Search results for: adult respiratory distress syndrome

Authors: M. A. Rahman, A. T. Ohta, H. V. Trinh, J. Hyvl

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Atmospheric pressure and oxygen (O2) concentration are critical life support parameters for human-occupied aerospace vehicles and habitats. Various medical conditions may require medical O2; for example, the American Medical Association has determined that commercial air travel exposes passengers to altitude-related hypoxia and gas expansion. It may cause some passengers to experience significant symptoms and medical complications during the flight, requiring supplemental medical-grade O2 to maintain adequate tissue oxygenation and prevent hypoxemic complications. Although supplemental medical grade O2 is a successful lifesaver for respiratory and cardiac failure, O2-enriched exhaled air can contain more than 95 % O2, increasing the likelihood of a fire. In an aerospace environment, a localized high concentration O2 bubble forms around a patient being treated for hypoxia, increasing the cabin O2 beyond the safe limit. To address this problem, this work describes a medical O2 delivery system that can reduce the O2 concentration from patient-exhaled O2-rich air to safe levels while maintaining the prescribed O2 administration to the patient. The O2 delivery system is designed to be a part of the medical O2 kit. The system uses cationic multimetallic cobalt complexes to reversibly, selectively, and stoichiometrically chemisorb O2 from the exhaled air. An air-release sub-system monitors the exhaled air, and as soon the O2 percentage falls below 21%, the air is released to the room air. The O2-enriched exhaled air is channeled through a layer of porous, thin-film heaters coated with the cobalt complex. The complex absorbs O2, and when saturated, the complex is heated to 100°C using the thin-film heater. Upon heating, the complex desorbs O2 and is once again ready to absorb or remove the excess O2 from exhaled air. The O2 absorption is a sub-second process, and desorption is a multi-second process. While heating at 0.685 °C/sec, the complex desorbs ~90% O2 in 110 sec. These fast reaction times mean that a simultaneous absorb/desorb process in the O2 delivery system will create a continuous absorption of O2. Moreover, the complex can concentrate O2 by a factor of 160 times that in air and desorb over 90% of the O2 at 100°C. Over 12 cycles of thermogravimetry measurement, less than 0.1% decrease in reversibility in O2 uptake was observed. The 1 kg complex can desorb over 20L of O2, so simultaneous O2 desorption by 0.5 kg of complex and absorption by 0.5 kg of complex can potentially continuously remove 9L/min O2 (~90% desorbed at 100°C) from exhaled air. The complex is synthesized and characterized for reversible O2 absorption and efficacy. The complex changes its color from dark brown to light gray after O2 desorption. In addition to thermogravimetric analysis, the O2 absorption/desorption cycle is characterized using optical imaging, showing stable color changes over ten cycles. The complex was also tested at room temperature in a low O2 environment in its O2 desorbed state, and observed to hold the deoxygenated state under these conditions. The results show the feasibility of using the complex for reversible O2 absorption in the proposed fire safe medical O2 delivery system.

Keywords: fire risk, medical oxygen, oxygen removal, reversible absorption

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Authors: Cecile Garry, Katia Rovira, Julie Brisson

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Autism Spectrum Disorder (ASD) is defined by a qualitative and quantitative impairment of social interaction. Indeed early intervention programs, such as the Early Start Denver Model (ESDM), aimed at encouraging the development of social skills. In classroom, the children need to be socially engaged to learn. Early intervention programs can thus be implemented in kindergarten schools. In these schools, ASD children have more opportunities to interact with their peers or adults than in elementary schools. However, the preschool children with ASD are less socially engaged than their typically developing peers in the classroom. They initiate, respond and maintain less the social interactions. In addition, they produce more responses than initiations. When they interact, the non verbal communication is more used than verbal or symbolic communication forms and they are more engaged with adults than with peers. Nevertheless, communicative patterns may vary according to the clinical profiles of ASD children. Indeed, the ASD children with better cognitive skills interact more with their peers and use more symbolic communication than the ASD children with a low cognitive level. ASD children with the less severe symptoms use more the verbal communication than ASD children with the more severe symptoms. Small groups and structured activities encourage coordinated joint engagement episodes in ASD children. Our goal is to evaluate ASD children’s social engagement development in class, with their peers or adults, during dyadic or group activities. Participants were 19 preschool children with ASD aged from 3 to 6 years old that benefited of an early intervention in special kindergarten schools. Severity of ASD symptoms was measured with the CARS at the beginning of the follow-up. Classroom situations of interaction were recorded during 10 minutes (5 minutes of dyadic interaction and 5 minutes of a group activity), every 2 months, during 10 months. Social engagement behaviors of children, including initiations, responses and imitation, directed to a peer or an adult, were then coded. The Observer software (Noldus) that allows to annotate behaviors was the coding system used. A double coding was conducted and revealed a good inter judges fidelity. Results show that ASD children were more often and longer socially engaged in dyadic than in groups situations. They were also more engaged with adults than with peers. Children with the less severe symptoms of ASD were more socially engaged in groups situations than children with the more severe symptoms of ASD. Then, ASD children with the less severe symptoms of ASD were more engaged with their peers than ASD children with the more severe symptoms of ASD. However, the engagement frequency increased during the 10 month of follow-up but only for ASD children with the more severe symptoms at the beginning. To conclude, these results highlighted the necessity of individualizing early intervention programs according to the clinical profile of the child.

Keywords: autism spectrum disorder, preschool children, developmental psychology, early interventions, social interactions

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Authors: D. O. Miranda, L. R. Azevedo, J. F. C. Cordeiro, A. H. Dos Santos, S. F. Lisboa, F. S. Guimarães, G. S. Bisson

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Background: The Colorectal cancer (CRC) is one of the most common cancers and the fourth leading cause of cancer death in the world. The prevalence of psychiatric-disorders among CRC patients, mainly depression, is high, resulting in impaired quality of life and side effects of primary treatment. High levels of proinflammatory cytokines at tumor microenvironment is a feature of CRC and the literature suggests that those mediators could contribute to the development of psychiatric disorders. Nevertheless, the ability of tumor-associated biological processes to affect the central nervous system (CNS) has only recently been explored in the context of symptoms of depression and is still not well understood. Therefore, the aim of the present study was to test the hypothesis that depressive-like behavior in an experimental model of CCR induced by N-methyl-N-nitro-N-nitrosoguanidine (MNNG) was correlated to proinflammatory profile in the periphery and in the brain. Methods: Colorectal carcinogenesis was induced in adult C57BL/6 mice (n=12) by administration of MNNG (5mg/kg, 0.1ml/intrarectal instillation) 2 times a week, for 2 week. Control group (n=12) received saline (0.1ml/intrarectal instillation). Eight weeks after beginning of MNNG administration animals were submitted to the forced swim test (FST) and the sucrose preference test for evaluation, respectively, of depressive- and anhedonia-like behaviors. After behavioral evaluation, the colon was collected and brain regions dissected (cortex-C, striatum-ST and hippocampus-HIP) for posterior evaluation of cytokine levels (IL-1β, IL-10, IL-17, and CX3CL1) by ELISA. Results: MNNG induced depressive-like behavior, represented by increased immobility time in the FST (Student t test, p < 0.05) and lower sucrose preference (Student t test, p < 0.05). Moreover, there were increased levels of IL-1β, IL-17 and CX3CL1 in the colonic tissue (Student t test, p < 0.05) and in the brain (IL-1 β in the ST and HIP, Student t test, p < 0.05; IL-17 and CX3CL1 in the C and HIP, p < 0.05). IL-10 levels, in contrast, were decreased in both the colon (p < 0.05) and the brain (C and HIP, p < 0.05). Conclusions: The results obtained in the present work support the notion that tumor growth induces neuroinflammation in stress-related brain regions and depressive-like behavior, which could be related to the high incidence of depression in colorectal carcinogenesis. This work have important clinical and research implications, taken into account that cytokine levels may be a marker promissory for the developing depression in CRC patients. New therapeutic strategies to assist in alleviating mental suffering in cancer patients might result from a better understanding of the role of cytokines in the pathophysiology of depression in these subjects.

Keywords: cytokines, brain, depression, colorectal cancer

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Authors: U. E. Mochalova, A. V. Demeneva, Shilkin A. G., J. Yu. Artiushina

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Objective. In medical ophthalmology OCT has been actively used in the last decade. It is a modern non-invasive method of high-precision hardware examination, which gives a detailed cross-sectional image of eye tissues structure with a high level of resolution, which provides in vivo morphological information at the microscopic level about corneal tissue, structures of the anterior segment, retina and optic nerve. The purpose of this study was to explore the possibility of using the OCT technology in complex ophthalmological examination in dogs and cats, to characterize the revealed pathological structural changes in corneal tissue in cats and dogs with some of the most common corneal diseases. Procedures. Optical coherence tomography of the cornea was performed in 112 animals: 68 dogs and 44 cats. In total, 224 eyes were examined. Pathologies of the organ of vision included: dystrophy and degeneration of the cornea, endothelial corneal dystrophy, dry eye syndrome, chronic superficial vascular keratitis, pigmented keratitis, corneal erosion, ulcerative stromal keratitis, corneal sequestration, chronic glaucoma and also postoperative period after performed keratoplasty. When performing OCT, we used certified medical devices: "Huvitz HOCT-1/1F», «Optovue iVue 80» and "SOCT Copernicus Revo (60)". Results. The results of a clinical study on the use of optical coherence tomography (OCT)of the cornea in cats and dogs, performed by the authors of the article in the complex diagnosis of keratopathies of variousorigins: endothelial corneal dystrophy, pigmented keratitis, chronic keratoconjunctivitis, chronic herpetic keratitis, ulcerative keratitis, traumatic corneal damage, sequestration of the cornea of cats, chronic keratitis, complicating the course of glaucoma. The characteristics of the OCT scans are givencorneas of cats and dogs that do not have corneal pathologies. OCT scans of various corneal pathologies in dogs and cats with a description of the revealed pathological changes are presented. Of great clinical interest are the data obtained during OCT of the cornea of animals undergoing keratoplasty operations using various forms of grafts. Conclusions. OCT makes it possible to assess the thickness and pathological structural changes of the corneal surface epithelium, corneal stroma and descemet membrane. We can measure them, determine the exact localization, and record pathological changes. Clinical observation of the dynamics of the pathological process in the cornea using OCT makes it possible to evaluate the effectiveness of drug treatment. In case of negative dynamics of corneal disease, it is necessary to determine the indications for surgical treatment (to assess the thickness of the cornea, the localization of its thinning zones, to characterize the depth and area of pathological changes). According to the OCT of the cornea, it is possible to choose the optimal surgical treatment for the patient, the technique and depth of optically constructive surgery (penetrating or anterior lamellar keratoplasty).; determine the depth and diameter of the planned microsurgical trepanation of corneal tissue, which will ensure good adaptation of the edges of the donor material.

Keywords: optical coherence tomography, corneal sequestration, optical coherence tomography of the cornea, corneal transplantation, cat, dog

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Authors: Ronald Croce, Timothy Quinn, John Miller

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Incomplete activation, or activation failure, of motor units during maximal voluntary contractions is often referred to as muscle inhibition (MI), and is defined as the inability of the central nervous system to maximally drive a muscle during a voluntary contraction. The purpose of the present study was to assess the interrelationship amongst peak torque (PT), muscle inhibition (MI; incomplete activation of motor units), and voluntary muscle activation (VMA) of the quadriceps’ muscle group as a function of knee angle and muscle length during maximal voluntary isometric contractions (MVICs). Nine young adult males (mean + standard deviation: age: 21.58 + 1.30 years; height: 180.07 + 4.99 cm; weight: 89.07 + 7.55 kg) performed MVICs in random order with the knee at 15, 55, and 95° flexion. MI was assessed using the interpolated twitch technique and was estimated by the amount of additional knee extensor PT evoked by the superimposed twitch during MVICs. Voluntary muscle activation was estimated by root mean square amplitude electromyography (EMGrms) and mechanomyography (MMGrms) of agonist (vastus medialis [VM], vastus lateralis [VL], and rectus femoris [RF]) and antagonist (biceps femoris ([BF]) muscles during MVICs. Data were analyzed using separate repeated measures analysis of variance. Results revealed a strong dependency of quadriceps’ PT (p < 0.001), MI (p < 0.001) and MA (p < 0.01) on knee joint position: PT was smallest at the most shortened muscle position (15°) and greatest at mid-position (55°); MI and MA were smallest at the most shortened muscle position (15°) and greatest at the most lengthened position (95°), with the RF showing the greatest change in MA. It is hypothesized that the ability to more fully activate the quadriceps at short compared to longer muscle lengths (96% contracted at 15°; 91% at 55°; 90% at 95°) might partly compensate for the unfavorable force-length mechanics at the more extended position and consequent declines in VMA (decreases in EMGrms and MMGrms muscle amplitude during MVICs) and force production (PT = 111-Nm at 15°, 217-NM at 55°, 199-Nm at 95°). Biceps femoris EMG and MMG data showed no statistical differences (p = 0.11 and 0.12, respectively) at joint angles tested, although there were greater values at the extended position. Increased BF muscle amplitude at this position could be a mechanism by which anterior shear and tibial rotation induced by high quadriceps’ activity are countered. Measuring and understanding the degree to which one sees MI and VMA in the QF muscle has particular clinical relevance because different knee-joint disorders, such ligament injuries or osteoarthritis, increase levels of MI observed and markedly reduced the capability of full VMA.

Keywords: electromyography, interpolated twitch technique, mechanomyography, muscle activation, muscle inhibition

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Authors: Debasish Nandy

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India’s foreign policy towards all of her neighbor countries is determinate on the basis of multi-dimensional factors. India’s relations with its South Asian neighbor can be classified into three categories. In the first category, there are four countries -Sri Lanka, Bangladesh, Nepal, and Afghanistan- whose bilateral relationships have encompassed cooperation, irritants, problems and crisis at different points in time. With Pakistan, the relationship has been perpetually adversarial. The third category includes Bhutan and Maldives whose relations are marked by friendship and cooperation, free of any bilateral problems. It is needless to say that Jawaharlal Nehru emphasized on friendly relations with the neighboring countries. The subsequent Prime Ministers of India especially I.K. Gujral had advocated in making of peaceful and friendly relations with the subcontinental countries. He had given a unique idea to foster bilateral relations with the neighbors. His idea is known as ‘Gujral Doctrine’. A dramatical change has been witnessed in Indian foreign policy since 1991.In the post-Cold War period, India’s national security has been vehemently threatened by terrorism, which originated from Pakistan-Afghanistan and partly Bangladesh. India has required a cooperative security, which can be made by mutual understanding among the South Asian countries. Additionally, the countries of South Asia need to evolve the concept of ‘Cooperative Security’ to explain the underlying logic of regional cooperation. According to C. Rajamohan, ‘cooperative security could be understood, as policies of governments, which see themselves as former adversaries or potential adversaries to shift from or avoid confrontationist policies.’ A cooperative security essentially reflects a policy of dealing peacefully with conflicts, not merely by abstention from violence or threats but by active engagement in negotiation, a search for practical solutions and with a commitment to preventive measures. Cooperative assumes the existence of a condition in which the two sides possess the military capabilities to harm each other. Establishing cooperative security runs into a complex process building confidence. South Asian nations often engaged with hostility to each other. Extra-regional powers have been influencing their powers in this region since a long time. South Asian nations are busy to purchase military equipment. In spite of weakened economic systems, these states are spending a huge amount of money for their security. India is the big power in this region in every aspect. The big states- small states syndrome is a negative factor in this respect. However, India will have to an initiative to extended ‘track II diplomacy’ or soft diplomacy for its security as well as the security of this region.Confidence building measures could help rejuvenate not only SAARC but also build trust and mutual confidence between India and its neighbors in South Asia. In this paper, I will focus on different aspects of India’s policy towards it, South-Asian neighbors. It will also be searched that how India is dealing with these countries by using a mixed type of diplomacy – both idealistic and realistic points of view. Security and cooperation are two major determinants of India’s foreign policy towards its South Asian neighbors.

Keywords: bilateral, diplomacy, infiltration, terrorism

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Authors: Ammar Asma, Bouafia Nabiha, Ben Cheikh Asma, Ezzi Olfa, Mahjoub Mohamed, Sma Nesrine, Chouchène Imed, Boussarsar Hamadi, Njah Mansour

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Background: Ventilator-associated pneumonia (VAP) is currently recognized as one of the most relevant causes of morbidity and mortality among intensive care unit (ICU) patients worldwide. Identifying modifiable risk factors for VAP could be helpful for future controlled interventional studies aiming at improving prevention of VAP. The purposes of this study were to determine the incidence and risk factors for VAP in in a Tunisian medical ICU. Materials / Methods: A retrospective case-control study design based on the prospective database collected over a 14-month period from September 15th, 2015 through November 15th, 2016 in an 8-bed medical ICU. Patients under ventilation for over 48 h were included. The number of cases was estimated by Epi-info Software with the power of statistical test equal to 90 %. Each case patient was successfully matched to two controls according to the length of mechanical ventilation (MV) before VAP for cases and the total length of MV in controls. VAP in the ICU was defined according to American Thoracic Society; Infectious Diseases Society of America guidelines. Early onset or late-onset VAP were defined whether the infectious process occurred within or after 96 h of ICU admission. Patients’ risk factors, causes of admission, comorbidities and respiratory specimens collected were reviewed. Univariate and multivariate analyses were performed to determine variables associated with VAP with a p-value < 0.05. Results: During the period study, a total of 169 patients under mechanical ventilation were considered, 34 patients (20.11%) developed at least one episode of VAP in the ICU. The incidence rate for VAP was 14.88/1000 ventilation days. Among these cases, 9 (26.5 %) were early-onset VAP and 25 (73.5 %) were late-onset VAP. It was a certain diagnosis in 66.7% of cases. Tracheal aspiration was positive in 80% of cases. Multi-drug resistant Acinerobacter baumanii was the most common species detected in cases; 67.64% (n=23). The rate of mortality out of cases was 88.23% (n= 30). In univariate analysis, the patients with VAP were statistically more likely to suffer from cardiovascular diseases (p=0.035) and prolonged duration of sedation (p=0.009) and tracheostomy (p=0.001), they also had a higher number of re-intubation (p=0.017) and a longer total time of intubation (p=0.012). Multivariate analysis showed that cardiovascular diseases (OR= 4.44; 95% IC= [1.3 - 14]; p=0.016), tracheostomy (OR= 4.2; 95% IC= [1.16 -15.12]; p= 0.028) and prolonged duration of sedation (OR=1.21; 95% IC= [1.07, 1.36]; p=0.002) were independent risk factors for the development of VAP. Conclusion: VAP constitutes a therapeutic challenge in an ICU setting, therefore; strategies that effectively prevent VAP are needed. An infection control-training program intended to all professional heath care in this unit insisting on bundles and elaboration of procedures are planned to reduce effectively incidence rate of VAP.

Keywords: case control study, intensive care unit, risk factors, ventilator associated pneumonia

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Authors: Rebecca Keyte, Helen Egan, Michail Mantzios

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It is well acknowledged that ongoing adherence is a major concern within CF. However recently literature has indicated that non-adherence should not be viewed just in terms of medical regimens. There are other damaging behaviours that some chronically ill patients engage in which can be viewed as a form of non-adherence, such as risky behaviours. Risky behaviours are a major concern within CF, as they can have adverse health effects on patients regardless of patients adherence to medical regimens. The risky behaviours this research is predominantly focusing on are smoking, excessive alcohol consumption, illicit drug use and risky sexual behaviour. This research investigates patient’s beliefs about their CF and the impact their CF has upon their life, exploring rationales for why some patients engage in risky behaviours. This research utilises qualitative semi-structured interviews taking an interpretive perspective. Twenty-four adult participants have been recruited (16 male, age range 19–66 yrs) from two UK regional CF centres, with a median FEV1 61.77% predicted. Participants were recruited via clinician guidance, with 13 participants identified by clinicians as partaking in risky behaviours. However, during the interviews 17 participants were identified as partaking in risky behaviours, illustrating that not all patients offer full disclosure of engagement in such behaviours to their clinicians. Preliminary findings illustrate a variety of reasons as to why some CF patients engage in risky behaviours, with many participants stating that one challenge in terms of living with CF is accepting their illness. Disclosure of illness was also an issue, the desire to be seen as ‘normal’ was important to many. It is often possible for CF patients to hide their illness as they do not always appear to be unwell. However, literature indicates a desire for normalcy can be accompanied with the engagement of normalised risky behaviours, enabling patients to retaliate against their illness identity. There was also evidence of a life-orientated perspective amongst participants, with some reporting that their desire for fun and enjoyment was the reason for why they were engaging in risky behaviours. Some participants did not acknowledge the impact their risky behaviours could have upon their CF, and others rationalised their continuation with the behaviours by suggesting that they were in fact beneficial to their health. There was an apparent lack of knowledge around the implications of risky behaviours, with participants indicating that they had not been informed of such potential consequences by their clinicians. Given the adverse health effects of risky behaviours within CF, more effective health promotion measures are needed to both reduce and more importantly prevent these behaviours. Due to the initiation of risky behaviours within the CF population commonly occurring during adolescence, the researcher now proposes to conduct semi-structured interviews with paediatric patients to investigate their awareness and beliefs towards risky behaviours. Overall, this research will highlight reasons why some CF patients engage in risky behaviours, in order to inform interventions aimed to prevent the initiation in risky behaviours by increasing patient awareness.

Keywords: cystic fibrosis, health beliefs, preliminary findings, risky health behaviours

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Authors: Rory Austin Clark

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This primary research represents an ongoing two year inductive mixed-methods project endeavouring to unravel the subjective reality of hyperconnected young adults in Western societies who have come of age with social media and smartphones. It is to be presented as well as analyzed and contextualized through a written master’s thesis as well as a documentary/mockumentary meshed with a Web 2.0 app providing the capacity for prosumer, 'audience 2.0' functionality. The media component seeks to explore not only thematic issues via real-life research interviews and fictional narrative but technical issues within the format relating to the quest for intimate, authentic connection as well as compelling dissemination of scholarly knowledge in an age of ubiquitous personalized daily digital media creation and consumption. The overarching hypothesis is that the aforementioned individuals process and make sense of their world, find shared meaning, and formulate notions-of-self in ways drastically different than pre-2007 via hyper-mediation-of-self and surroundings. In this pursuit, research questions have progressed from examining how young adult digital natives understand their use of social media to notions relating to the potential functionality of Web 2.0 for prosocial and altruistic engagement, on and offline, through the eyes of these individuals no longer understood as simply digital natives, but social media natives, and at the conclusion of that phase of research, as 'neoliberalized social media natives' (NSMN). This represents the two most potent macro factors in the paradigmatic shift in NSMS’s worldview, that they are not just children of social media, but of the palpable shift to neoliberal ways of thinking and being in the western socio-cultures since the 1980s, two phenomena that have a reflexive æffective relationship on their perception of figure and ground. This phase also resulted in the working hypothesis of 'social media comparison anxiety' and a nascent understanding of NSMN’s habitus and habitation in a subjective reality of fully converged online/offline worlds, where any phenomena originating in one realm in some way are, or at the very least can be, re-presented or have effect in the other—creating hyperreal reception. This might also be understood through a 'society as symbolic cyborg model', in which individuals have a 'digital essence'-- the entirety of online content that references a single person, as an auric living, breathing cathedral, museum, gallery, and archive of self of infinite permutations and rhizomatic entry and exit points.

Keywords: affect, hyperreal, neoliberalism, postmodernism, social media native, subjective reality, Web 2.0

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Authors: Christelle E. Chua, Alicia L. Ho

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The development of a panel of differential gene expression signatures has been of interest in the field of biomarker discovery for radiation exposure. In the absence of the availability of exposed human subjects, lymphocyte cell lines have often been used as a surrogate to human whole blood, when performing ex vivo irradiation studies. The extent of variation between different lymphocyte cell lines is currently unclear, especially with regard to the expression of extracellular miRNA. This study compares the expression profile of extracellular miRNA isolated from different lymphocyte cell lines. It also compares the profile of miRNA obtained when different exosome isolation kits are used. Lymphocyte cell lines were created using lymphocytes isolated from healthy adult males of similar racial descent (Chinese American and Chinese Singaporean) and immortalised with Epstein-Barr virus. The cell lines were cultured in exosome-free cell culture media for 72h and the cell culture supernatant was removed for exosome isolation. Two exosome isolation kits were used. Total exosome isolation reagent (TEIR, ThermoFisher) is a polyethylene glycol (PEG)-based exosome precipitation kit, while ExoSpin (ES, Cell Guidance Systems) is a PEG-based exosome precipitation kit that includes an additional size exclusion chromatography step. miRNA from the isolated exosomes were isolated using miRNEASY minikit (Qiagen) and analysed using nCounter miRNA assay (Nanostring). Principal component analysis (PCA) results suggested that the overall extracellular miRNA expression profile differed between the lymphocyte cell line originating from the Chinese American donor and the cell line originating from the Chinese Singaporean donor. As the gender, age and racial origins of both donors are similar, this may suggest that there are other genetic or epigenetic differences that account for the variation in extracellular miRNA gene expression in lymphocyte cell lines. However, statistical analysis showed that only 3 miRNA genes had a fold difference > 2 at p < 0.05, suggesting that the differences may not be of that great a significance as to impact overall conclusions drawn from different cell lines. Subsequent analysis using cell lines from other donors will give further insight into the reproducibility of results when difference cell lines are used. PCA results also suggested that the method of exosome isolation impacted the expression profile. 107 miRNA had a fold difference > 2 at p < 0.05. This suggests that the inclusion of an additional size exclusion chromatography step altered the subset of the extracellular vesicles that were isolated. In conclusion, these results suggest that extracellular miRNA can be isolated and analysed from exosomes derived from lymphocyte cell lines. However, care must be taken in the choice of cell line and method of exosome isolation used.

Keywords: biomarker, extracellular miRNA, isolation methods, lymphocyte cell line

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Authors: Weiam Daear, Patrick Lai, Elmar Prenner

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The human body offers many avenues that could be used for drug delivery. The pulmonary route, which is delivered through the lungs, presents many advantages that have sparked interested in the field. These advantages include; 1) direct access to the lungs and the large surface area it provides, and 2) close proximity to the blood circulation. The air-blood barrier of the alveoli is about 500 nm thick. The air-blood barrier consist of a monolayer of lipids and few proteins called the lung surfactant and cells. This monolayer consists of ~90% lipids and ~10% proteins that are produced by the alveolar epithelial cells. The two major lipid classes constitutes of various saturation and chain length of phosphatidylcholine (PC) and phosphatidylglycerol (PG) representing 80% of total lipid component. The major role of the lung surfactant monolayer is to reduce surface tension experienced during breathing cycles in order to prevent lung collapse. In terms of the pulmonary drug delivery route, drugs pass through various parts of the respiratory system before reaching the alveoli. It is at this location that the lung surfactant functions as the air-blood barrier for drugs. As the field of nanomedicine advances, the use of nanoparticles (NPs) as drug delivery vehicles is becoming very important. This is due to the advantages NPs provide with their large surface area and potential specific targeting. Therefore, studying the interaction of NPs with lung surfactant and whether they affect its stability becomes very essential. The aim of this research is to develop a biomimetic model of the human lung surfactant followed by a biophysical analysis of the interaction of polymeric NPs. This biomimetic model will function as a fast initial mode of testing for whether NPs affect the stability of the human lung surfactant. The model developed thus far is an 8-component lipid system that contains major PC and PG lipids. Recently, a custom made 16:0/16:1 PC and PG lipids were added to the model system. In the human lung surfactant, these lipids constitute 16% of the total lipid component. According to the author’s knowledge, there is not much monolayer data on the biophysical analysis of the 16:0/16:1 lipids, therefore more analysis will be discussed here. Biophysical techniques such as the Langmuir Trough is used for stability measurements which monitors changes to a monolayer's surface pressure upon NP interaction. Furthermore, Brewster Angle Microscopy (BAM) employed to visualize changes to the lateral domain organization. Results show preferential interactions of NPs with different lipid groups that is also dependent on the monolayer fluidity. Furthermore, results show that the film stability upon compression is unaffected, but there are significant changes in the lateral domain organization of the lung surfactant upon NP addition. This research is significant in the field of pulmonary drug delivery. It is shown that NPs within a certain size range are safe for the pulmonary route, but little is known about the mode of interaction of those polymeric NPs. Moreover, this work will provide additional information about the nanotoxicology of NPs tested.

Keywords: Brewster angle microscopy, lipids, lung surfactant, nanoparticles

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Authors: Nivedita T. Patil, M. T. Patil, M. S. Shashidhar, R. G. Gonnade

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Cyclitols are cycloalkane polyols which have raise attention since they have numerous biological and pharmaceutical properties. Among these, inositols are important cyclitols, which constitute a group of naturally occurring polyhydric alcohols. Myo, scyllo, allo, neo, D-chiro- are naturally occurring structural isomer of inositol while other four isomers (L-chiro, allo, epi-, and cis-inositol) are derived from myo-inositol by chemical synthesis. Myo-inositol, most abundant isomer, plays an important role in signal transduction process and for the treatment of type 2 diabetes, bacterial infections, stimulation of menstruation, ovulation in polycystic ovary syndrome, improvement of osteogenesis, and in treatment of neurological disorders. Considering the vast application of the derivatives, it becomes important to supply these compounds for further studies in quantitative amounts, but the synthesis of suitably protected chiral inositol derivatives is the key intermediates in most of the synthesis which is difficult. Chiral inositol derivatives could also be of interest to synthetic organic chemists as they could serve as potential starting materials for the synthesis of several natural products and their analogs. Thus, obtaining chiral myo-inositol derivatives in a more eco-friendly way is need for current inositol chemistry. Thus, the resolution of nonracemates by preferential crystallization of enantiomers has not been reported as a method for inositol derivatives. We are optimistic that this work might lead to the development of the two tosylate enantiomers as synthetic chiral pool molecules for organic synthesis. Resolution of racemic 4-O-benzyl 6-O-tosyl myo-inositol 1, 3, 5 orthoformate was successfully achieved on multigram scale by preferential crystallization, which is more scalable, eco-friendly method of separation than other reported methods. The separation of the conglomeric mixture of tosylate was achieved by suspending the mixture in ethyl acetate till the level of saturation is obtained. To this saturated clear solution was added seed crystal of the desired enantiomers. The filtration of the precipitated seed was carried out at its filtration window to get enantiomerically enriched tosylate, and the process was repeated alternatively. These enantiomerically enriched samples were recrystallized to get tosylate as pure enantiomers. The configuration of the resolved enantiomers was determined by converting it to previously reported dibenzyl ether myo-inositol, which is an important precursor for mono- and tetraphosphates. We have also developed a convenient and practical method for the preparation of enantiomeric 4-O and 6-O-allyl myo-inositol orthoesters by resolution of diastereomeric allyl dicamphante orthoesters on multigram scale. These allyl ethers can be converted to other chiral protected myo-inositol derivatives using routine synthetic transformations. The chiral allyl ethers can be obtained in gram quantities, and the methods are amenable to further scale-up due to the simple procedures involved. We believe that the work described enhances the pace of research to understand the intricacies of the myo-inositol cycle as the methods described provide efficient access to enantiomeric phosphoinositols, cyclitols, and their derivatives from the abundantly available myo-inositol as a starting material.

Keywords: cyclitols, diastereomers, enantiomers, myo-inositol, preferential crystallization, signal transduction

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Authors: Mike Wei, Mario Cedillo, Jiahui Lin, Carol Lorraine Storey-Johnson, Carla Boutin-Foster

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Purpose: The extent to which health insurance enrollment succeeds under the Affordable Care Act (ACA) rests heavily on the ability to reach the uninsured and motivate them to enroll. We sought to identify perceptions about the ACA among uninsured patients at a free clinic in New York City. Background: The ACA holds tremendous promise for reducing the number of uninsured Americans. As of April 2014, nearly 8 million people had signed up for health insurance through the Health Insurance Marketplace. Despite this early success, future and continued enrollment rests heavily on the degree of public awareness. Reaching eligible individuals and increasing their awareness and understanding remains a fundamental challenge to realizing the full potential of the ACA. Reaching out to uninsured patients who are seeking care through safety net facilities such as free clinics may provide important avenues for reaching potential enrollees. This project focuses on the experience at the free clinic at Weill Cornell Medical College, the Weill Cornell Community Clinic (WCCC), and seeks to understand perceptions about the ACA among its patient population. Methods: This was a cross-sectional study of all patients who visited the free clinic at Weill Cornell Medical College, the Weill Cornell Community Clinic, from July 2013 to May 2014. Patients who provided informed consent at their visit and completed a semi-structured questionnaire were included (N=62). The questionnaire comprised of questions about demographic characteristics and open-ended questions about their knowledge and perception of the impact of the ACA. Descriptive statistics were used to characterize the population demographics. Qualitative coding techniques were used for open-ended items. Results: Approximately one third of patients surveyed never had health insurance. Of the remaining 65%, 20% lost their insurance within the past year. Only 55% had heard about the ACA, and only 10% knew about the Health Benefits Exchange. Of those who had heard about the ACA, sentiments were tinged with optimistic misperceptions, such as “it will be free health care for all.” While optimistic, most of the responses focused on the economic implications of the ACA. Conclusions: These findings reveal the immense amount of misconception and lack of understanding with regards to the ACA. As such, the study highlights the need to educate and address the concerns of those who remain skeptical or uncertain about the implications of the ACA.

Keywords: Affordable Care Act, demographics, free clinics, underserved.

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Authors: Elena Maria Scalisi, Roberta Pecoraro, Martina Contino, Sara Ignoto, Carmelo Iaria, Santi Concetto Pavone, Gino Sorbello, Loreto Di Donato, Maria Violetta Brundo

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5G Technology is evolving to satisfy a variety of service requirements that may allow high data-rate connections (1Gbps) and lower latency times than current (<1ms). In order to support a high data transmission speed and a high traffic service for eMBB (enhanced mobile broadband) use cases, 5G systems have the characteristic of using different frequency bands of the radio wave spectrum (700 MHz, 3.6-3.8 GHz and 26.5-27.5 GHz), thus taking advantage of higher frequencies than previous mobile radio generations (1G-4G). However, waves at higher frequencies have a lower capacity to propagate in free space and therefore, in order to guarantee the capillary coverage of the territory for high reliability applications, it will be necessary to install a large number of repeaters. Following the introduction of this new technology, there has been growing concern over the past few months about possible harmful effects on human health. The aim of this preliminary study is to evaluate possible short term effects induced by 5G-millimeter waves on embryonic development and early life stages of Danio rerio by Z-FET. We exposed developing zebrafish at frequency of 27 GHz, with a standard pyramidal horn antenna placed at 15 cm far from the samples holder ensuring an incident power density of 10 mW/cm2. During the exposure cycle, from 6 h post fertilization (hpf) to 96 hpf, we measured a different morphological endpoints every 24 hours. Zebrafish embryo toxicity test (Z-FET) is a short term test, carried out on fertilized eggs of zebrafish and it represents an effective alternative to acute test with adult fish (OECD, 2013). We have observed that 5G did not reveal significant impacts on mortality nor on morphology because exposed larvae showed a normal detachment of the tail, presence of heartbeat, well-organized somites, therefore hatching rate was lower than untreated larvae even at 48 h of exposure. Moreover, the immunohistochemical analysis performed on larvae showed a negativity to the HSP-70 expression used as a biomarkers. This is a preliminary study on evaluation of potential toxicity induced by 5G and it seems appropriate to underline the importance that further studies would take, aimed at clarifying the probable real risk of exposure to electromagnetic fields.

Keywords: Biomarker of exposure, embryonic development, 5G waves, zebrafish embryo toxicity test

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Authors: Arif Tri Setyanto, Aditya Nanda Priyatama, Nugraha Arif Karyanta, Fadjri Kirana A., Afia Fitriani, Rini Setyowati, Moh.Abdul Hakim

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The Basic Health Research Report by the Ministry of Health (2018) shows an increase in the prevalence of mental health disorders in the adolescent and early adult age ranges. Supporting this finding, data on the psychological examination of the student health service unit at one State University recorded 115 cases of moderate and severe health problems in the period 2016 - 2019. More specifically, the highest number of cases was experienced by clients in the age range of 21-23 years or equivalent, with the mid-semester stage towards the end. Based on the distribution of cases experienced and the disorder becomes a psychological problem experienced by students. A total of 29% or the equivalent of 33 students experienced anxiety disorders, 25% or 29 students experienced problems ranging from mild to severe, as well as other classifications of disorders experienced, including adjustment disorders, family problems, academics, mood disorders, self-concept disorders, personality disorders, cognitive disorders, and others such as trauma and sexual disorders. Various mental health disorders have a significant impact on the academic life of students, such as low GPA, exceeding the limit in college, dropping out, disruption of social life on campus, to suicide. Based on literature reviews and best practices from universities in various countries, one of the effective ways to prevent and treat student mental health disorders is to implement a mental health monitoring system in universities. This study uses a participatory action research approach, with a sample of 423 from a total population of 32,112 students. The scale used in this study is the Beck Depression Inventory (BDI) to measure depression and the Taylor Minnesota Anxiety Scale (TMAS) to measure anxiety levels. This study aims to (1) develop a digital-based health monitoring system for students' mental health situations in the mental health category. , dangers, or those who have mental disorders, especially indications of symptoms of depression and anxiety disorders, and (2) implementing a mental health monitoring system in universities at the beginning and end of each semester. The results of the analysis show that from 423 respondents, the main problems faced by all coursework, such as thesis and academic assignments. Based on the scoring and categorization of the Beck Depression Inventory (BDI), 191 students experienced symptoms of depression. A total of 24.35%, or 103 students experienced mild depression, 14.42% (61 students) had moderate depression, and 6.38% (27 students) experienced severe or extreme depression. Furthermore, as many as 80.38% (340 students) experienced anxiety in the high category. This article will review this review of the student mental health service system on campus.

Keywords: monitoring system, mental health, psychological problems, students

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Authors: B. Grygalewicz, R. Woroniecka, J. Rygier, K. Borkowska, A. Labak, B. Nowakowska, B. Pienkowska-Grela

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Introduction: Chronic lymphocytic leukemia (CLL) is the most common form of adult leukemia in the Western world. Hemizygous and or homozygous loss at 13q14 occur in more than half of cases and constitute the most frequent chromosomal abnormality in CLL. It is believed that deletions 13q14 play a role in CLL pathogenesis. Two microRNA genes miR-15a and miR- 16-1 are targets of 13q14 deletions and plays a tumor suppressor role by targeting antiapoptotic BCL2 gene. Deletion size, as a single change detected in FISH analysis, has haprognostic significance. Patients with small deletions, without RB1 gene involvement, have the best prognosis and the longest overall survival time (OS 133 months). In patients with bigger deletion region, containing RB1 gene, prognosis drops to intermediate, like in patients with normal karyotype and without changes in FISH with overall survival 111 months. Aim: Precise delineation of 13q14 deletions regions in two groups of CLL patients, with mono- and biallelic deletions and qualifications of their prognostic significance. Methods: Detection of 13q14 deletions was performed by FISH analysis with CLL probe panel (D13S319, LAMP1, TP53, ATM, CEP-12). Accurate deletion size detection was performed by CGH Haematological Cancer and SNP array (8x60K). Results: Our investigated group of CLL patients with the 13q14 deletion, detected by FISH analysis, comprised two groups: 18 patients with monoallelic deletions and 18 patients with biallelic deletions. In FISH analysis, in the monoallelic group the range of cells with deletion, was 43% to 97%, while in biallelic group deletion was detected in 11% to 94% of cells. Microarray analysis revealed precise deletion regions. In the monoallelic group, the range of size was 348,12 Kb to 34,82 Mb, with median deletion size 7,93 Mb. In biallelic group discrepancy of total deletions, size was 135,27 Kb to 33,33 Mb, with median deletion size 2,52 Mb. The median size of smaller deletion regions on one copy chromosome 13 was 1,08 Mb while the average region of bigger deletion on the second chromosome 13 was 4,04 Mb. In the monoallelic group, in 8/18 deletion region covered RB1 gene. In the biallelic group, in 4/18 cases, revealed deletion on one copy of biallelic deletion and in 2/18 showed deletion of RB1 gene on both deleted 13q14 regions. All minimal deleted regions included miR-15a and miR-16-1 genes. Genetic results will be correlated with clinical data. Conclusions: Application of CGH microarrays technique in CLL allows accurately delineate the size of 13q14 deletion regions, what have a prognostic value. All deleted regions included miR15a and miR-16-1, what confirms the essential role of these genes in CLL pathogenesis. In our investigated groups of CLL patients with mono- and biallelic 13q14 deletions, patients with biallelic deletion presented smaller deletion sizes (2,52 Mb vs 7,93 Mb), what is connected with better prognosis.

Keywords: CLL, deletion 13q14, CGH microarrays, SNP array

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Authors: Azza El-Medany, Jamila El-Medany

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Residential and agricultural pesticide use is widespread in the world. Their extensive and indiscriminative use, in addition with their ability to interact with biological systems other than their primary targets constitute a health hazards to both humans and animals. The toxic effects of pesticides include alterations in metabolism; there is a lack of knowledge that organophosphates can cause pancreatic toxicity. The primary goal of this work is to study the effects of chronic exposure to Diazinon an organophosphate used in agriculture on pancreatic tissues and evaluate the ameliorating effect of Mesna as antioxidant on the toxicity of Diazinon on pancreatic tissues.40 adult male rats, their weight ranged between 300-350 g. The rats were classified into three groups; control (10 rats) was received corn oil at a dose of 1 0 mg/kg/day by gavage once a day for 2 months. Diazinon (15 rats) was received Diazinon at a dose of 10 mg/kg/day dissolved in corn oil by gavage once a day for 2 months. Treated group (15 rats), were received Mesna 180mg/kg once a week by gavage 15 minutes before administration of Diazinon for 2 months. At the end of the experiment, animals were anesthetized, blood samples were taken by cardiac puncture for glucose and insulin assays and pancreas was removed and divided into 3 portions; first portion for histopathological study; second portion for ultrastructural study; third portion for biochemical study using Elisa Kits including determination of malondialdehyde (MDA), tumor necrosis factor α (TNF-α), myeloperoxidase activity (MPO), interleukin 1β (IL-1β). A significant increase in the levels of MDA, TNF-α, MPO activity, IL-1β, serum glucose levels in the toxicated group with Diazinon were observed, while a significant reduction was noticed in GSH in serum insulin levels. After treatment with Mesna a significant reduction was observed in the previously mentioned parameters except that there was a significant rise in GSH in insulin levels. Histopathological and ultra-structural studies showed destruction in pancreatic tissues and β cells were the most affected cells among the injured islets as compared with the control group. The current study try to spot light about the effects of chronic exposure to pesticides on vital organs as pancreas also the role of oxidative stress that may be induced by them in evoking their toxicity. This study shows the role of antioxidant drugs in ameliorating or preventing the toxicity. This appears to be a promising approach that may be considered as a complementary treatment of pesticide toxicity.

Keywords: Diazinon, reduced glutathione, myeloperoxidase activity, tumor necrosis factor α, Mesna

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Authors: Thaminda Liyanage, Anuga Liyanage, Chamila Kurukulasuriya, Sidath Bandara

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Background: The burden of chronic kidney disease (CKD) is growing rapidly around the world, particularly in Asia. Over the last two decades Sri Lanka has experienced an epidemic of CKD with ever growing number of patients pursuing medical care due to CKD and its complications, specially in the “Mahaweli” river basin in north central region of the island nation. This was apparently a new form of CKD which was not attributable to conventional risk factors such as diabetes mellitus, hypertension or infection and widely termed as “CKD-unknown” or “CKDu”. In the past decade a number of small scale studies were conducted to determine the aetiology, prevalence and complications of CKDu in North Central region. These hospital-based studies did not provide an accurate estimate of the problem as merely 10% or less of the people with CKD are aware of their diagnosis even in developed countries with better access to medical care. Interestingly, similar observations were made on the changing epidemiology of obesity in the region but no formal study was conducted to date to determine the magnitude of obesity burden. Moreover, if increasing obesity in the region is associated with CKD epidemic is yet to be explored. Methods: We will conduct an area wide cross sectional survey among all adult residents of the “Mahaweli” development project area 5, in the North Central Province of Sri Lanka. We will collect relevant medical history, anthropometric measurements, blood and urine for hematological and biochemical analysis. We expect a participation rate of 75%-85% of all eligible participants. Participation in the study is voluntary, there will be no incentives provided for participation. Every analysis will be conducted in a central laboratory and data will be stored securely. We will calculate the prevalence of obesity and chronic kidney disease, overall and by stage using total number of participants as the denominator and report per 1000 population. The association of obesity and CKD will be assessed with regression models and will be adjusted for potential confounding factors and stratified by potential effect modifiers where appropriate. Results: This study will provide accurate information on the prevalence of obesity and CKD in the region. Furthermore, this will explore the association between obesity and CKD, although causation may not be confirmed. Conclusion: Obesity and CKD are increasingly recognized as major public health problems in Sri Lanka. Clearly, documenting the magnitude of the problem is the essential first step. Our study will provide this vital information enabling the government to plan a coordinated response to tackle both obesity and CKD in the region.

Keywords: BMI, Chronic Kidney Disease, obesity, Sri Lanka

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Authors: Sitira Williams, Georgina Cherry, Andrea Wright, Kevin Wells, Taran Rai, Richard Brown, Travis Street, Alasdair Cook

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Social media sources (50) were identified, keywords defined by veterinarians and organised into 6 topics known to be indicative of feline pruritus: body areas, behaviors, symptoms, diagnosis, and treatments. These were augmented using academic literature, a cat owner survey, synonyms, and Google Trends. The content was collected using a social intelligence solution, with keywords tagged and filtered. Data were aggregated and de-duplicated. SL content matching body areas, behaviors and symptoms were reviewed manually, and posts were marked relevant if: posted by a pet owner, identifying an itchy cat and not duplicated. A sub-set of 493 posts published from 2009-2022 was used for reflexive thematic analysis in NVIVO (Burlington, MA) to identify themes. Five themes were identified: allergy, pruritus, additional behaviors, unusual or undesirable behaviors, diagnosis, and treatment. Most (258) posts reported the cat was excessively licking, itching, and scratching. The majority were indoor cats and were less playful and friendly when itchy. Half of these posts did not indicate a known cause of pruritus. Bald spots and scabs (123) were reported, often causing swelling and fur loss, and 56 reported bumps, lumps, and dry patches. Other impacts on the cat’s quality of life were ear mites, cat self-trauma and stress. Seven posts reported their cats’ symptoms caused them ongoing anxiety and depression. Cats with food allergies to poultry (often chicken and beef) causing bald spots featured in 23 posts. Veterinarians advised switching to a raw food diet and/or changing their bowls. Some cats got worse after switching, leaving owners’ needs unmet. Allergic reactions to flea bites causing excessive itching, red spots, scabs, and fur loss were reported in 13 posts. Some (3) posts indicated allergic reactions to medication. Cats with seasonal and skin allergies, causing sneezing, scratching, headshaking, watery eyes, and nasal discharge, were reported 17 times. Eighty-five posts identified additional behaviors. Of these, 13 reported their cat’s burst pimple or insect bite. Common behaviors were headshaking, rubbing, pawing at their ears, and aggressively chewing. In some cases, bites or pimples triggered previously unseen itchiness, making the cat irritable. Twenty-four reported their cat had anxiety: overgrooming, itching, losing fur, hiding, freaking out, breathing quickly, sleeplessness, hissing and vocalising. Most reported these cats as having itchy skin, fleas, and bumps. Cats were commonly diagnosed with an ear infection, ringworm, acne, or kidney disease. Acne was diagnosed in cats with an allergy flare-up or overgrooming. Ear infections were diagnosed in itchy cats with mites or other parasites. Of the treatments mentioned, steroids were most frequently used, then anti-parasitics, including flea treatments and oral medication (steroids, antibiotics). Forty-six posts reported distress following poor outcomes after medication or additional vet consultations. SL provides veterinarians with unique insights. Verbatim comments highlight the detrimental effects of pruritus on pets and owner quality of life. This study demonstrates the need for veterinarians to communicate management and treatment options more effectively to relieve owner frustrations. Data analysis could be scaled up using machine learning for topic modeling.

Keywords: content analysis, feline, itch, pruritus, social media, thematic analysis, veterinary dermatology

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Authors: Karolina Wieczorek, Zofia Przypaśniak

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Climate change is a rapidly growing threat to global health, and part of the responsibility to combat it lies within the healthcare sector itself, including adequate education of future medical professionals. To mitigate the consequences, the General Medical Council (GMC) has equipped medical schools with a list of outcomes regarding sustainability teaching. Students are expected to analyze the impact of the healthcare sector’s emissions on climate change. The delivery of the related teaching content is, however, often inadequate and insufficient time is devoted for exploration of the topics. Teaching curricula lack in-depth exploration of the learning objectives. This study aims to assess the extent and characteristics of climate change and sustainability subjects teaching in the curriculum of a chosen UK medical school (Barts and The London School of Medicine and Dentistry). It compares the data to the national average scores from the Climate Change and Sustainability Teaching (C.A.S.T.) in Medical Education Audit to draw conclusions about teaching on a regional level. This is a single-center audit of the timetabled sessions of teaching in the medical course. The study looked at the academic year 2020/2021 which included a review of all non-elective, core curriculum teaching materials including tutorials, lectures, written resources, and assignments in all five years of the undergraduate and graduate degrees, focusing only on mandatory teaching attended by all students (excluding elective modules). The topics covered were crosschecked with GMC Outcomes for graduates: “Educating for Sustainable Healthcare – Priority Learning Outcomes” as gold standard to look for coverage of the outcomes and gaps in teaching. Quantitative data was collected in form of time allocated for teaching as proxy of time spent per individual outcomes. The data was collected independently by two students (KW and ZP) who have received prior training and assessed two separate data sets to increase interrater reliability. In terms of coverage of learning outcomes, 12 out of 13 were taught (with the national average being 9.7). The school ranked sixth in the UK for time spent per topic and second in terms of overall coverage, meaning the school has a broad range of topics taught with some being explored in more detail than others. For the first outcome 4 out of 4 objectives covered (average 3.5) with 47 minutes spent per outcome (average 84 min), for the second objective 5 out of 5 covered (average 3.5) with 46 minutes spent (average 20), for the third 3 out of 4 (average 2.5) with 10 mins pent (average 19 min). A disproportionately large amount of time is spent delivering teaching regarding air pollution (respiratory illnesses), which resulted in the topic of sustainability in other specialties being excluded from teaching (musculoskeletal, ophthalmology, pediatrics, renal). Conclusions: Currently, there is no coherent strategy on national teaching of climate change topics and as a result an unstandardized amount of time spent on teaching and coverage of objectives can be observed.

Keywords: audit, climate change, sustainability, education

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Authors: Charlotte K. Lee, Henrique R. N. Aguiar, Ralph Smith, James Baldock, Sam Botchey

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Background: Femoral neck stress fractures (FNSF) are uncommon, making up 1 to 7.2% of stress fractures in healthy subjects. FNSFs are prevalent in young women, military recruits, endurance athletes, and individuals with energy deficiency syndrome or female athlete triad. Presentation is often non-specific and is often misdiagnosed following the initial examination. There is limited research addressing the return–to–activity time after FNSF. Previous studies have demonstrated prognostic time predictions based on various imaging techniques. Here, (1) OxSport clinic FNSF practice standards are retrospectively reviewed, (2) FNSF cohort demographics are examined, (3) Regression models were used to predict return–to–activity prognosis and consequently determine bone stress risk factors. Methods: Patients with a diagnosis of FNSF attending Oxsport clinic between 01/06/2020 and 01/01/2020 were selected from the Rheumatology Assessment Database Innovation in Oxford (RhADiOn) and OxSport Stress Fracture Database (n = 14). (1) Clinical practice was audited against five criteria based on local and National Institute for Health Care Excellence guidance, with a 100% standard. (2) Demographics of the FNSF cohort were examined with Student’s T-Test. (3) Lastly, linear regression and Random Forest regression models were used on this patient cohort to predict return–to–activity time. Consequently, an analysis of feature importance was conducted after fitting each model. Results: OxSport clinical practice met standard (100%) in 3/5 criteria. The criteria not met were patient waiting times and documentation of all bone stress risk factors. Importantly, analysis of patient demographics showed that of the population with complete bone stress risk factor assessments, 53% were positive for modifiable bone stress risk factors. Lastly, linear regression analysis was utilized to identify demographic factors that predicted return–to–activity time [R2 = 79.172%; average error 0.226]. This analysis identified four key variables that predicted return-to-activity time: vitamin D level, total hip DEXA T value, femoral neck DEXA T value, and history of an eating disorder/disordered eating. Furthermore, random forest regression models were employed for this task [R2 = 97.805%; average error 0.024]. Analysis of the importance of each feature again identified a set of 4 variables, 3 of which matched with the linear regression analysis (vitamin D level, total hip DEXA T value, and femoral neck DEXA T value) and the fourth: age. Conclusion: OxSport clinical practice could be improved by more comprehensively evaluating bone stress risk factors. The importance of this evaluation is demonstrated by the population found positive for these risk factors. Using this cohort, potential bone stress risk factors that significantly impacted return-to-activity prognosis were predicted using regression models.

Keywords: eating disorder, bone stress risk factor, femoral neck stress fracture, vitamin D

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Authors: Onabajo Paul

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The testing and assessment of physical fitness of school-aged adolescents in Nigeria has been going on for several decades. Originally, these tests strictly focused on identifying health and physical fitness status and comparing the results of adolescents with others. There is a considerable interest in health and performance fitness of adolescents in which results attained are compared with criteria representing positive health rather than simply on score comparisons with others. Despite the fact that physical education program is being studied in secondary schools and physical activities are encouraged, it is observed that regular assessment of students’ fitness level and health status seems to be scarce or not being done in these schools. The purpose of the study was to assess the heath and performance fitness of adolescents in middle-income schools in Lagos State. A total number of 150 students were selected using the simple random sampling technique. Participants were measured on hand grip strength, sit-up, pacer 20 meter shuttle run, standing long jump, weight and height. The data collected were analyzed with descriptive statistics of means, standard deviations, and range and compared with fitness norms. It was concluded that majority 111(74.0%) of the adolescents achieved the healthy fitness zone, 33(22.0%) were very lean, and 6(4.0%) needed improvement according to the normative standard of Body Mass Index test. For muscular strength, majority 78(52.0%) were weak, 66(44.0%) were normal, and 6(4.0%) were strong according to the normative standard of hand-grip strength test. For aerobic capacity fitness, majority 93(62.0%) needed improvement and were at health risk, 36(24.0%) achieved healthy fitness zone, and 21(14.0%) needed improvement according to the normative standard of PACER test. Majority 48(32.0%) of the participants had good hip flexibility, 38(25.3%) had fair status, 27(18.0%) needed improvement, 24(16.0%) had very good hip flexibility status, and 13(8.7%) of the participants had excellent status. Majority 61(40.7%) had average muscular endurance status, 30(20.0%) had poor status, 29(18.3%) had good status, 28(18.7%) had fair muscular endurance status, and 2(1.3%) of the participants had excellent status according to the normative standard of sit-up test. Majority 52(34.7%) had low jump ability fitness, 47(31.3%) had marginal fitness, 31(20.7%) had good fitness, and 20(13.3%) had high performance fitness according to the normative standard of standing long jump test. Based on the findings, it was concluded that majority of the adolescents had better Body Mass Index status, and performed well in both hip flexibility and muscular endurance tests. Whereas majority of the adolescents performed poorly in aerobic capacity test, muscular strength and jump ability test. It was recommended that to enhance wellness, adolescents should be involved in physical activities and recreation lasting 30 minutes three times a week. Schools should engage in fitness program for students on regular basis at both senior and junior classes so as to develop good cardio-respiratory, muscular fitness and improve overall health of the students.

Keywords: adolescents, health-related fitness, performance-related fitness, physical fitness

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Authors: Mustafa Metin Donma, Orkide Donma

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Childhood obesity is an important clinical problem because it may lead to chronic diseases during the adulthood period of the individual. Obesity is a metabolic disease associated with low-grade inflammation. The liver occurs at the center of metabolic pathways. Adropin, fibroblast growth factor-21 (FGF-21), and fetuin-A are hepatokines. Due to the immense participation of the liver in glucose metabolism, these liver-derived factors may be associated with insulin resistance (IR), which is a phenomenon discussed within the scope of obesity problems. The aim of this study is to determine the concentrations of adropin, FGF-21, and fetuin-A in childhood obesity, to point out possible differences between the obesity groups, and to investigate possible associations among these three hepatokines in obese and morbidly obese children. A total of one hundred and thirty-two children were included in the study. Two obese groups were constituted. The groups were matched in terms of mean ± SD values of ages. Body mass index values of obese and morbidly obese groups were 25.0 ± 3.5 kg/m² and 29.8 ± 5.7 kg/m², respectively. Anthropometric measurements including waist circumference, hip circumference, head circumference, and neck circumference were recorded. Informed consent forms were taken from the parents of the participants. The ethics committee of the institution approved the study protocol. Blood samples were obtained after overnight fasting. Routine biochemical tests, including glucose- and lipid-related parameters, were performed. Concentrations of the hepatokines (adropin, FGF-21, fetuin A) were determined by enzyme-linked immunosorbent assay. Insulin resistance indices such as homeostasis model assessment for IR (HOMA-IR), alanine transaminase-to aspartate transaminase ratio (ALT/AST), diagnostic obesity notation model assessment laboratory index, diagnostic obesity notation model assessment metabolic syndrome index as well as obesity indices such as diagnostic obesity notation model assessment-II index, and fat mass index were calculated using the previously derived formulas. Statistical evaluation of the study data as well as findings of the study was performed by SPSS for Windows. Statistical difference was accepted significant when p is smaller than 0.05. Statistically significant differences were found for insulin, triglyceride, high-density lipoprotein cholesterol levels of the groups. A significant increase was observed for FGF-21 concentrations in the morbidly obese group. Higher adropin and fetuin-A concentrations were observed in the same group in comparison with the values detected in the obese group (p > 0.05). There was no statistically significant difference between the ALT/AST values of the groups. In all of the remaining IR and obesity indices, significantly increased values were calculated for morbidly obese children. Significant correlations were detected between HOMA-IR and each of the hepatokines. The highest one was the association with fetuin-A (r=0.373, p=0.001). In conclusion, increased levels observed in adropin, FGF-21, and fetuin-A have shown that these hepatokines possess increasing potential going from obese to morbid obese state. Out of the correlations found with the IR index, the most affected hepatokine was fetuin-A, the parameter possibly used as the indicator of the advanced obesity stage.

Keywords: adropin, fetuin A, fibroblast growth factor-21, insulin resistance, pediatric obesity

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Authors: S. F. Afzali, W. L. Wong

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The epizootic ulcerative syndrome (EUS) causes by the oomycete fungus, Aphanomyces invadans; known to be one of the infectious fish diseases for farmed and wild fishes in fresh and brackish-water from the Asia-pacific region, America and Africa. Although, EUS had been documented by the Office International des Epizooties (OIE) since 1995, hitherto, there is neither standard chemical agents that can be used for successful treatment of this destructive infection in the time of outbreak; nor available vaccine for prevention. Plant-based remedies in controlling fish diseases are gaining much attention recently as an alternative to chemical treatments, which possess negative effects to the environment and human. In present study, Sonneratia alba, a mangrove plant belongs to the Sonneratiaceae family, was screened in vitro and in vivo for its antifungal activity against A. invadans mycelium growth and its effects on fish innate immune system and disease resistant. The in vitro tests was performed using the disc diffusion methods with measurements of minimum inhibitory concentration (MIC) and inhibition zone. For in vivo study, the S. alba extract supplemented diets were administrated at 0.0, 1.0%, 3.0%, and 5.0% on healthy goldfish, Carassius auratus, which challenged with A. invadans zoospores (100 spores/ml). To compare the significant differences in the hematological and immunological parameters obtained from the experiments, the data were analysed using the SPSS. The methanol extract of S. alba effectively inhibited the mycelial growth of A. invadans at a minimum concentration of 1000 ppm for agar and filter paper diffusion experiments. In the agar diffusion test, 500 ppm of the extract inhibited the fungus mycelial growth up to 96 hours after exposure. The mycelial growth from the edge of the pre-inoculated A. invadans agar discs treated with S. alba extracts at concentrations of 100, 500 and 1000 ppm were 15, 8 and 0 mm respectively. The results of the filter paper disc test showed that the S. alba extract at its minimal inhibitory concentration (1000 ppm) has similar qualitative inhibitory effect as malachite green at 1 ppm and formalin at 250 ppm. According to the in vivo tests findings, in the infected fish fed with 3.0% and 5.0% supplementation diet, the numbers of white blood cell and myeloperoxidase activity significantly increased after the second week of treatment. Whilst the numbers of red blood cell significantly decreased in the infected fish fed with 0.0 and 1.0% supplementation diet. After the third week of feeding, significant increases in the total protein, albumin level, lysozyme activity were recorded in the infected fish fed with 3.0% and 5.0% supplementation diet. Also, the enriched diets increased the survival rate as compared to the untreated group that suffered from 90% mortality. The present study indicated that S. alba extract may inhibit the mycelial growth of A. invadans effectively, suggesting an alternative to other chemotherapeutic agents, which brought much environmental and health concerns to the public, for EUS treatment.

Keywords: fungal pathogen, goldfish, organic extract, treatment

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Authors: Diom Loreen Ndum, Omarine Njimanted

Abstract:

With increasing automation in clinical laboratories, the requirements for quality control materials have greatly increased in order to monitor daily performance. The constant use of commercial control material is not economically feasible for many developing countries because of non-availability or the high-cost of the materials. Therefore, preparation and use of in-house quality control serum will be a very cost-effective measure with respect to laboratory needs.The objective of this study was to determine the efficacy of in-house prepared pooled sera with respect to commercially acquired control sample for routine internal quality control at the Nkwen District Hospital Laboratory. This was an analytical study, serum was taken from leftover serum samples of 5 healthy adult blood donors at the blood bank of Nkwen District Hospital, which had been screened negative for human immunodeficiency virus (HIV), hepatitis C virus (HCV) and Hepatitis B antigens (HBsAg), and were pooled together in a sterile container. From the pooled sera, sixty aliquots of 150µL each were prepared. Forty aliquots of 150µL each of commercially acquired samples were prepared after reconstitution and stored in a deep freezer at − 20°C until it was required for analysis. This study started from the 9th June to 12th August 2022. Every day, alongside with commercial control sample, one aliquot of pooled sera was removed from the deep freezer and allowed to thaw before analyzed for the following parameters: blood urea, serum creatinine, aspartate aminotransferase (AST), alanine aminotransferase (ALT), potassium and sodium. After getting the first 20 values for each parameter of pooled sera, the mean, standard deviation and coefficient of variation were calculated, and a Levey-Jennings (L-J) chart established. The mean and standard deviation for commercially acquired control sample was provided by the manufacturer. The following results were observed; pooled sera had lesser standard deviation for creatinine, urea and AST than commercially acquired control samples. There was statistically significant difference (p<0.05) between the mean values of creatinine, urea and AST for in-house quality control when compared with commercial control. The coefficient of variation for the parameters for both commercial control and in-house control samples were less than 30%, which is an acceptable difference. The L-J charts revealed shifts and trends (warning signs), so troubleshooting and corrective measures were taken. In conclusion, in-house quality control sample prepared from pooled serum can be a good control sample for routine internal quality control.

Keywords: internal quality control, levey-jennings chart, pooled sera, shifts, trends, westgard rules

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Authors: Emily Schulz

Abstract:

The ancient Vedic culture from India had traditions that supported all aspects of health, including psychological health, and are relevant in the current era. These traditions have been compiled by Professor Dr. Purna, a rare Himalayan Master, into the Purna Health Management System (PHMS). The PHMS is a unique, holistic, and integrated approach to health management. It is comprised of four key factors: Health, Fitness, and Nutrition (HF&N), Life Balance (Stress Management) (LB-SM), Spiritual Growth and Development (SG&D); and Living in Harmony with the Natural Environment (LHWNE). The purpose of the PHMS is to give people the tools to take responsibility for managing their own holistic health and wellbeing. A study using a cross-sectional mixed-methods anonymous online survey was conducted during 2017-2018. Adult students of Professor Dr. Purna were invited to participate through announcements made at various events He held throughout the globe. Follow-up emails were sent with consenting language for interested parties and provided them with a link to the survey. Participation in the study was completely voluntary and no incentives were given to respond to the survey. The overall aim of the study was to investigate the effectiveness of implementation of the PHMS on practitioners' emotional balance. However, given the holistic nature of the PHMS, survey questions also inquired about participants’ physical health, stress level, ability to manage stress, and wellbeing using Likert scales. The survey also included some open-ended questions to gain an understanding of the participants’ experiences with the PHMS relative to their emotional balance. In total, 52 people out of 253 potential respondents participated in the study. Data were analyzed using nonparametric Spearman’s Rho correlation coefficient (rs) since the data were not on a normal distribution. Statistical significance was set at p < .05. Results of the study suggested that there are moderate to strong statistically significant relationships (p < .001) between participants' frequent implementation of each of the four key factors of the PHMS and self-reported mental/emotional health (HF&N rs = 0.42; LB-SM rs = 0.54; SG&D rs = 0.49; LHWNE rs = 0.45) Results also demonstrated statistically significant relationships (p < .001) between participants' frequent implementation of each of the four key factors of the PHMS and their self-reported ability to manage stress (HF&N rs = 0.44; LB-SM rs = 0.55; SG&D rs = 0.39; LHWNE rs = 0.55). Additionally, those who reported experiencing better physical health also reported better mental/emotional health (rs = 0.49, p < .001) and better ability to manage stress (rs = 0.46, p < .001). The findings of this study suggest that wisdom from the ancient Vedic culture may be useful for those working in the field of psychology and related fields who would like to assist clients in calming their mind and emotions and managing their stress levels.

Keywords: balanced emotions, balanced mind, stress management, Vedic philosophy

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Authors: Safaa S. Hassan, Mohammed H. Elbakry, Safwat A. Mangoura, Zainab M. Omar

Abstract:

Background: Liver fibrosis is the main reason for increased mortality in chronic liver disease. It has no standard treatment. Antioxidants from a variety of sources are capable of slowing or preventing oxidation of other molecules. Aim: to evaluate the hepatoprotective effect of vit. C, vit. E and gallic acid in comparison to silymarin in the rat model of carbon tetrachloride induced liver fibrosis and their possible mechanisms of action. Material& Methods: A total number of 60 adult male albino rats 160-200gm were divided into six equal groups; received subcutaneous (s.c) injection for 8 weeks. Group I: as control. Group II: received 1.5 mL/kg of CCL4 .Group III: CCL4 and co- treatment with silymarin 100mg/kg p.o. daily. Group IV: CCL4 and co-treatment with vit. C 50mg/kg p.o. daily. Group V: CCL4 and co-treatment with vit. E 200mg/kg. p.o. Group VI: CCL4 and co-treatment with Gallic acid 100mg/kg. p.o. daily. Liver was processed for histological and immunohistochemical examination. Levels of AST, ALT, ALP, reduced GSH, MDA, SOD and hydroxyproline concentration were measured and evaluated statistically. Results: Light and electron microscopic examination of liver of group II exhibited foci of altered cells with dense nuclei and vacuolated, granular cytoplasm, mononuclear cell infiltration in portal areas, profuse collagen fiber deposits were found around portal tract, more intense staining α-SMA-positive cells occupied most of the liver fibrosis tissue, electron lucent areas in the cytoplasm of the hepatocytes, margination of nuclear chromatin. Treatment by any of the antioxidants variably reduced the hepatic structural changes induced by CCL4. Biochemical analysis showed that carbon tetrachloride significantly increased the levels of serum AST, ALT, ALP, hepatic malondialdehyde and hydroxyproline content. Moreover, it decreased the activities of superoxide dismutase and glutathione. Treatment with silymarin, gallic acid, vit. C and vit. E decreased significantly the AST, ALT, and ALP levels in plasma, MDA and hydroxyproline and increased the activities of SOD and glutathione in liver tissue. The effect of administration of CCl4 was improved with the used antioxidants in variable degrees. The most efficient antioxidant was silymarin followed by gallic acid and vit. C then vit. E. It is possibly due to their antioxidant effect, free radical scavenging properties and the reduction of oxidant dependent activation and proliferation of HSCs. Conclusion: So these antioxidants can be a promising drugs candidate for ameliorating liver fibrosis better than the use of the drugs and their side effects.

Keywords: antioxidant, ccl4, gallic acid, liver fibrosis

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Authors: Ling-Ling E., Hong-Chen Liu, Dong-Sheng Wang, Fang Su, Xia Wu, Zhan-Ping Shi, Yan Lv, Jia-Zhu Wang

Abstract:

Objective: The objective of the present study is to evaluate the capacity of a tissue-engineered bone complex of recombinant human bone morphogenetic protein 2 (rhBMP-2) mediated dental pulp stem cells (DPSCs) and nano-hydroxyapatite/collagen/poly(L-lactide)(nHAC/PLA) to reconstruct critical-size alveolar bone defects in New Zealand rabbit. Methods: Autologous DPSCs were isolated from rabbit dental pulp tissue and expanded ex vivo to enrich DPSCs numbers, and then their attachment and differentiation capability were evaluated when cultured on the culture plate or nHAC/PLA. The alveolar bone defects were treated with nHAC/PLA, nHAC/PLA+rhBMP-2, nHAC/PLA+DPSCs, nHAC/PLA+DPSCs+rhBMP-2, and autogenous bone (AB) obtained from iliac bone or were left untreated as a control. X-ray and a polychrome sequential fluorescent labeling were performed post-operatively and the animals were sacrificed 12 weeks after operation for histological observation and histomorphometric analysis. Results: Our results showed that DPSCs expressed STRO-1 and vementin, and favoured osteogenesis and adipogenesis in conditioned media. DPSCs attached and spread well, and retained their osteogenic phenotypes on nHAC/PLA. The rhBMP-2 could significantly increase protein content, alkaline phosphatase (ALP) activity/protein, osteocalcin (OCN) content, and mineral formation of DPSCs cultured on nHAC/PLA. The X-ray graph, the fluorescent, histological observation and histomorphometric analysis showed that the nHAC/PLA+DPSCs+rhBMP-2 tissue-engineered bone complex had an earlier mineralization and more bone formation inside the scaffold than nHAC/PLA, nHAC/PLA+rhBMP-2 and nHAC/PLA+DPSCs, or even autologous bone. Implanted DPSCs contribution to new bone were detected through transfected eGFP genes. Conclutions: Our findings indicated that stem cells existed in adult rabbit dental pulp tissue. The rhBMP-2 promoted osteogenic capability of DPSCs as a potential cell source for periodontal bone regeneration. The nHAC/PLA could serve as a good scaffold for autologous DPSCs seeding, proliferation and differentiation. The tissue-engineered bone complex with nHAC/PLA, rhBMP-2, and autologous DPSCs might be a better alternative to autologous bone for the clinical reconstruction of periodontal bone defects.

Keywords: nano-hydroxyapatite/collagen/poly (L-lactide), dental pulp stem cell, recombinant human bone morphogenetic protein, bone tissue engineering, alveolar bone

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Authors: Perizat Aitmaganbet, Kerbez Kimatova, Gulmira Umarova

Abstract:

As a result of medical check-up conducted in the framework of this research study an evaluation of the health status of the population living in the oil-producing regions, namely Sarkul and Kenkiyak villages in Aktobe was examined. With the help of the Spearman correlation, the connection between the level of hazard chemical elements in the atmosphere and the health of population living in the regions of oil and gas industry was estimated. Background & Objective. The oil and gas resource-extraction industries play an important role in improving the economic conditions of the Republic of Kazakhstan, especially for the oil-producing administrative regions. However, environmental problems may adversely affect the health of people living in that area. Thus, the aim of the study is to evaluate the exposure to negative environmental factors of the adult population living in Sarkul and Kenkiyak villages, the oil and gas producing areas in the Aktobe region. Methods. After conducting medical check-up among the population of Sarkul and Kenkiyak villages. A single cross-sectional study was conducted. The population consisted of randomly sampled 372 adults (181 males and 191 females). Also, atmospheric air probes were taken to measure the level of hazardous chemical elements in the air. The nonparametric method of the Spearman correlation analysis was performed between the mean concentration of substances exceeding the Maximum Permissible Concentration and the classes of newly diagnosed diseases. Selection and analysis of air samples were carried out according to the developed research protocol; the qualitative-quantitative analysis was carried out on the Gas analyzer HANK-4 apparatus. Findings. The medical examination of the population identified the following diseases: the first two dominant were diseases of the circulatory and digestive systems, in the 3rd place - diseases of the genitourinary system, and the nervous system and diseases of the ear and mastoid process were on the fourth and fifth places. Moreover, significant pollution of atmospheric air by carbon monoxide (MPC-5,0 mg/m3), benzapyrene (MPC-1mg/m3), dust (MPC-0,5 mg/m3) and phenol (МРС-0,035mg/m3) were identified in places. Correlation dependencies between these pollutants of air and the diseases of the population were established, as a result of diseases of the circulatory system (r = 0,7), ear and mastoid process (r = 0,7), nervous system (r = 0,6) and digestive organs(r = 0,6 ); between the concentration of carbon monoxide and diseases of the circulatory system (r = 0.6), the digestive system(r = 0.6), the genitourinary system (r = 0.6) and the musculoskeletal system; between nitric oxide and diseases of the digestive system (r = 0,7) and the circulatory system (r = 0,6); between benzopyrene and diseases of the digestive system (r = 0,6), the genitourinary system (r = 0,6) and the nervous system (r = 0,4). Conclusion. The positive correlation was found between air pollution and the health of the population living in Sarkul and Kenkiyak villages. To enhance the reliability of the results we are going to continue this study further.

Keywords: atmospheric air, chemical substances, oil and gas, public health

Procedia PDF Downloads 103

Authors: Abdolamir Allameh, Shahnaz Esmaeili, Mina Allameh, Safoura Khajeniazi

Abstract:

Stem cells with therapeutic applications can be isolated from human placenta/umblical cord blood (UCB) as well as the cord tissue (UC). Stem cells in culture are vulnerable to oxidative stress, particularly when subjected to differentiation process. The aim of this study was to examine the chnages in the rate of oxidation that occurs to cellular macromolecules during hepatic differentiation of mononuclear cells (MSCs). In addition, the impact of the hepatic differentiation process of MSC on cellular and biological activity of the cells will be undertaken. For this purpose, first mononuclear cells (MNCs) were isolated from human UCB which was obtained from a healthy full-term infant. The cells were cultured at a density of 3×10⁵ cells/cm² in DMEM- low-glucose culture media supplemented with 20% FBS, 2 mM L-glutamine, 100 μg/ml streptomycin and 100 U/ml penicillin. Cell cultures were then incubated at 37°C in a humidified 5% CO₂ incubator. After removing non-adherent cells by replacing culture medium, fibroblast-like adherent cells were resuspended in 0.25% trypsin-EDTA and plated in 25 cm² flasks (1×10⁴/ml). Characterization of the MSCs was routinely done by observing their morphology and growth curve. MSCs were subjected to a 2-step hepatocyte differentiation protocol in presence of hepatocyte growth factor (HGF), dexamethazone (DEX) and oncostatin M (OSM). The hepatocyte-like cells derived from MSCs were checked every week for 3 weeks for changes in lipid peroxidation, protein carbonyl formation and DNA oxidation i.e., 8-hydroxy-2'-deoxyguanosine (8-OH-dG) assay. During the 3-week differentiation process of MSCs to hepatocyte-like cells we found that expression liver-specific markers such as albumin, was associated with increased levels of lipid peroxidation and protein carbonyl formation. Whereas, undifferentiated MSCs has relatively low levels of lipid peroxidation products. There was a significant increase ( p < 0.05) in lipid peroxidation products in hepatocytes on days 7, 14, and 21 of differentiation. Likewise, the level of protein carbonyls in the cells was elevated during the differentiation. The level of protein carbonyls measured in hepatocyte-like cells obtained 3 weeks after differentiation induction was estimated to be ~6 fold higher compared to cells recovered on day 7 of differentiation. On the contrary, there was a small but significant decrease in DNA damage marker (8-OH-dG) in hepatocytes recovered 3 weeks after differentiation onset. The level of 8-OHdG which was in consistent with formation of reactive oxygen species (ROS). In conclusion, this data suggest that despite the elevation in oxidation of lipid and protein molecules during hepatocyte development, the cells were normal in terms of DNA integrity, morphology, and biologically activity.

Keywords: adult stem cells, DNA integrity, free radicals, hepatic differentiation

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