Search results for: hypocaloric jelly
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 28

Search results for: hypocaloric jelly

28 Development of Low Calorie Jelly with Increased Content of Natural Compounds from Superfoods with No Added Sugar

Authors: Liana C. Salanță, Maria Tofană, Carmen R. Pop, Vlad Mureșan

Abstract:

The landscape of functional food is expanding very fast, due to the consumer interest for healthy natural products. Consumers nowadays demand healthy products that impart phytonutrients to encourage good health and well-being, prevent diseases, without sacrificing taste and texture. Candies are foodstuffs appreciated by all category of consumers. They are available in a range variety of forms (jellies, marshmallows, caramels, lollipops, etc.). Jelly is characterized by a gummy and chewy texture typically conferred by a hydrocolloid (gelatin, pectin). The purpose of this research was to obtain hypocaloric jelly (no added sugar) enriched with protein powder from acai, chia seeds and hemp, which are considered superfood. Peach and raspberry juice were used for obtaining functional jelly, due to the specific flavour, natural carbohydrate, natural pigments and vitamins (C, B1, PP, etc). Instead of classic hydrocolloids used in Romania for the industry of jelly, agar-agar was used in this study, due to its properties. Agar-agar is able to form gels in the aqueous medium, stronger than other gel-forming agents. High sugar concentrations or an acid environment (as is necessary with pectins) are not needed. In addition to its gelation properties, Agar-agar is considered to have important nutritional benefits, high content of fibre and has low calories. Six prototypes of jellies were obtained and evaluated by physicochemical, microbiological and sensorial analysis. For the textural profile analysis, the Brookfield CT3 Texture Analyzer, equipped with a 10kg load cell, was used. The results revealed that hypocaloric jelly can serve as a good source of bioactive compounds in the diet. The jelly is a convenient way of delivering potential health benefits of protein powder and agar-agar to a wide range of consumers.

Keywords: agar-agar, functional food, hypocaloric jelly, superfoods

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27 Analysis of Efficiency Production of Grass Black Jelly (Mesona palustris) in Double Scale

Authors: Irvan Adhin Cholilie, Susinggih Wijana, Yusron Sugiarto

Abstract:

The aim of this research is to compare the results of black grass jelly produced using laboratory scale and double scale. In this research, the production from the laboratory scale is using ingredients of 1 kg black grass jelly added with 5 liters of water, while the double scale is using 5 kg black grass jelly and 75 liters of water. The results of organoleptic tests performed by 30 panelists (general) to the sample gels of grass black powder produced from both of laboratory and double scale are not different significantly in color, odor, flavor, and texture. Proximate test results conducted in both of grass black jelly powder produced in laboratory scale and double scale also have no significant differences in all parameters. Grass black jelly powder from double scale contains water, carbohydrate, crude fiber, and yield in the amount of 12,25 %; 43,7 %; 5,89 %; and 16,28 % respectively. The results of the energy efficiency analysis by boiling, draining, evaporation, drying, and milling processes are 85,11 %; 76,97 %; 99,64 %; 99,99% and 99,39% respectively. The utility needs including water needs for each batch amounted 0.1 m3 and cost Rp 220,5 per batch, the electricity needs for each batch is 20.01 kWh and cost Rp 18569.28 per batch, and LPG needs for each batch is 30 kg costed Rp 234,000.00 so that the total cost spent for the process is Rp 252,789.78 .

Keywords: black grass jelly, powder, mass balance, energy balance, cost

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26 Optimization of Black Grass Jelly Formulation to Reduce Leaching and Increase Floating Rate

Authors: M. M. Nor, H. I. Sheikh, M. F. H. Hassan, S. Mokhtar, A. Suganthi, A. Fadhlina

Abstract:

Black grass jelly (BGJ) is a popular black jelly used in preparing various drinks and desserts. Food industries often use preservatives to maintain the physicochemical properties of foods, such as color and texture. These preservatives (e.g., phosphoric acid) are linked with deleterious health effects such as kidney disease. Using gelling agents, carrageenan, and gelatin to make BGJ could improve its physiochemical and textural properties. This study was designed to optimize BGJ-selected physicochemical and textural properties using carrageenan and gelatin. Various black grass jelly formulations (BGJF) were designed using an I-optimal mixture design in Design Expert® software. Data from commercial BGJ were used as a reference during the optimization process. The combination of carrageenan and gelatin added to the formulations was up to 14.38g (~5%), respectively. The results showed that adding 2.5g carrageenan and 2.5g gelatin at approximately 5g (~5%) effectively maintained most of the physiochemical properties with an overall desirability function of 0.81. This formulation was selected as the optimum black grass jelly formulation (OBGJF). The leaching properties and floating duration were measured on the OBGJF and commercial grass jelly for 20 min and 40 min, respectively. The results indicated that OBGJF showed significantly (p<0.0001) lower leaching rate and floating time (p<0.05). Hence, further optimization is needed to increase the floating duration of carrageenan and gelatin-based BGJ.

Keywords: cincau, Mesona chinensis, black grass jelly, carrageenan, gelatin

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25 Body Composition Response to Lower Body Positive Pressure Training in Obese Children

Authors: Basant H. El-Refay, Nabeel T. Faiad

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Background: The high prevalence of obesity in Egypt has a great impact on the health care system, economic and social situation. Evidence suggests that even a moderate amount of weight loss can be useful. Aim of the study: To analyze the effects of lower body positive pressure supported treadmill training, conducted with hypocaloric diet, on body composition of obese children. Methods: Thirty children aged between 8 and 14 years, were randomly assigned into two groups: intervention group (15 children) and control group (15 children). All of them were evaluated using body composition analysis through bioelectric impedance. The following parameters were measured before and after the intervention: body mass, body fat mass, muscle mass, body mass index (BMI), percentage of body fat and basal metabolic rate (BMR). The study group exercised with antigravity treadmill three times a week during 2 months, and participated in a hypocaloric diet program. The control group participated in a hypocaloric diet program only. Results: Both groups showed significant reduction in body mass, body fat mass and BMI. Only study group showed significant reduction in percentage of body fat (p = 0.0.043). Changes in muscle mass and BMR didn't reach statistical significance in both groups. No significant differences were observed between groups except for muscle mass (p = 0.049) and BMR (p = 0.042) favoring study group. Conclusion: Both programs proved effective in the reduction of obesity indicators, but lower body positive pressure supported treadmill training was more effective in improving muscle mass and BMR.

Keywords: children, hypocaloric diet, lower body positive pressure supported treadmill, obesity

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24 Development and Validation of HPLC Method on Determination of Acesulfame-K in Jelly Drink Product

Authors: Candra Irawan, David Yudianto, Ahsanu Nadiyya, Dewi Anna Br Sitepu, Hanafi, Erna Styani

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Jelly drink was produced from a combination of both natural and synthetic materials, such as acesulfame potassium (acesulfame-K) as synthetic sweetener material. Acesulfame-K content in jelly drink could be determined by High-Performance Liquid Chromatography (HPLC), but this method needed validation due to having a change on the reagent addition step which skips the carrez addition and comparison of mix mobile phase (potassium dihydrogen phosphate and acetonitrile) with ratio from 75:25 to 90:10 to be more efficient and cheap. This study was conducted to evaluate the performance of determination method for acesulfame-K content in the jelly drink by HPLC. The method referred to Deutsches Institut fur Normung European Standard International Organization for Standardization (DIN EN ISO):12856 (1999) about Foodstuffs, Determination of acesulfame-K, aspartame and saccharin. The result of the correlation coefficient value (r) on the linearity test was 0.9987 at concentration range 5-100 mg/L. Detection limit value was 0.9153 ppm, while the quantitation limit value was 1.1932 ppm. The recovery (%) value on accuracy test for sample concentration by spiking 100 mg/L was 102-105%. Relative Standard Deviation (RSD) value for precision and homogenization tests were 2.815% and 4.978%, respectively. Meanwhile, the comparative and stability tests were tstat (0.136) < ttable (2.101) and |µ1-µ2| (1.502) ≤ 0.3×CV Horwitz. Obstinacy test value was tstat < ttable. It can be concluded that the HPLC  method for the determination of acesulfame-K in jelly drink product by HPLC has been valid and can be used for analysis with good performance.

Keywords: acesulfame-K, jelly drink, HPLC, validation

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23 Effect of Varying Diets on Growth, Development and Survival of Queen Bee (Apis mellifera L.) in Captivity

Authors: Muhammad Anjum Aqueel, Zaighum Abbas, Mubasshir Sohail, Muhammad Abubakar, Hafiz Khurram Shurjeel, Abu Bakar Muhammad Raza, Muhammad Afzal, Sami Ullah

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Keeping in view the increasing demand, queen of Apis mellifera L. (Hymenoptera: Apidae) was reared artificially in this experiment at varying diets including royal jelly. Larval duration, pupal duration, weight, and size of pupae were evaluated at different diets including royal jelly. Queen larvae were raised by Doo Little grafting method. Four different diets were mixed with royal jelly and applied to larvae. Fructose, sugar, yeast, and honey were provided to rearing queen larvae along with same amount of royal jelly. Larval and pupal duration were longest (6.15 and 7.5 days, respectively) at yeast and shortest on honey (5.05 and 7.02 days, respectively). Heavier and bigger pupae were recorded on yeast (168.14 mg and 1.76 cm, respectively) followed by diets having sugar and honey. Due to production of heavier and bigger pupae, yeast was considered as best artificial diet for the growing queen larvae. So, in the second part of experiment, different amounts of yeast were provided to growing larvae along with fixed amount (0.5 g) of royal jelly. Survival rates of the larvae and queen bee were 70% and 40% in the 4-g food, 86.7% and 53.3% in the 6-g food, and 76.7% and 50% in the 8-g food. Weight of adult queen bee (1.459±0.191 g) and the number of ovarioles (41.7±21.3) were highest at 8 g of food. Results of this study are helpful for bee-keepers in producing fitter queen bees.

Keywords: apis melifera l, dietary effect, survival and development, honey bee queen

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22 Wound Healing Potential and Comparison of Mummy Substance Effect on Adipose and Wharton’s Jelly-Derived Mesenchymal Stem Cells Co-Cultured with Human Fibroblast

Authors: Sepideh Hassanpour Khodaei

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Background/Objectives: The purpose of this study is to evaluate the effect of mummy substances on two issues of proliferation and production of matrix protein synthesis in wound healing. Methods: The methodology used for this aim involves isolating mesenchymal stem cells and human fibroblasts procured at Pastor Institute, Iran. The cells were treated with mummy substances separately and co-cultured between ASCs and WJSCs, and fibroblasts. Proliferation was assessed by Ki67 method in monolayer conditions. Synthesis of components of extracellular matrix (ECM) such as collagen type I, type III, and fibronectin 1 (FN1) was determined by qPCR. Results: The effects of adipocyte stem cells (ASCs), Wharton Jelly Stem Cells (WJSCs), and Mummy material on fibroblast proliferation and migration were evaluated. The present finding underlined the importance of Mummy material, ASCs, and WJSCs in the proliferation and migration of fibroblast cells. Furthermore, the expression of collagen I, III, and FN1 was increased in the presence of the above material and cells. Conclusion: This study presented an effective in vitro method for the healing process. Hence, the prospect of utilizing Mummy material and stem cell-based therapies in wound healing as a therapeutic approach is promising.

Keywords: mummy material, wound healing, adipose tissue, Wharton’s jelly

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21 Survey of Potato Viral Infection Using Das-Elisa Method in Georgia

Authors: Maia Kukhaleishvili, Ekaterine Bulauri, Iveta Megrelishvili, Tamar Shamatava, Tamar Chipashvili

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Plant viruses can cause loss of yield and quality in a lot of important crops. Symptoms of pathogens are variable depending on the cultivars and virus strain. Selection of resistant potato varieties would reduce the risk of virus transmission and significant economic impact. Other way to avoid reduced harvest yields is regular potato seed production sampling and testing for viral infection. The aim of this study was to determine the occurrence and distribution of viral diseases according potato cultivars for further selection of virus-free material in Georgia. During the summer 2015- 2016, 5 potato cultivars (Sante, Laura, Jelly, Red Sonia, Anushka) at 5 different farms located in Akhalkalaki were tested for 6 different potato viruses: Potato virus A (PVA), Potato virus M (PVM), Potato virus S (PVS), Potato virus X (PVX), Potato virus Y (PVY) and potato leaf roll virus (PLRV). A serological method, Double Antibody Sandwich-Enzyme linked Immunosorbent Assay (DASELISA) was used at the laboratory to analyze the results. The result showed that PVY (21.4%) and PLRV (19.7%) virus presence in collected samples was relatively high compared to others. Researched potato cultivars except Jelly and Laura were infected by PVY with different concentrations. PLRV was found only in three potato cultivars (Sante, Jelly, Red Sonia) and PVM virus (3.12%) was characterized with low prevalence. PVX, PVA and PVS virus infection was not reported. It would be noted that 7.9% of samples were containing PVY/PLRV mix infection. Based on the results it can be concluded that PVY and PLRV infections are dominant in all research cultivars. Therefore significant yield losses are expected. Systematic, long-term control of potato viral infection, especially seed-potatoes, must be regarded as the most important factor to increase seed productivity.

Keywords: virus, potato, infection, diseases

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20 Effects of in Ovo Injection of Royal Jelly on Hatchability, One-Day Old Chickens Quality, Total Antioxidant Status and Blood Lipoproteins

Authors: Amin Adeli, Maryam Zarei

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Background and purpose: Royal jelly (RJ) is a natural product with anti-hyperlipidemic and antioxidant properties. In ovo administration of RJ may improve lipid profile and antioxidant properties. This study was conducted to evaluate, for first time, the effects of in ovo injection of the RJ on hatchability, one-day old chick quality, total antioxidant status and blood lipoproteins. Methods: 400 incubating eggs produced by Ross 308 strain (52 weeks of age in first stage of production) were prepared and assigned into 4 groups (n=100) and 4 replications per group (n=25). These 4 groups were injected by the following pattern: 1) 0.1 ml normal saline (control), 2) 0.1 mg RJ+0.1 ml normal saline, 3) 0.2 mg RJ+0.1 ml normal saline, and 4) 0.3 mg RJ+0.1 ml normal saline. Injections were performed using a laminar flow system Lipid profile, antioxidant properties, hatchability, and one-day old chicken quality were assessed. Results: The administration of RJ at concentration of 0.1increased the percentage of hatchability compared to concentration of 0.2 and control, significant differences have not been observed among groups for quality scores (P>0.05). The results showed that in ovo injection of the RJ did not have any significant effects on lipid profile; but administration of the RJ only decreased High-density lipoprotein (HDL cholesterol, HDL-C) (P<0.05). The results showed that injection of the RJ at concentration of 0.3 increased total antioxidant capacity (TAC) compared to control group (p<0.05). Injection of the RJ progressively increased gluthation peroxidase (GPx) activity (p<0.05). The results showed that injection of the RJ decreased superoxide dismutase (SOD) compared to control group (p<0.05). Conclusion: In ovo injection of the RJ at the highest concentration increased TAC and GPx, but it did not have significant effects on lipid profile. Future studies are needed to investigate the effects of the RJ on the above-mentioned mechanisms.

Keywords: antioxidant enzymes, chicken quality, hatchability, royal jelly

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19 Wharton's Jelly-Derived Mesenchymal Stem Cells Modulate Heart Rate Variability and Improve Baroreflex Sensitivity in Septic Rats

Authors: Cóndor C. José, Rodrigues E. Camila, Noronha L. Irene, Dos Santos Fernando, Irigoyen M. Claudia, Andrade Lúcia

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Sepsis induces alterations in hemodynamics and autonomic nervous system (ASN). The autonomic activity can be calculated by measuring heart rate variability (HRV) that represents the complex interplay between ASN and cardiac pacemaker cells. Wharton’s jelly mesenchymal stem cells (WJ-MSCs) are known to express genes and secreted factors involved in neuroprotective and immunological effects, also to improve the survival in experimental septic animals. We hypothesized, that WJ-MSCs present an important role in the autonomic activity and in the hemodynamic effects in a cecal ligation and puncture (CLP) model of sepsis. Methods: We used flow cytometry to evaluate WJ-MSCs phenotypes. We divided Wistar rats into groups: sham (shamoperated); CLP; and CLP+MSC (106 WJ-MSCs, i.p., 6 h after CLP). At 24 h post-CLP, we recorded the systolic arterial pressure (SAP) and heart rate (HR) over 20 min. The spectral analysis of HR and SAP; also the spontaneous baroreflex sensitivity (measure by bradycardic and tachycardic responses) were evaluated after recording. The one-way ANOVA and the post hoc Student– Newman– Keuls tests (P< 0.05) were used to data comparison Results: WJ-MSCs were negative for CD3, CD34, CD45 and HLA-DR, whereas they were positive for CD73, CD90 and CD105. The CLP group showed a reduction in variance of overall variability and in high-frequency power of HR (heart parasympathetic activity); furthermore, there is a low-frequency reduction of SAP (blood vessels sympathetic activity). The treatment with WJ-MSCs improved the autonomic activity by increasing the high and lowfrequency power; and restore the baroreflex sensitive. Conclusions: WJ-MSCs attenuate the impairment of autonomic control of the heart and vessels and might therefore play a protective role in sepsis. (Supported by FAPESP).

Keywords: baroreflex response, heart rate variability, sepsis, wharton’s jelly-derived mesenchymal stem cells

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18 Stereological Evaluation of Liver of Rabbit Fetuses After Transplantation of Human Wharton’s Jelly-Derived Mesenchymal Stromal/Stem Cells

Authors: Zahra Khodabandeh, Leila Rezaeian, Mohammad Amin Edalatmanesh, Asghar Mogheiseh, Nader Tanideh, Mehdi Dianatpour, Shahrokh Zare, Hossein Bordbar, Neda Baghban, Amin Tamadon

Abstract:

Background: In-utero xenotransplantation of stem cells in abnormal fetuses effectively treats several genetic illnesses. Objective: The current research aimed to evaluate structural and morphological alterations in the liver of rabbit fetuses following xenotransplantation of human Wharton’s jelly-derived mesenchymal stromal cells (hWJ-MSCs) using a stereological technique. Methods: hWJ-MSCs were isolated from the human umbilical cord, and their authenticity was established by flow cytometry and differentiation. At gestational day 14, the rabbits were anesthetized, and hWJ-MSCs were injected into the uteri of 24 fetuses. Twenty-two fetuses were born successfully. Ten rabbit liver specimens were prepared from injected fetuses, including eight rabbits on day three following birth and two rabbits on the 21st post-natal day. The non-injected fetuses were considered positive controls. The livers of the control and hWJ-MSCs-treated rabbits were fixed, processed, stained, and examined through stereological approaches. Results: In the hWJ-MSCs-treated group, the mean liver weight and volume increased by 42% and 78% compared to the control group. The total volume of the hepatocytes increased by 63% and that of sinusoids by threefold in the treated rabbits. The total volume of the central veins increased by 70%. The total number corresponding to hepatocytes in the experimental group increased by 112% compared to the rabbits in the control. The total volume of the hepatocyte nuclei in the experimental group increased by 117% compared to the rabbits in the control. Conclusion: After xenotransplantation of human MSCs, host tissue microenvironments (here, the rabbit liver) were altered, and these included quantitative factors corresponding to the liver tissue and hepatocyte morphometric indices.

Keywords: xenotransplantation, mesenchymal stromal, stem cell, Wharton ‘s jelly, liver

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17 Role of Human Wharton’s Jelly Mesenchymal Stem Cells Conditioned Media in Alleviating Kidney Injury via Inhibition of Renin-Angiotensin System in Diabetic Nephropathy

Authors: Pardis Abolghasemi, Benyamin Hatamsaz

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Background: Diabetic nephropathy is a serious health problem described by specific kidney structure and functional disturbance. Renoprotective effects of the stem cells secretase have been shown in many kidney diseases. The aim is to evaluate the capability of human Wharton’s jelly mesenchymal stem cells conditioned media (hWJMSCs-CM) to alleviate DN in streptozotocin (STZ)-induced diabetes. Methods: Diabetic nephropathy was induced by injection of STZ (60 mg/kg, IP) in twenty rats. Conditioned media was extracted from hWJMSCs at third passages. At week 8, diabetic rats were divided into two groups: treated (hWJMSCs-CM, 500 μl/rat for three weeks, IP) and not treated (DN). In the 11th week, three groups (control, DN and DN+hWJMSCs-CM) were kept in metabolic cages and urine was collected for 24h. Blood pressure (BP) and heart rate (HR) were continuously recorded. The serum samples were maintained for measuring BUN, Cr and angiotensin-converting enzyme (ACE) activity. The left kidney was kept at -80°C for ACE activity assessment. The right kidney and pancreas were used for histopathologic evaluation. Result: Diabetic nephropathy was detected by microalbuminuria and increased albumin/creatinine ratio, as well as the pancreas and renal structural disturbance. Glomerular filtration rate, BP and HR increased in the DN group. The ACE activity was elevated in the serum and kidneys of the DN group. Administration of hWJMSCs-CM modulated the renal functional and structural disturbance and decreased the ACE activity. Conclusion: Conditioned media was extracted from hWJMSCs may have a Renoprotective effect in diabetic nephropathy. This may happen through regulation of ACE activity and renin-angiotensin system inhibition.

Keywords: diabetic nephropathy, mesenchymal stem cells, immunomodulation, anti-inflammation

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16 Role of Honey Bees in Our Ecosystem

Authors: Akhter Hussain Najar

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Honey bees are the best insect grown on earth and are taking the lead in the global ecosystem. Life on earth depends upon honey bees pollinating each and every flower in its reign. The global economy is balanced by providing financial attribute due to the use of valuable resources generated from honey bees like honey, royal jelly, bee venom, propolis, bee wax even queens from parent colonies is now in demand. Livelihood stability is also maintained by the rearing of honey bees; beekeeping became a new professional work to be cared for by the experts. Indigenous methodologies are used to rear honey bees. The rearing of honey bees showed variation in different states like Uttar Pradesh, Rajasthan, Haryana, Punjab, Jammu, Kashmir, etc. The production of honey depends upon the area and the availability of crops and the colonies of beekeepers in these states. However, the kind of honey from Jammu and Kashmir, like Accacia and Solai, has different color and odour from the honey generated in the rest of the states. However, the nature and viscosity vary from state to state. But in the last few years, the honey from Jammu and Kashmir has given different shapes when kept at room temperature due to unconditional rainfall during honey peak season in J&K.

Keywords: ecosystem pollen, pollination, honey bee

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15 Effect of 8 Weeks of Intervention on Physical Fitness, Hepatokines, and Insulin Resistance in Obese Subjects

Authors: Adela Penesova, Zofia Radikova, Boris Bajer, Andrea Havranova, Miroslav Vlcek

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Background: The aim of our study was to compare the effect of intensified lifestyle intervention on insulin resistance (HOMA-IR), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and Fibroblast growth factor (FGF) 21 after 8 weeks of lifestyle intervention. Methods: A group of 43 obese patients (13M/30F; 43.0±12.4 years; BMI (body mass index) 31.2±6.3 kg/m2 participated in a weight loss interventional program (NCT02325804) following an 8-week hypocaloric diet (-30% energy expenditure) and physical activity 150 minutes/week. Insulin sensitivity was evaluated according to the homeostasis model assessment of insulin resistance (HOMA-IR) and insulin sensitivity indices according to Matsuda and Cederholm were calculated (ISImat and ISIced). Plasma ALT, AST, Fetuin-A, FGF 21, and physical fitness were measured. Results: The average reduction of body weight was 6.8±4.9 kg (0-15 kg; p=0.0006), accompanied with a significant reduction of body fat amount of fat mass (p=0.03), and waist circumference (p=0.02). Insulin sensitivity has been improved (IR HOMA 2.71±3.90 vs 1.24±0.83; p=0.01; ISIMat 6.64±4.38 vs 8.93±5.36 p ≤ 0.001). Total, LDL cholesterol, and triglycerides decreased (p=0.05, p=0.04, p=0.04, respectively). Physical fitness significantly improved after intervention (as measure VO2 max (maximal oxygen uptake) (p ≤ 0.001). ALT decreased significantly (0.44±0.26 vs post 0.33±0.18 ukat/l, p=0.004); however, AST not (pre 0.40±0.15 vs 0.35±0.09 ukat/l, p=0.07). Hepatokine Fetuin-A significantly decreased after intervention (43.1±10.8 vs 32.6±8.6 ng/ml, p < 0.001); however, FGF 21 levels tended to decrease (146±152 vs 132±164 pg/ml, p=0.07). Conclusion: 8-weeks of diet and physical activity intervention program in obese otherwise healthy subjects led to an improvement of insulin resistance parameters and liver marker profiles, as well as increased physical fitness. This study was supported by grants APVV 15-0228; VEGA 2/0161/16.

Keywords: obesity, diet, exercice, insulin sensitivity

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14 Up-Regulation of SCUBE2 Expression in Co-Cultures of Human Mesenchymal Stem Cell and Breast Cancer Cells

Authors: Hirowati Ali, Aisyah Ellyanti, Dewi Rusnita, Septelia Inawati Wanandi

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Stem cell has been known for its potency to be differentiated in many cells. Recently stem cell has been used for many treatment of degenerative medicine. It is still controversy whether stem cell can be used for therapy or these cells can activate cancer stem cell. SCUBE2 is a novel secreted and membrane-anchored protein which has been reported to its role in better prognosis and inhibition of cancer cell proliferation. Our study aims to observe whether stem cell can up-regulate SCUBE2 gene in MCF7 breast cancer cell line. We used in vitro study using MCF-7 cell treated with stem cell derived from placenta Wharton's jelly which has been known for its stemness and widely used. Our results showed that MCF-7 cell line grows up rapidly in 6-well culture dish. Stem cell was cultured in 6-well dish. After 50%-60% MCF-7 confluence, we co-cultured these cells with stem cells for 24 hours and 48 hours. We hypothesize SCUBE2 gene which is previously known for its higher expression in better prognosis of breast cancer, is up-regulated after stem cells addition in MCF7 culture dishes.

Keywords: breast cancer cells, inhibition of cancer cells, mesenchymal stem cells, SCUBE2

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13 Rejuvenation of Premature Ovarian Failure with Stem Cells/IVA Technique

Authors: Elham Vojoudi, Marzieh Mehrafza, Ahmad Hosseini, Azadeh Raofi, Maryam Najafi

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Premature ovarian failure (POF) has become one of the main causes of infertility in women of childbearing age and the incidence of this disorder is increasing year by year. In these patients, poor ovarian response (POR) to gonadotropins reflects a diminished ovarian reserve (DOR) that gives place to few follicles despite aggressive stimulation. Up to now, egg donation is the only way to resolve infertility problems in POF patients. Therefore, some novel aspects such as activating (Akt signaling pathway) and inhibiting (Hippo-signaling) elements have been identified as IVA procedure that promotes primordial follicle activation. In this study, we used the newly developed technique (combination of in vitro activation of dormant follicles (IVA) and stem cell therapy) to promote ovarian follicle growth much more efficiently than the natural, in vivo process for women with POF. Transplantation of Warton Jelly-MSCs to the ovaries of POF patients rescued overall ovarian function. Participants (10 patients) were followed up monthly for a period of six months by hormonal (AMH, FSH, LH and E2), clinical (resuming menstruation), and US (folliculometry) outcomes after a laparoscopic operation. In summary, IVA/WJ-MSC transplantation may provide an effective treatment for POF.

Keywords: POF, in vitro activation, stem cell therapy, infertility

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12 Behavioral Response of Bee Farmers to Climate Change in South East, Nigeria

Authors: Jude A. Mbanasor, Chigozirim N. Onwusiribe

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The enigma climate change is no longer an illusion but a reality. In the recent years, the Nigeria climate has changed and the changes are shown by the changing patterns of rainfall, the sunshine, increasing level carbon and nitrous emission as well as deforestation. This study analyzed the behavioural response of bee keepers to variations in the climate and the adaptation techniques developed in response to the climate variation. Beekeeping is a viable economic activity for the alleviation of poverty as the products include honey, wax, pollen, propolis, royal jelly, venom, queens, bees and their larvae and are all marketable. The study adopted the multistage sampling technique to select 120 beekeepers from the five states of Southeast Nigeria. Well-structured questionnaires and focus group discussions were adopted to collect the required data. Statistical tools like the Principal component analysis, data envelopment models, graphs, and charts were used for the data analysis. Changing patterns of rainfall and sunshine with the increasing rate of deforestation had a negative effect on the habitat of the bees. The bee keepers have adopted the Kenya Top bar and Langstroth hives and they establish the bee hives on fallow farmland close to the cultivated communal farms with more flowering crops.

Keywords: climate, farmer, response, smart

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11 Machine Learning Based Smart Beehive Monitoring System Without Internet

Authors: Esra Ece Var

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Beekeeping plays essential role both in terms of agricultural yields and agricultural economy; they produce honey, wax, royal jelly, apitoxin, pollen, and propolis. Nowadays, these natural products become more importantly suitable and preferable for nutrition, food supplement, medicine, and industry. However, to produce organic honey, majority of the apiaries are located in remote or distant rural areas where utilities such as electricity and Internet network are not available. Additionally, due to colony failures, world honey production decreases year by year despite the increase in the number of beehives. The objective of this paper is to develop a smart beehive monitoring system for apiaries including those that do not have access to Internet network. In this context, temperature and humidity inside the beehive, and ambient temperature were measured with RFID sensors. Control center, where all sensor data was sent and stored at, has a GSM module used to warn the beekeeper via SMS when an anomaly is detected. Simultaneously, using the collected data, an unsupervised machine learning algorithm is used for detecting anomalies and calibrating the warning system. The results show that the smart beehive monitoring system can detect fatal anomalies up to 4 weeks prior to colony loss.

Keywords: beekeeping, smart systems, machine learning, anomaly detection, apiculture

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10 Improvement of Cardiometabolic after 8 Weeks of Weight Loss Intervention

Authors: Boris Bajer, Andrea Havranova, Miroslav Vlcek, Richard Imrich, Adela Penesova

Abstract:

Lifestyle interventions can prevent the deterioration of impaired glucose tolerance to manifest type 2 diabetes, and also prevent cardiovascular diseases, as it showed many studies (the Finnish Diabetes Prevention Study, Diabetes Prevention Program (DPP), . the China Da Qing Diabetes Prevention Study, etc.) Therefore the aim of our study was to compare the effect of intensified lifestyle intervention on cardiometabolic parameters. Methods: It is an ongoing randomized interventional clinical study (NCT02325804) focused on the reduction of body weight/fat. Intervention: hypocaloric diet (30% restriction of calories) and physical activity 150 minutes/week. Before and after 8 weeks of intervention all patients underwent complete medical examination (measurement of physical fitness, resting metabolic rate (RMR), body composition analysis, oral glucose tolerance test, parameters of lipid metabolism, and other cardiometabolic risk factors. Results: So far 39 patients finished the intervention. The average reduction of body weight was 6,8 + 4,9 kg (0-15 kg; p=0,0006), accompanied with significant reduction of body fat percentage (p ≤ 0,0001), amount of fat mass (p=0,03), waist circumference (p=0.02). Amount of lean mass and RMR remained unchanged. Heart rate (p=0,02), systolic and diastolic blood pressure was reduced (p=0,01 p=0,02 resp.) as well as insulin sensitivity was improved. Lipid parameters also changed - cholesterol, LDL decreased (p=0,05, p=0,04 resp.), while triglycerides showed tendency to decrease (p=0,055). Liver function improved, alanine aminotrasnferase (ALT) were reduced (p=0,01). Physical fitness significantly improved (as measure VO2 max (p=0,02). Conclusion: Results of our study are in line with previous results about the beneficial effect of intensive lifestyle changes on the reduction of cardiometabolic risk factors and improvement of liver function. Supported by grants APVV 15-0228; VEGA 2/0161/16

Keywords: obesity, weight loss, diet lipids, blood pressure, liver enzymes

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9 Mediterranean Diet-Driven Changes in Gut Microbiota Decrease the Infiltration of Inflammatory Myeloid Cells into the Intestinal Tissue

Authors: Gema Gómez-Casado, Alba Rodríguez-Muñoz, Virginia Mela-Rivas, Pallavi Kompella, Francisco José Tinahones-Madueña, Isabel Moreno-Indias, Almudena Ortega-Gómez

Abstract:

Obesity is a high-priority health problem worldwide due to its high prevalence. The proportion of obese and overweight subjects in industrialized countries exceeds half of the population in most cases. Beyond the metabolic problem, obesity boosts inflammation levels in the organism. The gut microbiota, considered an organ by itself, controls a high variety of processes at a systemic level. In fact, the microbiota interacts closely with the immune system, being crucial in determining the maturation state of neutrophils, key effectors of the innate immune response. It is known that changes in the diet exert strong effects on the variety and activity of the gut microbiota. The effect that those changes have on the axis microbiota-immune response is an unexplored field. In this study, 10 patients with obesity (weight 114,3 ± 14,5Kg, BMI 40,47±3,66) followed a Mediterranean-hypocaloric diet for 3 months, reducing their initial weight by 12,71 ± 3%. A transplant of microbiota from these patients before and after the diet was performed into wild type “germ-free” mice (n=10/group), treated with antibiotics. Six weeks after the transplant, mice were euthanized, and the presence of cells from the innate immune system were analysed in different organs (bone marrow, blood, spleen, visceral adipose tissue, and intestine) by flow cytometry. No differences were observed in the number of myeloid cells in bone marrow, blood, spleen, or visceral adipose tissue of mice transplanted with patient’s microbiota before and after following the Mediterranean diet. However, the intestine of mice that received post-diet microbiota presented a marked decrease in the number of neutrophils (whose presence is associated with tissue inflammation), as well as macrophages. In line with these findings, intestine monocytes from mice with post-diet microbiota showed a less inflammatory profile (lower Ly6Gˡᵒʷ proportion of cells). These results point toward a decrease in the inflammatory state of the intestinal tissue, derived from changes in the gut microbiota, which occurred after a 3-month Mediterranean diet.

Keywords: obesity, nutrition, Mediterranean diet, gut microbiota, immune system

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8 Jelly and Beans: Appropriate Use of Ultrasound in Acute Kidney Injury

Authors: Raja Ezman Raja Shariff

Abstract:

Acute kidney injury (AKI) is commonly seen in inpatients, and places a great cost on the NHS and patients. Timely and appropriate management is both nephron sparing and potentially life-saving. Ultrasound scanning (USS) is a well-recognised method for stratifying patients. Subsequently, the NICE AKI guidance has defined groups in whom scanning is recommended within 6 hours of request (pyonephrosis), within 24 hours (obstruction/cause unknown), and in whom routine scanning isn't recommended (cause for AKI identified). The audit looks into whether Stockport NHS Trust USS practice was in line with such recommendations. The audit evaluated 92 patients with AKI who had USS, between 01/01/14 to 30/04/14. Data collection was divided into 2 parts. Firstly, radiology request cards and the online imaging software (PACS) were evaluated. Then, the electronic case notes (ADVANTIS) was evaluated further. Based on request cards, 10% of requests were for pyonephrosis. Only 33% were scanned within 6hours and a further 33% within 24hours. 75% were requested for possible obstructions and unknown cause collectively. Of those due to possible obstruction, 71% of patients were scanned within 24 hours. Of those with unknown cause, 50% were scanned within 24 hours. 15% of requests had a cause declared and so potentially did not require scanning. Evaluation of the patients’ notes suggested further interesting findings. Firstly, potentially 39% of patients had a known cause for AKI, therefore, did not need USS. Subsequently, the cohort of unknown cause and possible obstruction was collectively reduced to 45%. Alarmingly the patient cohort with possible pyonephrosis went up to 16%, suggesting an under-recognition of this life-threatening condition. We plan to highlight these findings within our institution and make changes to encourage more appropriate requesting and timely scanning. Time will tell if we manage to save or increase our costs in this cost-conscious NHS. Patient benefits, though, seem to be guaranteed.

Keywords: AKI, ARF, kidney, renal

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7 Shielding Engineered Islets with Mesenchymal Stem Cells Enhance Survival under Hypoxia by Inhibiting p38 MAPK

Authors: Bhawna Chandravanshi, Ramesh Bhonde

Abstract:

In the present study, we focused on the improvisation of islet survival in hypoxia. The Islet-like cell aggregates (ICAs) derived from Wharton's jelly mesenchymal stem cells (WJ-MSC) were cultured with and without WJ-MSC for 48h in hypoxia and normoxia and tested for their direct trophic effect on β cell survival. The WJ MSCs themselves secreted insulin upon glucose challenge and expressed the pancreatic markers at both transcription and translational level (C-peptide, Insulin, Glucagon and Glut 2). Direct contact of MSCs with ICAs facilitate the highest viability under hypoxia as evidenced by fluorescein diacetate/propidium iodide and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The cytokine analysis of the co-cultured ICAs revealed amplification of anti-inflammatory cytokine-like TGFβ and TNFα accompanied by depletion of pro-inflammatory cytokines. The increment in VEGF and PDGFa was also seen showing their ability to vascularize upon transplantation. This was further accompanied by reduction in total reactive oxygen species, nitric oxide, and super oxide ions and down-regulation of Caspase3, Caspase8, p53 and up regulation of Bcl2 confirming prevention of apoptosis in ICAs. There was a significant reduction in the expression of p38 protein in the presence of MSCs making the ICAs responsive to glucose. Taken together our data demonstrate for the first time that the WJ-MSC expressed pancreatic markers and their supplementation protected engineered islets against hypoxia, oxidative stress, and inflammatory cytokines by inhibiting p38 MAPK protein.

Keywords: hypoxia, islet-like cell aggregates, inflammatory cytokines, oxidative stress

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6 Chemical Composition and Characteristics of Organic Solvent Extracts from the Omani Seaweeds Melanothamnus Somalensis and Gelidium Omanense

Authors: Abdullah Al-Nassri, Ahmed Al-Alawi

Abstract:

Seaweeds are classified into three groups: red, green, and brown. Each group of seaweeds consists of several types that have differences in composition. Even at the species level, there are differences in some ingredients, although in general composition, they are the same. Environmental conditions, availability of nutrients, and maturity stage are the main reasons for composition differences. In this study, two red seaweed species, Melanothamnus somalensis & Gelidium omanense, were collected in September 2021 from Sadh (Dhofar governorate, Oman). Five organic solvents were used sequentially to achieve extraction. The solvents were applied in the following order: hexane, dichloromethane, ethyl acetate, acetone, and methanol. Preparative HPLC (PrepLC) was performed to fraction the extracts. The chemical composition was measured; also, total phenols, flavonoids, and tannins were investigated. The structure of the extracts was analyzed by Fourier-transform infrared spectroscopy (FTIR). Seaweeds demonstrated high differences in terms of chemical composition, total phenolic content (TPC), total flavonoid content (TFC), and total tannin content (TTC). Gelidium omanense showed high moisture content, lipid content and carbohydrates (9.8 ± 0.15 %, 2.29 ± 0.09 % and 70.15 ± 0.42 %, respectively) compared to Melanothamnus somalensis (6.85 ± 0.01 %, 2.05 ± 0.12 % and 52.7 ± 0.36 % respectively). However, Melanothamnus somalensis showed high ash content and protein (27.68 ± 0.40 % and 52.7 ± 0.36 % respectively) compared to Gelidium omanense (8.07 ± 0.39 % and 9.70 ± 0.22 % respectively). Melanothamnus somalensis showed higher elements and minerals content, especially sodium and potassium. This is attributed to the jelly-like structure of Melanothamnus somalensis, which allows storage of more solutes compared to the leafy-like structure of Gelidium omanense. Furthermore, Melanothamnus somalensis had higher TPC in all fractions except the hexane fraction than Gelidium omanense. Except with hexane, TFC in the other solvents’ extracts was significantly different between Gelidium omanense and Melanothamnus somalensis. In all fractions, except dichloromethane and ethyl acetate fractions, there were no significant differences in TTC between Gelidium omanense and Melanothamnus somalensis. FTIR spectra showed variation between fractions, which is an indication of different functional groups.

Keywords: chemical composition, organic extract, Omani seaweeds, biological activity, FTIR

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5 Comparison Study of 70% Ethanol Effect on Direct and Retrival Culture of Contaminated Umblical Cord Tissue for Expansion of Mesenchymal Stem Cells

Authors: Ganeshkumar, Ashika, Valavan, Ramesh, Thangam, Chirayu

Abstract:

MSCs are found in much higher concentration in the Wharton’s jelly compared to the umbilical cord blood, which is a rich source of hematopoietic stem cells. Umbilical cord tissue is collected at the time of birth; it is processed and stored in liquid nitrogen for future therapeutical purpose. The source of contamination might be either from vaginal tract of mother or from hospital environment or from personal handling during cord tissue sample collection. If the sample were contaminated, decontamination procedure will be done with 70% ethanol (1 minute) in order to avoid sample rejection. Ethanol is effective against a wide range of bacteria, protozoa and fungi and has low toxicity to humans. Among the 1954 samples taken for the study, 24 samples were found to be contaminated with microorganism. The organisms isolated from the positive samples were found to be E. coli, Stenotrophomonas maltophilia, Pseudomonas aueroginosa, Enterococcus fecalis, Acinetobacter bowmani, Staphylococcus epidermidis, Enterobacter cloacae, and Proteus mirabilis. Among these organisms 70% ethanol successfully eliminated E. coli, Enterococcus fecalis, Acinetobacter bowmani, Staphylococcus epidermidis, and Proteus mirabilis. 70% ethanol was unsuccessful in eliminating Stenotrophomonas maltophilia, Pseudomonas aueroginosa, and Enterobacter cloacae. Stenotrophomonas maltophilia and Pseudomonas aueroginosa have the ability to form biofilm that make them resistant to alcohol. Biofilm act as protective layer for bacteria and which protects them from host defense and antibiotic wash. Finally it was found 70% ethanol wash saved 58.3% cord tissue samples from rejection and it is ineffective against 41% of the samples. The contamination rate can be reduced by maintaining proper aseptic techniques during sample collection and processing.

Keywords: umblical cord tissue, decontamination, 70% ethanol effectiveness, contamination

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4 Cord Blood Hematopoietic Stem Cell Expansion Ability of Mesenchymal Stem Cells Isolated From Different Sources

Authors: Ana M. Lara, Manuela Llano, Felipe Gaitán, Rosa H. Bustos, Ana Maria Perdomo-Arciniegas, Ximena Bonilla

Abstract:

Umbilical cord blood is used as a source of progenitor and stem cells for the regeneration of the hematopoietic and immune system to treat patients with different hematological or non-hematological diseases. This stem cell source represents an advantage over the use of bone marrow or mobilized peripheral blood because it has a lower incidence rate of graft-versus-host disease, probably due to fewer immunological compatibility restrictions. However, its low cellular dose limits its use in pediatric patients. This work proposes the standardization of a cell expansion technique to compensate for the dose of infused cells through the ex-vivo manipulation of hematopoietic progenitor cells from umbilical cord blood before transplantation. The expansion model is carried out through co-cultures with mesenchymal stem cells (MSC) from bone marrow (BM) and less explored fetal tissues such as Wharton's jelly (WJ) and umbilical cord blood (UCB). Initially, a master cell bank of primary mesenchymal stem cells isolated from different sources was established and characterized following International Society of Cell Therapies (ISCT) indications. Additionally, we assessed the effect of a short 25 Gy cycle of gamma irradiation on cell cycle arrest of mesenchymal cells over the support capacity for the expansion of hematopoietic stem cells from umbilical cord blood was evaluated. The results show that co-cultures with MSC from WJ and UCB allow the cellular dose of HSPC to be maximized between 5 and 16 times having a similar support capacity as BM. In addition, was evaluated the hematopoietic stem progenitor cell's HSPC functionality through the evaluation of migration capacity, their differentiation capacity during culture time by flow cytometry to evaluate the expression of membrane markers associated with lineage-committed progenitors, their clonogenic potential, and the evaluation of secretome profile in the expansion process was evaluated. So far, the treatment with gamma irradiation maintains the hematopoietic support capacity of mesenchymal stem cells from the three sources studied compared to treatments without irradiation, favoring the use of fetal tissues that are generally waste to obtain mesenchymal cell lines for ex-vivo expansion systems. With the results obtained, a standardized protocol that will contribute to the development of ex-vivo expansion with MSC on a larger scale will be achieved, enabling its clinical use and expanding its application in adults.

Keywords: ex-vivo expansion, hematopoietic stem cells, hematopoietic stem cell transplantation, mesenchymal stem cells, umbilical cord blood

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3 A pilot Study of Umbilical Cord Mini-Clamp

Authors: Seng Sing Tan

Abstract:

Clamping of the umbilical cord after birth is widely practiced as a part of labor management. Further improvements were proposed to produce a smaller, lighter and more comfortable clamp while still maintaining current standards of clamping. A detachable holder was also developed to facilitate the clamping process. This pilot study on the efficacy of the mini-clamp was conducted to evaluate a tightness of the seal and a firm grip of the clamp on the umbilical cord. The study was carried out at National University Hospital, using 5 sets of placental cord. 18 samples of approximate 10 cm each were harvested. The test results showed that the mini-clamp was able to stop the flow through the cord after clamping without rupturing the cord. All slip tests passed with a load of 0.2 kg. In the pressure testing, 30kPa of saline was exerted into the umbilical veins. Although there was no physical sign of fluid leaking through the end secured by the mini-clamp, the results showed the pressure was not able to sustain the pressure set during the tests. 12 out of the 18 test samples have more than 7% of pressure drop in 30 seconds. During the pressure leak test, it was observed on several samples that when pressurized, small droplets of saline were growing on the outer surface of the cord lining membrane. It was thus hypothesized that the pressure drop was likely caused by the perfusion of the injected saline through the Wharton’s jelly and the cord lining membrane. The average pressure in the umbilical vein is roughly 2.67kPa (20 mmHg), less than 10% of 30kPa (~225mmHg), set for the pressure testing. As such, the pressure set could be over-specified, leading to undesirable outcomes. The development of the mini-clamp was an attempt to increase the comfort of newly born babies while maintaining the usability and efficacy of hospital grade umbilical cord clamp. The pressure leak in this study would be unfair to fully attribute it to the design and efficacy of the mini-clamp. Considering the unexpected leakage of saline through the umbilical membrane due to over-specified pressure exerted on the umbilical veins, improvements can definitely be made to the existing experimental setup to obtain a more accurate and conclusive outcome. If proven conclusive and effective, the mini-clamp with a detachable holder could be a smaller and potentially cheaper alternative to existing umbilical cord clamps. In addition, future clinical trials could be conducted to determine the user-friendliness of the mini-clamp and evaluate its practicality in the clinical setting by labor ward clinicians. A further potential improvement could be proposed on the sustainability factor of the mini-clamp. A biodegradable clamp would revolutionise the industry in this increasingly environmentally sustainability world.

Keywords: leak test, mini-clamp, slip test, umbilical cord

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2 Recognising and Managing Haematoma Following Thyroid Surgery: Simulation Teaching is Effective

Authors: Emily Moore, Dora Amos, Tracy Ellimah, Natasha Parrott

Abstract:

Postoperative haematoma is a well-recognised complication of thyroid surgery with an incidence of 1-5%. Haematoma formation causes progressive airway obstruction, necessitating emergency bedside haematoma evacuation in up to ¼ of patients. ENT UK, BAETS and DAS have developed consensus guidelines to improve perioperative care, recommending that all healthcare staff interacting with patients undergoing thyroid surgery should be trained in managing post-thyroidectomy haematoma. The aim was to assess the effectiveness of a hybrid simulation model in improving clinician’s confidence in dealing with this surgical emergency. A hybrid simulation was designed, consisting of a standardised patient wearing a part-task trainer to mimic a post-thyroidectomy haematoma in a real patient. The part-task trainer was an adapted C-spine collar with layers of silicone representing the skin and strap muscles and thickened jelly representing the haematoma. Both the skin and strap muscle layers had to be opened in order to evacuate the haematoma. Boxes have been implemented into the appropriate post operative areas (recovery and surgical wards), which contain a printed algorithm designed to assist in remembering a sequence of steps for haematoma evacuation using the ‘SCOOP’ method (skin exposure, cut sutures, open skin, open muscles, pack wound) along with all the necessary equipment to open the front of the neck. Small-group teaching sessions were delivered by ENT and anaesthetic trainees to members of the multidisciplinary team normally involved in perioperative patient care, which included ENT surgeons, anaesthetists, recovery nurses, HCAs and ODPs. The DESATS acronym of signs and symptoms to recognise (difficulty swallowing, EWS score, swelling, anxiety, tachycardia, stridor) was highlighted. Then participants took part in the hybrid simulation in order to practice this ‘SCOOP’ method of haematoma evacuation. Participants were surveyed using a Likert scale to assess their level of confidence pre- and post teaching session. 30 clinicians took part. Confidence (agreed/strongly agreed) in recognition of post thyroidectomy haematoma improved from 58.6% to 96.5%. Confidence in management improved from 27.5% to 89.7%. All participants successfully decompressed the haematoma. All participants agreed/strongly agreed, that the sessions were useful for their learning. Multidisciplinary team simulation teaching is effective at significantly improving confidence in both the recognition and management of postoperative haematoma. Hybrid simulation sessions are useful and should be incorporated into training for clinicians.

Keywords: thyroid surgery, haematoma, teaching, hybrid simulation

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1 Characterization of Surface Microstructures on Bio-Based PLA Fabricated with Nano-Imprint Lithography

Authors: D. Bikiaris, M. Nerantzaki, I. Koliakou, A. Francone, N. Kehagias

Abstract:

In the present study, the formation of structures in poly(lactic acid) (PLA) has been investigated with respect to producing areas of regular, superficial features with dimensions comparable to those of cells or biological macromolecules. Nanoimprint lithography, a method of pattern replication in polymers, has been used for the production of features ranging from tens of micrometers, covering areas up to 1 cm², down to hundreds of nanometers. Both micro- and nano-structures were faithfully replicated. Potentially, PLA has wide uses within biomedical fields, from implantable medical devices, including screws and pins, to membrane applications, such as wound covers, and even as an injectable polymer for, for example, lipoatrophy. The possibility of fabricating structured PLA surfaces, with structures of the dimensions associated with cells or biological macro- molecules, is of interest in fields such as cellular engineering. Imprint-based technologies have demonstrated the ability to selectively imprint polymer films over large areas resulting in 3D imprints over flat, curved or pre-patterned surfaces. Here, we compare nano-patterned with nano-patterned by nanoimprint lithography (NIL) PLA film. A silicon nanostructured stamp (provided by Nanotypos company) having positive and negative protrusions was used to pattern PLA films by means of thermal NIL. The polymer film was heated from 40°C to 60°C above its Tg and embossed with a pressure of 60 bars for 3 min. The stamp and substrate were demolded at room temperature. Scanning electron microscope (SEM) images showed good replication fidelity of the replicated Si stamp. Contact-angle measurements suggested that positive microstructuring of the polymer (where features protrude from the polymer surface) produced a more hydrophilic surface than negative micro-structuring. The ability to structure the surface of the poly(lactic acid), allied to the polymer’s post-processing transparency and proven biocompatibility. Films produced in this were also shown to enhance the aligned attachment behavior and proliferation of Wharton’s Jelly Mesenchymal Stem cells, leading to the observed growth contact guidance. The bacterial attachment patterns of some bacteria, highlighted that the nano-patterned PLA structure can reduce the propensity for the bacteria to attach to the surface, with a greater bactericidal being demonstrated activity against the Staphylococcus aureus cells. These biocompatible, micro- and nanopatterned PLA surfaces could be useful for polymer– cell interaction experiments at dimensions at, or below, that of individual cells. Indeed, post-fabrication modification of the microstructured PLA surface, with materials such as collagen (which can further reduce the hydrophobicity of the surface), will extend the range of applications, possibly through the use of PLA’s inherent biodegradability. Further study is being undertaken to examine whether these structures promote cell growth on the polymer surface.

Keywords: poly(lactic acid), nano-imprint lithography, anti-bacterial properties, PLA

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