solomon, David G. Amaral

Abstract:

Background: Autism symptoms are comprised of social-communication deficits and restricted/repetitive behaviors (RRB). The severity of these symptoms can change during childhood, with differences between boys and girls. From the literature, it was found that young autistic girls show a stronger tendency to decrease and a weaker tendency to increase their overall autism symptom severity levels compared to young autistic boys. It is not clear, however, which symptoms are driving these sex differences across childhood. In the current study, we evaluated the trajectories of independent autism symptoms across childhood and compared the patterns of change in such symptoms between boys and girls. Method: The study included 183 children diagnosed with autism (55 girls) evaluated three times across childhood, at ages 3, 6 and 11. We analyzed 22 independent items from the Autism Diagnostic Observation Scheudule-2 (ADOS-2), the gold-standard assessment tool for autism symptoms, each item representing a specific autism symptom. First, we used latent growth curve models to estimate the trajectories for the 22 ADOS-2 items for each child in the study. Second, we extracted the factor scores representing the individual slopes for each ADOS-2 item (i.e., slope representing that child’s change in that specific item). Third, we used factor analysis to identify common patterns of change among the ADOS-2 items, separately for boys and girls, i.e., which autism symptoms tend to change together and which change independently across childhood. Results: The best-emerging patterns for both boys and girls identified four common factors: three factors representative of changes in social-communication symptoms and one factor describing changes in RRB. Boys and girls showed the same pattern of change in RRB, with four items (e.g., speech abnormalities) changing together across childhood and three items (e.g., mannerisms) changing independently of other items. For social-communication deficits in boys, three factors were identified: the first factor included six items representing initiating and engaging in social-communication (e.g., quality of social overtures, conversation), the second factor included five items describing responsive social-communication (e.g., response to name) and the third factor included three items related to different aspects of social-communication (e.g., level of language). Girls’ social-communications deficits also loaded onto three factors: the first factor included five items (e.g., unusual eye contact), the second factor included six items (e.g., quality of social response), and the third factor included four items (e.g., showing). Some items showed similar patterns of change for both sexes (e.g., responsive joint attention), while other items showed differences (e.g., shared enjoyment). Conclusions: Girls and boys had different patterns of change in autism symptom severity across childhood. For RRB, both sexes showed similar patterns. For social-communication symptoms, however, there were both similarities and differences between boys and girls in the way symptoms changed over time. The strongest patterns of change were identified for initiating and engaging in social communication for boys and responsive social communication for girls.

Keywords: autism spectrum disorder, autism symptom severity, symptom trajectories, sex differences

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Authors: Itzel Amaro-Estrada, Eduardo Vergara-Rivera, Virginia Juarez-Flores, Mayra Cobaxin-Cardenas, Rosa Estela Quiroz, Jesus F. Preciado, Sergio Rodriguez-Camarillo

Abstract:

Bovine anaplasmosis is an infectious, tick-borne disease caused mainly by Anaplasma marginale; typical signs include anemia, fever, abortion, weight loss, decreased milk production, jaundice, and potentially death. Sick bovine can recover when antibiotics are administered; however, it usually remains as carrier for life, being a risk of infection for susceptible cattle. Anaplasma marginale is an obligate intracellular Gram-negative bacterium with genetic composition highly diverse among geographical isolates. There are currently no vaccines fully effective against bovine anaplasmosis; therefore, the economic losses due to disease are present. Vaccine formulation became a hard task for several pathogens as Anaplasma marginale, but peptide-based vaccines are an interesting proposal way to induce specific responses. Phage-displayed peptide libraries have been proved one of the most powerful technologies for identifying specific ligands. Screening of these peptides libraries is also a tool for studying interactions between proteins or peptides. Thus, it has allowed the identification of ligands recognized by polyclonal antiserums, and it has been successful for the identification of relevant epitopes in chronic diseases and toxicological conditions. Protective immune response to bovine anaplasmosis includes high levels of immunoglobulins subclass G2 (IgG2) but not subclass IgG1. Therefore, IgG2 from the serum of protected bovine can be useful to identify ligands, which can be part of an immunogen for cattle. In this work, phage display random peptide library Ph.D. ™ -12 was incubating with IgG2 or blood sera of immunized bovines against A. marginale as targets. After three rounds of biopanning, several candidates were selected for additional analysis. Subsequently, their reactivity with sera immunized against A. marginale, as well as with positive and negative sera to A. marginale was evaluated by immunoassays. A collection of recognized peptides tested by ELISA was generated. More than three hundred phage-peptides were separately evaluated against molecules which were used during panning. At least ten different peptides sequences were determined from their nucleotide composition. In this approach, three phage-peptides were selected by their binding and affinity properties. In the case of the development of vaccines or diagnostic reagents, it is important to evaluate the immunogenic and antigenic properties of the peptides. Immunogenic in vitro and in vivo behavior of peptides will be assayed as synthetic and as phage-peptide for to determinate their vaccine potential. Acknowledgment: This work was supported by grant SEP-CONACYT 252577 given to I. Amaro-Estrada.

Keywords: bovine anaplasmosis, peptides, phage display, veterinary vaccines

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