Search results for: shRNA-library screening

Authors: Alex Eldo Simon, Abhishek Yadav

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This study aimed to evaluate the utility of postmortem computed tomography (CT) and heart weight measurements in the assessment of cardiomegaly in cases of sudden death due to cardiac origin by comparing the results of these two diagnostic methods. The study retrospectively analyzed postmortem computed tomography (PMCT) data from 54 cases of sudden natural death and compared the findings with those of the autopsy. The study involved measuring the cardiothoracic ratio (CTR) from coronal computed tomography (CT) images and determining the actual cardiac weight by weighing the heart during the autopsy. The inclusion criteria for the study were cases of sudden death suspected to be caused by cardiac pathology, while exclusion criteria included death due to unnatural causes such as trauma or poisoning, diagnosed natural causes of death related to organs other than the heart, and cases of decomposition. Sensitivity, specificity, and diagnostic accuracy were calculated, and to evaluate the accuracy of using the cardiothoracic ratio (CTR) to detect an enlarged heart, the study generated receiver operating characteristic (ROC) curves. The cardiothoracic ratio (CTR) is a radiological tool used to assess cardiomegaly by measuring the maximum cardiac diameter in relation to the maximum transverse diameter of the chest wall. The clinically used criteria for CTR have been modified from 0.50 to 0.57 for use in postmortem settings, where abnormalities can be detected by comparing CTR values to this threshold. A CTR value of 0.57 or higher is suggestive of hypertrophy but not conclusive. Similarly, heart weight is measured during the traditional autopsy, and a cardiac weight greater than 450 grams is defined as hypertrophy. Of the 54 cases evaluated, 22 (40.7%) had a cardiothoracic ratio (CTR) ranging from > 0.50 to equal 0.57, and 12 cases (22.2%) had a CTR greater than 0.57, which was defined as hypertrophy. The mean CTR was calculated as 0.52 ± 0.06. Among the 54 cases evaluated, the weight of the heart was measured, and the mean was calculated as 369.4 ± 99.9 grams. Out of the 54 cases evaluated, 12 were found to have hypertrophy as defined by PMCT, while only 9 cases were identified with hypertrophy in traditional autopsy. The sensitivity and specificity of the test were calculated as 55.56% and 84.44%, respectively. The sensitivity of the hypertrophy test was found to be 55.56% (95% CI: 26.66, 81.12¹), the specificity was 84.44% (95% CI: 71.22, 92.25¹), and the diagnostic accuracy was 79.63% (95% CI: 67.1, 88.23¹). The limitation of the study was a low sample size of only 54 cases, which may limit the generalizability of the findings. The comparison of the cardiothoracic ratio with heart weight in this study suggests that PMCT may serve as a screening tool for medico-legal autopsies when performed by forensic pathologists. However, it should be noted that the low sensitivity of the test (55.5%) may limit its diagnostic accuracy, and therefore, further studies with larger sample sizes and more diverse populations are needed to validate these findings.

Keywords: PMCT, virtopsy, CTR, cardiothoracic ratio

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Authors: Manohar Salla, Mark S. Butler, Ruby Pelingon, Geraldine Kaeslin, Daniel E. Croker, Janet C. Reid, Jong Min Baek, Paul V. Bernhardt, Elizabeth M. J. Gillam, Matthew A. Cooper, Avril A. B. Robertson

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MCC950 is a potent and selective inhibitor of the NOD-like receptor pyrin domain-containing protein 3 (NLRP3) inflammasome that shows early promise for treatment of inflammatory diseases. The identification of major metabolites of lead molecule is an important step during drug development process. It provides an information about the metabolically labile sites in the molecule and thereby helping medicinal chemists to design metabolically stable molecules. To identify major metabolites of MCC950, the compound was incubated with human liver microsomes and subsequent analysis by (+)- and (−)-QTOF-ESI-MS/MS revealed a major metabolite formed due to hydroxylation on 1,2,3,5,6,7-hexahydro-s-indacene moiety of MCC950. This major metabolite can lose two water molecules and three possible regioisomers were synthesized. Co-elution of major metabolite with each of the synthesized compounds using HPLC-ESI-SRM-MS/MS revealed the structure of the metabolite (±) N-((1-hydroxy-1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-4-(2-hydroxypropan-2-yl)furan-2-sulfonamide. Subsequent synthesis of individual enantiomers and coelution in HPLC-ESI-SRM-MS/MS using a chiral column revealed the metabolite was R-(+)- N-((1-hydroxy-1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-4-(2-hydroxypropan-2-yl)furan-2-sulfonamide. To study the possible cytochrome P450 enzyme(s) responsible for the formation of major metabolite, MCC950 was incubated with a panel of cytochrome P450 enzymes. The result indicated that CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C18, CYP2C19, CYP2J2 and CYP3A4 are most likely responsible for the formation of the major metabolite. The biological activity of the major metabolite and the other synthesized regioisomers was also investigated by screening for for NLRP3 inflammasome inhibitory activity and cytotoxicity. The major metabolite had 170-fold less inhibitory activity (IC50-1238 nM) than MCC950 (IC50-7.5 nM). Interestingly, one regioisomer had shown nanomolar inhibitory activity (IC50-232 nM). However, no evidence of cytotoxicity was observed with any of these synthesized compounds when tested in human embryonic kidney 293 cells (HEK293) and human liver hepatocellular carcinoma G2 cells (HepG2). These key findings give an insight into the SAR of the hexahydroindacene moiety of MCC950 and reveal a metabolic soft spot which could be blocked by chemical modification.

Keywords: Cytochrome P450, inflammasome, MCC950, metabolite, microsome, NLRP3

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Authors: Josephine Muchiri, Makena Muriithi

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Adverse mental health consequences are experienced by those that have been touched by Intimate Partner Violence (IPV), whether directly or indirectly. These negative effects are felt not only in the short term but in years to come. It is important to examine the prevalence and co-occurrence of mental disorders in order to provide strategic interventions for women who have experienced IPV. The aim of this study was to examine the prevalence and comorbidity of post-traumatic stress disorder (PTSD), Depression, and Anxiety among women who had experienced intimate Partner violence in two selected informal settlements in Nairobi County, Kenya. Participants were 116 women (15-60 years) selected through purposive and snowball sampling from the low social, economic settlements (Kawangware and Kibera) in Nairobi, Kenya. A social demographic questionnaire and the Woman Abuse Screening Tool (WAST) were used to collect data on intimate partner violence experiences. The PTSD Checklist for DSM-5 (PCL-5), Beck’s Depression Inventory, and the Beck’s Anxiety Inventory assessed for post-traumatic stress disorder, depression, and anxiety, respectively. Data analysis was conducted using the Statistical Package for Social Sciences (SPSS) version 29, utilizing descriptive and correlation analyses. Findings indicated that the women had undergone various forms of abuse from their intimate partners, which were physical abuse 111(92.5%), sexual abuse 70(88.6%), and verbal abuse 92(93.9%). The prevalence of the mental disorders was PTSD 47(32.4%); M= 44.11, S.D =14.67, depression was the highest at n=131(90.3%; M=33.37±9.98) with the levels of depression having varying prevalence rates where severe depression had the highest representation [moderate: n= 35; 24.1%, severe: n=69 (47.6%) and extremely severe: n=27(18.6%)]. Anxiety had the second highest prevalence of n=99 (68.8%; M= 28.55±13.63) with differing prevalence rates in the levels of anxiety which were normal anxiety: 45(31.3%), moderate anxiety n=62(43.1%) and severe anxiety: n=37(25.7%). Regarding comorbidities, the Pearson correlation test showed that there was a significant (p=0.000) positive relationship between PTSD and depression (r=0.379; p=.000), PTSD and anxiety (r=0.624; p=.000), and depression and anxiety (r=0.386; p=.000) such that increase in one disorder concomitantly led to increase of the other two disorders; hence comorbidity of the three disorders was ascertained. Conclusion: The study asserted the adverse impacts of IPV on women’s mental well-being, where the prevalence of PTSD, depression, and anxiety was established. Almost all the women had depressive symptoms; whereas more than half had anxiety and slightly more than a third had PTSD. Regarding the severity levels of anxiety and depression, almost half of the women with depression had severe depression whereas moderate anxiety was more prevalent for those with anxiety. The three disorders were found to co-occur where comorbidities of PTSD and anxiety had the highest probability of co-occurrence. It is thus recommended that mental health interventions with a focus on the three disorders be offered for women undergoing IPV.

Keywords: anxiety, comorbidity, depression, intimate partner violence, post-traumatic stress disorder

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Authors: Emílie Kučerová, Tereza Veverková, Marina Morozovová, Eva Kudová, Jitka Viktorová

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The blood-brain barrier (BBB) is a key element regulating the transport of substances between the blood and the central nervous system (CNS). The BBB protects the CNS from potentially harmful substances and maintains a suitable environment for nervous activity in the CNS, but at the same time, it represents a significant obstacle to the entry of drugs into the CNS. Pharmacoresistant epilepsy is a form of epilepsy that cannot be suppressed using two (or more) appropriately chosen antiepileptic drugs. In many cases, pharmacoresistant epilepsy is characterized by an increased concentration of efflux pumps on the luminal sides of the endothelial cells that form the BBB and an increased number of drug-metabolizing enzymes in the BBB cells, thereby preventing the effective transport of antiepileptic drugs into the CNS. Currently, a number of scientific groups are focusing on the preparation and improvement of BBB models in vitro in order to study cell interactions or transport mechanisms. However, in pathological conditions such as pharmacoresistant epilepsy, there are changes in BBB structure, and current BBB models are insufficient for related research. Our goal is to develop a suitable BBB model for pharmacoresistant epilepsy in vitro and use it to test the transfer of potential antiepileptic drugs. This model is created by co-culturing immortalized human cerebral microvascular endothelial cells, human vascular pericytes and immortalized human astrocytes. The BBB in vitro is cultivated in the form of a 2D transwell model and the integrity of the barrier is verified by measuring transendothelial electrical resistance (TEER). From the current results, a contact cell arrangement with the cultivation of endothelial cells on the upper side of the insert and the co-cultivation of astrocytes and pericytes on the lower side of the insert is selected as the most promising for BBB model cultivation. The pharmacoresistance of the BBB model is achieved by long-term cultivation of endothelial cells in an increasing concentration of selected antiepileptic drugs, which should lead to increased production of efflux pumps and drug-metabolizing enzymes. The pharmacoresistant BBB model in vitro will be further used for the screening of substances that could act both as antiepileptics and at the same time as inhibitors of efflux pumps in endothelial cells. This project was supported by the Technology Agency of the Czech Republic (TACR), Personalized Medicine: Translational research towards biomedical applications, No. TN02000109 and by the Academy of Sciences of the Czech Republic (AS CR) – grant RVO 61388963.

Keywords: antiepileptic drugs, blood-brain barrier, efflux transporters, pharmacoresistance

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Authors: Zhaozhang Sun

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Health communication is essential in promoting healthy lifestyles, preventing unhealthy behaviours, managing disease conditions, and eventually reducing health disparities. Nowadays, social media provides unprecedented opportunities for enhancing health communication for both healthcare providers and people with health conditions, including self-management of chronic conditions such as diabetes. Meanwhile, a special group of active social media users have started playing a pivotal role in providing health ‘solutions’. Such individuals are often referred to as ‘influencers’ because of their ‘central’ position in the online communication system and the persuasive effect their actions and advice may have on audiences' health-related knowledge, attitudes, confidence and behaviours. Work on social media influencers (SMIs) has gained much attention in a specific research field of “influencer marketing”, which mainly focuses on emphasising the use of SMIs to promote or endorse brands’ products and services in the business. Yet to date, a lack of well-studied and empirical evidence has been conducted to guide the exploration of health-related social media influencers. The failure to investigate health-related SMIs can significantly limit the effectiveness of communicating health on social media. Therefore, this article presents a study to identify key diabetes-related SMIs in the UK and the potential implications of information provided by identified social media influencers on their audiences’ diabetes-related knowledge, attitudes and behaviours to bridge the research gap that exists in linking work on influencers in marketing to health communication. The multidisciplinary theories and methods in social media, communication, marketing and diabetes have been adopted, seeking to provide a more practical and promising approach to investigate the potential of social media influencers in health communication. Twitter was chosen as the social media platform to initially identify health influencers and the Twitter API academic was used to extract all the qualitative data. Health-related Influencer Identification Model was developed based on social network analysis, analytic hierarchy process and other screening criteria. Meanwhile, a two-section English-version online questionnaire has been developed to explore the potential implications of social media influencers’ (SMI’s) diabetes-related narratives on the health-related knowledge, attitudes and behaviours (KAB) of their audience. The paper is organised as follows: first, the theoretical and research background of health communication and social media influencers was discussed. Second, the methodology was described by illustrating the model for the identification of health-related SMIs and the development process of the SMIKAB instrument, followed by the results and discussions. The limitations and contributions of this study were highlighted in the summary.

Keywords: health communication, Interdisciplinary research, social media influencers, diabetes management

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Authors: Lesley Northrop, Justin DeGrazia, Jessica Greenwood

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ASPIRA Labs now offers an en-suit and ready-to-implement technology transfer solution to enable labs and hospitals that lack the resources to build it themselves to offer in-house genetic testing. This unique platform employs a patented Molecular Inversion Probe (MIP) technology that combines the specificity of a hybrid capture protocol with the ease of an amplicon-based protocol and utilizes an advanced bioinformatics analysis pipeline based on machine learning. To demonstrate its efficacy, two independent genetic tests were validated on this technology transfer platform: expanded carrier screening (ECS) and hereditary cancer testing (HC). The analytical performance of ECS and HC was validated separately in a blinded manner for calling three different types of variants: SNVs, short indels (typically, <50 bp), and large indels/CNVs defined as multi-exonic del/dup events. The reference set was constructed using samples from Coriell Institute, an external clinical genetic testing laboratory, Maine Molecular Quality Controls Inc. (MMQCI), SeraCare and GIAB Consortium. Overall, the analytical performance showed a sensitivity and specificity of >99.4% for both ECS and HC in detecting SNVs. For indels, both tests reported specificity of 100%, and ECS demonstrated a sensitivity of 100%, whereas HC exhibited a sensitivity of 96.5%. The bioinformatics pipeline also correctly called all reference CNV events resulting in a sensitivity of 100% for both tests. No additional calls were made in the HC panel, leading to a perfect performance (specificity and F-measure of 100%). In the carrier panel, however, three additional positive calls were made outside the reference set. Two of these calls were confirmed using an orthogonal method and were re-classified as true positives leaving only one false positive. The pipeline also correctly identified all challenging carrier statuses, such as positive cases for spinal muscular atrophy and alpha-thalassemia, resulting in 100% sensitivity. After confirmation of additional positive calls via long-range PCR and MLPA, specificity for such cases was estimated at 99%. These performance metrics demonstrate that this tech-transfer solution can be confidently internalized by clinical labs and hospitals to offer mainstream ECS and HC as part of their test catalog, substantially increasing access to quality germline genetic testing for labs of all sizes and resources levels.

Keywords: clinical genetics, genetic testing, molecular genetics, technology transfer

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Authors: Allulu Yousef Alturki, Raghad Abdullah Alshafi, Sara Abdulaziz Alghashem, Sahar Saleh Alghamdi, Rasha Saad Suliman, Zeyad Alehaideb, Rizwan Ali

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Cancer is recognized as a worldwide public health concern. Therefore, there is a continuous quest to discover new effective medications with less side-effects. In recent years, researchers have shown an increased interest in medicinal plants as several plant species have shown promising biological activities. Thus, we seek to investigate three medicinal herbs that are commonly-found in the Middle Easternregion and yet have not been explored in depth, including plants belonging to the Rhamnaceae, Polygonaceae, and Apocynaceaeplant families. Initially, we investigated using three types of cancer cell lines for breast, colorectal, and liver cancers. We performed high Content Imaging (HCI)-Apoptosis Assay and ApoTox-Glo™ Triplex Assay on KAIMRC2 and HCT8 cell lines. The highest activity of HCI-Apoptosis Assay was with Calligonumcomosum and Ziziphusnummularia in ethanol, followed by Calotropis procera and Ziziphusnummularia in ethyl acetate. The IC50values for the families of Rhamnaceae, Polygonaceae, and Apocynaceae in HepG2 and HCT8 cell lines ranged from 0.089 to 9.84mg/mL and 0.080to 15.08mg/mL, respectively. Further screening was conducted on an additional two cell lines, namely the MDA-MB-231 and KAIMRC2, for selected seven extracts with the highest activity having IC50values ranged from 0.058 to0.51mg/mL and 0.029 to0.19mg/mL, respectively. Continuous scientific investigations to isolate and characterize the potent bioactive phytochemical(s) are warranted. Funding: The authors acknowledge financial support from King Abdullah International Medical Research Center (KAIMRC), Ministry of National Guard Health Affairs, Riyadh, Kingdom of Saudi Arabia. Institutional Review Board Statement: The study was approved by the Institutional Review Board of the Institutional Review Board of King Abdullah International Medical Research Center (SP21R/463/12, 24 January 2022). Acknowledgments: The authors want to express their gratitude to the College of Pharmacy (COP) at King Saud bin Abdulaziz University for Health Sciences (KSAU-HS) and King Abdullah International Medical Research Center (KAIMRC) for their continued support.

Keywords: rhamnaceae, polygonaceae, apocynaceae, natural products

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Authors: Zachary Muthii Rukenya, James Mbaria, Peter Mbaabu, Kiama Stephen Gitahi, Ronald Onzago

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Aloe turkanensis is one of the widely used medicinal shrub and in Kenya the plant is mainly found in Baringo, Isiolo, Laikipia, Turkana and West Pokot Counties where it is used in ethno-medicine and ethno-veterinary medicine. The Turkana community uses the plant products to manage malaria, wounds, stomach ache, constipation, pain, skin infection, poultry diseases and retained afterbirth in cows. This evaluated the efficacy and safety of the plant obtained from Turkana County, Kenya. Preliminary data on the use of the plant in the county was collected through observation, photographing and interviews. A sample of the whole plant was harvested in Natira sublocation, in ex-Turkana west district in February 2012 after identification by Aloe-working group herbalists who voluntarily provided information on its medicinal uses. Botanical identification was done at Kenya Forest Research Centre in Karura where voucher specimen was deposited. Cold maceration using 70% methanol and distilled water was used for extraction. Bioassays were to determine the effects of the plant extracts on brine shrimp and selected bacterial and fungal cultures. The extracts were tested in-vitro activity against standard cultures of B. cereus (ATCC 11778), S. aureus (ATCC25923), P. aeroginosa (ATCC 27853), E. coli (ATCC 25922) and a human infections clinical isolate of C. albicans. The extracts of Aloe turkanensis inhibited the growth B. cereus (100-200 mg/ml), S. aureus (50-100 mg/ml), P. aeroginosa (200mg/ml), E. coli (400mg/ml) while C. albicans was not affected. The extracts also inhibited the growth of S. aureus and B. cereus with mean diameters of inhibition zones being 19.75±1 mm and 18.5±05 mm reapectively. Phytochemical screening showed the presence of alkaloids, tarpenoids, steroids, quinones, saponins and tannins in the plant extracts. The extract was found to be non-toxic at a concentration of 1000µg/ml with a 100% survival of Brine Shrimp larva. It was concluded that Aloe turkanensis growing the study area has metabolites that inhibit the growth of microorganisms and is however, there is need for further studies to validate the in-vivo bioactivity of the plant and more generate adequate toxicological data.to support conservation, value chain addition of its products and widespread use as a herbal remedy.

Keywords: Aloe turkanensis, bioactivity, cultivated, human infections

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Authors: S. Cheng, C. Catallo

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Introduction: Canada's healthcare and social services systems are failing high risk, vulnerable older adults. Care for vulnerable older Canadians (65 and older) is not optimal in Canada. It does not address the care needs of vulnerable, high risk adults using a holistic approach. Given the growing aging population, and the care needs for seniors with complex conditions is one of the highest in Canada's health care system, there is a sense of urgency to optimize care. Integration of health and social services is an emerging trend in Canada when compared to European countries. There is no common and universal understanding of healthcare and social services integration within the country. Consequently, a clear understanding and definition of integrated health and social services are absent in Canada. Objectives: A study was undertaken to develop a case definition for integrated health and social care initiatives that serve older adults, which was then tested against three Canadian integrated initiatives. Methodology: A limited literature review was undertaken to identify common characteristics of integrated health and social care initiatives that serve older adults, and comprised both scientific and grey literature, in order to develop a case definition. Three Canadian integrated initiatives that are located in the province of Ontario, were identified using an online search and a screening process. They were surveyed to determine if the literature-based integration definition applied to them. Results: The literature showed that there were 24 common healthcare and social services integration characteristics that could be categorized into ten themes: 1) patient-care approach; 2) program goals; 3) measurement; 4) service and care quality; 5) accountability and responsibility; 6) information sharing; 7) Decision-making and problem-solving; 8) culture; 9) leadership; and 10) staff and professional interaction. The three initiatives showed agreement on all the integration characteristics except for those characteristics associated with healthcare and social care professional interaction, collaborative leadership and shared culture. This disagreement may be due to several reasons, including the existing governance divide between the healthcare and social services sectors within the province of Ontario that has created a ripple effect in how professions in the two different sectors interact. In addition, the three initiatives may be at maturing levels of integration, which may explain disagreement on the characteristics associated with leadership and culture. Conclusions: The development of a case definition for healthcare and social services integration that incorporates common integration characteristics can act as a useful instrument in identifying integrated healthcare and social services, particularly given the emerging and evolutionary state of this phenomenon within Canada.

Keywords: Canada, case definition, healthcare and social services integration, integration, seniors health, services delivery

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Authors: Muhammad Bilal, Amelia Jane Llyod, Habsah Mohamad, Jia Hui Wong, Abdul Aziz Mohamed Yusoff, Jafri Malin Abdullah, Jingli Zhang

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Epilepsy is one of the major brain afflictions occurs with uncontrolled excitation of cortex; disturbed 50 million of world’s population. About 25 percent of patients subjected to adverse effects from antiepileptic drugs (AEDs) such as depression, nausea, tremors, gastrointestinal symptoms, osteoporosis, dizziness, weight change, drowsiness, fatigue are commonly observed indications; therefore, new drugs are required to cure epilepsy. GABA is principle inhibitory neurotransmitter, control excitation of the brain. Mutation or dysfunction of GABA receptor is one of the primary causes of epilepsy, which is confirmed from many acquired models of epilepsy like traumatic brain injury, kindling, and status epilepticus models of epilepsy. GABA receptor has 3 distinct types such as GABA (A), GABA (B), GABA(C).GABA (A) receptor has 20 different subunits, α1β2γ2 subunits composition of GABA (A) receptor is the most used combination of subunits for screening of compounds against epilepsy. We expressed α1β2γ2s subunits of GABA (A) Receptor in Xenopus leavis oocytes and examined the enhancement potential of 4-Hydroxybenzoic acid compound on GABA (A) receptor via two-electrode voltage clamp current recording technique. Bamboo shoots are the young, tender offspring of bamboo, which are usually harvested after a cultivating period of 2 weeks. Proteins, acids, fat, starch, carbohydrate, fatty acid, vitamin, dietary fiber, and minerals are the major constituent found systematically in bamboo shoots. These shoots reported to have anticancer, antiviral, antibacterial activity, also possess antioxidant properties due to the presence of phenolic compounds. Student t-test analysis suggested that 4- hydroxybenzoic acid positively allosteric GABA (A) receptor, increased normalized current amplitude to 1.0304±0.0464(p value 0.032) compared with vehicle. 4-Hydrobenzoic acid, a compound from Dendrocalamus Asper bamboo shoot gives new insights for future studies on bamboo shoots with motivation for extraction of more compounds to investigate their effects on human and rodents against epilepsy, insomnia, and anxiety.

Keywords: α1β2γ2S, antiepileptic, bamboo shoots, epilepsy GABA (A) receptor, two-microelectrode voltage clamp, xenopus laevis oocytes

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Authors: Mariam Dianat Sabet Gilani, Lars M. Blank, Birgitta E. Ebert

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Plant-derived lupane-type, pentacyclic triterpenoids are biologically active compounds that are highly interesting for applications in medical, pharmaceutical, and cosmetic industries. Due to the low abundance of these valuable compounds in their natural sources, and the environmentally harmful downstream process, alternative production methods, such as microbial cell factories, are investigated. Engineered Saccharomyces cerevisiae strains, harboring the heterologous genes for betulinic acid synthesis, can produce up to 2 g L-1 triterpenoids, showing high potential for large-scale production of triterpenoids. One limitation of the microbial synthesis is the intracellular product accumulation. It not only makes cell disruption a necessary step in the downstream processing but also limits productivity and product yield per cell. To overcome these restrictions, the aim of this study is to develop an in-situ extraction method, which extracts triterpenoids into a second organic phase. Such a continuous or sequential product removal from the biomass keeps the cells in an active state and enables extended production time or biomass recycling. After screening of twelve different solvents, selected based on product solubility, biocompatibility, as well as environmental and health impact, isopropyl myristate (IPM) was chosen as a suitable solvent for in-situ product removal from S. cerevisiae. Impedance-based single-cell analysis and off-gas measurement of carbon dioxide emission showed that cell viability and physiology were not affected by the presence of IPM. Initial experiments demonstrated that after the addition of 20 vol % IPM to cultures in the stationary phase, 40 % of the total produced triterpenoids were extracted from the cells into the organic phase. In future experiments, the application of IPM in a repeated batch process will be tested, where IPM is added at the end of each batch run to remove triterpenoids from the cells, allowing the same biocatalysts to be used in several sequential batch steps. Due to its high biocompatibility, the amount of IPM added to the culture can also be increased to more than 20 vol % to extract more than 40 % triterpenoids in the organic phase, allowing the cells to produce more triterpenoids. This highlights the potential for the development of a continuous large-scale process, which allows biocatalysts to produce intracellular products continuously without the necessity of cell disruption and without limitation of the cell capacity.

Keywords: betulinic acid, biocompatible solvent, in-situ extraction, isopropyl myristate, process development, secondary metabolites, triterpenoids, yeast

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Authors: A. Mulry, C. Kealey, D. B. Brady

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Planktonic bacteria can form biofilms which are microbial aggregates embedded within a matrix of extracellular polymeric substances (EPS). They can be found attached to abiotic or biotic surfaces. Biofilms are responsible for oral diseases such as dental caries, gingivitis and the progression of periodontal disease. Biofilms can resist 500 to 1000 times the concentration of biocides and antibiotics used to kill planktonic bacteria. Biofilm development on oral surfaces involves four stages, initial attachment, early development, maturation and dispersal of planktonic cells. The Minimum Inhibitory Concentration (MIC) was determined using a range of saturated and unsaturated fatty acids using the resazurin assay, followed by serial dilution and spot plating on BHI agar plates to establish the Minimum Bactericidal Concentration (MBC). Log reduction of bacteria was also evaluated for each fatty acid. The Minimum Biofilm Inhibition Concentration (MBIC) was determined using crystal violet assay in 96 well plates on forming and pre-formed S. mutans biofilms using BHI supplemented with 1% sucrose. Saturated medium-chain fatty acids Octanoic (C8.0), Decanoic (C10.0) and Undecanoic acid (C11.0) do not display strong antibiofilm properties; however, Lauric (C12.0) and Myristic (C14.0) display moderate antibiofilm properties with 97.83% and 97.5% biofilm inhibition with 1000 µM respectively. Monounsaturated, Oleic acid (C18.1) and polyunsaturated large chain fatty acids, Linoleic acid (C18.2) display potent antibiofilm properties with biofilm inhibition of 99.73% at 125 µM and 100% at 65.5 µM, respectively. Long-chain polyunsaturated Omega-3 fatty acids α-Linoleic (C18.3), Eicosapentaenoic Acid (EPA) (C20.5), Docosahexaenoic Acid (DHA) (C22.6) have displayed strong antibiofilm efficacy from concentrations ranging from 31.25-250µg/ml. DHA is the most promising antibiofilm agent with an MBIC of 99.73% with 15.625µg/ml. This may be due to the presence of six double bonds and the structural orientation of the fatty acid. To conclude, fatty acids displaying the most antimicrobial activity appear to be medium or long-chain unsaturated fatty acids containing one or more double bonds. Most promising agents include Omega-3-fatty acids Linoleic, α-Linoleic, EPA and DHA, as well as Omega-9 fatty acid Oleic acid. These results indicate that fatty acids have the potential to be used as antimicrobials and antibiofilm agents against S. mutans. Future work involves further screening of the most potent fatty acids against a range of bacteria, including Gram-positive and Gram-negative oral pathogens. Future work will involve incorporating the most effective fatty acids onto dental implant devices to prevent biofilm formation.

Keywords: antibiofilm, biofilm, fatty acids, S. mutans

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Authors: M. Gowri, E. K. Girija, V. Ganesh

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Background and Significance: Among the dental infections, inflammation and infection of the root canal are common among all age groups. Currently, the management of root canal infections involves cleaning the canal with powerful irrigants followed by intracanal medicament application. Though these treatments have been in vogue for a long time, root canal failures do occur. Treatment for root canal infections is limited due to the anatomical complexity in terms of small micrometer volumes and poor penetration of drugs. Thus, infections of the root canal seem to be a challenge that demands development of new agents that can eradicate C. albicans. Methodology: In the present study, we synthesized and screened silver-curcumin nanoparticle against Candida albicans. Detailed molecular studies were carried out with silver-curcumin nanoparticle on C. albicans pathogenicity. Morphological cell damage and antibiofilm activity of silver-curcumin nanoparticle on C. albicans was studied using scanning electron microscopy (SEM). Biochemical evidence for membrane damage was studied using flow cytometry. Further, the antifungal activity of silver-curcumin nanoparticle was evaluated in an ex vivo dentinal tubule infection model. Results: Screening data showed that silver-curcumin nanoparticle was active against C. albicans. Silver-curcumin nanoparticle exerted time kill effect and post antifungal effect. When used in combination with fluconazole or nystatin, silver-curcumin nanoparticle revealed a minimum inhibitory concentration (MIC) decrease for both drugs used. In-depth molecular studies with silver-curcumin nanoparticle on C. albicans showed that silver-curcumin nanoparticle inhibited yeast to hyphae (Y-H) conversion. Further, SEM images of C. albicans showed that silver-curcumin nanoparticle caused membrane damage and inhibited biofilm formation. Biochemical evidence for membrane damage was confirmed by increased propidium iodide (PI) uptake in flow cytometry. Further, the antifungal activity of silver-curcumin nanoparticle was evaluated in an ex vivo dentinal tubule infection model, which mimics human tooth root canal infection. Confocal laser scanning microscopy studies showed eradication of C. albicans and reduction in colony forming unit (CFU) after 24 h treatment in the infected tooth samples in this model. Conclusion: The results of this study can pave the way for developing new antifungal agents with well deciphered mechanisms of action and can be a promising antifungal agent or medicament against root canal infection.

Keywords: C. albicans, ex vivo dentine model, inhibition of biofilm formation, root canal infection, yeast to hyphae conversion inhibition

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Authors: Badryah Alshehri, Kevin Gormley, Gillian Prue, Karen McCutcheon

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In a disaster event, sharing patient information between the pre-hospital Emergency Medical Services (EMS) and Emergency Department (ED) hospitals is a complex process during which important information may be altered or lost due to poor communication. The aim of this study was to critically discuss the current evidence base in relation to communication between pre- EMS hospital and ED hospital professionals by the use of Information and Communication Systems (ICT). This study followed the systematic approach; six electronic databases were searched: CINAHL, Medline, Embase, PubMed, Web of Science, and IEEE Xplore Digital Library were comprehensively searched in January 2018 and a second search was completed in April 2020 to capture more recent publications. The study selection process was undertaken independently by the study authors. Both qualitative and quantitative studies were chosen that focused on factors that are positively or negatively associated with coordinated communication between pre-hospital EMS and ED teams in a disaster event. These studies were assessed for quality, and the data were analyzed according to the key screening themes which emerged from the literature search. Twenty-two studies were included. Eleven studies employed quantitative methods, seven studies used qualitative methods, and four studies used mixed methods. Four themes emerged on communication between EMTs (pre-hospital EMS and ED staff) in a disaster event using the ICT. (1) Disaster preparedness plans and coordination. This theme reported that disaster plans are in place in hospitals, and in some cases, there are interagency agreements with pre-hospital and relevant stakeholders. However, the findings showed that the disaster plans highlighted in these studies lacked information regarding coordinated communications within and between the pre-hospital and hospital. (2) Communication systems used in the disaster. This theme highlighted that although various communication systems are used between and within hospitals and pre-hospitals, technical issues have influenced communication between teams during disasters. (3) Integrated information management systems. This theme suggested the need for an integrated health information system that can help pre-hospital and hospital staff to record patient data and ensure the data is shared. (4) Disaster training and drills. While some studies analyzed disaster drills and training, the majority of these studies were focused on hospital departments other than EMTs. These studies suggest the need for simulation disaster training and drills, including EMTs. This review demonstrates that considerable gaps remain in the understanding of the communication between the EMS and ED hospital staff in relation to response in disasters. The review shows that although different types of ICTs are used, various issues remain which affect coordinated communication among the relevant professionals.

Keywords: emergency medical teams, communication, information and communication technologies, disaster

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Authors: Obiageli Helen Ezeh, Chuks Clement Ezeh

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Background: It is estimated that more than 330 million persons are living with HIV-infection globally and in Nigeria about 3.4 persons are living with the infection, with an annual death rate of 180,000. Psychological morbidity often accompany chronic illnesses and may be associated with substance abuse, poor health seeking behavior and adherence to treatment program; it may worsen existing health problems and the overall quality of life. Until the burden is effectively identified, intervention cannot be planned. Until there is a cure, the goal is to manage and cope effectively with HIV-infection. Little if any studies have been done in this area in the North West geo-political zone of Nigeria. The study would help to identify high risk groups and prevent the progression and spread of the infection. Aim: To identify HIV-infected persons with psychological morbidity, accessing HIV- clinic at Shika Hospital, Zaria, Kaduna State; and analyze their socio-demographic profile. Methods: A cross sectional descriptive study was carried out to assess and analyze the socio-demographic characteristics of HIV-infected persons attending Shika hospital Zaria Nigeria, who screened positive for psychological morbidity. A total of 109 HIV-infected persons receiving HAART at Shika clinic, Zaria, Kaduna State, Nigeria, were administered questionnaires, the General Health Questionnaire (GHQ-12)measuring psychological morbidity and socio-demographic data. The participants ranged in age between 18 and 75 years. Results: Data were analyzed using SPSS software 15. Both descriptive and inferential Statistics were performed on the data. Results indicate a total prevalent rate of psychological morbidity of 78 percent among participants. Of this, about 16.2 percent were severely distressed, 25.1 percent moderately distressed and 36.7percent were mildly distressed. More females (65 percent of those with psychological morbidity) were found to be distressed than their male (55 percent) counterparts. It was (44 percent) for patients whose HIV-infection was of relatively shorter duration(2-4 years) than those of longer duration(5-9 years; and 10 years/above). The age group (21-30 years) was the most affected (35 percent). The rate was also 55 percent for Christians and 45 percent for Muslims. For married patients with partners it was 20 percent and for singles 30 percent; for the widowed (12 percent) and divorced (38 percent). At the level of tribal/ethnic groups, it was 13 percent for Ibos, 22 percent for Yorubas, 27 percent for Hausas and 33 percent for all the other minority tribes put together. Conclusion/Recommendation: The study has been able to identify the presence of psychological morbidity among HIV-infected persons as high and analyze the socio-demographic factors associated with it as significant. Periodic screening of HIV-infected persons for psychological morbidity and psychosocial intervention was recommended.

Keywords: socio-demographics, psychological morbidities, HIV-Infection, HAART

Procedia PDF Downloads 229

Authors: Lakmini Wijesooriya, Vicki Chalker, Jessica Day, Priyantha Perera, N. P. Sunil-Chandra

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M. pneumoniae has been identified as an etiology for exacerbation of asthma (EQA), although viruses play a major role in EOA. M. pneumoniae infection is treated empirically with macrolides, and its antibiotic sensitivity is not detected routinely. Characterization of the organism by genotyping and determination of macrolide resistance is important epidemiologically as it guides the empiric antibiotic treatment. To date, there is no such characterization of M. pneumoniae performed in Sri Lanka. The present study describes the characterization of M. pneumoniae detected from a child with EOA following a screening of 100 children with EOA. Of the hundred children with EOA, M. pneumoniae was identified only in one child by Real-Time polymerase chain reaction (PCR) test for identifying the community-acquired respiratory distress syndrome (CARDS) toxin nucleotide sequences. The M. pneumoniae identified from this patient underwent detection of macrolide resistance via conventional PCR, amplifying and sequencing the region of the 23S rDNA gene that contains single nucleotide polymorphisms that confer resistance. Genotyping of the isolate was performed via nested Multilocus Sequence Typing (MLST) in which eight (8) housekeeping genes (ppa, pgm, gyrB, gmk, glyA, atpA, arcC, and adk) were amplified via nested PCR followed by gene sequencing and analysis. As per MLST analysis, the M. pneumoniae was identified as sequence type 14 (ST14), and no mutations that confer resistance were detected. Resistance to macrolides in M. pneumoniae is an increasing problem globally. Establishing surveillance systems is the key to informing local prescriptions. In the absence of local surveillance data, antibiotics are started empirically. If the relevant microbiological samples are not obtained before antibiotic therapy, as in most occasions in children, the course of antibiotic is completed without a microbiological diagnosis. This happens more frequently in therapy for M. pneumoniae which is treated with a macrolide in most patients. Hence, it is important to understand the macrolide sensitivity of M. pneumoniae in the setting. The M. pneumoniae detected in the present study was macrolide sensitive. Further studies are needed to examine a larger dataset in Sri Lanka to determine macrolide resistance levels to inform the use of macrolides in children with EOA. The MLST type varies in different geographical settings, and it also provides a clue to the existence of macrolide resistance. The present study enhances the database of the global distribution of different genotypes of M. pneumoniae as this is the first such characterization performed with the increased number of samples to determine macrolide resistance level in Sri Lanka. M. pneumoniae detected from a child with exacerbation of asthma in Sri Lanka was characterized as ST14 by MLST and no mutations that confer resistance were detected.

Keywords: mycoplasma pneumoniae, Sri Lanka, characterization, macrolide resistance

Procedia PDF Downloads 157

Authors: Yunjia Yang, Peng Li, Gordon Xu, Timothy Mahony, Bing Zhang, Neena Mitter, Karishma Mody

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Sheep flystrike is one of the most economically important diseases affecting the Australian sheep and wool industry (>356M/annually). Currently, control of Lucillia cuprina relies almost exclusively on chemicals controls, and the parasite has developed resistance to nearly all control chemicals used in the past. It is, therefore, critical to develop an alternative solution for the sustainable control and management of flystrike. RNA interference (RNAi) technologies have been successfully explored in multiple animal industries for developing parasites controls. This research project aims to develop a RNAi based biological control for sheep blowfly. Double-stranded RNA (dsRNA) has already proven successful against viruses, fungi, and insects. However, the environmental instability of dsRNA is a major bottleneck for successful RNAi. Bentonite polymer (BenPol) technology can overcome this problem, as it can be tuned for the controlled release of dsRNA in the gut challenging pH environment of the blowfly larvae, prolonging its exposure time to and uptake by target cells. To investigate the potential of BenPol technology for dsRNA delivery, four different BenPol carriers were tested for their dsRNA loading capabilities, and three of them were found to be capable of affording dsRNA stability under multiple temperatures (4°C, 22°C, 40°C, 55°C) in sheep serum. Based on stability results, dsRNA from potential targeted genes was loaded onto BenPol carriers and tested in larvae feeding assays, three genes resulting in knockdowns. Meanwhile, a primary blowfly embryo cell line (BFEC) derived from L. cuprina embryos was successfully established, aim for an effective insect cell model for testing RNAi efficacy for preliminary assessments and screening. The results of this study establish that the dsRNA is stable when loaded on BenPol particles, unlike naked dsRNA rapidly degraded in sheep serum. The stable nanoparticle delivery system offered by BenPol technology can protect and increase the inherent stability of dsRNA molecules at higher temperatures in a complex biological fluid like serum, providing promise for its future use in enhancing animal protection.

Keywords: lucilia cuprina, primary cell line establishment, RNA interference, insect cell transfection

Procedia PDF Downloads 51

Authors: Sugandha Asthana, Geetika Vajpayee, Pratibha Kumari, Shanthy Sundaram

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An economically important disease of wheat in North Western region of India is Karnal Bunt caused by smut fungus Tilletia indica. This fungal pathogen spreads by air, soil and seed borne sporodia at the time of flowering, which ultimately leads to partial bunting of wheat kernels with fishy odor and taste to wheat flour. It has very serious effects due to quarantine measures which have to be applied for grain exports. Chemical fungicides such as mercurial compounds and Propiconazole applied to the control of Karnal bunt have been only partially successful. Considering the harmful effects of chemical fungicides on man as well as environment, many countries are developing biological control as the superior substitute to chemical control. Repeated use of fungicides can be responsible for the development of resistance in fungal pathogens against certain chemical compounds. The present investigation is based on the isolation and evaluation of antifungal properties of some isolated (from natural manure) and commercial bacterial strains against Tilletia indica. Total 23 bacterial isolates were obtained and antagonistic activity of all isolates and commercial bacterial strains (Bacillus subtilis MTCC8601, Bacillus pumilus MTCC 8743, Pseudomonas aeruginosa) were tested against T. indica by dual culture plate assay (pour plate and streak plate). Test for the production of antifungal volatile organic compounds (VOCs) by antagonistic bacteria was done by sealed plate method. Amongst all s1, s3, s5, and B. subtilis showed more than 80% inhibition. Production of extracellular hydrolytic enzymes such as protease, beta 1, 4 glucanase, HCN and ammonia was studied for confirmation of antifungal activity. s1, s3, s5 and B. subtilis were found to be the best for protease activity and s5 and B. subtilis for beta 1, 4 glucanase activity. Bacillus subtilis was significantly effective for HCN whereas s3, s5 and Bacillus subtilis for ammonia production. Isolates were identified as Pseudomonas aeruginosa (s1) and B. licheniformis (s3, s5) by various biochemical assays and confirmed by16s rRNA sequencing. Use of microorganisms or their secretions as biocontrol agents to avoid plant diseases is ecologically safe and may offer long term of protection to crop. The above study reports the promising effects of these strains in better pathogen free crop production and quality maintenance as well as prevention of the excessive use of synthetic fungicides.

Keywords: antagonistic, antifungal, biocontrol, Karnal bunt

Procedia PDF Downloads 255

Authors: Razvan Mercut, Mihaela Ionescu, Vlad Parvanescu, Razvan Ghita, Tudor-Gabriel Caragea, Cristina Simionescu, Marius-Eugen Ciurea

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Introduction: Over the past years, the incidence of non-melanoma skin cancer (NMSC) has continuously increased, being one of the most commonly diagnosed carcinomasofthe cephalic extremity. NMSC regroups basal cell carcinoma (BCC), squamous cell carcinoma (SCC), Merkel cell carcinoma, cutaneous lymphoma, and sarcoma. The most common forms are BCC and SCC, both still implying a significant level of morbidity due to local invasion (especially BCC), even if the overall death rates are declining. The objective of our study was the evaluation of clinical and histological aspects of NMSC for a group of patients with BCC and SCC, from Craiova, a south-western major city in Romania. Materialand method: Our study lot comprised 65 patients, with an almost equal distribution of sexes, and ages between 23-91 years old (mean value±standard deviation62.61±16.67), all treated within the Clinic of Plastic Surgery and Reconstructive Microsurgery, Clinical Emergency County Hospital Craiova, Romania, between 2019-2020. In order to determine the main morphological characteristics of both studied cancers, we used paraffin embedding techniques, with various staining methods:hematoxylin-eosin, Masson's trichrome stain with aniline blue, and Periodic acid-schiffAlcian Blue. The statistical study was completed using Microsoft Excel (Microsoft Corp., Redmond, WA, USA), with XLSTAT (Addinsoft SARL, Paris, France). Results: The overall results of our study indicate that BCC accounts for 67.69% of all NMSC forms; SCC covers 27.69%, while 4.62% are representedby other forms. The most frequent site is the nose for BCC (27.69%, 18 patients), being followed by preauricular regions, forehead, and periorbital areas. For patients with SCC, tumors were mainly located at lips level (66.67%, 12 patients). The analysis of NMSC histological forms indicated that nodular BCC is predominant (45.45%, 20 patients), as well as ulcero-vegetant SCC (38.89%, 7 patients). We have not identified any topographic characteristics or NMSC forms significantly related to age or sex. Conclusions: The most frequent NMSC form identified for our study lot was BCC. The preferred location was the nose for BCC. For SCC, the oral cavity is the most frequent anatomical site, especially the lips level. Nodular BCC and ulcero-vegetant SCC were the most commonly identified histological types. Our findings emphasize the need for periodic screening, in order to improve prevention and early treatment for these malignancies.

Keywords: non-melanoma skin cancer, basal cell carcinoma, squamous cell carcinoma, histological

Procedia PDF Downloads 156

Authors: Maryam M. Bashir, Luai A. Ahmed, Meera R. Alshamsi, Sara Almahrooqi, Taif Alyammahi, Shooq A. Alshehhi, Waad I. Alhammadi, Fatima H. Alhammadi, Hind A. Alhosani, Rami H. Al-Rifai, Fatma Al-Maskari

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Obesity in the Middle East and North Africa (MENA) region has exponentially increased over the past five decades due to rapid urbanization and unhealthy lifestyle changes. It has been well established that overweight and obesity increase the risk of non-communicable diseases (NCDs) and are the leading cause of mortality and economic burden locally, and globally. In the United Arab Emirates (UAE), there is a growing epidemic of obesity and other chronic diseases like type 2 diabetes mellitus and cardiovascular diseases. Prevalence of overweight and obesity in UAE range up to 70% depending on the group being studied. Hence, there is a need to explore their patterns in the country for more targeted and responsive interventions. Our study aimed to explore the patterns of overweight and obesity and some self-reported chronic diseases among university students in Abu Dhabi, the capital city of UAE. A validated online self-administered questionnaire was used to collect data from UAE University (UAEU) students, 18years and above, from August to September 2021. Students’ characteristics were summarized using appropriate descriptive statistics. Overweight, obesity and self-reported chronic diseases were described and compared between male and female students using chi-square and t tests. Other associated factors were also explored in relation to overweight and obesity. All analyses were conducted using STATA statistical software version 16.1 (StataCorp LLC, College Station, TX, USA). 902 students participated in the study. 79.8% were females and mean age was 21.90 ± 5.19 years. Majority of the respondents were undergraduate students (80.71%). The prevalence of self-reported chronic diseases was 22.95%. Obesity (BMI≥30kg/m2), Diabetes Mellitus, and Asthma/Allergies were the commonest diseases (12.48%, 4.21% & 3.22%, respectively). Approximately 5% of the students reported more than one chronic disease. Out of the 833 participating students who had complete weight and height data, prevalence of overweight and obesity was 34.81% (22.33% and 12.48%, respectively). More than half of the male students (54.36%) were overweight or obese. This is significantly higher than in female students (30.56%, p=0.001). Overweight/obesity when compared to normal weight is associated with increasing mean age [23.40 vs 21.01, respectively (p=0.001)]. In addition to gender and age, being married [57.63% vs 31.05% (p=0.001)], being a postgraduate student [51.59% vs 30.92% (p=0.001)] and having two or more chronic diseases [65.85% vs 33.21% (p=0.001)] were also significantly associated with overweight/obesity. Our study showed that almost a quarter of the participating university students reported at least one chronic disease. Obesity was the commonest and more than 1 in 3 students were either overweight or obese. This shows the need for intensive health promotion and screening programs on obesity and other chronic diseases to meet the health needs of these students. This study is also a basis for further research, especially qualitative, to explore the relevant risk factors and risk groups for more targeted interventions.

Keywords: chronic disease, obesity, overweight, students, United Arab Emirates

Procedia PDF Downloads 98

Authors: Yuliya Huseva

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To investigate the contents of tears to determine latent blood. Methods: Tear samples from 72 women were studied with the microscopy of tears aspirated with a capillary and stained by Nocht and with a chemical method of test strips with chromogen. Statistical data processing was carried out using statistical packages Statistica 10.0 for Windows, calculation of Pearson's chi-square test, Yule association coefficient, the method of determining sensitivity and specificity. Results:, In 30.6% (22) of tear samples erythrocytes were revealed microscopically. Correlations between the presence of erythrocytes in the tear and the phase of the menstrual cycle has been discovered. In the follicular phase of the cycle, erythrocytes were found in 59.1% (13) people, which is significantly more (x2=4.2, p=0.041) compared to the luteal phase - in 40.9% (9) women. In the first seven days of the follicular phase of the menstrual cycle the erythrocytes were predominanted of in the tears of women examined testifies in favour of the vicarious bleeding from the mucous membranes of extragenital organs in sync with menstruation. Of the other cellular elements in tear samples with latent haemolacria, neutrophils prevailed - in 45.5% (10), while lymphocytes were less common - in 27.3% (6), because neutrophil exudation is accompanied by vasodilatation of the conjunctiva and the release of erythrocytes into the conjunctival cavity. It was found that the prognostic significance of the chemical method was 0.53 of the microscopic method. In contrast to microscopy, which detected blood in tear samples from 30.6% (22) of women, blood was detected chemically in tears of 16.7% (12). An association between latent haemolacria and endometriosis was found (k=0.75, p≤0.05). Microscopically, in the tears of patients with endometriosis, erythrocytes were detected in 70% of cases, while in healthy women without endometriosis - in 25% of cases. The proportion of women with erythrocytes in tears, determined by a chemical method, was 41.7% among patients with endometriosis, which is significantly more (x2=6.5, p=0.011) than 11.7% among women without endometriosis. The data obtained can be explained by the etiopathogenesis of the extragenital endometriosis which is caused by hematogenous spread of endometrial tissue into the orbit. In endometriosis, erythrocytes are found against the background of accumulations of epithelial cells. In the tear samples of 4 women with endometriosis, glandular cuboidal epithelial cells, morphologically similar to endometrial cells, were found, which may indicate a generalization of the disease. Conclusions: Single erythrocytes can normally be found in the tears, their number depends on the phase of the menstrual cycle, increasing in the follicular phase. Erythrocytes found in tears against the background of accumulations of epitheliocytes and their glandular atypia may indicate a manifestation of extragenital endometriosis. Both used methods (microscopic and chemical) are informative in revealing latent haemolacria. The microscopic method is more sensitive, reveals intact erythrocytes, and besides, it provides information about other cells. At the same time, the chemical method is faster and technically simpler, it determines the presence of haemoglobin and its metabolic products, and can be used as a screening.

Keywords: tear, blood, microscopy, epitheliocytes

Procedia PDF Downloads 97

Authors: Marlin Mubarak

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Background: Endometriosis is a complex disabling disease affecting young females in the reproductive period mainly. The aim of this project is to generate a diagnostic model to predict severity and stage of endometriosis prior to Laparoscopic surgery. This will help to improve the pre-operative diagnostic accuracy of stage 3 & 4 endometriosis and as a result, refer relevant women to a specialist centre for complex Laparoscopic surgery. The model is based on the British Society of Gynaecological Endoscopy (BSGE) pain questionnaire, clinical examination and ultrasound scan. Design: This is a prospective, observational, study, in which women completed the BSGE pain questionnaire, a BSGE requirement. Also, as part of the routine preoperative assessment patient had a routine ultrasound scan and when recto-vaginal and deep infiltrating endometriosis was suspected an MRI was performed. Setting: Luton & Dunstable University Hospital. Patients: Symptomatic women (n = 56) scheduled for laparoscopy due to pelvic pain. The age ranged between 17 – 52 years of age (mean 33.8 years, SD 8.7 years). Interventions: None outside the recognised and established endometriosis centre protocol set up by BSGE. Main Outcome Measure(s): Sensitivity and specificity of endometriosis diagnosis predicted by symptoms based on BSGE pain questionnaire, clinical examinations and imaging. Findings: The prevalence of diagnosed endometriosis was calculated to be 76.8% and the prevalence of advanced stage was 55.4%. Deep infiltrating endometriosis in various locations was diagnosed in 32/56 women (57.1%) and some had DIE involving several locations. Logistic regression analysis was performed on 36 clinical variables to create a simple clinical prediction model. After creating the scoring system using variables with P < 0.05, the model was applied to the whole dataset. The sensitivity was 83.87% and specificity 96%. The positive likelihood ratio was 20.97 and the negative likelihood ratio was 0.17, indicating that the model has a good predictive value and could be useful in predicting advanced stage endometriosis. Conclusions: This is a hypothesis-generating project with one operator, but future proposed research would provide validation of the model and establish its usefulness in the general setting. Predictive tools based on such model could help organise the appropriate investigation in clinical practice, reduce risks associated with surgery and improve outcome. It could be of value for future research to standardise the assessment of women presenting with pelvic pain. The model needs further testing in a general setting to assess if the initial results are reproducible.

Keywords: deep endometriosis, endometriosis, minimally invasive, MRI, ultrasound.

Procedia PDF Downloads 320

Authors: Sophie Oakes-Rogers, Di Bailey, Karen Slade

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Female offenders are a uniquely vulnerable group, who are at high risk of suicide. Whilst the prevention of self-harm and suicide remains a key global priority, we need to better understand the relationship between these challenging behaviours that constitute a pressing problem, particularly in environments designed to prioritise safety and security. Method choice is unlikely to be random, and is instead influenced by a range of cultural, social, psychological and environmental factors, which change over time and between countries. A key aspect of self-harm and suicide in women receiving forensic care is the lack of free access to methods. At a time where self-harm and suicide rates continue to rise internationally, understanding the role of these influencing factors and the impact of current suicide prevention strategies on the use of near-lethal methods is crucial. This poster presentation will present findings from 25 interviews and 3 focus groups, which enlisted a Participatory Action Research approach to explore the differences between self-harming and suicidal behavior. A key element of this research was using the lived experiences of women receiving forensic care from one forensic pathway in the UK, and the staffs who care for them, to discuss the role of near-lethal self-harm (NLSH). The findings and suggestions from the lived accounts of the women and staff will inform a draft assessment tool, which better assesses the risk of suicide based on the lethality of methods. This tool will be the first of its kind, which specifically captures the needs of women receiving forensic services. Preliminary findings indicate women engage in NLSH for two key reasons and is determined by their history of self-harm. Women who have a history of superficial non-life threatening self-harm appear to engage in NLSH in response to a significant life event such as family bereavement or sentencing. For these women, suicide appears to be a realistic option to overcome their distress. This, however, differs from women who appear to have a lifetime history of NLSH, who engage in such behavior in a bid to overcome the grief and shame associated with historical abuse. NLSH in these women reflects a lifetime of suicidality and indicates they pose the greatest risk of completed suicide. Findings also indicate differences in method selection between forensic provisions. Restriction of means appears to play a role in method selection, and findings suggest it causes method substitution. Implications will be discussed relating to the screening of female forensic patients and improvements to the current suicide prevention strategies.

Keywords: forensic mental health, method substitution, restriction of means, suicide

Procedia PDF Downloads 155

Authors: Ying Li, Rui Hu, Shizhen Chen, Xin Zhou, Yunhuang Yang

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Prostate cancer is one of the leading causes of cancer-related death among men. Prostate-specific antigen (PSA), a specific product of prostatic epithelial cells, is an important indicator of prostate cancer. Though PSA is not a specific serum biomarker for the screening of prostate cancer, it is recognized as an indicator for prostate cancer recurrence and response to therapy for patient’s post-prostatectomy. Since radical prostatectomy eliminates the source of PSA production, serum PSA levels fall below 50 pg/mL, and may be below the detection limit of clinical immunoassays (current clinical immunoassay lower limit of detection is around 10 pg/mL). Many clinical studies have shown that intervention at low PSA levels was able to improve patient outcomes significantly. Therefore, ultra-sensitive and precise assays that can accurately quantify extremely low levels of PSA (below 1-10 pg/mL) will facilitate the assessment of patients for the possibility of early adjuvant or salvage treatment. Currently, the commercially available ultra-sensitive ELISA kit (not used clinically) can only reach a detection limit of 3-10 pg/mL. Other platforms developed by different research groups could achieve a detection limit as low as 0.33 pg/mL, but they relied on sophisticated instruments to get the final readout. Herein we report a microfluidic platform for point-of-care (POC) detection of PSA with a detection limit of 0.5 pg/mL and without the assistance of any equipment. This platform is based on a previously reported volumetric-bar-chart chip (V-Chip), which applies platinum nanoparticles (PtNPs) as the ELISA probe to convert the biomarker concentration to the volume of oxygen gas that further pushes the red ink to form a visualized bar-chart. The length of each bar is used to quantify the biomarker concentration of each sample. We devised a long reading channel V-Chip (LV-Chip) in this work to achieve a wide detection window. In addition, LV-Chip employed a unique enzyme-free ELISA probe that enriched PtNPs significantly and owned 500-fold enhanced catalytic ability over that of previous V-Chip, resulting in a significantly improved detection limit. LV-Chip is able to complete a PSA assay for five samples in 20 min. The device was applied to detect PSA in 50 patient serum samples, and the on-chip results demonstrated good correlation with conventional immunoassay. In addition, the PSA levels in finger-prick whole blood samples from healthy volunteers were successfully measured on the device. This completely stand-alone LV-Chip platform enables convenient POC testing for patient follow-up in the physician’s office and is also useful in resource-constrained settings.

Keywords: point-of-care detection, microfluidics, PSA, ultra-sensitive

Procedia PDF Downloads 89

Authors: Leonard Tiong, Sonit Singh, Kevin Ho Shon, Sarah Lewis

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Investigation into the use of artificial intelligence in radiology continues to develop at a rapid rate. During the coronavirus pandemic, the combination of an exponential increase in chest x-rays and unpredictable staff shortages resulted in a huge strain on the department's workload. There is a World Health Organisation estimate that two-thirds of the global population does not have access to diagnostic radiology. Therefore, there could be demand for a program that could detect acute changes in imaging compatible with infection to assist with screening. We generated a conventional neural network and tested its efficacy in recognizing changes compatible with coronavirus infection. Following ethics approval, a deidentified set of 77 normal and 77 abnormal chest x-rays in patients with confirmed coronavirus infection were used to generate an algorithm that could train, validate and then test itself. DICOM and PNG image formats were selected due to their lossless file format. The model was trained with 100 images (50 positive, 50 negative), validated against 28 samples (14 positive, 14 negative), and tested against 26 samples (13 positive, 13 negative). The initial training of the model involved training a conventional neural network in what constituted a normal study and changes on the x-rays compatible with coronavirus infection. The weightings were then modified, and the model was executed again. The training samples were in batch sizes of 8 and underwent 25 epochs of training. The results trended towards an 85.71% true positive/true negative detection rate and an area under the curve trending towards 0.95, indicating approximately 95% accuracy in detecting changes on chest X-rays compatible with coronavirus infection. Study limitations include access to only a small dataset and no specificity in the diagnosis. Following a discussion with our programmer, there are areas where modifications in the weighting of the algorithm can be made in order to improve the detection rates. Given the high detection rate of the program, and the potential ease of implementation, this would be effective in assisting staff that is not trained in radiology in detecting otherwise subtle changes that might not be appreciated on imaging. Limitations include the lack of a differential diagnosis and application of the appropriate clinical history, although this may be less of a problem in day-to-day clinical practice. It is nonetheless our belief that implementing this program and widening its scope to detecting multiple pathologies such as lung masses will greatly assist both the radiology department and our colleagues in increasing workflow and detection rate.

Keywords: artificial intelligence, COVID, neural network, machine learning

Procedia PDF Downloads 66

Authors: Daniel Todorov, Anton Hinkov, Petya Angelova, Kalina Shishkova, Venelin Tsvetkov, Stoyan Shishkov

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Human herpes viruses (HHV) are ubiquitous pathogens with a pandemic spread across the globe. HHV type 1 is the main causative agent of cold sores and fever blisters around the mouth and on the face, whereas HHV type 2 is generally responsible for genital herpes outbreaks. The treatment of both viruses is more or less successful with antivirals from the nucleoside analogues group. Their wide application increasingly leads to the emergence of resistant mutants In the past, medicinal plants have been used to treat a number of infectious and non-infectious diseases. Their diversity and ability to produce the vast variety of secondary metabolites according to the characteristics of the environment give them the potential to help us in our warfare with viral infections. The variable chemical characteristics and complex composition is an advantage in the treatment of herpes since the emergence of resistant mutants is significantly complicated. The screening process is difficult due to the lack of standardization. That is why it is especially important to follow the mechanism of antiviral action of plants. On the one hand, it may be expected to interact with its compounds, resulting in enhanced antiviral effects, and the most appropriate environmental conditions can be chosen to maximize the amount of active secondary metabolites. During our study, we followed the activity of various plant extracts on the viral replication cycle as well as their effect on the extracellular virion. We obtained our results following the logical sequence of the experimental settings - determining the cytotoxicity of the extracts, evaluating the overall effect on viral replication and extracellular virion.During our research, we have screened a variety of plant extracts for their antiviral activity against both virus replication and the virion itself. We investigated the effect of the extracts on the individual stages of the viral replication cycle - viral adsorption, penetration and the effect on replication depending on the time of addition. If there are positive results in the later experiments, we had studied the activity over viral adsorption, penetration and the effect of replication according to the time of addition. Our results indicate that some of the extracts from the Lamium album have several targets. The first stages of the viral life cycle are most affected. Several of our active antiviral agents have shown an effect on extracellular virion and adsorption and penetration processes. Our research over the last decade has shown several curative antiviral plants - some of which are from the Lamiacea family. The rich set of active ingredients of the plants in this family makes them a good source of antiviral preparation.

Keywords: human herpes virus, antiviral activity, Lamium album, Nepeta nuda

Procedia PDF Downloads 134

Authors: Adam Aboalkaz, Rana Shamoon, Duncan Meikle, James Lewis

Abstract:

Background and aims: In orthopaedic theatres, radiological screening during operations is a commonly used and useful technique to visualise and guide the operating surgeon. Within any theatre using ionising radiation, it is imperative that the use of protective equipment and the wearing of a dosimeter at all times. 1. To assess compliance with use of protective equipment during orthopaedic procedures involving ionising radiation. 2. To assess the radiation risk knowledge of staff members regularly present in an orthopaedic theatre of a national major trauma centre, in accordance to the ionising radiation regulation (2000) guidelines. Method: We conducted an Observational study of 21 operations at the University Hospital of Wales, which is a major trauma centre, recording the compliance with use of protective equipment (lead aprons and thyroid shields) and dosimeters. The observations were performed sporadically over a two week period to ensure that all staff in monitored operating theatres were not aware of the ongoing study, as to avoid bias. A questionnaire testing the knowledge of trainees and staff within the orthopaedic department was given following completion of the initial phase of the study, with 19 responses. The questions were based on knowledge of ionising radiation exposure and monitoring. The questions also tested the general staff knowledge of what equipment should be worn and where to locate such equipment. Results: This study found that only 25% of staff members were wearing thyroid protectors when less than 1 meter from the radiation source and only 50% were wearing appropriate lead aprons whilst in this same vicinity. The study also showed that 0% of all staff members used a dosimeter whilst in an area of radiation exposure. From the distributed questionnaires, only 40% of staff understood where to stand whilst radiation was being used, and only 25% of staff knew where to find protective equipment. Conclusion: Overall our audit showed poor compliance with regards to the National and local policies, due to lack of awareness of the policy and lack of basic ionising radiation exposure knowledge. It was evident from the observational study and questionnaire that staff were not fully aware of what equipment should be worn, where to find such equipment and did not appreciate that the distance from the ionising radiation source altered its exposure effect. This lack of knowledge may affect the staff health and safety after long term exposure. Changes to clinical practice: From the outcome of this study, we managed to drastically increase awareness of ionising radiation within the orthopaedic department. A mandatory teaching session on the safety of ionising radiation has been incorporated into the orthopaedic induction week for all staff. The dosimeters have been moved to a visible location within the trauma operating theatre and all staff made aware of where to find protective equipment.

Keywords: audit, ionising radiation, observational study, protection

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Authors: M. Naimi, M. B. Khaled

Abstract:

The aim of the present work was to test in vitro inhibition of food pathogens and spoilage bacteria by crude bacteriocins from autochthonous lactic acid bacteria. Thirty autochthonous lactic acid bacteria isolated previously, belonging to the genera: Lactobacillus, Carnobacterium, Lactococcus, Vagococcus, Streptococcus, and Pediococcus, have been screened by an agar spot test and a well diffusion assay against Gram-positive and Gram-negative harmful bacteria: Bacillus cereus, Bacillus subtilis ATCC 6633, Escherichia coli ATCC 8739, Salmonella typhimurium ATCC 14028, Staphylococcus aureus ATCC 6538, and Pseudomonas aeruginosa under conditions means to reduce lactic acid and hydrogen peroxide effect to select bacteria with high bacteriocinogenic potential. Furthermore, crude bacteriocins semiquantification and heat sensitivity to different temperatures (80, 95, 110°C, and 121°C) were performed. Another exploratory test concerning the response of St. aureus ATCC 6538 to the presence of crude bacteriocins was realized. It has been observed by the agar spot test that fifteen candidates were active toward Gram-positive targets strains. The secondary screening demonstrated an antagonistic activity oriented only against St. aureus ATCC 6538, leading to the selection of five isolates: Lm14, Lm21, Lm23, Lm24, and Lm25 with a larger inhibition zone compared to the others. The ANOVA statistical analysis reveals a small variation of repeatability: Lm21: 0.56%, Lm23: 0%, Lm25: 1.67%, Lm14: 1.88%, Lm24: 2.14%. Conversely, slight variation was reported in terms of inhibition diameters: 9.58± 0.40, 9.83± 0.46, and 10.16± 0.24 8.5 ± 0.40 10 mm for, Lm21, Lm23, Lm25, Lm14and Lm24, indicating that the observed potential showed a heterogeneous distribution (BMS = 0.383, WMS = 0.117). The repeatability coefficient calculated displayed 7.35%. As for the bacteriocins semiquantification, the five samples exhibited production amounts about 4.16 for Lm21, Lm23, Lm25 and 2.08 AU/ml for Lm14, Lm24. Concerning the sensitivity the crude bacteriocins were fully insensitive to heat inactivation, until 121°C, they preserved the same inhibition diameter. As to, kinetic of growth , the µmax showed reductions in pathogens load for Lm21, Lm23, Lm25, Lm14, Lm24 of about 42.92%, 84.12%, 88.55%, 54.95%, 29.97% in the second trails. Inversely, this pathogen growth after five hours displayed differences of 79.45%, 12.64%, 11.82%, 87.88%, 85.66% in the second trails, compared to the control. This study showed potential inhibition to the growth of this food pathogen, suggesting the possibility to improve the hygienic food quality.

Keywords: exploratory test, lactic acid bacteria, crude bacteriocins, spoilage, pathogens

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Authors: Jamal Alkoteesh, Mohammed Zeki, Mouza Alnaqbi

Abstract:

Objective: To evaluate pregnancy outcomes, complications and neonatal outcomes in women who had previously undergone uterine arterial embolization. Design: Retrospective study. In this study, most women opted for UFE as a fertility treatment after failure of myomectomy or in vitro fertilization, or because hysterectomy was the only suggested option. Background. Myomectomy is the standard approach in patients with fibroids desiring a future pregnancy. However, myomectomy may be difficult in cases of numerous interstitial and/or submucous fibroids.In these cases, UFE has the advantage of embolizing all fibroids in one procedure. This procedure is an accepted nonsurgical treatment for symptomatic uterine fibroids. Study Methods: A retrospective study of 210 patients treated with UFE for symptomatic uterine fibroids between 2011-2016 was performed. UFE was performed using ((PVA; Embozen, Beadblock) (500-900 µm in diameter). Pregnancies were identified using screening questionnaires and the study database. Of the 210 patients who received UFE treatment, 35 women younger than the age of 40 wanted to conceive and had been unable. All women in our study were advised to wait six months or more after UFE before attempting to become pregnant, of which the reported time range before attempting to conceive was seven to 33 months (average 20 months). RESULTS: In a retrospective chart review of patients younger than the age of 40 (35 patients,18 patients reported 23 pregnancies, of which five were miscarriages. Two more pregnancies were complicated by premature labor. Of the 23 pregnancies, 16 were normal full-term pregnancies, 15 women had conceived once, and four had become pregnant twice. The remaining patients did not conceive. In the study, there was no reported intrauterine growth retardation in the prenatal period, fetal distress during labor, or problems related to uterine integrity. Two patients reported minor problems during pregnancy that were borderline oligohydramnios and low-lying placenta. In the cohort of women who did conceive, overall, 16 out of 18 births proceeded normally without any complications (86%). Eight women delivered by cesarean section, and 10 women had normal vaginal delivery. In this study of 210 women, UFE had a fertility rate of 47%. Our group of 23 pregnancies was small, but did confirm successful pregnancy after UFE. The 45.7% pregnancy rate in women below the age of 40 years old who completed a term pregnancy compares favorably with women who underwent myomectomy via other method. Of the women in the cohort who did conceive, subsequent birth proceeded normally (86%). Conclusion: Pregnancy after UFE is well-documented. The risks of infertility following embolization, premature menopause, and hysterectomy are small, as is the radiation exposure during embolization. Fertility rates appear similar to patients undergoing myomectomy.UFE should not be contraindicated in patients who want to conceive and they should be able to choose between surgical options and UFE.

Keywords: fibroid, pregnancy, therapeutic embolization, uterine artery

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Authors: Freja Rydahl Rasmussen, Naja Villadsen, Stig Koust

Abstract:

Air purifiers have become widely implemented in a wide range of settings, including households, schools, institutions, and hospitals, as they tackle the pressing issue of indoor air pollution. With their ability to enhance indoor air quality and create healthier environments, air purifiers are particularly vital when ventilation options are limited. These devices incorporate a diverse array of technologies, including HEPA filters, active carbon filters, UV-C light, photocatalytic oxidation, and ionizers, each designed to combat specific pollutants and improve air quality within enclosed spaces. However, the safety of air purifiers has not been investigated thoroughly, and many questions still arise when applying them. Certain air purification technologies, such as UV-C light or ionization, can unintentionally generate undesirable byproducts that can negatively affect indoor air quality and health. It is well-established that these technologies can inadvertently generate nanoparticles or convert common gaseous compounds into harmful ones, thus exacerbating air pollution. However, the formation of byproducts can vary across products, necessitating further investigation. There is a particular concern about the formation of the carcinogenic substance formaldehyde from common gases like acetone. Many air purifiers use mechanical filtration to remove particles, dust, and pollen from the air. Filters need to be replaced periodically for optimal efficiency, resulting in an additional cost for end-users. Currently, there are no guidelines for filter lifespan, and replacement recommendations solely rely on manufacturers. A market screening revealed that manufacturers' recommended lifespans vary greatly (from 1 month to 10 years), and there is a need for general recommendations to guide consumers. Activated carbon filters are used to adsorb various types of chemicals that can pose health risks or cause unwanted odors. These filters have a certain capacity before becoming saturated. If not replaced in a timely manner, the adsorbed substances are likely to be released from the filter through off-gassing or losing adsorption efficiency. The goal of this study is to investigate the lifespan of filters as well as investigate the potentially harmful effects of air purifiers. Understanding the lifespan of filters used in air purifiers and the potential formation of harmful byproducts is essential for ensuring their optimal performance, guiding consumers in their purchasing decisions, and establishing industry standards for safer and more effective air purification solutions. At this time, a selection of air purifiers has been chosen, and test methods have been established. In the following 3 months, the tests will be conducted, and the results will be ready for presentation later.

Keywords: air purifiers, activated carbon filters, byproducts, clean air, indoor air quality

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