Search results for: obese mice
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 837

Search results for: obese mice

87 An Inquiry on Imaging of Soft Tissues in Micro-Computed Tomography

Authors: Matej Patzelt, Jana Mrzilkova, Jan Dudak, Frantisek Krejci, Jan Zemlicka, Zdenek Wurst, Petr Zach, Vladimir Musil

Abstract:

Introduction: Micro-CT is well used for examination of bone structures and teeth. On the other hand visualization of the soft tissues is still limited. The goal of our study was to elaborate methodology for soft tissue samples imaging in micro-CT. Methodology: We used organs of rats and mice. We either did a preparation of the organs and fixation in contrast solution or we did cannulation of blood vessels and their injection for imaging of the vascular system. First, we scanned native specimens, then we created corrosive specimens by resins. In the next step, we injected vascular system either by Aurovist contrast agent or by Exitron. In the next step, we focused on soft tissues contrast increase. We scanned samples fixated in Lugol solution, samples fixated in pure ethanol and in formaldehyde solution. All used methods were afterwards compared. Results: Native specimens did not provide sufficient contrast of the tissues in any of organs. Corrosive samples of the blood stream provided great contrast and details; on the other hand, it was necessary to destroy the organ. Further examined possibility was injection of the AuroVist contrast that leads to the great bloodstream contrast. Injection of Exitron contrast agent comparing to Aurovist did not provide such a great contrast. The soft tissues (kidney, heart, lungs, brain, and liver) were best visualized after fixation in ethanol. This type of fixation showed best results in all studied tissues. Lugol solution had great results in muscle tissue. Fixation by formaldehyde solution showed similar quality of contrast in the tissues like ethanol. Conclusion: Before imaging, we need to, first, determinate which structures of the soft tissues we want to visualize. In the case of the bloodstream, the best was AuroVist and corrosive specimens. Muscle tissue is best visualized by Lugol solution. In the case of the organs containing cavities, like kidneys or brain, the best way was ethanol fixation.

Keywords: experimental imaging, fixation, micro-CT, soft tissues

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86 Effect of Lifestyle Modification for Two Years on Obesity and Metabolic Syndrome Components in Elementary Students: A Community-Based Trial

Authors: Bita Rabbani, Hossein Chiti, Faranak Sharifi, Saeedeh Mazloomzadeh

Abstract:

Background: Lifestyle modifications, especially improving nutritional patterns and increasing physical activity, are the most important factors in preventing obesity and metabolic syndrome in children and adolescents. For this purpose, the following interventional study was designed to investigate the effects of educational programs for students, as well as changes in diet and physical activity, on obesity and components of the metabolic syndrome. Methods: This study is part of an interventional research project (elementary school) conducted on all students of Sama schools in Zanjan and Abhar in three levels of elementary, middle, and high school, including 1000 individuals in Zanjan (intervention group) and 1000 individuals (control group) in Abhar in 2011. Interventions were based on educating students, teachers, and parents, changes in food services, and physical activity. We primarily measured anthropometric indices, fasting blood sugar, lipid profiles, and blood pressure and completed standard nutrition and physical activity questionnaires. Also, blood insulin levels were randomly measured in a number of students. Data analysis was done by SPSS software version 16.0. Results: Overall, 589 individuals (252 male, 337 female) entered the case group, and 803 individuals (344 male, 459 female) entered the control group. After two years of intervention, mean waist circumference (63.8 ± 10.9) and diastolic BP (63.8 ± 10.4) were significantly lower; however, mean systolic BP (10.1.0 ± 12.5), food score (25.0 ± 5.0) and drinking score (12.1 ± 2.3) were higher in the intervention group (p<0.001). Comparing components of metabolic syndrome between the second year and at time of recruitment within the intervention group showed that although number of overweight/obese individuals, individuals with hypertriglyceridemia and high LDL increased, abdominal obesity, high BP, hyperglycemia, and insulin resistance decreased (p<0.001). On the other hand, in the control group, number of individuals with high BP increased significantly. Conclusion: The prevalence of abdominal obesity and hypertension, which are two major components of metabolic syndrome, are much higher in our study than in other regions of country. However, interventions for modification of diet and increase in physical activity are effective in lowering their prevalence.

Keywords: metabolic syndrome, obesity, life style, nutrition, hypertension

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85 A Qualitative Study on Cyberbullying and Traditional Bullying among Taiwanese High School Students

Authors: Chia-Wen Wang, Patou Masika Musumari, Teeranee Techasrivichien, S. Pilar Suguimoto, Chang-Chuan Chan, Masako Ono-Kihara, Masahiro Kihara

Abstract:

Background: In recent years, a particular form of bullying, referred to as 'cyberbullying' has emerged along with the rapid expansion of the Internet, social network services (SNSs) and smart phones. Many Asian countries, including Taiwan, are faced with both the cyberbullying and the traditional form of bullying. This study aims to explore Taiwanese adolescents’ experiences, perceptions and opinions regarding cyberbullying and traditional bullying through the perspective of victim, perpetrator, or witness. Method: This is a qualitative study using face-to-face in-depth interviews guided by a semi-structured questionnaire among high school students -aged 16 to 18 years- in Taipei, Taiwan. The participants were recruited through convenience sampling from five high schools between June and November 2016. Interviews were digitally recorded, transcribed, and analyzed using the thematic analysis approach. Results: Forty-eight participants were recruited, of which, 14 (29.2%) reported had ever experienced bullying. Specifically, 7 participants (14.6%) reported had ever been victims of cyberbullying, 1 (2%) had been victims of traditional bullying, and 6 (12.5%) had been victims of both cyber and traditional bullying. The majority (70.8%) reported had ever witnessed acts of bullying; however, none of the participants recognized had ever been a perpetrator of bullying. Cyberbullying mostly happens on social media (Facebook and Instagram) or LINE instant messaging application, and included upload and sharing of degrading pictures and videos of victims, as well as gossip and mean messages by the perpetrators. The anonymous and public nature of social media groups in schools made it easier to perpetrate bullying. The victim of traditional bullying reported being the target of verbal attack because of his physical appearance. Regardless of the type of bullying, victims reported feeling bad, angry, or depressed as a result of being bullied. Witnesses of both cyber- and traditional bullying cited physical appearance (e.g. having the big/flat bust or big butt, or overweight or obese) and disability as the most reasons of being a bullying victim. Conclusion: Both cyberbullying and traditional bullying had negative emotional and psychological impacts on victims. This study warrants further research to assess the extent of this phenomenon and understand the characteristics of perpetrators, victims, and witnesses to inform the design of tailored interventions using appropriate channels of dissemination.

Keywords: cyberbullying, traditional bullying, social media, adolescents

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84 Agreement between Basal Metabolic Rate Measured by Bioelectrical Impedance Analysis and Estimated by Prediction Equations in Obese Groups

Authors: Orkide Donma, Mustafa M. Donma

Abstract:

Basal metabolic rate (BMR) is widely used and an accepted measure of energy expenditure. Its principal determinant is body mass. However, this parameter is also correlated with a variety of other factors. The objective of this study is to measure BMR and compare it with the values obtained from predictive equations in adults classified according to their body mass index (BMI) values. 276 adults were included into the scope of this study. Their age, height and weight values were recorded. Five groups were designed based on their BMI values. First group (n = 85) was composed of individuals with BMI values varying between 18.5 and 24.9 kg/m2. Those with BMI values varying from 25.0 to 29.9 kg/m2 constituted Group 2 (n = 90). Individuals with 30.0-34.9 kg/m2, 35.0-39.9 kg/m2, > 40.0 kg/m2 were included in Group 3 (n = 53), 4 (n = 28) and 5 (n = 20), respectively. The most commonly used equations to be compared with the measured BMR values were selected. For this purpose, the values were calculated by the use of four equations to predict BMR values, by name, introduced by Food and Agriculture Organization (FAO)/World Health Organization (WHO)/United Nations University (UNU), Harris and Benedict, Owen and Mifflin. Descriptive statistics, ANOVA, post-Hoc Tukey and Pearson’s correlation tests were performed by a statistical program designed for Windows (SPSS, version 16.0). p values smaller than 0.05 were accepted as statistically significant. Mean ± SD of groups 1, 2, 3, 4 and 5 for measured BMR in kcal were 1440.3 ± 210.0, 1618.8 ± 268.6, 1741.1 ± 345.2, 1853.1 ± 351.2 and 2028.0 ± 412.1, respectively. Upon evaluation of the comparison of means among groups, differences were highly significant between Group 1 and each of the remaining four groups. The values were increasing from Group 2 to Group 5. However, differences between Group 2 and Group 3, Group 3 and Group 4, Group 4 and Group 5 were not statistically significant. These insignificances were lost in predictive equations proposed by Harris and Benedict, FAO/WHO/UNU and Owen. For Mifflin, the insignificance was limited only to Group 4 and Group 5. Upon evaluation of the correlations of measured BMR and the estimated values computed from prediction equations, the lowest correlations between measured BMR and estimated BMR values were observed among the individuals within normal BMI range. The highest correlations were detected in individuals with BMI values varying between 30.0 and 34.9 kg/m2. Correlations between measured BMR values and BMR values calculated by FAO/WHO/UNU as well as Owen were the same and the highest. In all groups, the highest correlations were observed between BMR values calculated from Mifflin and Harris and Benedict equations using age as an additional parameter. In conclusion, the unique resemblance of the FAO/WHO/UNU and Owen equations were pointed out. However, mean values obtained from FAO/WHO/UNU were much closer to the measured BMR values. Besides, the highest correlations were found between BMR calculated from FAO/WHO/UNU and measured BMR. These findings suggested that FAO/WHO/UNU was the most reliable equation, which may be used in conditions when the measured BMR values are not available.

Keywords: adult, basal metabolic rate, fao/who/unu, obesity, prediction equations

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83 Beneficial Effects of Curcumin against Stress Oxidative and Mitochondrial Dysfunction Induced by Trinitrobenzene Sulphonic Acid in Colon

Authors: Souad Mouzaoui, Bahia Djerdjouri

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Oxidative stress is one of the main factors involved in the onset and chronicity of inflammatory bowel disease (IBD). In this study, we investigated the beneficial effects of a potent natural antioxidant, curcumin (Cur) on colitis and mitochondrial dysfunction in trinitrobenzene sulfonic acid (TNBS)-induced colitis in mice. Rectal instillation of the chemical irritant TNBS (30 mg kg-1) induced the disruption of distal colonic architecture and a massive inflammatory cells influx to the mucosa and submucosa layers. Under these conditions, daily administration of Cur (25 mg kg-1) efficiently decreased colitis scores in the inflamed distal colon by reducing leukocyte infiltrate as attested by reduced myeloperoxidase (MPO) activity. Moreover, the levels of nitrite, an end product of inducible NO synthase activity (iNOS) and malonyl dialdehyde (MDA), a marker of lipid peroxidation increased in a time depending manner in response to TNBS challenge. Conversely, the markers of the antioxidant pool, reduced glutathione (GSH) and catalase activity (CAT) were drastically reduced. Cur attenuated oxidative stress markers and partially restored CAT and GSH levels. Moreover, our results expanded the effect of Cur on TNBS-induced colonic mitochondrial dysfunction. In fact, TNBS induced mitochondrial swelling and lipids peroxidation. These events reflected in the opening of mitochondrial transition pore and could be an initial indication in the cascade process leading to cell death. TNBS inhibited also mitochondrial respiratory activity, caused overproduction of mitochondrial superoxide anion (O2-.) and reduced level of mitochondrial GSH. Nevertheless, Cur reduced the extent of mitochondrial oxidative stress induced by TNBS and restored colonic mitochondrial function. In conclusion, our results showed the critical role of oxidative stress in TNBS-induced colitis. They highlight the role of colonic mitochondrial dysfunction induced by TNBS, as a potential source of oxidative damages. Due to its potent antioxidant properties, Cur opens a promising therapeutic approach against oxidative inflammation in IBD.

Keywords: colitis, curcumin, mitochondria, oxidative stress, TNBS

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82 Role of Onion Extract for Neuro-Protection in Experimental Stroke Model

Authors: Richa Shri, Varinder Singh, Kundan Singh Bora, Abhishek Bhanot, Rahul Kumar, Amit Kumar, Ravinder Kaur

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The term ‘neuroprotection’ means preserving/salvaging function and structure of neurons. Neuroprotection is an adjunctive treatment option for neurodegenerative disorders. Oxidative stress is considered a major culprit in neurodegenerative disorders; hence, management strategies include use of antioxidants. Our search for a neuroprotective agent began with Allium cepa L. or onions, (family Amaryllidaceae) - a potent antioxidant. We have investigated the neuroprotective potential of onions in experimental models of ischemic stroke, diabetic neuropathy, neuropathic pain, and dementia. In pre and post-ischemic stroke model, the methanol extract of outer scales of onion bulbs (MEOS) prevented memory loss and motor in-coordination; reduced oxidative stress and cerebral infarct size. This also prevented and ameliorated diabetic neuropathy in mice. The MEOS was fractionated to yield a flavonoid rich fraction (FRF) that successfully reversed ischemia-reperfusion induced neuronal damage, thereby demonstrating that the flavonoids are responsible for the activity. The FRF effectively ameliorated chronic constriction induced neuropathic pain in rats. The FRF was subjected to bioactivity-guided fractionated. It was seen that FRF is more effective as compared to the isolated components probably due to synergism among the constituents (i.e., quercetin and quercetin glucosides) in the FRF. The outer scales of onion bulbs have great potential for prevention as well as for treatment of neuronal disorders. Red onions, with higher amounts of flavonoids as compared to the white onions, produced more significant neuroprotection. Thus, the standardized FRF from the waste material of a commonly used vegetable, especially the red variety, may be developed as a valuable neuroprotective agent.

Keywords: Allium cepa, antioxidant activity, flavonoid rich fraction, neuroprotection

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81 Pooled Analysis of Three School-Based Obesity Interventions in a Metropolitan Area of Brazil

Authors: Rosely Sichieri, Bruna K. Hassan, Michele Sgambato, Barbara S. N. Souza, Rosangela A. Pereira, Edna M. Yokoo, Diana B. Cunha

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Obesity is increasing at a fast rate in low and middle-income countries where few school-based obesity interventions have been conducted. Results of obesity prevention studies are still inconclusive mainly due to underestimation of sample size in cluster-randomized trials and overestimation of changes in body mass index (BMI). The pooled analysis in the present study overcomes these design problems by analyzing 4,448 students (mean age 11.7 years) from three randomized behavioral school-based interventions, conducted in public schools of the metropolitan area of Rio de Janeiro, Brazil. The three studies focused on encouraging students to change their drinking and eating habits over one school year, with monthly 1-h sessions in the classroom. Folders explaining the intervention program and suggesting the participation of the family, such as reducing the purchase of sodas were sent home. Classroom activities were delivered by research assistants in the first two interventions and by the regular teachers in the third one, except for culinary class aimed at developing cooking skills to increase healthy eating choices. The first intervention was conducted in 2005 with 1,140 fourth graders from 22 public schools; the second, with 644 fifth graders from 20 public schools in 2010; and the last one, with 2,743 fifth and sixth graders from 18 public schools in 2016. The result was a non-significant change in BMI after one school year of positive changes in dietary behaviors associated with obesity. Pooled intention-to-treat analysis using linear mixed models was used for the overall and subgroup analysis by BMI status, sex, and race. The estimated mean BMI changes were from 18.93 to 19.22 in the control group and from 18.89 to 19.19 in the intervention group; with a p-value of change over time of 0.94. Control and intervention groups were balanced at baseline. Subgroup analyses were statistically and clinically non-significant, except for the non-overweight/obese group with a 0.05 reduction of BMI comparing the intervention with control. In conclusion, this large pooled analysis showed a very small effect on BMI only in the normal weight students. The results are in line with many of the school-based initiatives that have been promising in relation to modifying behaviors associated with obesity but of no impact on excessive weight gain. Changes in BMI may require great changes in energy balance that are hard to achieve in primary prevention at school level.

Keywords: adolescents, obesity prevention, randomized controlled trials, school-based study

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80 Multi-Scale Modelling of the Cerebral Lymphatic System and Its Failure

Authors: Alexandra K. Diem, Giles Richardson, Roxana O. Carare, Neil W. Bressloff

Abstract:

Alzheimer's disease (AD) is the most common form of dementia and although it has been researched for over 100 years, there is still no cure or preventive medication. Its onset and progression is closely related to the accumulation of the neuronal metabolite Aβ. This raises the question of how metabolites and waste products are eliminated from the brain as the brain does not have a traditional lymphatic system. In recent years the rapid uptake of Aβ into cerebral artery walls and its clearance along those arteries towards the lymph nodes in the neck has been suggested and confirmed in mice studies, which has led to the hypothesis that interstitial fluid (ISF), in the basement membranes in the walls of cerebral arteries, provides the pathways for the lymphatic drainage of Aβ. This mechanism, however, requires a net reverse flow of ISF inside the blood vessel wall compared to the blood flow and the driving forces for such a mechanism remain unknown. While possible driving mechanisms have been studied using mathematical models in the past, a mechanism for net reverse flow has not been discovered yet. Here, we aim to address the question of the driving force of this reverse lymphatic drainage of Aβ (also called perivascular drainage) by using multi-scale numerical and analytical modelling. The numerical simulation software COMSOL Multiphysics 4.4 is used to develop a fluid-structure interaction model of a cerebral artery, which models blood flow and displacements in the artery wall due to blood pressure changes. An analytical model of a layer of basement membrane inside the wall governs the flow of ISF and, therefore, solute drainage based on the pressure changes and wall displacements obtained from the cerebral artery model. The findings suggest that an active role in facilitating a reverse flow is played by the components of the basement membrane and that stiffening of the artery wall during age is a major risk factor for the impairment of brain lymphatics. Additionally, our model supports the hypothesis of a close association between cerebrovascular diseases and the failure of perivascular drainage.

Keywords: Alzheimer's disease, artery wall mechanics, cerebral blood flow, cerebral lymphatics

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79 Evaluation of Alpha-Glucosidase Inhibitory Effect of Two Plants from Brazilian Cerrado

Authors: N. A. P. Camaforte, P. M. P. Vareda, L. L. Saldanha, A. L. Dokkedal, J. M. Rezende-Neto, M. R. Senger, F. P. Silva-Jr, J. R. Bosqueiro

Abstract:

Diabetes mellitus is a disease characterized by deficiency of insulin secretion and/or action which results in hyperglycemia. Nowadays, acarbose is a medicine used by diabetic people to inhibit alpha-glucosidases leading to the decreasing of post-feeding glycaemia, but with low effectiveness and many side effects. Medicinal plants have been used for the treatment of many diseases including diabetes and their action occurs through the modulation of insulin-depending processes, pancreas regeneration or inhibiting glucose absorption by the intestine. Previous studies in our laboratory showed that the treatment using two crude extracts of plants from Brazilian cerrado was able to decrease fasting blood glucose and improve glucose tolerance in streptozotocin-diabetic mice. Because of this and the importance of the search for new alternatives to decrease the hyperglycemia, we decided to evaluate the inhibitory action of two plants from Brazilian cerrado - B.H. and Myrcia bella. The enzymatic assay was performed in 50 µL of final volume using pancreatic α-amylase and maltase together with theirs commercial substrates. The inhibition potency (IC50) was determined by the incubation of eight different concentrations of both extracts and the enzymes for 5 minutes at 37ºC. After, the substrate was added to start the reaction. Glucosidases assay was evaluated measuring the quantity of p-nitrophenol in 405 nmin 384 wells automatic reader. The in vitro assay with the extracts of B.H. and M. bella showed an IC50 of 28,04µg/mL and 16,93 µg/mL for α-amilase, and 43,01µg/mL and 17 µg/mL for maltase, respectively. M. bella extract showed a higher inhibitory activity for those enzymes than B.H. extract. The crude extracts tested showed a higher inhibition rate to α-amylase, but were less effective against maltase in comparison to acarbose (IC50 36µg/mL and 9 µg/mL, respectively). In conclusion, the crude extract of B.H. and M. bella showed a potent inhibitory effect against α-amylase and showed promising results to the possible development of new medicines to treat diabetes with less or even without side effects.

Keywords: alfa-glucosidases, diabetes mellitus, glycaemia, medicinal plants

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78 Association of Maternal Diet Quality Indices and Dietary Patterns during Lactation and the Growth of Exclusive Breastfed Infant

Authors: Leila Azadbakht, Maedeh Moradi, Mohammad Reza Merasi, Farzaneh Jahangir

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Maternal dietary intake during lactation might affect the growth rate of an exclusive breastfed infant. The present study was conducted to evaluate the effect of maternal dietary patterns and quality during lactation on the growth of the exclusive breastfed infant. Methods: 484 healthy lactating mothers with their infant were enrolled in this study. Only exclusive breastfed infants were included in this study which was conducted in Iran. Dietary intake of lactating mothers was assessed using a validated and reliable semi-quantitative food frequency questionnaire. Diet quality indices such as alternative Healthy eating index (HEI), Dietary energy density (DED), and adherence to Mediterranean dietary pattern score, Nordic and dietary approaches to stop hypertension (DASH) eating pattern were created. Anthropometric features of infant (weight, height, and head circumference) were recorded at birth, two and four months. Results: Weight, length, weight for height and head circumference of infants at two months and four months age were mostly in the normal range among those that mothers adhered more to the HEI in lactation period (normal weight: 61%; normal height: 59%). The prevalence of stunting at four months of age among those whose mothers adhered more to the HEI was 31% lower than those with the least adherence to HEI. Mothers in the top tertiles of HEI score had the lowest frequency of having underweight infants (18% vs. 33%; P=0.03). Odds ratio of being overweight or obese at four months age was the lowest among those infants whose mothers adhered more to the HEI (OR: 0.67 vs 0.91; Ptrend=0.03). However, there was not any significant association between adherence of mothers to Mediterranean diet as well as DASH diet and Nordic eating pattern and the growth of infants (none of weight, height or head circumference). Infant weight, length, weight for height and head circumference at two months and four months did not show significant differences among different tertile categories of mothers’ DED. Conclusions: Higher diet quality indices and more adherence of lactating mother to HEI (as an indicator of diet quality) may be associated with better growth indices of the breastfed infant. However, it seems that DED of the lactating mother does not affect the growth of the breastfed infant. Adherence to the different dietary patterns such as Mediterranean, DASH or Nordic among mothers had no different effect on the growth indices of the infants. However, higher diet quality indices and more adherence of lactating mother to HEI may be associated with better growth indices of the breastfed infant. Breastfeeding is a complete way that is not affected much by the dietary patterns of the mother. However, better diet quality might be associated with better growth.

Keywords: breastfeeding, growth, infant, maternal diet

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77 ROCK Signaling and Radio Resistance: The Association and the Effect

Authors: P. Annapurna, Cecil Ross, Sudhir Krishna, Sweta Srivastava

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Irradiation plays a pivotal role in cervical cancer treatment, however some tumors exhibit resistance to therapy while some exhibit relapse, due to better repair and enhanced resistance mechanisms operational in their cells. The present study aims to understand the signaling mechanism operational in resistance phenotype and in the present study we report the role of Rho GTPase associated protein kinase (ROCK) signaling in cervical carcinoma radio-resistance. ROCK signaling has been implicated in several tumor progressions and is important for DNA repair. Irradiation of spheroid cultures of SiHa cervical carcinoma derived cell line at 6Gy resulted in generation of resistant cells in vitro which had better clonogenic abilities and formed larger and more colonies, in soft agar colony formation assay, as compared to the non-irradiated cells. These cells also exhibited an enhanced motility phenotype. Cell cycle profiling showed the cells to be blocked in G2M phase with enhanced pCDC2 levels indicating onset of possible DNA repair mechanism. Notably, 3 days post-irradiation, irradiated cells showed increased ROCK2 translocation to the nucleus with enhanced protein expression as compared to the non-irradiated cells. Radio-sensitization of the resistant cells was enhanced using Y27632, an inhibitor to ROCK signaling. The treatment of resistant cells with Y27632 resulted in increased cell death upon further irradiation. This observation has been confirmed using inhibitory antibodies to ROCK1/2. Result show that both ROCK1/2 have a functional contribution in radiation resistance of cervical cancer cells derived from cell lines. Interestingly enrichment of stem like cells (Hoechst negative cells) was also observed upon irradiation and these cells were markedly sensitive to Y27632 treatment. Our results thus suggest the role of ROCK signaling in radio-resistance in cervical carcinoma. Further studies with human biopsies, mice models and mechanistic of ROCK signaling in the context of radio-resistance will clarify the role of this molecule further and allow for therapeutics development.

Keywords: cervical carcinoma, radio-resistance, ROCK signaling, cancer treatment

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76 Coherent Optical Tomography Imaging of Epidermal Hyperplasia in Vivo in a Mouse Model of Oxazolone Induced Atopic Dermatitis

Authors: Eric Lacoste

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Laboratory animals are currently widely used as a model of human pathologies in dermatology such as atopic dermatitis (AD). These models provide a better understanding of the pathophysiology of this complex and multifactorial disease, the discovery of potential new therapeutic targets and the testing of the efficacy of new therapeutics. However, confirmation of the correct development of AD is mainly based on histology from skin biopsies requiring invasive surgery or euthanasia of the animals, plus slicing and staining protocols. However, there are currently accessible imaging technologies such as Optical Coherence Tomography (OCT), which allows non-invasive visualization of the main histological structures of the skin (like stratum corneum, epidermis, and dermis) and assessment of the dynamics of the pathology or efficacy of new treatments. Briefly, female immunocompetent hairless mice (SKH1 strain) were sensitized and challenged topically on back and ears for about 4 weeks. Back skin and ears thickness were measured using calliper at 3 occasions per week in complement to a macroscopic evaluation of atopic dermatitis lesions on back: erythema, scaling and excoriations scoring. In addition, OCT was performed on the back and ears of animals. OCT allows a virtual in-depth section (tomography) of the imaged organ to be made using a laser, a camera and image processing software allowing fast, non-contact and non-denaturing acquisitions of the explored tissues. To perform the imaging sessions, the animals were anesthetized with isoflurane, placed on a support under the OCT for a total examination time of 5 to 10 minutes. The results show a good correlation of the OCT technique with classical HES histology for skin lesions structures such as hyperkeratosis, epidermal hyperplasia, and dermis thickness. This OCT imaging technique can, therefore, be used in live animals at different times for longitudinal evaluation by repeated measurements of lesions in the same animals, in addition to the classical histological evaluation. Furthermore, this original imaging technique speeds up research protocols, reduces the number of animals and refines the use of the laboratory animal.

Keywords: atopic dermatitis, mouse model, oxzolone model, histology, imaging

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75 Opportunities of an Industrial City in the Leisure Tourism

Authors: E. Happ, A. Albert Tóth

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The aim of the research is to investigate the forms of the demands of leisure tourism in a West-Hungarian industrial city, Győr. Today, Győr is still a traditional industrial city, its industry is mainly based on vehicle industry, but the role of tourism is increasing in the life of the city as well. Because of the industrial nature and the strong economy of the city, the ratio of business tourists is high. It can be stated that MICE tourism is dominating in Győr. Developments of the last decade can help the city with new tourism products to increase the leisure tourism. The new types of tourism – besides business tourism – can help the providers to increase the occupancy rates and the demand at the weekends. The research demonstrates the theoretical background of the topic, and it shows the present situation of the tourism in Győr with secondary data. The secondary research contains statistical data from the Hungarian Statistical Office and the city council, and it is based on the providers’ data. The next part of the paper shows the potential types of leisure tourism with the help of primary research. The primary research contains the results of an online questionnaire with a sample of 1000 potential customers. It is completed with 10 in-depth interviews with tourism experts, who explained their opinions about the opportunities of leisure tourism in Győr from the providers’ side. The online questionnaire was filled out in spring 2017 by customers, who have already stayed in Győr or plan to visit the city. At the same time in-depth interviews were made with hotel managers, head of touristic institutions and employees at the council. Based on the research it can be stated that the touristic supply of Győr allows the increase of the leisure tourism ratio in the city. Primarily, the cultural and health tourism show potential development, but the supply side of touristic services can be developed in order to increase the number of guest nights. The tourism marketing needs to be strengthened in the city, and a distinctive marketing activity - from other cities - is needed as well. To conclude, although Győr is an industrial city, it has a transforming industrial part, and tourism is also strongly present in its economy. Besides the leading role of business tourism, different types of leisure tourism have the opportunity to take place in the city.

Keywords: business tourism, Győr, industrial city, leisure tourism, touristic demand

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74 Insulin-Producing Cells from Adult Human Bone Marrow Mesenchymal Stem Cells Control Chemically-Induced Diabetes in Dogs

Authors: Maha Azzam, Mahmoud Gabr, Mahmoud Zakaria, Ayman Refaie, Amani Ismail, Sherry Khater, Sylvia Ashamallah, Mohamed Ghoniem

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Evidence was provided that human bone marrow-derived mesenhymal stem cells (HBM-MSCs) could be differentiated to form insulin-producing cells (IPCs). Transplantation of these cells was able to cure chemically-induced diabetes in nude mice. The efficacy of these cells to control diabetes in large animals was carried out to evaluate the sufficient number of cells needed/Kg body weight and to determine the functional longevity in vivo. Materials/Methods: Ten male mongrel dogs weighing 15-20 Kg were used in this study. Diabetes was chemically-induced in 7 dogs by a mixture of alloxan and streptozotocin. Three non-diabetic served as normal controls. Differentiated HBM-MSCs (5 million/Kg) were encapsulated in theracyte capsules and transplanted beneath the rectus sheath. Each dog received 2 capsules. One dog died 4 days postoperative from inhalation pneumonia. The remaining 6 dogs were followed up for 6-18 months. Results: Four dogs became normoglycemic within 6-8 weeks with normal glucose tolerance curves providing evidence that the transplanted cells were glucose-sensitive and insulin-responsive. In the remaining 2 dogs, fasting blood glucose was reduced but did not reach euglycemic levels. The sera of all transplanted dogs contained human insulin and c-peptide but negligible levels of canine insulin. When the HBM-MSCs loaded capsules were removed, rapid return of diabetic state was noted. The harvested capsules were examined by immunofluorescence. IPCs were seen and co-expression of with c-peptide was confirmed. Furthermore, all the pancreatic endocrine genes were expressed by the transplanted cells. Conclusions: This study provided evidence that theracyte capsules could protect the xenogenic HBM-MSCs from the host immune response. This is an important issue when clinical stem cell therapy is considered for definitive treatment for T1DM.

Keywords: diabetes, mesenchymal stem cells, dogs, Insulin-producing cells

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73 Liver Regeneration of Small in situ Injury

Authors: Ziwei Song, Junjun Fan, Jeremy Teo, Yang Yu, Yukun Ma, Jie Yan, Shupei Mo, Lisa Tucker-Kellogg, Peter So, Hanry Yu

Abstract:

Liver is the center of detoxification and exposed to toxic metabolites all the time. It is highly regenerative after injury, with the ability to restore even after 70% partial hepatectomy. Most of the previous studies were using hepatectomy as injury models for liver regeneration study. There is limited understanding of small-scale liver injury, which can be caused by either low dose drug consumption or hepatocyte routine metabolism. Although these small in situ injuries do not cause immediate symptoms, repeated injuries will lead to aberrant wound healing in liver. Therefore, the cellular dynamics during liver regeneration is critical for our understanding of liver regeneration mechanism. We aim to study the liver regeneration of small-scale in situ liver injury in transgenic mice labeling actin (Lifeact-GFP). Previous studies have been using sample sections and biopsies of liver, which lack real-time information. In order to trace every individual hepatocyte during the regeneration process, we have developed and optimized an intravital imaging system that allows in vivo imaging of mouse liver for consecutive 5 days, allowing real-time cellular tracking and quantification of hepatocytes. We used femtosecond-laser ablation to make controlled and repeatable liver injury model, which mimics the real-life small in situ liver injury. This injury model is the first case of its kind for in vivo study on liver. We found that small-scale in situ liver injury is repaired by the coordination of hypertrophy and migration of hepatocytes. Hypertrophy is only transient at initial phase, while migration is the main driving force to complete the regeneration process. From cellular aspect, Akt/mTOR pathway is activated immediately after injury, which leads to transient hepatocyte hypertrophy. From mechano-sensing aspect, the actin cable, formed at apical surface of wound proximal hepatocytes, provides mechanical tension for hepatocyte migration. This study provides important information on both chemical and mechanical signals that promote liver regeneration of small in situ injury. We conclude that hypertrophy and migration play a dominant role at different stages of liver regeneration.

Keywords: hepatocyte, hypertrophy, intravital imaging, liver regeneration, migration

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72 Knowledge State of Medical Students in Morocco Regarding Metabolic Dysfunction Associated with Non-alcoholic Fatty Liver Disease (MASLD)

Authors: Elidrissi Laila, El Rhaoussi Fatima-Zahra, Haddad Fouad, Tahiri Mohamed, Hliwa Wafaa, Bellabah Ahmed, Badre Wafaa

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Introduction: Metabolic Dysfunction Associated with Non-Alcoholic Fatty Liver Disease (MASLD), formerly known as Non-Alcoholic Fatty Liver Disease (NAFLD), is the leading cause of chronic liver disease. The cardiometabolic risk factors associated with MASLD represent common health issues and significant public health challenges. Medical students, being active participants in the healthcare system and a young demographic, are particularly relevant for understanding this entity to prevent its occurrence on a personal and collective level. The objective of our study is to assess the level of knowledge among medical students regarding MASLD, its risk factors, and its long-term consequences. Materials and Methods: We conducted a descriptive cross-sectional study using an anonymous questionnaire distributed through social media over a period of 2 weeks. Medical students from various faculties in Morocco answered 22 questions about MASLD, its etiological factors, diagnosis, complications, and principles of treatment. All responses were analyzed using the Jamovi software. Results: A total of 124 students voluntarily provided complete responses. 59% of our participants were in their 3rd year, with a median age of 21 years. Among the respondents, 27% were overweight, obese, or diabetic. 83% correctly answered more than half of the questions, and 77% believed they knew about MASLD. However, 84% of students were unaware that MASLD is the leading cause of chronic liver disease, and 12% even considered it a rare condition. Regarding etiological factors, overweight and obesity were mentioned in 93% of responses, and type 2 diabetes in 84%. 62% of participants believed that type 1 diabetes could not be implicated in MASLD. For 83 students, MASLD was considered a diagnosis of exclusion, while 41 students believed that a biopsy was mandatory for diagnosis. 12% believed that MASLD did not lead to long-term complications, and 44% were unaware that MASLD could progress to hepatocellular carcinoma. Regarding treatment, 85% included weight loss, and 19% did not consider diabetes management as a therapeutic approach for MASLD. At the end of the questionnaire, 89% of the students expressed a desire to learn more about MASLD and were invited to access an informative sheet through a hyperlink. Conclusion: MASLD represents a significant public health concern due to the prevalence of its risk factors, notably the obesity pandemic, which is widespread among the young population. There is a need for awareness about the seriousness of this emerging and long-underestimated condition among young future physicians.

Keywords: MASLD, medical students, obesity, diabetes

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71 EverPro as the Missing Piece in the Plant Protein Portfolio to Aid the Transformation to Sustainable Food Systems

Authors: Aylin W Sahin, Alice Jaeger, Laura Nyhan, Gregory Belt, Steffen Münch, Elke K. Arendt

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Our current food systems cause an increase in malnutrition resulting in more people being overweight or obese in the Western World. Additionally, our natural resources are under enormous pressure and the greenhouse gas emission increases yearly with a significant contribution to climate change. Hence, transforming our food systems is of highest priority. Plant-based food products have a lower environmental impact compared to their animal-based counterpart, representing a more sustainable protein source. However, most plant-based protein ingredients, such as soy and pea, are lacking indispensable amino acids and extremely limited in their functionality and, thus, in their food application potential. They are known to have a low solubility in water and change their properties during processing. The low solubility displays the biggest challenge in the development of milk alternatives leading to inferior protein content and protein quality in dairy alternatives on the market. Moreover, plant-based protein ingredients often possess an off-flavour, which makes them less attractive to consumers. EverPro, a plant-protein isolate originated from Brewer’s Spent Grain, the most abundant by-product in the brewing industry, represents the missing piece in the plant protein portfolio. With a protein content of >85%, it is of high nutritional value, including all indispensable amino acids which allows closing the protein quality gap of plant proteins. Moreover, it possesses high techno-functional properties. It is fully soluble in water (101.7 ± 2.9%), has a high fat absorption capacity (182.4 ± 1.9%), and a foaming capacity which is superior to soy protein or pea protein. This makes EverPro suitable for a vast range of food applications. Furthermore, it does not cause changes in viscosity during heating and cooling of dispersions, such as beverages. Besides its outstanding nutritional and functional characteristics, the production of EverPro has a much lower environmental impact compared to dairy or other plant protein ingredients. Life cycle assessment analysis showed that EverPro has the lowest impact on global warming compared to soy protein isolate, pea protein isolate, whey protein isolate, and egg white powder. It also contributes significantly less to freshwater eutrophication, marine eutrophication and land use compared the protein sources mentioned above. EverPro is the prime example of sustainable ingredients, and the type of plant protein the food industry was waiting for: nutritious, multi-functional, and environmentally friendly.

Keywords: plant-based protein, upcycled, brewers' spent grain, low environmental impact, highly functional ingredient

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70 Food for Health: Understanding the Importance of Food Safety in the Context of Food Security

Authors: Carmen J. Savelli, Romy Conzade

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Background: Access to sufficient amounts of safe and nutritious food is a basic human necessity, required to sustain life and promote good health. Food safety and food security are therefore inextricably linked, yet the importance of food safety in this relationship is often overlooked. Methodologies: A literature review and desk study were conducted to examine existing frameworks for discussing food security, especially from an international perspective, to determine the entry points for enhancing considerations for food safety in national and international policies. Major Findings: Food security is commonly understood as the state when all people at all times have physical, social and economic access to sufficient, safe and nutritious food to meet their dietary needs and food preferences for an active and healthy life. Conceptually, food security is built upon four pillars including food availability, access, utilization and stability. Within this framework, the safety of food is often wrongly assumed as a given. However, in places where food supplies are insufficient, coping mechanisms for food insecurity are primarily focused on access to food without considerations for ensuring safety. Under such conditions, hygiene and nutrition are often ignored as people shift to less nutritious diets and consume more potentially unsafe foods, in which chemical, microbiological, zoonotic and other hazards can pose serious, acute and chronic health risks. While food supplies might be safe and nutritious, if consumed in quantities insufficient to support normal growth, health and activity, the result is hunger and famine. Recent estimates indicate that at least 842 million people, or roughly one in eight, still suffer from chronic hunger. Even if people eat enough food that is safe, they will become malnourished if the food does not provide the proper amounts of micronutrients and/or macronutrients to meet daily nutritional requirements, resulting in under- or over-nutrition. Two billion people suffer from one or more micronutrient deficiencies and over half a billion adults are obese. Access to sufficient amounts of nutritious food is not enough. If food is unsafe, whether arising from poor quality supplies or inadequate treatment and preparation, it increases the risk of foodborne infections such as diarrhoea. 70% of diarrhoea episodes occurring annually in children under five are due to biologically contaminated food. Conclusions: An integrated approach is needed where food safety and nutrition are systematically introduced into mainstream food system policies and interventions worldwide in order to achieve health and development goals. A new framework, “Food for Health” is proposed to guide policy development and requires all three aspects of food security to be addressed in balance: sufficiency, nutrition and safety.

Keywords: food safety, food security, nutrition, policy

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69 Application of Compressed Sensing and Different Sampling Trajectories for Data Reduction of Small Animal Magnetic Resonance Image

Authors: Matheus Madureira Matos, Alexandre Rodrigues Farias

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Magnetic Resonance Imaging (MRI) is a vital imaging technique used in both clinical and pre-clinical areas to obtain detailed anatomical and functional information. However, MRI scans can be expensive, time-consuming, and often require the use of anesthetics to keep animals still during the imaging process. Anesthetics are commonly administered to animals undergoing MRI scans to ensure they remain still during the imaging process. However, prolonged or repeated exposure to anesthetics can have adverse effects on animals, including physiological alterations and potential toxicity. Minimizing the duration and frequency of anesthesia is, therefore, crucial for the well-being of research animals. In recent years, various sampling trajectories have been investigated to reduce the number of MRI measurements leading to shorter scanning time and minimizing the duration of animal exposure to the effects of anesthetics. Compressed sensing (CS) and sampling trajectories, such as cartesian, spiral, and radial, have emerged as powerful tools to reduce MRI data while preserving diagnostic quality. This work aims to apply CS and cartesian, spiral, and radial sampling trajectories for the reconstruction of MRI of the abdomen of mice sub-sampled at levels below that defined by the Nyquist theorem. The methodology of this work consists of using a fully sampled reference MRI of a female model C57B1/6 mouse acquired experimentally in a 4.7 Tesla MRI scanner for small animals using Spin Echo pulse sequences. The image is down-sampled by cartesian, radial, and spiral sampling paths and then reconstructed by CS. The quality of the reconstructed images is objectively assessed by three quality assessment techniques RMSE (Root mean square error), PSNR (Peak to Signal Noise Ratio), and SSIM (Structural similarity index measure). The utilization of optimized sampling trajectories and CS technique has demonstrated the potential for a significant reduction of up to 70% of image data acquisition. This result translates into shorter scan times, minimizing the duration and frequency of anesthesia administration and reducing the potential risks associated with it.

Keywords: compressed sensing, magnetic resonance, sampling trajectories, small animals

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68 The Relationship between Anthropometric Obesity Indices and Insulin in Children with Metabolic Syndrome

Authors: Mustafa M. Donma, Orkide Donma

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The number of indices developed for the evaluation of obesity both in adults and pediatric population is ever increasing. These indices are also used in cases with metabolic syndrome (MetS), mostly the ultimate form of morbid obesity. Aside from anthropometric measurements, formulas constituted using these parameters also find clinical use. These formulas can be listed as two groups; being weight-dependent and –independent. Some are extremely sophisticated equations and their clinical utility is questionable in routine clinical practice. The aim of this study is to compare presently available obesity indices and find the most practical one. Their associations with MetS components were also investigated to determine their capacities in differential diagnosis of morbid obesity with and without MetS. Children with normal body mass index (N-BMI) and morbid obesity were recruited for this study. Three groups were constituted. Age- and sex- dependent BMI percentiles for morbid obese (MO) children were above 99 according to World Health Organization tables. Of them, those with MetS findings were evaluated as MetS group. Children, whose values were between 85 and 15 were included in N-BMI group. The study protocol was approved by the Ethics Committee of the Institution. Parents filled out informed consent forms to participate in the study. Anthropometric measurements and blood pressure values were recorded. Body mass index, hip index (HI), conicity index (CI), triponderal mass index (TPMI), body adiposity index (BAI), body shape index (ABSI), body roundness index (BRI), abdominal volume index (AVI), waist-to-hip ratio (WHR) and waist circumference+hip circumference/2 ((WC+HC)/2) were the formulas examined within the scope of this study. Routine biochemical tests including fasting blood glucose (FBG), insulin (INS), triglycerides (TRG), high density lipoprotein-cholesterol (HDL-C) were performed. Statistical package program SPSS was used for the evaluation of study data. p<0.05 was accepted as the statistical significance degree. Hip index did not differ among the groups. A statistically significant difference was noted between N-BMI and MetS groups in terms of ABSI. All the other indices were capable of making discrimination between N-BMI-MO, N-BMI- MetS and MO-MetS groups. No correlation was found between FBG and any obesity indices in any groups. The same was true for INS in N-BMI group. Insulin was correlated with BAI, TPMI, CI, BRI, AVI and (WC+HC)/2 in MO group without MetS findings. In MetS group, the only index, which was correlated with INS was (WC+HC)/2. These findings have pointed out that complicated formulas may not be required for the evaluation of the alterations among N-BMI and various obesity groups including MetS. The simple easily computable weight-independent index, (WC+HC)/2, was unique, because it was the only index, which exhibits a valuable association with INS in MetS group. It did not exhibit any correlation with other obesity indices showing associations with INS in MO group. It was concluded that (WC+HC)/2 was pretty valuable practicable index for the discrimination of MO children with and without MetS findings.

Keywords: children, insulin, metabolic syndrome, obesity indices

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67 Inquiry on Regenerative Tourism in an Avian Destination: A Case Study of Kaliveli in Tamil Nadu, India

Authors: Anu Chandran, Reena Esther Rani

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Background of the Study: Dotted with multiple Unique Destination Prepositions (UDPs), Tamil Nadu is an established tourism brand as regards leisure, MICE, culture, and ecological flavors. Albeit, the enchanting destination possesses distinctive attributes and resources yet to be tapped for better competitive advantage. Being a destination that allures an incredible variety of migratory birds, Tamil Nadu is deemed to be an ornithologist’s paradise. This study primarily explores the prospects of developing Kaliveli, recognized as a bird sanctuary in the Tindivanam forest division of the Villupuram district in the State. Kaliveli is an ideal nesting site for migratory birds and is currently apt for a prospective analysis of regenerative tourism. Objectives of the study: This research lays an accent on avian tourism as part and parcel of sustainable tourism ventures. The impacts of projects like the Ornithological Conservation Centre on tourists have been gauged in the present paper. It maps the futuristic proactive propositions linked to regenerative tourism on the site. How far technological innovations can do a world of good in Kaliveli through Artificial Intelligence, Smart Tourism, and similar latest coinages to entice real eco-tourists, have been conceptualized. The experiential dimensions of resource stewardship as regards facilitating tourists’ relish the offerings in a sustainable manner is at the crux of this work. Methodology: Modeled as a case study, this work tries to deliberate on the impact of existing projects attributed to avian fauna in Kalveli. Conducted in the qualitative research design mode, the case study method was adopted for the processing and presentation of study results drawn by applying thematic content analysis based on the data collected from the field. Result and discussion: One of the key findings relates to the kind of nature trails that can be a regenerative dynamic for eco-friendly tourism in Kaliveli. Field visits have been conducted to assess the niche tourism aspects which could be incorporated with the regenerative tourism model to be framed as part of the study.

Keywords: regenerative tourism, Kaliveli bird sanctuary, sustainable development, resource Stewardship, Ornithology, Avian Fauna

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66 In vitro and in vivo Anticancer Activity of Nanosize Zinc Oxide Composites of Doxorubicin

Authors: Emma R. Arakelova, Stepan G. Grigoryan, Flora G. Arsenyan, Nelli S. Babayan, Ruzanna M. Grigoryan, Natalia K. Sarkisyan

Abstract:

Novel nanosize zinc oxide composites of doxorubicin obtained by deposition of 180 nm thick zinc oxide film on the drug surface using DC-magnetron sputtering of a zinc target in the form of gels (PEO+Dox+ZnO and Starch+NaCMC+Dox+ZnO) were studied for drug delivery applications. The cancer specificity was revealed both in in vitro and in vivo models. The cytotoxicity of the test compounds was analyzed against human cancer (HeLa) and normal (MRC5) cell lines using MTT colorimetric cell viability assay. IC50 values were determined and compared to reveal the cancer specificity of the test samples. The mechanistic study of the most active compound was investigated using Flow cytometry analyzing of the DNA content after PI (propidium iodide) staining. Data were analyzed with Tree Star FlowJo software using cell cycle analysis Dean-Jett-Fox module. The in vivo anticancer activity estimation experiments were carried out on mice with inoculated ascitic Ehrlich’s carcinoma at intraperitoneal introduction of doxorubicin and its zinc oxide compositions. It was shown that the nanosize zinc oxide film deposition on the drug surface leads to the selective anticancer activity of composites at the cellular level with the range of selectivity index (SI) from 4 (Starch+NaCMC+Dox+ZnO) to 200 (PEO(gel)+Dox+ZnO) which is higher than that of free Dox (SI = 56). The significant increase in vivo antitumor activity (by a factor of 2-2.5) and decrease of general toxicity of zinc oxide compositions of doxorubicin in the form of the above mentioned gels compared to free doxorubicin were shown on the model of inoculated Ehrlich's ascitic carcinoma. Mechanistic studies of anticancer activity revealed the cytostatic effect based on the high level of DNA biosynthesis inhibition at considerable low concentrations of zinc oxide compositions of doxorubicin. The results of studies in vitro and in vivo behavior of PEO+Dox+ZnO and Starch+NaCMC+Dox+ZnO composites confirm the high potential of the nanosize zinc oxide composites as a vector delivery system for future application in cancer chemotherapy.

Keywords: anticancer activity, cancer specificity, doxorubicin, zinc oxide

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65 Gold Nanoparticle Conjugated with Andrographolide Ameliorates Viper Venom-Induced Inflammatory Response and Organ Toxicity in Animal Model

Authors: Sourav Ghosh, Antony Gomes

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Since 1894 anti-snake venom serum (ASVS) is the only available treatment against snake envenomation, although there are many side effects and limitations. The need for a supportive treatment was felt for a long time to overcome the side effects and limitations of ASVS. Andrographolide conjugated with gold nanoparticle (A-GNP) has been found to antagonize viper venom-induced local damages. The present study was aimed to study the protective efficacy of A-GNP against Viper venom-induced inflammatory response and organ toxicity in animal model. Ethical clearance was obtained from animal experiments. Physico-chemical characterization of A-GNP was done by DLS (diameter and zeta potential), FE-SEM and XRD. Swiss albino male mice were divided into 4 groups: Gr.1-Sham control, Gr.2- Russell’s Viper venom (RVV) control, Gr.3- andrographolide treated and Gr.4- A-GNP treated. The 1/5th minimum lethal dose of RVV (500µg/kg, s.c.) was induced in animals of group 2, 3 & 4 animals, followed by treatment with andrographolide (100mg/kg, i.p.) and A-GNP (100mg/kg, i.v.) in group 3 & 4 animals, respectively. Blood was collected after 18 h, serum was prepared, and inflammatory markers (IL 1β, 6, 17a, 10, TNF α) and biochemical markers (AST, ACP, LDH, urea, creatinine) were assessed. Values were expressed as mean±SEM (n=4), one way ANOVA was done, P<0.05 was considered as statistically significant. DLS size showed the hydrodynamic diameter of A-GNP to be 230-260nm with polydispersity index of 0.103 and zeta potential was -18.32mV. XRD data confirmed the presence of crystalline gold in A-GNP, and FESEM indicated the presence of nearly spherical particle with size18-24nm.Treatment with A-GNP significantly decreased viper venom-induced proinflammatory markers (IL 1β, 6, 17, TNF α) increased anti-inflammatory markers (IL 10) and decreased organ toxicity markers (AST, ACP, LDH, urea, creatinine) in animal model. Venom neutralization efficacy of A-GNP was > andrographolide, which confirmed the increased efficacy of andrographolide after gold nanoparticle conjugation. Venom neutralization by A-GNP was due to anti-oxidant/anti-inflammatory activity of andrographolide, which showed increased efficacy after gold nanoparticle tagging. Thus, A-GNP may serve as a supportive therapy in snake-bite (against inflammatory response and organ toxicity) subject to further detail studies.

Keywords: andrographolide, gold nanoparticle, inflammatory response, organ toxicity, snake venom, snake venom neutralization, viper venom

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64 Human Rabies Survivors in India: Epidemiological, Immunological and Virological Studies

Authors: Madhusudana S. N., Reeta Mani, Ashwini S. Satishchandra P., Netravati, Udhani V., Fiaz A., Karande S.

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Rabies is an acute encephalitis which is considered 100% fatal despite occasional reports of survivors. However, in recent times more cases of human rabies survivors are being reported. In the last 5 years, there are six laboratories confirmed human rabies survivors in India alone. All cases were children below 15 years and all contracted the disease by dog bites. All of them also had received the full or partial course of rabies vaccination and 4 out of 6 had also received rabies immunoglobulin. All cases were treated in intensive care units in hospitals at Bangalore, Mumbai, Chandigarh, Lucknow and Goa. We report here the results of immunological and virological studies conducted at our laboratory on these patients. The clinical samples that were obtained from these patients were Serum, CSF, nuchal skin biopsy and saliva. Serum and CSF samples were subjected to standard RFFIT for estimation of rabies neutralizing antibodies. Skin biopsy, CSF and saliva were processed by TaqMan real-time PCR for detection of viral RNA. CSF, saliva and skin homogenates were also processed for virus isolation by inoculation of suckling mice. The PBMCs isolated from fresh blood was subjected to ELISPOT assay to determine the type of immune response (Th1/Th2). Both CSF and serum were also investigated for selected cytokines by Luminex assay. The level of antibodies to virus G protein and N protein were determined by ELISA. All survivors had very high titers of RVNA in serum and CSF 100 fold higher than non-survivors and vaccine controls. A five-fold rise in titer could be demonstrated in 4 out of 6 patients. All survivors had a significant increase in antibodies to G protein in both CSF and serum when compared to non-survivors. There was a profound and robust Th1 response in all survivors indicating that interferon gamma could play an important factor in virus clearance. We could isolate viral RNA in only one patient four years after he had developed symptoms. The partial N gene sequencing revealed 99% homology to species I strain prevalent in India. Levels of selected cytokines in CSF and serum did not reveal any difference between survivors and non-survivors. To conclude, survival from rabies is mediated by virus-specific immune responses of the host and clearance of rabies virus from CNS may involve the participation of both Th2 and Th1 immune responses.

Keywords: rabies, rabies treatment, rabies survivors, immune reponse in rabies encephalitis

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63 Effect of Internet Addiction on Dietary Behavior and Lifestyle Characteristics among University Students

Authors: Hafsa Kamran, Asma Afreen, Zaheer Ahmed

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Internet addiction, an emerging mental health disorder from last two decades, is manifested by the inability in the controlled use of internet leading to academics, social, physiological and/or psychological difficulties. The present study aimed to assess the levels of internet addiction among university students in Lahore and to explore the effects of internet addiction on their dietary behavior and lifestyle. It was an analytical cross-sectional study. Data was collected from October to December 2016 from students of four universities selected through two-stage sampling method. The numbers of participants were 500 and 13 questionnaires were rejected due to incomplete information. Levels of Internet Addiction (IA) were calculated using Young Internet Addiction Test (YIAT). Data was also collected on students’ demographics, lifestyle factors and dietary behavior using self-reported questionnaire. Data was analyzed using SPSS (version 21). Chi-square test was applied to evaluate the relationship between variables. Results of the study revealed that 10% of the population had severe internet addiction while moderate Internet Addiction was present in 42%. High prevalence was found among males (11% vs. 8%), private sector university students (p = 0.008) and engineering students (p = 0.000). The lifestyle habits of internet addicts were significantly of poorer quality than normal users (p = 0.05). Internet addiction was found associated with lesser physically activity (p = 0.025), had shorter duration of physical activity (p = 0.016), had more disorganized sleep pattern (p = 0.023), had less duration of sleep (p = 0.019), reported being more tired and sleepy in class (p = 0.033) and spending more time on internet as compared to normal users. Severe and moderate internet addicts also found to be more overweight and obese than normal users (p = 0.000). The dietary behavior of internet addicts was significantly poorer than normal users. Internet addicts were found to skip breakfast more than a normal user (p = 0.039). Common reasons for meal skipping were lack of time and snacking between meals (p = 0.000). They also had increased meal size (p = 0.05) and habit of snacking while using the internet (p = 0.027). Fast food (p = 0.016) and fried items (p = 0.05) were most consumed snacks, while carbonated beverages (p = 0.019) were most consumed beverages among internet addicts. Internet Addicts were found to consume less than recommended daily servings of dairy (p = 0.008) and fruits (p = 0.000) and more servings of meat group (p = 0.025) than their no internet addict counterparts. In conclusion, in this study, it was demonstrated that internet addicts have unhealthy dietary behavior and inappropriate lifestyle habits. University students should be educated regarding the importance of balanced diet and healthy lifestyle, which are critical for effectual primary prevention of numerous chronic degenerative diseases. Furthermore, it is necessary to raise awareness concerning adverse effects of internet addiction among youth and their parents.

Keywords: dietary behavior, internet addiction, lifestyle, university students

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62 Evaluation of Mito-Uncoupler Induced Hyper Metabolic and Aggressive Phenotype in Glioma Cells

Authors: Yogesh Rai, Saurabh Singh, Sanjay Pandey, Dhananjay K. Sah, B. G. Roy, B. S. Dwarakanath, Anant N. Bhatt

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One of the most common signatures of highly malignant gliomas is their capacity to metabolize more glucose to lactic acid than normal brain tissues, even under normoxic conditions (Warburg effect), indicating that aerobic glycolysis is constitutively upregulated through stable genetic or epigenetic changes. However, oxidative phosphorylation (OxPhos) is also required to maintain the mitochondrial membrane potential for tumor cell survival. In the process of tumorigenesis, tumor cells during fastest growth rate exhibit both high glycolytic and high OxPhos. Therefore, metabolically reprogrammed cancer cells with combination of both aerobic glycolysis and altered OxPhos develop a robust metabolic phenotype, which confers a selective growth advantage. In our study, we grew the high glycolytic BMG-1 (glioma) cells with continuous exposure of mitochondrial uncoupler 2, 4, dinitro phenol (DNP) for 10 passages to obtain a phenotype of high glycolysis with enhanced altered OxPhos. We found that OxPhos modified BMG (OPMBMG) cells has similar growth rate and cell cycle distribution but high mitochondrial mass and functional enzymatic activity than parental cells. In in-vitro studies, OPMBMG cells showed enhanced invasion, proliferation and migration properties. Moreover, it also showed enhanced angiogenesis in matrigel plug assay. Xenografted tumors from OPMBMG cells showed reduced latent period, faster growth rate and nearly five folds reduction in the tumor take in nude mice compared to BMG-1 cells, suggesting that robust metabolic phenotype facilitates tumor formation and growth. OPMBMG cells which were found radio-resistant, showed enhanced radio-sensitization by 2-DG as compared to the parental BMG-1 cells. This study suggests that metabolic reprogramming in cancer cells enhances the potential of migration, invasion and proliferation. It also strengthens the cancer cells to escape the death processes, conferring resistance to therapeutic modalities. Our data also suggest that combining metabolic inhibitors like 2-DG with conventional therapeutic modalities can sensitize such metabolically aggressive cancer cells more than the therapies alone.

Keywords: 2-DG, BMG, DNP, OPM-BMG

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61 The Coaching on Lifestyle Intervention (CooL): Preliminary Results and Implementation Process

Authors: Celeste E. van Rinsum, Sanne M. P. L. Gerards, Geert M. Rutten, Ien A. M. van de Goor, Stef P. J. Kremers

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Combined lifestyle interventions have shown to be effective in changing and maintaining behavioral lifestyle changes and reducing overweight and obesity. A lifestyle coach is expected to promote lifestyle changes in adults related to physical activity and diet. The present Coaching on Lifestyle (CooL) study examined participants’ physical activity level, dietary behavioral, and motivational changes immediately after the intervention and at 1.5 years after baseline. In CooL intervention a lifestyle coach coaches individuals from eighteen years and older with (a high risk of) obesity in group and individual sessions. In addition a process evaluation was conducted in order to examine the implementation process and to be able to interpret the changes within the participants. This action-oriented research has a pre-post design. Participants of the CooL intervention (N = 200) completed three questionnaires: at baseline, immediately after the intervention (on average after 44 weeks), and at 1.5 years after baseline. T-tests and linear regressions were conducted to test self-reported changes in physical activity (IPAQ), dietary behaviors, their quality of motivation for physical activity (BREQ-3) and for diet (REBS), body mass index (BMI), and quality of life (EQ-5D-3L). For the process evaluation, we used individual and group interviews, observations and document analyses to gain insight in the implementation process (e.g. the recruitment) and how the intervention was valued by the participants, lifestyle coaches, and referrers. The study is currently ongoing and therefore the results presented here are preliminary. On average, the participants that finished the intervention and those that have completed the long-term measurement improved their level of vigorous-intense physical activity, sedentary behavior, sugar-sweetened beverage consumption and BMI. Mixed results were observed in motivational regulation for physical activity and nutrition. Moreover, an improvement on the quality of life dimension anxiety/depression was found, also in the long-term. All the other constructs did not show significant change over time. The results of the process evaluation have shown that recruitment of clients was difficult. Participants evaluated the intervention positively and the lifestyle coaches have continuously adapted the structure and contents of the intervention throughout the study period, based on their experiences and feedback from research. Preliminary results indicate that the CooL-intervention may have beneficial effects on overweight and obese participants in terms of energy balance-related behaviors, weight reduction, and quality of life. Recruitment of participants and embedding the position of the lifestyle coach in traditional care structures is challenging.

Keywords: combined lifestyle intervention, effect evaluation, lifestyle coaching, process evaluation, overweight, the Netherlands

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60 Cobalamin, Folate and Metabolic Syndrome Parameters in Pediatric Morbid Obesity and Metabolic Syndrome

Authors: Mustafa M. Donma, Orkide Donma

Abstract:

Obesity is known to be associated with many clinically important diseases including metabolic syndrome (MetS). Vitamin B12 plays essential roles in fat and protein metabolisms and its cooperation with vitamin B9 is well-known. The aim of this study is to investigate the possible contributions as well as associations of these micronutrients upon obesity and MetS during childhood. A total of 128 children admitted to Namik Kemal University, Medical Faculty, Department of Pediatrics Outpatient Clinics were included into the scope of this study. The mean age±SEM of 92 morbid obese (MO) children and 36 with MetS were 118.3±3.8 months and 129.5±6.4 months, respectively (p > 0.05). The study was approved by Namık Kemal University, Medical Faculty Ethics Committee. Written informed consent forms were obtained from the parents. Demographic features and anthropometric measurements were recorded. WHO BMI-for age percentiles were used. The values above 99 percentile were defined as MO. Components of MetS [waist circumference (WC), fasting blood glucose (FBG), triacylglycerol (TRG), high density lipoprotein cholesterol (HDL-Chol), systolic pressure (SP), diastolic pressure (DP)] were determined. Routine laboratory tests were performed. Serum vitamin B12 concentrations were measured using electrochemiluminescence immunoassay. Vitamin B9 was analyzed by an immunoassay analyzer. Values for vitamin B12 < 148 pmol/L, 148-221 pmol/L, > 221 pmol/L were accepted as low, borderline and normal, respectively. Vitamin B9 levels ≤ 4 mcg/L defined deficiency state. Statistical evaluations were performed by SPSSx Version 16.0. p≤0.05 was accepted as statistical significance level. Statistically higher body mass index (BMI), WC, hip circumference (C) and neck C were calculated in MetS group compared to children with MO. No difference was noted for head C. All MetS components differed between the groups (SP, DP p < 0.001; WC, FBG, TRG p < 0.01; HDL-Chol p < 0.05). Significantly decreased vitamin B9 and vitamin B12 levels were detected (p < 0.05) in children with MetS. In both groups percentage of folate deficiency was 5.5%. No cases were below < 148 pmol/L. However, in MO group 14.3% and in MetS group 22.2% of the cases were of borderline status. In MO group B12 levels were negatively correlated with BMI, WC, hip C and head C, but not with neck C. WC, hip C, head C and neck C were all negatively correlated with HDL-Chol. None of these correlations were observed in the group of children with MetS. Strong positive correlation between FBG and insulin as well as strong negative correlation between TRG and HDL-Chol detected in MO children were lost in MetS group. Deficiency state end-products of both B9 and B12 may interfere with the expected profiles of MetS components. In this study, the alterations in MetS components affected vitamin B12 metabolism and also its associations with anthropometric body measurements. Further increases in vitamin B12 and vitamin B9 deficiency in MetS associated with the increased vitamin B12 as well as vitamin B9 deficiency metabolites may add to MetS parameters.

Keywords: children, cobalamin, folate, metabolic syndrome, obesity

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59 Effect of a Synthetic Platinum-Based Complex on Autophagy Induction in Leydig TM3 Cells

Authors: Ezzati Givi M., Hoveizi E., Nezhad Marani N.

Abstract:

Platinum-based anticancer therapeutics are the most widely used drugs in clinical chemotherapy but have major limitations and various side effects in clinical applications. Gonadotoxicity and sterility is one of the most common complications for cancer survivors, which seem to be drug-specific and dose-related. Therefore, many efforts have been dedicated to discovering a new structure of platinum-based anticancer agents with improved therapeutic index, fewer side effects. In this regard, new Pt(II)-phosphane complexes containing heterocyclic thionate ligands (PCTL) have been synthesized, which show more potent antitumor activities in comparison to cisplatin. Cisplatin, the best leading metal-based antitumor drug in the field, induces testicular toxicity on Leydig and Sertoli cells leading to serious side effects such as azoospermia and infertility. Therefore in the present study, we aimed to investigate the cytotoxicity effect of PCTL on mice TM4 Sertoli cells with particular emphasis on the role of autophagy in comparison to cisplatin. In this study, an MTT assay was performed to evaluate the IC50 of PCTL and to analyze the TM3 Leydig cell's viability. Cells morphology was evaluated via invert microscope and Changing in morphology for nuclei swelling or autophagic vacuoles formation were assessed by DAPI and MDC staining. Testosterone production in the culture medium was measured using an ELISA kit. Finally, the expression of Autophagy-related genes, Atg5, Beclin1 and p62, were analyzed by qPCR. Based on the obtained results by MTT, the IC50 value of PCTL was 50 μM in TM3 cells and cytotoxic effects was in a dose- and time-dependent manner. Cells morphological changes investigated by inverted microscopy, DAPI, and MDC staining which showed the cytotoxic concentrations of PCTL was significantly higher than cisplatin in the treated TM3 Leydig cells. The results of PCR showed a lack of expression of the p62, Atg5 and Beclin1 gene in TM3 cells treated with PCTL in comparison to cisplatin and control groups. It should be noted that the effects of 25 μM PCTL concentration on TM3 cells have been associated with increased testosterone production and secretion, which requires further study to explain the possible causes and involved molecular mechanisms. The results of the study showed that the PCTL had less-lethal effects on TM3 cells in comparison to cisplatin and probably did not induce autophagy in TM3 cells.

Keywords: platinum-based anticancer agents, cisplatin, Leydig TM3 cells, autophagy

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58 The Valuable Triad of Adipokine Indices to Differentiate Pediatric Obesity from Metabolic Syndrome: Chemerin, Progranulin, Vaspin

Authors: Mustafa M. Donma, Orkide Donma

Abstract:

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

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

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