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Authors: Katarzyna Drela, Miroslaw Wielgos, Mikolaj Wrobel, Barbara Lukomska

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Mesenchymal stem cells (MSCs) have a great potential in regenerative medicine. Since the initial number of isolated MSCs is limited, in vitro propagation is often required to reach sufficient numbers of cells for therapeutic applications. During long-term culture MSCs may undergo genetic or epigenetic alterations that subsequently increase the probability of spontaneous malignant transformation. Thus, factors that influence genomic stability of MSCs following long-term expansions need to be clarified before cultured MSCs are employed for clinical application. The aim of our study was to investigate the potential for spontaneous transformation of human neonatal cord blood (HUCB-MSCs) and adult bone marrow (BM-MSCs) derived MSCs. Materials and Methods: HUCB-MSCs and BM-MSCs were isolated by standard Ficoll gradient centrifugations method. Isolated cells were initially plated in high density 106 cells per cm2. After 48 h medium were changed and non-adherent cells were removed. The malignant transformation of MSCs in vitro was evaluated by morphological changes, proliferation rate, ability to enter cell senescence, the telomerase expression and chromosomal abnormality. Proliferation of MSCs was analyzed with WST-1 reduction method and population doubling time (PDT) was calculated at different culture stages. Then the expression pattern of genes characteristic for mesenchymal or epithelial cells, as well as transcriptions factors were examined by RT-PCR. Concomitantly, immunocytochemical analysis of gene-related proteins was employed. Results: Our studies showed that MSCs from all bone marrow isolations ultimately entered senescence and did not undergo spontaneous malignant transformation. However, HUCB-MSCs from one of the 15 donors displayed an increased proliferation rate, failed to enter senescence, and exhibited an altered cell morphology. In this sample we observed two different cell phenotypes: one mesenchymal-like exhibited spindle shaped morphology and express specific mesenchymal surface markers (CD73, CD90, CD105, CD166) with low proliferation rate, and the second one with round, densely package epithelial-like cells with significantly increased proliferation rate. The PDT of epithelial-like populations was around 1day and 100% of cells were positive for proliferation marker Ki-67. Moreover, HUCB-MSCs showed a positive expression of human telomerase reverse transcriptase (hTERT), cMYC and exhibit increased number of CFU during the long-term culture in vitro. Furthermore, karyotype analysis revealed chromosomal abnormalities including duplications. Conclusions: Our studies demonstrate that HUCB-MSCs are susceptible to spontaneous malignant transformation during long-term culture. Spontaneous malignant transformation process following in vitro culture has enormous effect on the biosafety issues of future cell-based therapies and regenerative medicine regimens.

Keywords: mesenchymal stem cells, spontaneous, transformation, long-term culture

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Authors: M. Salvador, J. C. Martinez-Garcia, A. Moyano, M. C. Blanco-Lopez, M. Rivas

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Biosensors play a crucial role in the detection of molecules nowadays due to their advantages of user-friendliness, high selectivity, the analysis in real time and in-situ applications. Among them, Lateral Flow Immunoassays (LFIAs) are presented among technologies for point-of-care bioassays with outstanding characteristics such as affordability, portability and low-cost. They have been widely used for the detection of a vast range of biomarkers, which do not only include proteins but also nucleic acids and even whole cells. Although the LFIA has traditionally been a positive/negative test, tremendous efforts are being done to add to the method the quantifying capability based on the combination of suitable labels and a proper sensor. One of the most successful approaches involves the use of magnetic sensors for detection of magnetic labels. Bringing together the required characteristics mentioned before, our research group has developed a biosensor to detect biomolecules. Superparamagnetic nanoparticles (SPNPs) together with LFIAs play the fundamental roles. SPMNPs are detected by their interaction with a high-frequency current flowing on a printed micro track. By means of the instant and proportional variation of the impedance of this track provoked by the presence of the SPNPs, quantitative and rapid measurement of the number of particles can be obtained. This way of detection requires no external magnetic field application, which reduces the device complexity. On the other hand, the major limitations of LFIAs are that they are only qualitative or semiquantitative when traditional gold or latex nanoparticles are used as color labels. Moreover, the necessity of always-constant ambient conditions to get reproducible results, the exclusive detection of the nanoparticles on the surface of the membrane, and the short durability of the signal are drawbacks that can be advantageously overcome with the design of magnetically labeled LFIAs. The approach followed was to coat the SPIONs with a specific monoclonal antibody which targets the protein under consideration by chemical bonds. Then, a sandwich-type immunoassay was prepared by printing onto the nitrocellulose membrane strip a second antibody against a different epitope of the protein (test line) and an IgG antibody (control line). When the sample flows along the strip, the SPION-labeled proteins are immobilized at the test line, which provides magnetic signal as described before. Preliminary results using this practical combination for the detection and quantification of the Prostatic-Specific Antigen (PSA) shows the validity and consistency of the technique in the clinical range, where a PSA level of 4.0 ng/mL is the established upper normal limit. Moreover, a LOD of 0.25 ng/mL was calculated with a confident level of 3 according to the IUPAC Gold Book definition. Its versatility has also been proved with the detection of other biomolecules such as troponin I (cardiac injury biomarker) or histamine.

Keywords: biosensor, lateral flow immunoassays, point-of-care devices, superparamagnetic nanoparticles

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Authors: Alvarez M., Chapela M. J., Balboa E., Rubianes D., Sinde E., Fernandez de Ana C., Rodríguez-Blanco A.

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The gut microbiota plays an important role as gut inflammation could contribute to colorectal cancer development; however, this role is still not fully understood, and tools able to prevent this progression are yet to be developed. The main objective of this study was to monitor the effects of a mushroom extracts formulation in gut microbial community composition of an Azoxymethane (AOM)/Dextran sodium sulfate (DSS) mice model of inflammation-linked carcinogenesis. For the in vivo study, 41 adult male mice of the C57BL / 6 strain were obtained. 36 of them have been induced in a state of colon carcinogenesis by a single intraperitoneal administration of AOM at a dose of 12.5 mg/kg; the control group animals received instead of the same volume of 0.9% saline. DSS is an extremely toxic polysaccharide sulfate that causes chronic inflammation of the colon mucosa, favoring the appearance of severe colitis and the production of tumors induced by AOM. Induction by AOM/DSS is an interesting platform for chemopreventive intervention studies. This time the model was used to monitor gut microbiota changes as a result of supplementation with a specific mushroom extracts formulation previously shown to have prebiotic activity. The animals have been divided into three groups: (i) Cancer + mushroom extracts formulation experimental group: to which the MicoDigest2.0 mushroom extracts formulation developed by Hifas da Terra S.L has been administered dissolved in drinking water at an estimated concentration of 100 mg / ml. (ii) Control group of animals with Cancer: to which normal water has been administered without any type of treatment. (iii) Control group of healthy animals: these are the animals that have not been induced cancer or have not received any treatment in drinking water. This treatment has been maintained for a period of 3 months, after which the animals were sacrificed to obtain tissues that were subsequently analyzed to verify the effects of the mushroom extract formulation. A microbiological analysis has been carried out to compare the microbial communities present in the intestines of the mice belonging to each of the study groups. For this, the methodology of massive sequencing by molecular analysis of the 16S gene has been used (Ion Torrent technology). Initially, DNA extraction and metagenomics libraries were prepared using the 16S Metagenomics kit, always following the manufacturer's instructions. This kit amplifies 7 of the 9 hypervariable regions of the 16S gene that will then be sequenced. Finally, the data obtained will be compared with a database that makes it possible to determine the degree of similarity of the sequences obtained with a wide range of bacterial genomes. Results obtained showed that, similarly to certain natural compounds preventing colorectal tumorigenesis, a mushroom formulation enriched the Firmicutes and Proteobacteria phyla and depleted Bacteroidetes. Therefore, it was demonstrated that the consumption of the mushroom extracts’ formulation developed could promote the recovery of the microbial balance that is disrupted in the mice model of carcinogenesis. More preclinical and clinical studies are needed to validate this promising approach.

Keywords: carcinogenesis, microbiota, mushroom extracts, inflammation

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Authors: Sonia Butalia, Huong Luu, Alexis Guigue, Karen J. B. Martins, Khanh Vu, Scott W. Klarenbach

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Obesity-related health conditions impose a substantial economic burden on payers due to increased healthcare use. Estimates of healthcare resource use and costs associated with obesity-related comorbidities are needed to inform policies and interventions targeting these conditions. Methods: Adults living with obesity were identified (a procedure-related body mass index code for class 2/3 obesity between 2012 and 2019 in Alberta, Canada; excluding those with bariatric surgery), and outcomes were compared over 1-year (2019/2020) between those who had and did not have specific obesity-related comorbidities. The probability of using a healthcare service (based on the odds ratio of a zero [OR-zero] cost) was compared; 95% confidence intervals (CI) were reported. Logistic regression and a generalized linear model with log link and gamma distribution were used for total healthcare cost comparisons ($CDN); cost ratios and estimated cost differences (95% CI) were reported. Potential socio-demographic and clinical confounders were adjusted for, and incremental cost differences were representative of a referent case. Results: A total of 220,190 adults living with obesity were included; 44% had hypertension, 25% had osteoarthritis, 24% had type-2 diabetes, 17% had cardiovascular disease, 12% had insulin resistance, 9% had chronic back pain, and 4% of females had polycystic ovarian syndrome (PCOS). The probability of hospitalization, ED visit, and ambulatory care was higher in those with a following obesity-related comorbidity versus those without: chronic back pain (hospitalization: 1.8-times [OR-zero: 0.57 [0.55/0.59]] / ED visit: 1.9-times [OR-zero: 0.54 [0.53/0.56]] / ambulatory care visit: 2.4-times [OR-zero: 0.41 [0.40/0.43]]), cardiovascular disease (2.7-times [OR-zero: 0.37 [0.36/0.38]] / 1.9-times [OR-zero: 0.52 [0.51/0.53]] / 2.8-times [OR-zero: 0.36 [0.35/0.36]]), osteoarthritis (2.0-times [OR-zero: 0.51 [0.50/0.53]] / 1.4-times [OR-zero: 0.74 [0.73/0.76]] / 2.5-times [OR-zero: 0.40 [0.40/0.41]]), type-2 diabetes (1.9-times [OR-zero: 0.54 [0.52/0.55]] / 1.4-times [OR-zero: 0.72 [0.70/0.73]] / 2.1-times [OR-zero: 0.47 [0.46/0.47]]), hypertension (1.8-times [OR-zero: 0.56 [0.54/0.57]] / 1.3-times [OR-zero: 0.79 [0.77/0.80]] / 2.2-times [OR-zero: 0.46 [0.45/0.47]]), PCOS (not significant / 1.2-times [OR-zero: 0.83 [0.79/0.88]] / not significant), and insulin resistance (1.1-times [OR-zero: 0.88 [0.84/0.91]] / 1.1-times [OR-zero: 0.92 [0.89/0.94]] / 1.8-times [OR-zero: 0.56 [0.54/0.57]]). After fully adjusting for potential confounders, the total healthcare cost ratio was higher in those with a following obesity-related comorbidity versus those without: chronic back pain (1.54-times [1.51/1.56]), cardiovascular disease (1.45-times [1.43/1.47]), osteoarthritis (1.36-times [1.35/1.38]), type-2 diabetes (1.30-times [1.28/1.31]), hypertension (1.27-times [1.26/1.28]), PCOS (1.08-times [1.05/1.11]), and insulin resistance (1.03-times [1.01/1.04]). Conclusions: Adults with obesity who have specific disease-related health conditions have a higher probability of healthcare use and incur greater costs than those without specific comorbidities; incremental costs are larger when other obesity-related health conditions are not adjusted for. In a specific referent case, hypertension was costliest (44% had this condition with an additional annual cost of $715 [$678/$753]). If these findings hold for the Canadian population, hypertension in persons with obesity represents an estimated additional annual healthcare cost of $2.5 billion among adults living with obesity (based on an adult obesity rate of 26%). Results of this study can inform decision making on investment in interventions that are effective in treating obesity and its complications.

Keywords: administrative data, healthcare cost, obesity-related comorbidities, real world evidence

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Authors: Emily Rowe

Abstract:

Introduction: Balance assessments can be used to help evaluate a person’s risk of falls, determine causes of balance deficits and inform intervention decisions. It is widely accepted that instrumented quantitative analysis can be more reliable and specific than semi-qualitative ordinal scales or itemised scoring methods. However, the uptake of quantitative methods is hindered by expense, lack of portability, and set-up requirements. During stepping, foot placement is actively coordinated with the body centre of mass (COM) kinematics during pre-initiation. Based on this, the potential to use COM velocity just prior to foot off and foot placement error as an outcome measure of dynamic balance is currently being explored using complex 3D motion capture. Inertial sensors and pressure mats might be more practical technologies for measuring these parameters in clinical settings. Objective: The aim of this study was to test the criterion validity and test-retest reliability of a synchronised inertial sensor and pressure mat-based approach to measure foot placement error and COM velocity while stepping. Methods: Trials were held with 15 healthy participants who each attended for two sessions. The trial task was to step onto one of 4 targets (2 for each foot) multiple times in a random, unpredictable order. The stepping target was cued using an auditory prompt and electroluminescent panel illumination. Data was collected using 3D motion capture and a combined inertial sensor-pressure mat system simultaneously in both sessions. To assess the reliability of each system, ICC estimates and their 95% confident intervals were calculated based on a mean-rating (k = 2), absolute-agreement, 2-way mixed-effects model. To test the criterion validity of the combined inertial sensor-pressure mat system against the motion capture system multi-factorial two-way repeated measures ANOVAs were carried out. Results: It was found that foot placement error was not reliably measured between sessions by either system (ICC 95% CIs; motion capture: 0 to >0.87 and pressure mat: <0.53 to >0.90). This could be due to genuine within-subject variability given the nature of the stepping task and brings into question the suitability of average foot placement error as an outcome measure. Additionally, results suggest the pressure mat is not a valid measure of this parameter since it was statistically significantly different from and much less precise than the motion capture system (p=0.003). The inertial sensor was found to be a moderately reliable (ICC 95% CIs >0.46 to >0.95) but not valid measure for anteroposterior and mediolateral COM velocities (AP velocity: p=0.000, ML velocity target 1 to 4: p=0.734, 0.001, 0.000 & 0.376). However, it is thought that with further development, the COM velocity measure validity could be improved. Possible options which could be investigated include whether there is an effect of inertial sensor placement with respect to pelvic marker placement or implementing more complex methods of data processing to manage inherent accelerometer and gyroscope limitations. Conclusion: The pressure mat is not a suitable alternative for measuring foot placement errors. The inertial sensors have the potential for measuring COM velocity; however, further development work is needed.

Keywords: dynamic balance, inertial sensors, portable, pressure mat, reliability, stepping, validity, wearables

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Authors: Faride Peña, Teresita Castillo, Concepción Campo

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Sexual violence, and particularly child sexual abuse, is a serious problem all over the world, México included. Given its importance, there are several preventive and care programs done by the government and the civil society all over the country but most of them are developed in urban areas even though these problems are especially serious in rural areas. Yucatán, a state in southern México, occupies one of the first places in child sexual abuse. Considering the above, the University Unit of Clinical Research and Victimological Attention (UNIVICT) of the Autonomous University of Yucatan, designed, implemented and is currently evaluating the program named “Sharing Mouth to Mouth: My Body, Nobody Can Touch It”, a program to prevent child sexual abuse in rural communities of Yucatán, México. Its aim was to develop skills for the detection of risk situations, providing protection strategies and mechanisms for prevention through culturally relevant psycho-educative strategies to increase personal resources in children, in collaboration with parents, teachers, police and municipal authorities. The diagnosis identified that a particularly vulnerable population were children between 4 and 10 years. The program run during 2015 in primary schools in the municipality whose inhabitants are mostly Mayan. The aim of this paper is to present its evaluation in terms of its effectiveness and efficiency. This evaluation included documental analysis of the work done in the field, psycho-educational and recreational activities with children, evaluation of knowledge by participating children and interviews with parents and teachers. The results show high efficiency in fulfilling the tasks and achieving primary objectives. The efficiency shows satisfactory results but also opportunity areas that can be resolved with minor adjustments to the program. The results also show the importance of including culturally relevant strategies and activities otherwise it minimizes possible achievements. Another highlight is the importance of participatory action research in preventive approaches to child sexual abuse since by becoming aware of the importance of the subject people participate more actively; in addition to design culturally appropriate strategies and measures so that the proposal may not be distant to the people. Discussion emphasizes the methodological implications of prevention programs (convenience of using participatory action research (PAR), importance of monitoring and mediation during implementation, developing detection skills tools in creative ways using psycho-educational interactive techniques and working assessment issued by the participants themselves). As well, it is important to consider the holistic character this type of program should have, in terms of incorporating social and culturally relevant characteristics, according to the community individuality and uniqueness, consider type of communication to be used and children’ language skills considering that there should be variations strongly linked to a specific cultural context.

Keywords: child sexual abuse, evaluation, PAR, prevention

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Authors: Mariia A. Parfenenko, Ilya S. Dantsev, Sergei V. Bochenkov, Natalia V. Vinogradova, Olga S. Groznova, Victoria Yu. Voinova

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Introduction: Autism is the most common neurodevelopmental disorder. Autism is characterized by difficulties in social interaction and adherence to stereotypic behavioral patterns and frequently co-occurs with epilepsy, intellectual disabilities, connective tissue disorders, and other conditions. CHD8 codes for chromodomain-helicase-DNA-binding protein 8 - a chromatin remodeler that regulates cellular proliferation and neurodevelopment in embryogenesis. CHD8 is one of the genes most frequently involved in autism. Patients and methods: 2 unrelated male patients, P3 and P12, aged 3 and 12 years old, underwent whole genome sequencing, which determined that they both had different likely pathogenic variants, both previously undescribed in literature. Sanger sequencing later determined that P12 inherited the variant from his affected mother. Results: P3 and P12 presented with autism, a developmental delay, ataxia, sleep disorders, overgrowth, and macrocephaly, as well as other clinical features typically present in patients with causative variants in CHD8. The mother of P12 also has autistic traits, as well as ataxia, hypotonia, sleep disorders, and other symptoms. However, P3 and P12 also have different cardiac abnormalities. P3 had signs of a repolarization disorder: a flattened T wave in the III and aVF derivations and a negative T wave in the V1-V2 derivations. He also had structural valve anomalies with associated regurgitation, local contractility impairment of the left ventricular, and diastolic dysfunction of the right ventricle. Meanwhile, P12 had Wolff-Parkinson-White syndrome and underwent radiofrequency ablation at the age of 2 years. At the time of observation, P12 had mild sinus arrhythmia and an incomplete right bundle branch block, as well as arterial hypertension. Discussion: Cardiac abnormalities were not previously reported in patients with causative variants in CHD8. The underlying mechanism for the formation of those abnormalities is currently unknown. However, the two hypotheses are either a disordered interaction with CHD7 – another chromodomain remodeler known to be directly involved in the cardiophenotype of CHARGE syndrome – a rare condition characterized by coloboma, heart defects and growth abnormalities, or the disrupted functioning of CHD8 as an A-Kinase Anchoring Protein, which are known to modulate cardiac function. Conclusion: We observed 2 unrelated autistic males with likely pathogenic variants in CHD8 that presented with typical symptoms of CHD8-related neurodevelopmental disorder, as well as cardiac abnormalities. Cardiac abnormalities have, until now, been considered uncharacteristic for patients with causative variants in CHD8. Further accumulation of data, including experimental evidence of the involvement of CHD8 in heart formation, will elucidate the mechanism underlying the cardiophenotype of those patients. Acknowledgements: Molecular genetic testing of the patients was made possible by the Charity Fund for medical and social genetic aid projects «Life Genome.»

Keywords: autism spectrum disorders, chromodomain-helicase-DNA-binding protein 8, neurodevelopmental disorder, cardio phenotype

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Authors: Shulamit Sternin, Raymond M. McKie, Carter Winberg, Robb N. Travers, Terry P. Humphreys, Elke D. Reissing

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Issues surrounding sexual consent negotiation have become a major topic of societal concern. The majority of current research focuses on the complexities of sexual consent negotiations and the multitude of nuanced issues that surround the consent obtainment of heterosexual adults in post-secondary educational institutions. To date, the only study that has addressed sexual consent negotiation behaviour in same-sex relationships focused on the extent to which individuals used a variety of different verbal and nonverbal sexual consent behaviours to initiate or respond to sexual activity. The results were consistent with trends found within heterosexual individuals; thus, suggesting that the current understanding of sexual consent negotiation, which is grounded in heterosexual research, can serve as a strong foundation for further exploration of sexual consent negotiation within same-sex relationships populations. The current study quantitatively investigated the differences between heterosexual men and gay and bisexual men (GBM) in their understanding of sexual consent negotiation. Exploring how the perceptions of GBM differ from heterosexual males provides insight into some of the unique challenges faced by GBM. Data were collected from a sample of 252 heterosexual men and 314 GBM from Canada, the United States, and Western Europe. Participants responded to the question, 'do you think sexual consent and sex negotiation is different for heterosexual men compared to gay men? If so, how?' by completed an online survey. Responses were analysed following Braun & Clarke’s (2006) six phase thematic analysis guidelines. Inter-rater coding was validated using Cohen’s Kappa value and was calculated at (ϰ = 0.84), indicating a very strong level of agreement between raters. The final thematic structure yielded four major themes: understanding of sexual interaction, unique challenges, scripted role, and universal consent. Respondents spoke to their understanding of sexual interaction, believing GBM sexual consent negotiation to be faster and more immediate. This was linked to perceptions of emotional attachment and the idea that sexual interaction and emotional involvement were distinct and separate processes in GBM sexual consent negotiation, not believed to be the case in heterosexual interactions. Unique challenges such as different protection concerns, role declaration, and sexualization of spaces were understood to hold differing levels of consideration for heterosexual men and GBM. The perception of a clearly defined sexual script for GBM was suggested as a factor that may create ambiguity surrounding sexual consent negotiation, which in turn holds significant implications on unwanted sexual experiences for GBM. Broadening the scope of the current understanding of sexual consent negotiation by focusing on heterosexual and GBM population, the current study has revealed variations in perception of sexual consent negotiation between these two populations. These differences may be understood within the context of sexual scripting theory and masculinity gender role theory. We suggest that sexual consent negotiation is a health risk factor for GBM that has not yet been adequately understood and addressed. Awareness of the perceptions that surround the sexual consent negotiation of both GBM and heterosexual men holds implications on public knowledge, which in turn can better inform policy making, education, future research, and clinical treatment.

Keywords: sexual consent, negotiation, heterosexual men, GBM, sexual script

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Authors: Ippei Torii, Kaoruko Ohtani, Takahito Niwa, Naohiro Ishii

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This paper shows developing an objectivity index using the measurement of subtle eyeball movement to diagnose autism. The developmentally disabled assessment varies, and the diagnosis depends on the subjective judgment of professionals. Therefore, a supplementary inspection method that will enable anyone to obtain the same quantitative judgment is needed. The diagnosis are made based on a comparison of the time of gazing an object in the conventional autistic study, but the results do not match. First, we divided the pupil into four parts from the center using measurements of subtle eyeball movement and comparing the number of pixels in the overlapping parts based on an afterimage. Then we developed the objective evaluation indicator to judge non-autistic and autistic people more clearly than conventional methods by analyzing the differences of subtle eyeball movements between the right and left eyes. Even when a person gazes at one point and his/her eyeballs always stay fixed at that point, their eyes perform subtle fixating movements (ie. tremors, drifting, microsaccades) to keep the retinal image clear. Particularly, the microsaccades link with nerves and reflect the mechanism that process the sight in a brain. We converted the differences between these movements into numbers. The process of the conversion is as followed: 1) Select the pixel indicating the subject's pupil from images of captured frames. 2) Set up a reference image, known as an afterimage, from the pixel indicating the subject's pupil. 3) Divide the pupil of the subject into four from the center in the acquired frame image. 4) Select the pixel in each divided part and count the number of the pixels of the overlapping part with the present pixel based on the afterimage. 5) Process the images with precision in 24 - 30fps from a camera and convert the amount of change in the pixels of the subtle movements of the right and left eyeballs in to numbers. The difference in the area of the amount of change occurs by measuring the difference between the afterimage in consecutive frames and the present frame. We set the amount of change to the quantity of the subtle eyeball movements. This method made it possible to detect a change of the eyeball vibration in numerical value. By comparing the numerical value between the right and left eyes, we found that there is a difference in how much they move. We compared the difference in these movements between non-autistc and autistic people and analyzed the result. Our research subjects consists of 8 children and 10 adults with autism, and 6 children and 18 adults with no disability. We measured the values through pasuit movements and fixations. We converted the difference in subtle movements between the right and left eyes into a graph and define it in multidimensional measure. Then we set the identification border with density function of the distribution, cumulative frequency function, and ROC curve. With this, we established an objective index to determine autism, normal, false positive, and false negative.

Keywords: subtle eyeball movement, autism, microsaccade, pursuit eye movements, ROC curve

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Authors: Bakytzhan Bimbetov, Abay Zhangabilov, Saule Aitbaeva, Galymzhan Meirambekov

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Mare’s milk (saumal) contains in total about 40 biological components necessary for the human body. The most significant among them are amino acids, fats, carbohydrates, enzymes (lysozyme, amylase), more minerals and vitamins which are well balanced with each other. In Kazakhstan, Company "Eurasia Invest Ltd.” produces a freeze-dried saumal in form of powder by the use of modern German innovative technology by means of evaporating at low temperature (-35°C) with an appropriate pasteurization. Research of freeze-dried biomilk for the qualitative content showed that main ingredients of freshly drown milk are being preserved. We are currently studying medical and dietary properties of freeze-dried mare's milk for diseases of the digestive system, including for nonalcoholic steatohepatitis (NASH) and liver cirrhosis (LC) viral etiology. The studied group consisted of 14 patients with NASH, and 7 patients with LC viral etiology of Class A severity degree as per Child-Pugh. Patients took freeze-dried saumal, preliminary dissolved in boiled warm water (24 g. powder per 200 ml water) 3-4 times a day for a month in conjunction with basic therapy. The results were compared to a control group (11 patients with NASH and LC) who received only basic therapy without mare’s milk. Results of preliminary research showed an improvement of subjective and objective conditions of all patients, but more significant improvement of clinical symptoms and syndromes were observed in the treatment group compared to the control one. Patients with NASH significantly over time compared to the beginning of therapy decreased asthenic and dyspeptic syndromes (p<0,01). Hepatomegaly, identified on the basis of ultrasound prior to treatment was observed in 92,8±2,4% of patients, and after combination therapy hepatomegaly the rate decreased by 14,3%, amounting to 78,5±2,8%. Patients with LC also noted the improvement of asthenic (p<0,01) and dyspeptic (p<0,05) syndromes and hemorrhagic syndrome (nosebleeds and bleeding gums when brushing your teeth, p<0,05), and jaundice. Laboratory study also showed improvement in the research group, but more significant changes were observed in the experimental group. Group of patients with NASH showed a significant improvement of index in cytolysis in conjunction with a combination therapy (p<0,05). In the control group, these indicators were also improved, but they were not statistically reliable (p>0,05). Markers of liver failure were additionally studied during the study of laboratory parameters in patients with liver cirrhosis, in particular, bilirubin, albumin and prothrombin index (PTI). Combined therapy with the use of basic treatment and mare's milk showed a significant improvement in cytolysis and bilirubin (p<0,05). In our opinion, a very important and interesting fact is that, in conjunction with basic therapy, the use of mare's milk revealed an improvement of liver function in the form of normalized PTI and albumin in patients with liver cirrhosis viral etiology. Results of this work have shown therapeutic efficiency of the use of mare's milk in complex treatment of patients with liver disease and require further in-depth study.

Keywords: liver cirrhosis, non-alcohol steatohepatitis, saumal, mare’s milk

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Authors: David Dolbow, Daniel Credeur, Mujtaba Rahimi, Dobrivoje Stokic, Jennifer Lemacks, Andrew Courtner

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Introduction: Obesity is at epidemic proportions in the spinal cord injury (SCI) population (66-75%), as individuals who suffer from paralysis undergo a dramatic decrease in muscle mass and a dramatic increase in adipose deposition. Obesity is a major public health concern which includes a doubling of the risk of heart disease, stroke and type II diabetes mellitus. It has been demonstrated that physical activity, and especially HIIT, can promote a healthy body composition and decrease the risk cardiometabolic disease in the able-bodied population. However, SCI typically limits voluntary exercise to the arms, but a high prevalence of shoulder pain in persons with chronic SCI (60-90%) can cause increased arm exercise to be problematic. Functional electrical stimulation (FES) cycling has proven to be a safe and effective way to exercise paralyzed leg muscles in clinical and home settings, saving the often overworked arms. Yet, HIIT-FES cycling had not been investigated prior to the current study. The purpose of this study was to investigate the body composition changes with combined HIIT-FES cycling and nutritional counseling on individuals with SCI. Design: A matched (level of injury, time since injury, body mass index) and controlled trail. Setting: University exercise performance laboratory. Subjects: Ten individuals with chronic SCI (C5-T9) ASIA impairment classification (A & B) were divided into the treatment group (n=5) for 30 minutes of HIIT-FES cycling 3 times per week for 8 weeks and nutritional counseling over the phone for 30 minutes once per week for 8 weeks and the control group (n=5) who received nutritional counseling only. Results: There was a statistically significant difference between the HIIT-FES group and the control group in mean body fat percentage change (-1.14 to +0.24) respectively, p = .030). There was also a statistically significant difference between the HIIT-FES and control groups in mean change in legs lean mass (+0.78 kg to -1.5 kg) respectively, p = 0.004. There was a nominal decrease in weight, BMI, total fat mass and a nominal increase in total lean mass for the HIIT-FES group over the control group. However, these changes were not found to be statistically significant. Additionally, there was a nominal decrease in the mean blood glucose levels for both groups 101.8 to 97.8 mg/dl for the HIIT-FES group and 94.6 to 93 mg/dl for the Nutrition only group, however, neither were found to be statistically significant. Conclusion: HIIT-FES cycling combined with nutritional counseling can provide healthful body composition changes including decreased body fat percentage in just 8 weeks. Future study recommendations include a greater number of participants, a primer electrical stimulation exercise program to better ready participants for HIIT-FES cycling and a greater volume of training above 30 minutes, 3 times per week for 8 weeks.

Keywords: body composition, functional electrical stimulation cycling, high-intensity interval training, spinal cord injury

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

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Obesity in childhood establishes a ground for adulthood obesity. Especially morbid obesity is an important problem for the children because of the associated diseases such as diabetes mellitus, cancer and cardiovascular diseases. In this study, body mass index (BMI), body fat ratios, anthropometric measurements and ratios were evaluated together with different laboratory indices upon evaluation of obesity in morbidly obese (MO) children. Children with nutritional problems participated in the study. Written informed consent was obtained from the parents. Study protocol was approved by the Ethics Committee. Sixty-two MO girls aged 129.5±35.8 months and 75 MO boys aged 120.1±26.6 months were included into the scope of the study. WHO-BMI percentiles for age-and-sex were used to assess the children with those higher than 99^th as morbid obesity. Anthropometric measurements of the children were recorded after their physical examination. Bio-electrical impedance analysis was performed to measure fat distribution. Anthropometric ratios, body fat ratios, Index-I and Index-II as well as insulin sensitivity indices (ISIs) were calculated. Girls as well as boys were binary grouped according to homeostasis model assessment-insulin resistance (HOMA-IR) index of <2.5 and >2.5, fasting glucose to insulin ratio (FGIR) of <6 and >6 and quantitative insulin sensitivity check index (QUICKI) of <0.33 and >0.33 as the frequently used cut-off points. They were evaluated based upon their BMIs, arms, legs, trunk, whole body fat percentages, body fat ratios such as fat mass index (FMI), trunk-to-appendicular fat ratio (TAFR), whole body fat ratio (WBFR), anthropometric measures and ratios [waist-to-hip, head-to-neck, thigh-to-arm, thigh-to-ankle, height/2-to-waist, height/2-to-hip circumference (C)]. SPSS/PASW 18 program was used for statistical analyses. p≤0.05 was accepted as statistically significance level. All of the fat percentages showed differences between below and above the specified cut-off points in girls when evaluated with HOMA-IR and QUICKI. Differences were observed only in arms fat percent for HOMA-IR and legs fat percent for QUICKI in boys (p≤ 0.05). FGIR was unable to detect any differences for the fat percentages of boys. Head-to-neck C was the only anthropometric ratio recommended to be used for all ISIs (p≤0.001 for both girls and boys in HOMA-IR, p≤0.001 for girls and p≤0.05 for boys in FGIR and QUICKI). Indices which are recommended for use in both genders were Index-I, Index-II, HOMA/BMI and log HOMA (p≤0.001). FMI was also a valuable index when evaluated with HOMA-IR and QUICKI (p≤0.001). The important point was the detection of the severe significance for HOMA/BMI and log HOMA while they were evaluated also with the other indices, FGIR and QUICKI (p≤0.001). These parameters along with Index-I were unique at this level of significance for all children. In conclusion, well-accepted ratios or indices may not be valid for the evaluation of both genders. This study has emphasized the limiting properties for boys. This is particularly important for the selection process of some ratios and/or indices during the clinical studies. Gender difference should be taken into consideration for the evaluation of the ratios or indices, which will be recommended to be used particularly within the scope of obesity studies.

Keywords: anthropometry, childhood obesity, gender, insulin sensitivity index

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Authors: Meenu S. Babu, K. Jayasankara Reddy

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Depression is a serious mental health concern that leads to significant distress and dysfunction in an individual. Due to the high physical, psychological, social, and economic burden it causes, it is important to study various bio-psycho-social factors that influence the onset, course, duration, intensity of depressive symptoms. The study aims to explore relationship between autobiographical memory (AM) functions, perceived control over stressful events and depressive symptoms. AM functions and perceived control were both found to be protective factors for individuals against depression and were both modifiable to predict better behavioral and affective outcomes. An extensive review of literatur, with a systematic search on Google Scholar, JSTOR, Science Direct and Springer Journals database, was conducted for the purpose of this review paper. These were used for all the aforementioned databases. The time frame used for the search was 2010-2021. An additional search was conducted with no time bar to map the development of the theoretical concepts. The relevant studies with quantitative, qualitative, experimental, and quasi- experimental research designs were included for the review. Studies including a sample with a DSM- 5 or ICD-10 diagnosis of depressive disorders were excluded from the study to focus on the behavioral patterns in a non-clinical population. The synthesis of the findings that were obtained from the review indicates there is a significant relationship between cognitive variables of AM functions and perceived control and depressive symptoms. AM functions were found to be have significant effects on once sense of self, interpersonal relationships, decision making, self- continuity and were related to better emotion regulation and lower depressive symptoms. Not all the components of AM function were equally significant in their relationships with various depressive symptoms. While self and directive functions were more related to emotion regulation, anhedonia, motivation and hence mood and affect, the social function was related to perceived social support and social engagement. Perceived control was found to be another protective cognitive factor that provides individuals a sense of agency and control over one’s life outcomes which was found to be low in individuals with depression. This was also associated to the locus of control, competency beliefs, contingency beliefs and subjective well being in individuals and acted as protective factors against depressive symptoms. AM and perceived control over stressful events serve adaptive functions, hence it is imperative to study these variables more extensively. They can be imperative in planning and implementing therapeutic interventions to foster these cognitive protective factors to mitigate or alleviate depressive symptoms. Exploring AM as a determining factor in depressive symptoms along with perceived control over stress creates a bridge between biological and cognitive factors underlying depression and increases the scope of developing a more eclectic and effective treatment plan for individuals. As culture plays a crucial role in AM functions as well as certain aspects of control such as locus of control, it is necessary to study these variables keeping in mind the cultural context to tailor culture/community specific interventions for depression.

Keywords: autobiographical memories, autobiographical memory functions, perceived control, depressive symptoms, depression, young adults

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Authors: Monisha Elumalai, Joana Guerreiro, Joana Carvalho, Marta Prado

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DNA biosensors popularity has been increasing over the past few years. Traditional analytical techniques tend to require complex steps and expensive equipment however DNA biosensors have the advantage of getting simple, fast and economic. Additionally, the combination of DNA biosensors with nanomaterials offers the opportunity to improve the selectivity, sensitivity and the overall performance of the devices. DNA biosensors are based on oligonucleotides as sensing elements. These oligonucleotides are highly specific to complementary DNA sequences resulting in the hybridization of the strands. DNA biosensors are not only an advantage in the clinical field but also applicable in numerous research areas such as food analysis or environmental control. Zebra Mussels (ZM), Dreissena polymorpha are invasive species responsible for enormous negative impacts on the environment and ecosystems. Generally, the detection of ZM is made when the observation of adult or macroscopic larvae's is made however at this stage is too late to avoid the harmful effects. Therefore, there is a need to develop an analytical tool for the early detection of ZM. Here, we present a portable plasmonic biosensor for the detection of environmental DNA (eDNA) released to the environment from this invasive species. The plasmonic DNA biosensor combines gold nanoparticles, as transducer elements, due to their great optical properties and high sensitivity. The detection strategy is based on the immobilization of a short base pair DNA sequence on the nanoparticles surface followed by specific hybridization in the presence of a complementary target DNA. The hybridization events are tracked by the optical response provided by the nanospheres and their surrounding environment. The identification of the DNA sequences (synthetic target and probes) to detect Zebra mussel were designed by using Geneious software in order to maximize the specificity. Moreover, to increase the optical response enzyme amplification of DNA might be used. The gold nanospheres were synthesized and characterized by UV-visible spectrophotometry and transmission electron microscopy (TEM). The obtained nanospheres present the maximum localized surface plasmon resonance (LSPR) peak position are found to be around 519 nm and a diameter of 17nm. The DNA probes modified with a sulfur group at one end of the sequence were then loaded on the gold nanospheres at different ionic strengths and DNA probe concentrations. The optimal DNA probe loading will be selected based on the stability of the optical signal followed by the hybridization study. Hybridization process leads to either nanoparticle dispersion or aggregation based on the presence or absence of the target DNA. Finally, this detection system will be integrated into an optical sensing platform. Considering that the developed device will be used in the field, it should fulfill the inexpensive and portability requirements. The sensing devices based on specific DNA detection holds great potential and can be exploited for sensing applications in-loco.

Keywords: ZM DNA, DNA probes, nicking enzyme, gold nanoparticles

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Authors: Lauren Dhugga, Meena Parameswaran, David Blundell, Abbas Khushnood

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Context: A multidisciplinary weekly safety briefing was implemented at the Paediatric Cardiothoracic Unit at the Freeman Hospital in Newcastle-upon-Tyne. It is a tertiary referral centre with a quarternary cardiac paediatric intensive care unit and provides complexed care including heart and lung transplants, mechanical support and advanced heart failure assessment. Aim: The aim of this briefing is to provide a structured platform of communication, in an effort to improve efficiency, safety, and patient care. Problem: The paediatric cardiothoracic unit is made up of a vast multidisciplinary team including doctors, intensivists, anaesthetists, surgeons, specialist nurses, echocardiogram technicians, physiotherapists, psychologists, dentists, and dietitians. It provides care for children with congenital and acquired cardiac disease and is one of only two units in the UK to offer paediatric heart transplant. The complexity of cases means that there can be many teams involved in providing care to each patient, and frequent movement of children between ward, high dependency, and intensive care areas. Currently, there is no structured forum for communicating important information across the department, for example, staffing shortages, prescribing errors and significant events. Strategy: An initial survey questioning the need for better communication found 90% of respondents agreed that they could think of an incident that had occurred due to ineffective communication, and 85% felt that incident could have been avoided had there been a better form of communication. Lastly, 80% of respondents felt that a weekly 60 second safety briefing would be beneficial to improve communication within our multidisciplinary team. Based on those promising results, a weekly 60 second safety briefing was implemented to be conducted on a Monday morning. The safety briefing covered four key areas (SAFE): staffing, awareness, fix and events. This was to highlight any staffing gaps, any incident reports to be learned from, any issues that required fixing and any events including teachings for the week ahead. The teams were encouraged to email suggestions or issues to be raised for the week or to approach in person with information to add. The safety briefing was implemented using change theory. Effect: The safety briefing has been trialled over 6 weeks and has received a good buy in from staff across specialties. The aim is to embed this safety briefing into a weekly meeting using the PDSA cycle. There will be a second survey in one month to assess the efficacy of the safety briefing and to continue to improve the delivery of information. The project will be presented at the next clinical governance briefing to attract wider feedback and input from across the trust. Lessons: The briefing displays promise as a tool to improve vigilance and communication in a busy multi-disciplinary unit. We have learned about how to implement quality improvement and about the culture of our hospital - how hierarchy influences change. We demonstrate how to implement change through a grassroots process, using a junior led briefing to improve the efficiency, safety, and communication in the workplace.

Keywords: briefing, communication, safety, team

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Authors: Nicole Selvi Hill, Archchana Radhakrishnan

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Introduction: Traumatic brain injury (TBI) is a leading cause of mortality among trauma patients. In the management of TBI, the main principle is avoiding cerebral ischemia, as this is a strong determiner of neurological outcomes. The use of vasoactive drugs, such as vasopressin, has an important role in maintaining cerebral perfusion pressure to prevent secondary brain injury. Current guidelines do not suggest a preferred vasoactive drug to administer in the management of TBI, and there is a paucity of information on the therapeutic potential of vasopressin following TBI. Vasopressin is also an endogenous anti-diuretic hormone (AVP), and pathways mediated by AVP play a large role in the underlying pathological processes of TBI. This creates an overlap of discussion regarding the therapeutic potential of vasopressin following TBI. Currently, its popularity lies in vasodilatory and cardiogenic shock in the intensive care setting, with increasing support for its use in haemorrhagic and septic shock. Methodology: This is a review article based on a literature review. An electronic search was conducted via PubMed, Cochrane, EMBASE, and Google Scholar. The aim was to identify clinical studies looking at the therapeutic administration of vasopressin in severe traumatic brain injury. The primary aim was to look at the neurological outcome of patients. The secondary aim was to look at surrogate markers of cerebral perfusion measurements, such as cerebral perfusion pressure, cerebral oxygenation, and cerebral blood flow. Results: Eight papers were included in the final number. Three were animal studies; five were human studies, comprised of three case reports, one retrospective review of data, and one randomised control trial. All animal studies demonstrated the benefits of vasopressors in TBI management. One animal study showed the superiority of vasopressin in reducing intracranial pressure and increasing cerebral oxygenation over a catecholaminergic vasopressor, phenylephrine. All three human case reports were supportive of vasopressin as a rescue therapy in catecholaminergic-resistant hypotension. The retrospective review found vasopressin did not increase cerebral oedema in TBI patients compared to catecholaminergic vasopressors; and demonstrated a significant reduction in the requirements of hyperosmolar therapy in patients that received vasopressin. The randomised control trial results showed no significant differences in primary and secondary outcomes between TBI patients receiving vasopressin versus those receiving catecholaminergic vasopressors. Apart from the randomised control trial, the studies included are of low-level evidence. Conclusion: Studies favour vasopressin within certain parameters of cerebral function compared to control groups. However, the neurological outcomes of patient groups are not known, and animal study results are difficult to extrapolate to humans. It cannot be said with certainty whether vasopressin’s benefits stand above usage of other vasoactive drugs due to the weaknesses of the evidence. Further randomised control trials, which are larger, standardised, and rigorous, are required to improve knowledge in this field.

Keywords: catecholamines, cerebral perfusion pressure, traumatic brain injury, vasopressin, vasopressors

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Authors: Fatima Meirkhankyzy Shaizadina, Alua Oralovna Omarova, Praskovya Mikhailovna Britskaya, Nessipkul Oryntayevna Alysheva

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In recent years in the Republic of Kazakhstan there have been registered outbreaks of measles among the population. The objective of work was the analysis of outbreak of measles in 2014 among the population of North-Kazakhstan region of the Republic of Kazakhstan. For the analysis of the measles outbreak descriptive and analytical research, techniques were used and threshold levels of morbidity were calculated. The increase of incidence was noted from March to July. The peak was registered in May and made 9.0 per 100000 population. High rates were registered in April – 5.7 per 100000 population, and in June and July they made 5.7 and 3.1 respectively. Duration of the period of increase made 5 months. The analysis of monthly incidence of measles revealed spring and summer seasonality. Across the territory it was established that 69.2% of cases were registered in the city, 29.1% in rural areas and 1.7% of cases were brought in from other regions of Kazakhstan. The registered cases and threshold values of measles during the outbreak revealed that from 12 to 24 week, and also during the 40th week the cases exceeding the threshold levels are registered. Thus, for example, for the analyzed 1 week the number of the revealed patients made 4, which exceeds the calculated threshold value (3) by 33.3%. The data exceeding the threshold values confirm the emergence of a disease outbreak or the beginning of epidemic rise in morbidity. Epidemic rise in incidence of the population of North-Kazakhstan region was observed throughout 2014. The risk group includes 0-4 year-old children, who made 22.7%, 15-19 year-olds – 25.6%, 20-24 year-olds – 20.9%. The analysis of measles cases registration by gender revealed that women are registered 1.1 times more often than men. The ratio of women to men made 1:0.87. In social and professional groups often ill are unorganized children – 23.3% and students – 19.8%. Studying clinical manifestations of measles in the hospitalized patients, the typical beginning of a disease with expressed intoxication symptoms – weakness, sickliness was established. In individual cases expressed intoxication symptoms, hemorrhagic and dyspeptic syndromes, complications in the form of overlay of a secondary bacterial infection, which defined high severity of the illness, were registered both in adults and in children. The average duration of stay of patients in the hospital made 6.9 days. The average duration of time between date of getting the disease and date of delivery of health care made 3.6 days. Thus, the analysis of monthly incidence of measles revealed spring and summer seasonality, the peak of which was registered in May. Urban dwellers are ill more often (69.2%), while in rural areas people are ill more rarely (29.1%). Throughout 2014 an epidemic rise in incidence of the population of North-Kazakhstan region was observed. Risk group includes: children under 4 – 22.7%, 15-19 year-olds – 25.6%, 20-24 year-olds – 20.9%. The ratio of women and men made 1:0.87. The typical beginning of a disease in all hospitalized with the expressed intoxication symptoms – weakness, sickliness was established.

Keywords: epidemiological analysis, measles, morbidity, outbreak

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Authors: Aline Woine, Moïra Mikolajczak, Virginie Dardier, Isabelle Roskam

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Background information: Becoming a parent is said to be the happiest event one can ever experience in one’s life. This popular (and almost absolute) truth–which no reasonable and decent human being would ever dare question on pain of being singled out as a bad parent–contrasts with the nuances that reality offers. Indeed, while many parents do thrive in their parenting role, some others falter and become progressively overwhelmed by their parenting role, ineluctably caught in a spiral of exhaustion. Parental burnout (henceforth PB) sets in when parental demands (stressors) exceed parental resources. While it is now generally acknowledged that PB affects the parent’s behavior in terms of neglect and violence toward their offspring, little is known about the impact that the syndrome might have on the children’s internalized (anxious and depressive symptoms, somatic complaints, etc.) and/or externalized (irritability, violence, aggressiveness, conduct disorder, oppositional disorder, etc.) behaviors. Furthermore, at the time of writing, to our best knowledge, no research has yet tested the reverse effect, namely, that of the child's internalized and/or externalized behaviors on the onset and/or maintenance of parental burnout symptoms. Goals and hypotheses: The present pioneering research proposes to fill an important gap in the existing literature related to PB by investigating the bidirectional effect between PB and the child’s internalized and/or externalized behaviors. Relying on a cross-lagged longitudinal study with three waves of data collection (4 months apart), our study tests a transactional model with bidirectional and recursive relations between observed variables and at the three waves, as well as autoregressive paths and cross-sectional correlations. Methods: As we write this, wave-two data are being collected via Qualtrics, and we expect a final sample of about 600 participants composed of French-speaking (snowball sample) and English-speaking (Prolific sample) parents. Structural equation modeling is employed using Stata version 17. In order to retain as much statistical power as possible, we use all available data and therefore apply the maximum likelihood with a missing value (mlmv) as the method of estimation to compute the parameter estimates. To limit (in so far is possible) the shared method variance bias in the evaluation of the child’s behavior, the study relies on a multi-informant evaluation approach. Expected results: We expect our three-wave longitudinal study to show that PB symptoms (measured at T1) raise the occurrence/intensity of the child’s externalized and/or internalized behaviors (measured at T2 and T3). We further expect the child’s occurrence/intensity of externalized and/or internalized behaviors (measured at T1) to augment the risk for PB (measured at T2 and T3). Conclusion: Should our hypotheses be confirmed, our results will make an important contribution to the understanding of both PB and children’s behavioral issues, thereby opening interesting theoretical and clinical avenues.

Keywords: exhaustion, structural equation modeling, cross-lagged longitudinal study, violence and neglect, child-parent relationship

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Authors: Nkeiruka Elsie Ezomike, Chinwe L. Onyekonwu, Anthony N. Ikefuna, Bede C. Ibe

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Superficial fungal infections (SFIs) are one of the common cutaneous infections that affect children worldwide. They may lead to school absenteeism or school drop-out and hence setback in the education of the child. Community-based studies in any locality are good reflections of the health conditions within that area. There is a dearth of information in the literature about SFI among primary school children in Enugu. This study aimed to determine the clinicomycological pattern of SFIs among primary school children in rural and urban communities in Enugu. This was a comparative descriptive cross-sectional study among primary school children in Awgu (rural) and Enugu North (urban) Local Government Areas (LGAs). Subjects' selection was made over 6 months using a multi-stage sampling method. Information such as age, sex, parental education, and occupation were collected using questionnaires. Socioeconomic classes of the children were determined using the classification proposed by Oyedeji et al. The samples were collected from subjects with SFIs. Potassium hydroxide tests were done on the samples. The samples that tested positive were cultured for SFI by inoculating onto Sabouraud's dextrose chloramphenicol actidione agar. The characteristics of the isolates were identified according to their morphological features using Mycology Online, Atlas 2000, and Mycology Review 2003. Equal numbers of children were recruited from the two LGAs. A total of 1662 pupils were studied. The mean ages of the study subjects were 9.03 ± 2.10years in rural and 10.46 ± 2.33years in urban communities. The male to female ratio was 1.6:1 in rural and 1:1.1 in urban communities. The personal hygiene of the children was significantly related to the presence of SFIs. The overall prevalence of SFIs among the study participants was 45%. In the rural, the prevalence was 29.6%, and in the urban prevalence was 60.4%. The types of SFIs were tinea capitis (the commonest), tinea corporis, pityriasis Versicolor, tinea unguium, and tinea manuum with prevalence rates lower in rural than urban communities. The clinical patterns were gray patch and black dot type of non-inflammatory tinea capitis, kerion, tinea corporis with trunk and limb distributions, and pityriasis Versicolor with face, trunk and limb distributions. Gray patch was the most frequent pattern of SFI seen in rural and urban communities. Black dot type was more frequent in rural than urban communities. SFIs were frequent among children aged 5 to 8years in rural and 9 to 12 years in urban communities. SFIs were commoner in males in the rural, whereas female dominance was observed in the urban. SFIs were more in children from low social class and those with poor hygiene. Trichophyton tonsurans and Trichophyton soudanese were the common mycological isolates in rural and urban communities, respectively. In conclusion, SFIs were less prevalent in rural than in urban communities. Trichophyton species were the most common fungal isolates in the communities. Health education of mothers and their children on SFI and good personal hygiene will reduce the incidence of SFIs.

Keywords: clinicomycological pattern, communities, primary school children, superficial fungal infections

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Authors: Paulina Clara Appiah, Kofi Issah, Timothy Letsa, Kennedy Nartey, Amanua Chinbuah, Adoma Dwomo-Fokuo, Jacqeline G. Asibey

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Background: The Every Newborn Action Plan provides evidence–based interventions to end preventable deaths in high burden countries. Brong Ahafo Region is one of ten regions in Ghana with less than half of its district hospitals having sick newborn units. Facility-based neonatal care is not prioritized and under-funded, and there is also inadequate knowledge and competence to manage the sick. The aim of this intervention was to make available in–patient care for sick newborns in all 19 district hospitals through the strengthening of facility-based systems. Methods: With the development and dissemination of the National Newborn Strategy and Action Plan 2014-2018, the country was able to attract PATH which provided the region with basic resuscitation equipment, supported hospital providers’ capacity building in Helping Babies Breathe, Essential Care of Every Baby, Infection Prevention and Management and held a symposia on managing the sick newborn. Newborn advocacy was promoted through newborn champions at the facility and community levels. Hospital management was then able to mobilize resources from communities, corporate organizations and from internally generated funds; created or expanded sick newborn care units and provided essential medicines and equipment. Kangaroo Mother Care was initiated in 6 hospitals. Pediatric specialist outreach services initiated comprised telephone consultations, teaching ward rounds and participating in perinatal death audits meetings. Newborn data capture and management was improved through the provision and training on the use of standard registers provided from the national level. Results: From February 2015 to November 2017, hospitals with sick newborn units increased from 7 to 19 (37%-100%). 180 pieces each of newborn ventilation bags and masks size 0, 1 and penguin suction bulbs were distributed to the hospitals, in addition to 20 newborn mannequin sets and 90 small clinical reminder posters. 802 providers (96.9%) were trained in resuscitation, of which 96% were successfully followed up in 6 weeks, 91% in 6 months and 80% in 12 months post-training. 53 clinicians (65%) were trained and mentored to manage sick newborns. 56 specialist teaching ward rounds were conducted. Data completeness improved from 92.6% - 99.9%. Availability of essential medicines improved from 11% to 100%. Number of hospital cots increased from 116 to 248 (214%). Cot occupancy rate increased from 57.4% to 92.5%. Hospitals with phototherapy equipment increased from 0 to 12 (63%). Hospitals with incubators increased from 1 to 12 (5%-63%). Newborn deaths among admissions reduced from 6.3% to 5.4%. Conclusion: Access to in-patient care increased significantly. Newborn advocacy successfully mobilized resources required for strengthening facility –based systems.

Keywords: facility-based systems, Ghana, in-patient care, newborn advocacy

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Authors: D. Dixon, J. Nicol, J. A. Coulter, E. Harrison

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The ease with which they can be functionalized, combined with their excellent biocompatibility, make gold nanoparticles (AuNPs) ideal candidates for various applications in nanomedicine. Indeed several promising treatments are currently undergoing human clinical trials (CYT-6091 and Auroshell). A successful nanoparticle treatment must first evade the immune system, then accumulate within the target tissue, before enter the diseased cells and delivering the payload. In order to create a clinically relevant drug delivery system, contrast agent or radiosensitizer, it is generally necessary to functionalize the AuNP surface with multiple groups; e.g. Polyethylene Glycol (PEG) for enhanced stability, targeting groups such as antibodies, peptides for enhanced internalization, and therapeutic agents. Creating and characterizing the biological response of such complex systems remains a challenge. The two commonly used methods to attach multiple groups to the surface of AuNPs are the creation of a mixed monolayer, or by binding groups to the AuNP surface using a bi-functional PEG linker. While some excellent in-vitro and animal results have been reported for both approaches further work is necessary to directly compare the two methods. In this study AuNPs capped with both PEG and a Receptor Mediated Endocytosis (RME) peptide were prepared using both mixed monolayer and PEG linker approaches. The PEG linker used was SH-PEG-SGA which has a thiol at one end for AuNP attachment, and an NHS ester at the other to bind to the peptide. The work builds upon previous studies carried out at the University of Ulster which have investigated AuNP synthesis, the influence of PEG on stability in a range of media and investigated intracellular payload release. 18-19nm citrate capped AuNPs were prepared using the Turkevich method via the sodium citrate reduction of boiling 0.01wt% Chloroauric acid. To produce PEG capped AuNPs, the required amount of PEG-SH (5000Mw) or SH-PEG-SGA (3000Mw Jenkem Technologies) was added, and the solution stirred overnight at room temperature. The RME (sequence: CKKKKKKSEDEYPYVPN, Biomatik) co-functionalised samples were prepared by adding the required amount of peptide to the PEG capped samples and stirring overnight. The appropriate amounts of PEG-SH and RME peptide were added to the AuNP to produce a mixed monolayer consisting of approximately 50% PEG and 50% RME. The PEG linker samples were first fully capped with bi-functional PEG before being capped with RME peptide. An increase in diameter from 18-19mm for the ‘as synthesized’ AuNPs to 40-42nm after PEG capping was observed via DLS. The presence of PEG and RME peptide on both the mixed monolayer and PEG linker co-functionalized samples was confirmed by both FTIR and TGA. Bi-functional PEG linkers allow the entire AuNP surface to be capped with PEG, enabling in-vitro stability to be achieved using a lower molecular weight PEG. The approach also allows the entire outer surface to be coated with peptide or other biologically active groups, whilst also offering the promise of enhanced biological availability. The effect of mixed monolayer versus PEG linker attachment on both stability and non-specific protein corona interactions was also studied.

Keywords: nanomedicine, gold nanoparticles, PEG, biocompatibility

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Authors: Natalia Lukasik, Ewa Wagner-Wysiecka

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Development of selective, fast-responsive, and economical sensors for diverse ions detection and determination is one of the most extensively studied areas due to its importance in the field of clinical, environmental and industrial analysis. Among chemical sensors, vast popularity has gained ionophore-based optical sensors, where the generated analytical signal is a consequence of the molecular recognition of ion by the ionophore. Change of color occurring during host-guest interactions allows for quantitative analysis and for 'naked-eye' detection without the need of using sophisticated equipment. An example of application of such sensors is colorimetric detection of iron(III) cations. Iron as one of the most significant trace elements plays roles in many biochemical processes. For these reasons, the development of reliable, fast, and selective methods of iron ions determination is highly demanded. Taking all mentioned above into account a chromogenic amide derivative of 3,4-dihydroxybenzoic acid was synthesized, and its ability to iron(III) recognition was tested. To the best of authors knowledge (according to chemical abstracts) the obtained ligand has not been described in the literature so far. The catechol moiety was introduced to the ligand structure in order to mimic the action of naturally occurring siderophores-iron(III)-selective receptors. The ligand–ion interactions were studied using spectroscopic methods: UV-Vis spectrophotometry and infrared spectroscopy. The spectrophotometric measurements revealed that the amide exhibits affinity to iron(III) in dimethyl sulfoxide and fully aqueous solution, what is manifested by the change of color from yellow to green. Incorporation of the tested amide into a polymeric matrix (cellulose triacetate) ensured effective recognition of iron(III) at pH 3 with the detection limit 1.58×10⁻⁵ M. For the obtained sensor material parameters like linear response range, response time, selectivity, and possibility of regeneration were determined. In order to evaluate the effect of the size of the sensing material on iron(III) detection nanospheres (in the form of nanoemulsion) containing the tested amide were also prepared. According to DLS (dynamic light scattering) measurements, the size of the nanospheres is 308.02 ± 0.67 nm. Work parameters of the nanospheres were determined and compared with cellulose triacetate-based material. Additionally, for fast, qualitative experiments the test strips were prepared by adsorption of the amide solution on a glass microfiber material. Visual limit of detection of iron(III) at pH 3 by the test strips was estimated at the level 10⁻⁴ M. In conclusion, reported here amide derived from 3,4- dihydroxybenzoic acid proved to be an effective candidate for optical sensing of iron(III) in fully aqueous solutions. N. L. kindly acknowledges financial support from National Science Centre Poland the grant no. 2017/01/X/ST4/01680. Authors thank for financial support from Gdansk University of Technology grant no. 032406.

Keywords: ion-selective optode, iron(III) recognition, nanospheres, optical sensor

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Authors: Johnson C. Y. Pang, Amy S. N. Fu, Ryan K. L. Lee, Allan C. L. Fu

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Dry needling (DN) is one of the puncturing methods that involves the insertion of needles into the tender spots of the human body without the injection of any substance. DN has long been used to treat the patient with knee pain caused by knee osteoarthritis (KOA) and patellofemoral pain syndrome (PFPS), but the effectiveness is still inconsistent. This study aimed to conduct a systematic review and meta-analysis to assess the intervention methods and effects of DN with and without ultrasound guidance for treating pain and dysfunctions in people with KOA and PFPS. Design: This systematic review adhered to the PRISMA reporting guidelines. The registration number of the study protocol published in the PROSPERO database was CRD42021221419. Six electronic databases were searched manually through CINAHL Complete (1976-2020), Cochrane Library (1996-2020), EMBASE (1947-2020), Medline (1946-2020), PubMed (1966-2020), and Psychinfo (1806-2020) in November 2020. Randomized controlled trials (RCTs) and controlled clinical trials were included to examine the effects of DN on knee pain, including KOA and PFPS. The key concepts included were: DN, acupuncture, ultrasound guidance, KOA, and PFPS. Risk of bias assessment and qualitative analysis were conducted by two independent reviewers using the PEDro score. Results: Fourteen articles met the inclusion criteria, and eight of them were high-quality papers in accordance with the PEDro score. There were variations in the techniques of DN. These included the direction, depth of insertion, number of needles, duration of stay, needle manipulation, and the number of treatment sessions. Meta-analysis was conducted on eight articles. DN group showed positive short-term effects (from immediate after DN to less than 3 months) on pain reduction for both KOA and PFPS with the overall standardized mean difference (SMD) of -1.549 (95% CI=-0.588 to -2.511); with great heterogeneity (P=0.002, I²=96.3%). In subgroup analysis, DN demonstrated significant effects in pain reduction on PFPS (p < 0.001) that could not be found in subjects with KOA (P=0.302). At 3-month post-intervention, DN also induced significant pain reduction in both subjects with KOA and PFPS with an overall SMD of -0.916 (95% CI=-0.133 to -1.699, and great heterogeneity (P=0.022, I²=95.63%). Besides, DN induced significant short-term improvement in function with the overall SMD=6.069; 95% CI=8.595 to 3.544; with great heterogeneity (P<0.001, I²=98.56%) when analyzed was conducted on both KOA and PFPS groups. In subgroup analysis, only PFPS showed a positive result with SMD=6.089, P<0.001; while KOA showed statistically insignificant with P=0.198 in short-term effect. Similarly, at 3-month post-intervention, significant improvement in function after DN was found when the analysis was conducted in both groups with the overall SMD=5.840; 95% CI=9.252 to 2.428; with great heterogeneity (P<0.001, I²=99.1%), but only PFPS showed significant improvement in sub-group analysis (P=0.002, I²=99.1%). Conclusions: The application of DN in KOA and PFPS patients varies among practitioners. DN is effective in reducing pain and dysfunction at short-term and 3-month post-intervention in individuals with PFPS. To our best knowledge, no study has reported the effects of DN with ultrasound guidance on KOA and PFPS. The longer-term effects of DN on KOA and PFPS are waiting for further study.

Keywords: dry needling, knee osteoarthritis, patellofemoral pain syndrome, ultrasound guidance

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Authors: Yi Jen Wang, Yu Ju Chen

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Background: Heart rate variability (HRV) biofeedback can promote autonomic nervous function, sleep quality and reduce psychological stress. In HRV biofeedback training, it is hoped that through the guidance of machine video or audio, the patient can breathe slowly according to his own heart rate changes so that the heart and lungs can achieve resonance, thereby promoting the related effects of autonomic nerve function; while, it is also pointed out that if slow breathing of 6 times per minute can also guide the case to achieve the effect of cardiopulmonary resonance. However, there is no relevant research to explore the comparison of the effectiveness of cardiopulmonary resonance by using video or audio HRV biofeedback training and metronome-guided slow breathing. Purpose: To compare the promotion of autonomic nervous function performance between using HRV biofeedback and slow breathing guided by a metronome. Method: This research is a kind of experimental design with convenient sampling; the cases are randomly divided into the heart rate variability biofeedback training group and the slow breathing training group. The HRV biofeedback training group will conduct HRV biofeedback training in a four-week laboratory and use the home training device for autonomous training; while the slow breathing training group will conduct slow breathing training in the four-week laboratory using the mobile phone APP breathing metronome to guide the slow breathing training, and use the mobile phone APP for autonomous training at home. After two groups were enrolled and four weeks after the intervention, the autonomic nervous function-related performance was repeatedly measured. Using the chi-square test, student’s t-test and other statistical methods to analyze the results, and use p <0.05 as the basis for statistical significance. Results: A total of 27 subjects were included in the analysis. After four weeks of training, the HRV biofeedback training group showed significant improvement in the HRV indexes (SDNN, RMSSD, HF, TP) and sleep quality. Although the stress index also decreased, it did not reach statistical significance; the slow breathing training group was not statistically significant after four weeks of training, only sleep quality improved significantly, while the HRV indexes (SDNN, RMSSD, TP) all increased. Although HF and stress indexes decreased, they were not statistically significant. Comparing the difference between the two groups after training, it was found that the HF index improved significantly and reached statistical significance in the HRV biofeedback training group. Although the sleep quality of the two groups improved, it did not reach that level in a statistically significant difference. Conclusion: HRV biofeedback training is more effective in promoting autonomic nervous function than slow breathing training, but the effects of reducing stress and promoting sleep quality need to be explored after increasing the number of samples. The results of this study can provide a reference for clinical or community health promotion. In the future, it can also be further designed to integrate heart rate variability biological feedback training into the development of AI artificial intelligence wearable devices, which can make it more convenient for people to train independently and get effective feedback in time.

Keywords: autonomic nervous function, HRV biofeedback, heart rate variability, slow breathing

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Authors: Salman Imran, Chris Healey

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Traditionally, medical care of children in England and Wales starts from primary care with a referral to secondary care paediatricians who may not investigate further. Many primary care doctors do not undergo a paediatric rotation/exposure in training. As a result, there are many who have not acquired the necessary skills to manage children hence increasing hospital referral. With the current demand on hospitals in the National Health Service managing more problems in the community is needed. One way of handling this is to set up clinics, meetings and huddles in GP surgeries where professionals involved (general practitioner, paediatrician, health visitor, community nurse, dietician, school nurse) come together and share information which can help improve communication and care. The increased awareness and education that paediatricians can impart in this way will help boost confidence for primary care professionals to be able to be more self-sufficient. This has been tried successfully in other regions e.g., St. Mary’s Hospital in London but is crucial for a more rural setting like ours. The primary aim of this project would be to educate specifically GP’s and generally all other health professionals involved. Additional benefits would be providing care nearer home, increasing patient’s confidence in their local surgery, improving communication and reducing unnecessary patient flow to already stretched hospital resources. Methods: This was done as a plan do study act cycle (PDSA). Three clinics were delivered in different practices over six months where feedback from staff and patients was collected. Designated time for teaching/discussion was used which involved some cases from the actual clinics. Both new and follow up patients were included. Two clinics were conducted by a paediatrician and nurse whilst the 3rd involved paediatrician and local doctor. The distance from hospital to clinics varied from two miles to 22 miles approximately. All equipment used was provided by primary care. Results: A total of 30 patients were seen. All patients found the location convenient as it was nearer than the hospital. 70-90% clearly understood the reason for a change in venue. 95% agreed to the importance of their local doctor being involved in their care. 20% needed to be seen in the hospital for further investigations. Patients felt this to be a more personalised, in-depth, friendly and polite experience. Local physicians felt this to be a more relaxed, familiar and local experience for their patients and they managed to get immediate feedback regarding their own clinical management. 90% felt they gained important learning from the discussion time and the paediatrician also learned about their understanding and gaps in knowledge/focus areas. 80% felt this time was valuable for targeted learning. Equipment, information technology, and office space could be improved for the smooth running of any future clinics. Conclusion: The acute paediatric outpatient clinic can be successfully established in primary care facilities. Careful patient selection and adequate facilities are important. We have demonstrated a further step in the reduction of patient flow to hospitals and upskilling primary care health professionals. This service is expected to become more efficient with experience.

Keywords: clinics, education, paediatricians, primary care

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Authors: Ripon Sarkar, Kabita Chaterjee, Ananya Barui

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Smoking is one of the leading causes of oral cancer. Cigarette smoke affects various cellular parameters and alters molecular metabolism of cells. Epithelial cells losses their cytoskeleton structure, membrane integrity, cellular polarity that subsequently initiates the process of epithelial cells to mesenchymal transition due to long exposure of cigarette smoking. It changes the normal cellular metabolic activity which induces oxidative stress and enhances the reactive oxygen spices (ROS) formation. Excessive ROS and associated oxidative stress are considered to be a driving force in alteration in cellular phenotypes, polarity distribution and mitochondrial metabolism. Noninvasive assessment of such parameters plays essential role in development of routine screening system for early diagnosis of oral cancer. Electrical cell-substrate impedance sensing (ECIS) is one of such method applied for detection of cellular membrane impedance which can be correlated to cell membrane integrity. Present study intends to explore the alteration in cellular impedance along with the expression of cellular polarity molecules and cytoskeleton distributions in oral epithelial cells of habitual smokers and to correlate the outcome to that of clinically diagnosed oral leukoplakia and oral squamous cell carcinoma patients. Total 80 subjects were categorized into four study groups: nonsmoker (NS), cigarette smoker (CS), oral leukoplakia (OLPK) and oral squamous cell carcinoma (OSCC). Cytoskeleton distribution was analyzed by staining of actin filament and generation of ROS was measured using assay kit using standard protocol. Cell impedance was measured through ECIS method at different frequencies. Expression of E-cadherin and protease-activated receptor (PAR) proteins were observed through immune-fluorescence method. Distribution of actin filament is well organized in NS group however; distribution pattern was grossly varied in CS, OLPK and OSCC. Generation of ROS was low in NS which subsequently increased towards OSCC. Expressions of E-cadherin and change in cellular electrical impedance in different study groups indicated the hallmark of cancer progression from NS to OSCC. Expressions of E-cadherin, PAR protein, and cell impedance were decreased from NS to CS and farther OSCC. Generally, the oral epithelial cells exhibit apico-basal polarity however with cancer progression these cells lose their characteristic polarity distribution. In this study expression of polarity molecule and ECIS observation indicates such altered pattern of polarity among smoker group. Overall the present study monitored the alterations in intracellular ROS generation and cell metabolic function, membrane integrity in oral epithelial cells in cigarette smokers. Present study thus has clinical significance, and it may help in developing a noninvasive technique for early diagnosis of oral cancer amongst susceptible individuals.

Keywords: cigarette smoking, early oral cancer detection, electric cell-substrate impedance sensing, noninvasive screening

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Authors: Hanan Begali, Shahi Dost, Annett Ziegler, Markus Cornberg, Maria-Esther Vidal, Anke R. M. Kraft

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Chronic hepatitis B virus (HBV) infection can be treated with nucleot(s)ide analog (NA), for example, which inhibits HBV replication. However, they have hardly any influence on the functional cure of HBV, which is defined by hepatitis B surface antigen (HBsAg) loss. NA needs to be taken life-long, which is not available for all patients worldwide. Additionally, NA-treated patients are still at risk of developing cirrhosis, liver failure, or hepatocellular carcinoma (HCC). Although each patient has the same components of the immune system, immune responses vary between patients. Therefore, a deeper understanding of the immune response against HBV in different patients is necessary to understand the parameters leading to HBV cure and to use this knowledge to optimize HBV therapies. This requires seamless integration of an enormous amount of diverse and fine-grained data from viral markers, e.g., hepatitis B core-related antigen (HBcrAg) and hepatitis B surface antigen (HBsAg). The data integration system relies on the assumption that profiling human immune systems requires the analysis of various variables (e.g., demographic data, treatments, pre-existing conditions, immune cell response, or HLA-typing) rather than only one. However, the values of these variables are collected independently. They are presented in a myriad of formats, e.g., excel files, textual descriptions, lab book notes, and images of flow cytometry dot plots. Additionally, patients can be identified differently in these analyses. This heterogeneity complicates the integration of variables, as data management techniques are needed to create a unified view in which individual formats and identifiers are transparent when profiling the human immune systems. The proposed study (HBsRE) aims at integrating heterogeneous data sets of 87 chronically HBV-infected patients, e.g., clinical data, immune cell response, and HLA-typing, with knowledge encoded in biomedical ontologies and open-source databases into a knowledge-driven framework. This new technique enables us to harmonize and standardize heterogeneous datasets in the defined modeling of the data integration system, which will be evaluated in the knowledge graph (KG). KGs are data structures that represent the knowledge and data as factual statements using a graph data model. Finally, the analytic data model will be applied on top of KG in order to develop a deeper understanding of the immune profiles among various patients and to evaluate factors playing a role in a holistic profile of patients with HBsAg level loss. Additionally, our objective is to utilize this unified approach to stratify patients for new effective treatments. This study is developed in the context of the project “Transforming big data into knowledge: for deep immune profiling in vaccination, infectious diseases, and transplantation (ImProVIT)”, which is a multidisciplinary team composed of computer scientists, infection biologists, and immunologists.

Keywords: chronic hepatitis B infection, immune response, knowledge graphs, ontology

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Authors: M. Koksal, T. Ozyazici, E. Gurdal, M. Yarım, E. Demirpolat, M. B. Y. Aycan

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The discovery of new drugs in cancer chemotherapy is still a major topic because of severe side effects, selectivity problems and resistance development potential of existing drugs. In recent years, combined anticancer therapies or multi-acting drugs are clinically preferred over traditional cytotoxic treatment, with the aim of avoiding resistance and toxic side effects. Arrangement of multi-acting targets can be carried out either by combination of several drugs with different mechanisms or by usage of a single chemical compound capable of regulating several targets of a disease with multiple factors. In literature, several pyrimidine and piperazine derivatives have been involved in the structure of many compounds which have been used as chemotherapeutic agents along with wide clinical applications. The aim of this study is to combine pyrimidine and piperazine core structures to research and develop novel piperazinylpyrimidine derivatives with selective cytotoxicity over cancer cells. In this study, a group of novel 6-fluorophenyl-3-[2-(substitutedpiperazinyl)ethyl] hexahydropyrimidine-2,4-dione derivatives designed to observe the desired anticancer activity due to pyrimidine and piperazine based scaffolds. Target compounds were obtained by the reaction of appropriate piperazine derivatives and 6-(2/4-fluorophenyl)-3-(2-chloroethyl)hexahydropyrimidine-2,4-dione. The synthetic pathway of 6-(2/4-fluorophenyl)-3-(2-chloroethyl)hexahydropyrimidine-2,4-dione was started with Rodionov reaction using aldehyde, malonic acid and ammonium acetate in ethanol. Isolated β-fluorophenyl-β-amino acids were treated with 2-chloroethylisocyanate in the presence of an aqueous sodium hydroxide solution at room temperature to yield the sodium salts of the corresponding ureido acids. By addition of a mineral acid, ureido acids were precipitated. Later, these ureido acids were refluxed in thionyl chloride to give the 6-(2/4-fluorophenyl)-3-(2-chloroethyl)hexahydropyrimidine-2,4-di-one which were furthermore treated with secondary amines. Structures of purified compounds were characterized with IR, 1H-NMR, 13C-NMR, mass spectroscopies and elemental analysis. All of the compounds gave satisfactory analytical and spectroscopic data, which were in full accordance with their depicted structures. In IR spectra of the compounds, N-H group was seen at 3230-3213 cm⁻¹. C-H was seen at 3100-2820 cm⁻¹ and C=O vibrational peaks were observed approximately at 1725 and 1665 cm⁻¹ in accordance with literature. In the NMR spectra of target compounds, the methylene protons of piperazine give two separate multiplet peaks around 3.5 and 4.5 ppm representing the successful N-alkylation of the structure. The cytotoxic activity of the synthesized compounds was investigated on human bronchial epithelial (BEAS 2B), lung (A549), colon adenocarcinoma (COLO205) and breast (MCF7) cell lines, by means of sulphorhodamine B (SRB) assays in triplicate. IC₅₀ values of the screened derivatives were found in range of 11.8-78 µM. This project was supported by The Scientific and Technological Research Council of Turkey (TUBITAK, Project no: 215S157).

Keywords: cytotoxicity, hexahydropyrimidine, piperazine, sulphorhodamine B assay

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Authors: Yollande Sènan Djivoh, Dominique de Jaeger

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Background: Abdominal exercises are commonly practised nowadays. Specific techniques of abdominal muscles strengthening like hypopressive exercises have recently emerged and their practice is encouraged against the practice of Curl-up especially in postpartum. The acute and the training effects of these exercises did not allow to advise one exercise to the detriment of another. However, physiotherapists remain reluctant to perform Curl-up with postpartum women because of its potential harmful effect on the pelvic floor. Design: This study was a randomized control trial registered under the number PACTR202110679363984. Objective: to observe the training effect of two experimental protocols (Curl-up versus Drawing-in+Diaphragmatic aspiration) on the abdominal wall (interrecti distance, rectus and transversus abdominis thickness, abdominal strength) in Beninese postpartum women. Pelvic floor function (tone, endurance, urinary incontinence) will be assessed to evaluate potential side effects of exercises on the pelvic floor. Method: Postpartum women diagnosed with diastasis recti were randomly assigned to one of three groups (Curl-up, Drawingin+Diaphragmatic aspiration and control). Abdominal and pelvic floor parameters were assessed before and at the end of the 6-week protocol. The interrecti distance and the abdominal muscles thickness were assessed by ultrasound and abdominal strength by dynamometer. Pelvic floor tone and strength were assessed with Biofeedback and urinary incontinence was quantified by pad test. To compare the results between the three groups and the two measurements, a two-way Anova test with repeated measures was used (p<0.05). When interaction was significant, a posthoc using Student t test, with Bonferroni correction, was used to compare the three groups regarding the difference (end value minus initial value). To complete these results, a paired Student t test was used to compare in each group the initial and end values. Results: Fifty-eight women participated in this study, divided in three groups with similar characteristics regarding their age (29±5 years), parity (2±1 children), BMI (26±4 kg/m2 ), time since the last birth (10±2 weeks), weight of their baby at birth (330±50 grams). Time effect and interaction were significant (p<0.001) for all abdominal parameters. Experimental groups improved more than control group. Curl-up group improved more (p=0.001) than Drawing-in+Diaphragmatic aspiration group regarding the interrecti distance (9.3±4.2 mm versus 6.6±4.6 mm) and abdominal strength (20.4±16.4 Newton versus 11.4±12.8 Newton). Drawingin+Diaphragmatic aspiration group improved (0.8±0.7 mm) more than Curl-up group (0.5±0.7 mm) regarding the transversus abdominis thickness (p=0.001). Only Curl-up group improved (p<0.001) the rectus abdominis thickness (1.5±1.2 mm). For pelvic floor parameters, both experimental groups improved (p=0.01) except for tone which improved (p=0.03) only in Drawing-in+Diaphragmatic aspiration group from 19.9±4.1 cmH2O to 22.2±4.5 cmH2O. Conclusion: Curl-up was more efficient to improve abdominal function than Drawingin+Diaphragmatic aspiration. However, these exercises are complementary. None of them degraded the pelvic floor, but Drawing-in+Diaphragmatic aspiration improved further the pelvic floor function. Clinical implications: Curl-up, Drawing-in and Diaphragmatic aspiration can be used for the management of abdominal function in postpartum women. Exercises must be chosen considering the specific needs of each woman’s abdominal and pelvic floor function.

Keywords: curl-up, drawing-in, diaphragmatic aspiration, hypopressive exercise, postpartum women

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Authors: Z. Yousefniayejahr, S. Bolognini, A. Bonini, C. Campobasso, N. Poma, F. Vivaldi, M. Di Luca, A. Tavanti, F. Di Francesco

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Pathogenic bacteria represent a threat to healthcare systems and the food industry because their rapid detection remains challenging. Electrochemical biosensors are gaining prominence as a novel technology for the detection of pathogens due to intrinsic features such as low cost, rapid response time, and portability, which make them a valuable alternative to traditional methodologies. These sensors use biorecognition elements that are crucial for the identification of specific bacteria. In this context, bacteriophages are promising tools for their inherent high selectivity towards bacterial hosts, which is of fundamental importance when detecting bacterial pathogens in complex biological samples. In this study, we present the development of a low-cost and portable sensor based on the Zeno phage for the rapid detection of Staphylococcus aureus. Screen-printed gold electrodes functionalized with the Zeno phage were used, and electrochemical impedance spectroscopy was applied to evaluate the change of the charge transfer resistance (Rct) as a result of the interaction with S. aureus MRSA ATCC 43300. The phage-based biosensor showed a linear range from 101 to 104 CFU/mL with a 20-minute response time and a limit of detection (LOD) of 1.2 CFU/mL under physiological conditions. The biosensor’s ability to recognize various strains of staphylococci was also successfully demonstrated in the presence of clinical isolates collected from different geographic areas. Assays using S. epidermidis were also carried out to verify the species-specificity of the phage sensor. We only observed a remarkable change of the Rct in the presence of the target S. aureus bacteria, while no substantial binding to S. epidermidis occurred. This confirmed that the Zeno phage sensor only targets S. aureus species within the genus Staphylococcus. In addition, the biosensor's specificity with respect to other bacterial species, including gram-positive bacteria like Enterococcus faecium and the gram-negative bacterium Pseudomonas aeruginosa, was evaluated, and a non-significant impedimetric signal was observed. Notably, the biosensor successfully identified S. aureus bacterial cells in a complex matrix such as a nasal swab, opening the possibility of its use in a real-case scenario. We diluted different concentrations of S. aureus from 108 to 100 CFU/mL with a ratio of 1:10 in the nasal swap matrices collected from healthy donors. Three different sensors were applied to measure various concentrations of bacteria. Our sensor indicated high selectivity to detect S. aureus in biological matrices compared to time-consuming traditional methods, such as enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), and radioimmunoassay (RIA), etc. With the aim to study the possibility to use this biosensor to address the challenge associated to pathogen detection, ongoing research is focused on the assessment of the biosensor’s analytical performances in different biological samples and the discovery of new phage bioreceptors.

Keywords: electrochemical impedance spectroscopy, bacteriophage, biosensor, Staphylococcus aureus

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