Search results for: high phosphate containing foods and beverages

Authors: Olivia Bory Devisme, Denis Genon-Catalot, Frederic Alicalapa, Pierre-Olivier Lucas De Peslouan, Jean-Pierre Chabriat

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In the context of climate change, it is essential that island territories gain energy autonomy. Currently mostly dependent on fossil fuels, the island of Reunion lo- cated in the Indian Ocean nevertheless has a high potential for solar energy. As the market for photovoltaic panels has been growing in recent years, the issues of energy losses linked to the multiple conversions from direct current to alternating current are emerging. In order to quantify these advantages and disadvantages by a comparative study, this document present the measurements carried out on a direct current test bench, particularly for lighting, ventilation, air condi- tioning and office equipment for the tertiary sector. All equipment is supplied with DC power from energy produced by photovoltaic panels. A weather sta- tion, environmental indoor sensors, and drivers are also used to control energy. Self-consumption is encouraged in order to manage different priorities between user consumption and energy storage in a lithium iron phosphate battery. The measurements are compared to a conventional electrical architecture (DC-AC- DC) for energy consumption, equipment overheating, cost, and life cycle analysis.

Keywords: DC microgrids, solar energy, smart buildings, storage

Procedia PDF Downloads 143

Authors: Nisha Govender, Siju Senan, Zeti-Azura Hussein, Wickneswari Ratnam

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Jatropha curcas, a well-described bioenergy crop has been extensively accepted as future fuel need especially in tropical regions. Ideal planting material required for large-scale plantation is still lacking. Breeding programmes for improved J. curcas varieties are rendered difficult due to limitations in genetic diversity. Using a combined transcriptome and physiological data, we investigated the molecular and physiological differences in high and low yielding Jatropha curcas to address plausible heritable variations underpinning these differences, in regard to photosynthesis, a key metabolism affecting yield potentials. A total of 6 individual Jatropha plant from 4 accessions described as high and low yielding planting materials were selected from the Experimental Plot A, Universiti Kebangsaan Malaysia (UKM), Bangi. The inflorescence and shoots were collected for transcriptome study. For the physiological study, each individual plant (n=10) from the high and low yielding populations were screened for agronomic traits, chlorophyll content and stomatal patterning. The J. curcas transcriptomes are available under BioProject PRJNA338924 and BioSample SAMN05827448-65, respectively Each transcriptome was subjected to functional annotation analysis of sequence datasets using the BLAST2Go suite; BLASTing, mapping, annotation, statistical analysis and visualization Large-scale phenotyping of the number of fruits per plant (NFPP) and fruits per inflorescence (FPI) classified the high yielding Jatropha accessions with average NFPP =60 and FPI > 10, whereas the low yielding accessions yielded an average NFPP=10 and FPI < 5. Next generation sequencing revealed genes with differential expressions in the high yielding Jatropha relative to the low yielding plants. Distinct differences were observed in transcript level associated to photosynthesis metabolism. DEGs collection in the low yielding population showed comparable CAM photosynthetic metabolism and photorespiration, evident as followings: phosphoenolpyruvate phosphate translocator chloroplastic like isoform with 2.5 fold change (FC) and malate dehydrogenase (2.03 FC). Green leaves have the most pronounced photosynthetic activity in a plant body due to significant accumulation of chloroplast. In most plants, the leaf is always the dominant photosynthesizing heart of the plant body. Large number of the DEGS in the high-yielding population were found attributable to chloroplast and chloroplast associated events; STAY-GREEN chloroplastic, Chlorophyllase-1-like (5.08 FC), beta-amylase (3.66 FC), chlorophyllase-chloroplastic-like (3.1 FC), thiamine thiazole chloroplastic like (2.8 FC), 1-4, alpha glucan branching enzyme chloroplastic amyliplastic (2.6FC), photosynthetic NDH subunit (2.1 FC) and protochlorophyllide chloroplastic (2 FC). The results were parallel to a significant increase in chlorophyll a content in the high yielding population. In addition to the chloroplast associated transcript abundance, the TOO MANY MOUTHS (TMM) at 2.9 FC, which code for distant stomatal distribution and patterning in the high-yielding population may explain high concentration of CO2. The results were in agreement with the role of TMM. Clustered stomata causes back diffusion in the presence of gaps localized closely to one another. We conclude that high yielding Jatropha population corresponds to a collective function of C3 metabolism with a low degree of CAM photosynthetic fixation. From the physiological descriptions, high chlorophyll a content and even distribution of stomata in the leaf contribute to better photosynthetic efficiency in the high yielding Jatropha compared to the low yielding population.

Keywords: chlorophyll, gene expression, genetic variation, stomata

Procedia PDF Downloads 215

Authors: Deog-Gyu Seo, Jin-Soo Ahn

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Objectives: Recently, the development of adhesive primers on stable bonding between zirconia and resin cement has been on the increase. The bond strength of zirconia-resin cement can be effectively increased with the treatment of primer composed of the adhesive monomer that can chemically bond with the oxide layer, which forms on the surface of zirconia. 10-methacryloyloxydecyl dihydrogen phosphate (10-MDP) that contains phosphate ester and acidic monomer 4-methacryloxyethyl trimellitic anhydride(4-META) have been suggested as monomers that can form chemical bond with the surface oxide layer of zirconia. Also, these suggested monomers have proved to be effective zirconia surface treatment for bonding to resin cement. The purpose of this study is to evaluate the effects of primer treatment on the bond strength of Zirconia-resin cement by using three different kinds of primers on the market. Methods: Zirconia blocks were prepared into 60 disk-shaped specimens by using a diamond saw. Specimens were divided into four different groups: first three groups were treated with zirconiaLiner(Sun Medical Co., Ltd., Furutaka-cho, Moriyama, Shiga, Japan), Alloy primer (Kuraray Noritake Dental Inc., Sakaju, Kurashiki, Okayama, Japan), and Universal primer (Tokuyama dental Corp., Taitou, Taitou-ku, Tokyo, Japan) respectively. The last group was the control with no surface treatment. Dual cured resin cement (Biscem, Bisco Inc., Schaumburg, IL, USA) was luted to each group of specimens. And then, shear bond strengths were measured by universal tesing machine. The significance of the result was statistically analyzed by one-way ANOVA and Tukey test. The failure sites in each group were inspected under a magnifier. Results: Mean shear bond strength were 0.60, 1.39, 1.03, 1.38 MPa for control, Zirconia Liner (ZL), Alloy primer (AP), Universal primer (UP), respectively. Groups with application of each of the three primers showed significantly higher shear bond strength compared to the control group (p < 0.05). Among the three groups with the treatment, ZL and UP showed significantly higher shear bond strength than AP (p < 0.05), and there were no significant differences in mean shear bond strength between ZL and UP (p < 0.05). While the most specimens of control groups showed adhesive failure (80%), the most specimens of three primer-treated groups showed cohesive or mixed failure (80%).

Keywords: primer, resin cement, shear bond strength, zirconia

Procedia PDF Downloads 184

Authors: Beata Bilska, Marzena Tomaszewska, Danuta Kolozyn-Krajewska

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Food loss and food waste are a global problem of the modern economy. The research undertaken aimed to analyze how food is handled in catering establishments when it comes to food waste and to demonstrate the main ways of management with foods/dishes not served to consumers. A survey study was conducted from January to June 2019. The selection of catering establishments participating in the study was deliberate. The study included establishments located only in Mazowieckie Voivodeship (Poland). Forty-two completed questionnaires were collected. In some questions, answers were based on a 5-point scale of 1 to 5 (from "always" / "every day" to "never"). The survey also included closed questions with a suggested cafeteria of answers. The respondents stated that in their workplaces, dishes served cold and hot ready meals are discarded every day or almost every day (23.7% and 20.5% of answers respectively). A procedure most frequently used for dealing with dishes not served to consumers on a given day is their storage at a cool temperature until the following day. In the research, 1/5 of respondents admitted that consumers "always" or "usually" leave uneaten meals on their plates, and over 41% "sometimes" do so. It was found additionally that food not used in the foodservice sector is most often thrown into a public container for rubbish. Most often thrown into the public container (with communal trash) were: expired products (80.0%), plate waste (80.0%) and inedible products (fruit and vegetable peels, eggshells) (77.5%). Most frequently into the container dedicated only to food waste were thrown out used deep-frying oil (62.5%). 10% of respondents indicated that inedible products in their workplaces are allocated for animal feeds. Food waste in the foodservice sector remains an insufficiently studied issue, as owners of these objects are often unwilling to disclose data about the subject. Incorrect ways of management with foods not served to consumers were observed. There is a need to develop educational activities for employees and management in the context of food waste management in the foodservice sector.

Keywords: food waste, inedible products, plate waste, used deep-frying oil

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Authors: Nurudeen Afolami Ayeni, Kasali Adewale Bello

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Study of the effect of dye-resin complexation on the degree of dye absorption were carried out using Procion Blue MX-R to dye cotton fabric in the presence hexamethylol melamine (MR6) and its phosphate derivative (MPR4) for resination. The highest degree of dye exhaustion was obtained at 400C for 1 hour with the resinated fabric showing more affinity for the dye than the ordinary fibre. Improved fastness properties was recorded which show a relatively higher stability of dye-resin complex formed in the fibre.

Keywords: affinity, cotton, dyeing, reactive dye, resination

Procedia PDF Downloads 289

Authors: Tatyane Lopes de Freitas, Antonio Diogo S. Vieira, Susana Marta Isay Saad, Maria Ines Genovese

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The fruit processing industries generate a large volume of residues to produce juices, pulps, and jams. These residues, or by-products, consisting of peels, seeds, and pulps, are routinely discarded. Fruits are rich in bioactive compounds, including polyphenols, which have positive effects on health. Dry residues from two fruits, acerola (M. emarginata D. C.) and orange (C. sinensis), were characterized in relation to contents of ascorbic acid, minerals, total dietary fibers, moisture, ash, lipids, proteins, and carbohydrates, and also high performance liquid chromatographic profile of flavonoids, total polyphenols and proanthocyanidins contents, and antioxidant capacity by three different methods (Ferric reducing antioxidant power assay-FRAP, Oxygen Radical Absorbance Capacity-ORAC, 1,1-diphenyl-2-picrylhydrazil (DPPH) radical scavenging activity). Acerola by-products presented the highest acid ascorbic content (605 mg/100 g), and better antioxidant capacity than orange by-products. The dry residues from acerola demonstrated high contents of proanthocyanidins (617 µg CE/g) and total polyphenols (2525 mg gallic acid equivalents - GAE/100 g). Both presented high total dietary fiber (above 60%) and protein contents (acerola: 10.4%; orange: 9.9%), and reduced fat content (acerola: 1.6%; orange: 2.6%). Both residues showed high levels of potassium, calcium, and magnesium, and were considered sources of these minerals. With acerola by-product, four formulations of probiotics fermented milks were produced: F0 (without the addition of acerola residue (AR)), F2 (2% AR), F5 (5% AR) and F10 (10% AR). The physicochemical characteristics of the fermented milks throughout of storage were investigated, as well as the impact of in vitro simulated gastrointestinal conditions on flavonoids and probiotics. The microorganisms analyzed maintained their populations around 8 log CFU/g during storage. After the gastric phase of the simulated digestion, the populations decreased, and after the enteric phase, no colonies were detected. On the other hand, the flavonoids increased after the gastric phase, maintaining or suffering small decrease after enteric phase. Acerola by-products powder is a valuable ingredient to be used in functional foods because is rich in vitamin C, fibers and flavonoids. These flavonoids appear to be highly resistant to the acids and salts of digestion.

Keywords: acerola, orange, by-products, fermented milk

Procedia PDF Downloads 110

Authors: Theresa MacNeil-Kelly

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Blue Zones are particular regions of the world with a high prevalence of centenarians who share common characteristics, lifestyles and environments. There are currently only five recognized Blue Zones, two of which include Okinawa, Japan and the Nicoya Peninsula in Costa Rica. Individuals living in these areas tend to have positive outlooks on life, utilize daily movement, rely on strong social support groups, and eat little to no processed foods. The current research sought to further understand how centenarians living in Okinawa and in the Nicoya Peninsula utilize Blue Zone lifestyle elements in their daily living habits. To accomplish this, the author traveled to both Okinawa and the Nicoya Peninsula, Costa Rica, and interviewed several centenarians, paying particular attention to lifestyle choices and their effects on the aging process. Thematic analysis was used to analyze interview responses, and several themes emerged, such as the importance of family, friends, faith/spirituality, mindfulness, nutrition and daily movement as key foundations to aging in healthy and productive ways. Suggestions for ways to implement these habits globally was also discussed.

Keywords: aging, blue zones, centenarians, nicoya peninsula, okinawa

Procedia PDF Downloads 240

Authors: Chica Riska Ashari, Yoli Farradika

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Background: Nutritional problems arise due to food security problems in the family, such as the ability of families to obtain food which is common in poor people due to lack of economic access to buy food. For this reason, it is hoped that there will be actions or behaviors that can be taken to fulfill their food or known as the Food Coping Strategy. The purpose of this study is to identify the eating patterns and Food Coping strategies of household students of prof. DR. HAMKA Muhammadiyah University Jakarta during the covid-19 pandemic. Methods: This study is a quantitative observational study with a cross-sectional approach. The dependent variable in this study is food coping strategies and eating patterns. The location of this research is Prof. DR. Hamka Muhammadiyah University. The population in this study were all students of Prof. DR. HAMKA Muhammadiyah University. The sampling technique is purposive sampling. The minimum number of samples in this study is 97 people with a response rate or drop out an estimate of 10%, so the total number of samples was 107 people. Statistical analysis with descriptive analysis. Results: The results showed that most of the food coping strategies were carried out by the students of the household of Prof. DR. HAMKA Muhammadiyah University, were buying the cheaper food (91.6%), then changing the priority of buying food (75.7%) and household members who carry out this food coping strategy are mothers (59.8%) then followed by students themselves (57, 9%). The diet of most students at the Prof. DR. HAMKA Muhammadiyah University in a day was fond of consuming sugar and foods containing sugar (candy, sugar, honey, sweet drinks) (98.1%) then eggs (97.2%). Conclusion: Food coping strategies are mostly used by households students at Prof. DR. HAMKA Muhammadiyah University who were buying the cheaper food and the member who did this behavior the most were the mothers. The diet of most students at Prof. DR. HAMKA Muhammadiyah University in a day was fond of consuming sugar and foods containing sugar (candy, sugar, honey, sweet drinks).

Keywords: behavior, eating patterns, food coping strategies, food security, students

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Authors: Seyed Hassan Nikookar, Mahmoud Fazeli-Dinan, Seyyed Payman Ziapour, Ahmad-Ali Enayati

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Background: Mosquitoes look for all kinds of aquatic habitats for laying eggs. Characteristics of water habitats are important factors in determining whether a mosquito can survive and successfully completed their developmental stages. Physicochemical factors can display an important role in vector control programs. This investigate determined whether physicochemical factors differ between habitats can be effective in the larvae density in Mazandaran province. Methods: Larvae were collected by the standard dipper up to 350 ml for 15-20 minutes from fixed habitats in 16 villages of 30 townships, the specimens identified by morphological key. Water samples were collected during larval collection and were evaluated for temperature (°C), acidity (pH), turbidity (NTU), electrical conductivity (μS/cm), alkalinity (mg/l), total hardness (mg/l), nitrate (mg/l), chloride (mg/l), phosphate (mg/l), sulfate (mg/l) in selected habitats using standard methods. Spearman Correlation coefficient was used for analyze data. Results: Totally 7566 mosquito larvae of three genera and 15 species were collected of fixed habitats. Cx. pipiens was the dominant species except in villages of Tileno, Zavat, Asad Abad, Shah Mansur Mahale which An. maculipennis, Cx. torrentium were as the predominant species. Turbidity in Karat Koti, Chloride in Al Tappeh, nitrate, phosphate and sulfate in Chalmardi, electrical conductivity, alkalinity, total hardness in Komishan villages were significantly higher than other villages (P < 0.05). There were a significant positive correlation between Cx. pipiens and Electrical conductivity, Alkalinity, Total hardness, Chloride, Cx. tritaeniorhynchus and Chloride, whereas a significant negative correlation observed between Sulfate and Cx. perexiguss. Conclusion: The correlations observed between physicochemical factor and larval density, possibly can confirm the effect of these parameters on the breeding activities of mosquitoes, and could probability facilitate larval control programs by the handwork of such factors.

Keywords: anopheles, culex, culiseta, physicochemical, habitats, larvae density, correlation

Procedia PDF Downloads 244

Authors: Delgado-Meza M., Minor-Pérez H.

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Protease is the generic name of enzymes that hydrolyze proteins. These are classified in the subgroup EC3.4.11-99X of the classification enzymes. In food technology the proteolysis is used to modify functional and nutritional properties of food, and in some cases this proteolysis may cause food spoilage. In general, seafood and rainbow trout have accelerated decomposition process once it has done its capture, due to various factors such as the endogenous enzymatic activity that can result in loss of structure, shape and firmness, besides the release of amino acid precursors of biogenic amines. Some studies suggest the use of protease inhibitors from legume as biological regulators of proteolytic activity. The enzyme inhibitors are any substance that reduces the rate of a reaction catalyzed by an enzyme. The objective of this study was to evaluate the reduction of the proteolytic activity of enzymes in extracts of rainbow trout with protease inhibitors obtained from chickpea flour. Different proportions of rainbow trout enzyme extract (75%, 50% and 25%) and extract chickpea enzyme inhibitors were evaluated. Chickpea inhibitors were obtained by mixing 5 g of flour in 30 mL of pH 7.0 phosphate buffer. The sample was centrifuged at 8000 rpm for 10 min. The supernatant was stored at -15°C. Likewise, 20 g of rainbow trout were ground in 20 mL of phosphate buffer solution at pH 7.0 and the mixture was centrifuged at 5000 rpm for 20 min. The supernatant was used for the study. In each treatment was determined the specific enzymatic activity with the technique of Kunitz, using hemoglobin as substrate for the enzymes acid fraction and casein for basic enzymes. Also biuret protein was quantified for each treatment. The results showed for fraction of basic enzymes in the treatments evaluated, that were inhibition of endogenous enzymatic activity. Inhibition values compared to control were 51.05%, 56.59% and 59.29% when the proportions of endogenous enzymes extract rainbow trout were 75%, 50% and 25% and the remaining volume used was extract with inhibitors. Treatments with acid enzymes showed no reduction in enzyme activity. In conclusion chickpea flour reduced the endogenous enzymatic activity of rainbow trout, which may favor its application to increase the half-life of this food. The authors acknowledge the funding provided by the CONACYT for the project 131998.

Keywords: rainbouw trout, enzyme inhibitors, proteolysis, enzyme activity

Procedia PDF Downloads 398

Authors: R. B. B. Samantha Ramachandra, L. D. J. Upul Senarath, S. H. Padmal De Silva

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Dietary pattern largely differs over countries and even within a country, it shows cultural differences. The dietary pattern changes the energy consumption and micronutrient intake, directly affects the pregnancy outcome. The dietary diversity was used as an indirect measure to assess micronutrient adequacy for pregnant mothers in this study. The study was conducted as a baseline survey with the objective of designing an intervention to improve the dietary diversity of pregnant mothers in Sri Lanka. The survey was conducted in Kalutara district of Sri Lanka in 2015 among 769 pregnant mothers at different gestational ages. Dietary diversity questionnaire developed by Food and Agricultural Organization’s (FAO) Food and Nutrition technical Assistance (FANTA) II project, recommended for cross-country use with adaptations was used for data collection. Trained data collectors met pregnant mothers at field ante-natal clinic and questioned on last 24hr dietary recall with portion size and coded food items to identify the diversity. Pregnant mothers were identified from randomly selected 21 clusters of public health midwife areas. 81.5% mothers (n=627) in the sample had been registered at Public Health Midwife (PHM) before 8 weeks of gestation. 24.4% of mothers were with low starting BMI and 22.7% mothers were with high starting BMI. 47.6% (n=388) mothers had abstained from at least one food item during the pregnancy. The food group with the highest consumption was rice (98.4%) followed by sugar (89.9%). 76.1% mothers had consumed milk, 73% consumed fish and sea foods. Consumption of green leaves was 52% and Vit A rich foods consumed only by 49% mothers. Animal organs, flesh meat and egg all showed low prevalence as 4.7%, 21.6% and 20% respectively. Consumption of locally grown roots, nut, legumes all showed very low prevalence. Consumption of 6 or more food groups was considered as good dietary diversity (DD), 4 to 5 food groups as moderate diversity and 3 or less food groups as poor diversity by FAO FANTA II project. 42.1% mothers demonstrated good DD while another 42.1% recorded moderate diversity. Working mothers showed better DD (51.6%, n=82/159) compared to housewives in the sample (chi = 10.656a,. df=2, p=0.005). The good DD showed gradual improvement from 43.1% to 55.5% along the poorest to richest wealth index (Chi=48.045, df=8 and p=0.000). DD showed significant association with the ethnicity and Moors showed the lowest DD. DD showed no association with the home gardening even though where better diversity expected among those who have home gardening (p=0.548). Sri Lanka is a country where many food items can be grown in the garden and semi-urban setting have adequate space for gardening. Many Sri Lankan mothers do not add homegrown items in their meal. At the same time, their consumption of animal food shows low prevalence. The DD of most of the mothers being either moderate or low (58%) may result from inadequate micro nutrient intake during pregnancy. It is recommended that adding green leaves, locally grown vegetables, roots, nuts and legumes can help increasing the DD of Sri Lankan mothers at low cost.

Keywords: dietary diversity, pregnant mothers, micro-nutrient, food groups

Procedia PDF Downloads 148

Authors: N. E. Gallagher, C. E. Neville, N. Lyner, J. Yarnell, C. C. Patterson, J. E. Gallacher, Y. Ben-Shlomo, A. Fehily, J. V. Woodside

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Hearing loss is highly prevalent in older people and can reduce quality of life substantially. Emerging research suggests that potentially modifiable risk factors, including risk factors previously related to cardiovascular disease risk, may be associated with a decreased or increased incidence of hearing loss. This has prompted investigation into the possibility that certain nutrients, foods or dietary patterns may also be associated with incidence of hearing loss. The aim of this study was to determine any associations between dietary patterns and hearing loss in men enrolled in the Caerphilly study. The Caerphilly prospective cohort study began in 1979-1983 with recruitment of 2512 men aged 45-59 years. Dietary data was collected using a self-administered, semi-quantitative, 56-item food frequency questionnaire (FFQ) at baseline (1979-1983), and 7-day weighed food intake (WI) in a 30% sub-sample, while pure-tone unaided audiometric threshold was assessed at 0.5, 1, 2 and 4 kHz, between 1984 and 1988. Principal components analysis (PCA) was carried out to determine a posteriori dietary patterns and multivariate linear and logistic regression models were used to examine associations with hearing level (pure tone average (PTA) of frequencies 0.5, 1, 2 and 4 kHz in decibels (dB)) for linear regression and with hearing loss (PTA>25dB) for logistic regression. Three dietary patterns were determined using PCA on the FFQ data- Traditional, Healthy, High sugar/Alcohol avoider. After adjustment for potential confounding factors, both linear and logistic regression analyses showed a significant and inverse association between the Healthy pattern and hearing loss (P<0.001) and linear regression analysis showed a significant association between the High sugar/Alcohol avoider pattern and hearing loss (P=0.04). Three similar dietary patterns were determined using PCA on the WI data- Traditional, Healthy, High sugar/Alcohol avoider. After adjustment for potential confounding factors, logistic regression analyses showed a significant and inverse association between the Healthy pattern and hearing loss (P=0.02) and a significant association between the Traditional pattern and hearing loss (P=0.04). A Healthy dietary pattern was found to be significantly inversely associated with hearing loss in middle-aged men in the Caerphilly study. Furthermore, a High sugar/Alcohol avoider pattern (FFQ) and a Traditional pattern (WI) were associated with poorer hearing levels. Consequently, the role of dietary factors in hearing loss remains to be fully established and warrants further investigation.

Keywords: ageing, diet, dietary patterns, hearing loss

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Authors: Sarina Sulaiman, N.Khairudin , P.Jamal, M.Z. Alam, Zaki Zainudin, S. Azmi

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In this study, fish bone waste was used as a new catalyst for biodiesel production. Instead of discarding the fish bone waste, it will be utilized as a source for catalyst that can provide significant benefit to the environment. Also, it can be substitute as a calcium oxide source instead of using eggshell, crab shell and snail shell. The XRD and SEM analysis proved that calcined fish bone contains calcium oxide, calcium phosphate and hydroxyapatite. The catalyst was characterized using Scanning Electron Microscope (SEM) and X-ray Diffraction (XRD).

Keywords: calcinations, fish bone, transesterification, waste catalyst

Procedia PDF Downloads 275

Authors: Heba Al-Kelesh

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Day after day, high-grade iron ores are consumed. Because of the strong global demand for iron and steel, it has necessitated the utilization of various low-grade iron ores, which are not suitable for direct exploitation in the iron industry. The low-grade ores cannot be dressed using traditional mineral processing methods because of complicated mineral compositions. The present work is aimed to investigate the reducibility of some Egyptian iron ores and concentrates by conditions emulate different blast furnace areas. Representative specimens are collected from El-Gedida–Baharia oasis, Eastern South Aswan, and Eastern desert-wadi Kareem (EDC). Some mineralogical and morphological characterizations are executed. The reactivity arrangement of green samples is Baharia>Aswan>EDC. The presence of magnetite decreased reactivity of EDC. The reducibility of the Aswan sample is lower than Baharia due to the presence of agglomerated metallic grain surrounded by semi-melted phases. Specimens are annealed at 1000ᵒC for 3 hours. After firing, the reducibility of Aswan becomes the lowest due to the formation of fayalite and calcium phosphate phases. The relative attitude for green and fired samples reduced at different conditions are studied. For thermal and top areas, the reactivity of fired samples is greater than green ones, which were confirmed by morphological examinations.

Keywords: reducibility, low grade, iron industry, blast furnace

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Authors: Z. Herceg, V. Stulic, T. Vukusic, A. Rezek Jambrak

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Gas phase plasma treatment is a new nonthermal technology used for food and water decontamination. In this study, we have investigated influence of the gas phase plasma treatment on yeast cells of S. cerevisiae. Sample was composed of 10 mL of yeast suspension and 190 mL of 0.01 M NaNO₃ with a medium conductivity of 100 µS/cm. Samples were treated in a glass reactor with a point- to-plate electrode configuration (high voltage electrode-titanium wire in the gas phase and grounded electrode in the liquid phase). Air or argon were injected into the headspace of the reactor at the gas flow of 5 L/min. Frequency of 60, 90 and 120 Hz, time of 5 and 10 min and positive polarity were defined parameters. Inactivation was higher with the applied higher frequency, longer treatment time and injected argon. Inactivation was not complete which resulted in complete recovery. Cellular leakage (260 nm and 280 nm) was higher with a longer treatment time and higher frequency. Leakage at 280 nm which defines a leakage of proteins was higher than leakage at 260 nm which defines a leakage of nucleic acids. The authors would like to acknowledge the support by Croatian Science Foundation and research project 'Application of electrical discharge plasma for preservation of liquid foods'.

Keywords: Saccharomyces cerevisiae, inactivation, gas-phase plasma treatment, cellular leakage

Procedia PDF Downloads 179

Authors: McKenna Halverson, Allison Karpyn

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Introduction: The COVID-19 pandemic posed unprecedented challenges for families with young children in the United States. The Special Supplemental Nutrition Program for Women, Infants, and Children (WIC), a federal nutrition assistance program that provides low-income mothers and young children with access to healthy foods (e.g., infant formula, milk, and peanut butter), mitigated some financial challenges for families. However, the U.S. experienced a national infant formula shortage and rising inflation rates during the pandemic, which likely impacted WIC participants’ shopping experiences and well-being. As such, this study aimed to characterize how the COVID-19 pandemic and related events impacted Delaware WIC participants’ in-store benefit redemption experiences and overall well-being. Method: The authors conducted semi-structured interviews with 51 WIC participants in Wilmington, Delaware. Survey measures included demographic questions and open-ended questions regarding participants’ experiences with WIC benefit redemption during the COVID-19 pandemic. Data were analyzed using a hybrid inductive and deductive coding approach. Findings: The COVID-19 pandemic significantly impacted WIC participants’ shopping experiences and well-being. Specifically, participants were forced to alter their shopping behaviors to account for rising food prices (e.g., used coupons, bought less food, used food banks). Additionally, WIC participants experienced significant distress during the national infant formula shortage resulting from difficulty finding formula to feed their children. Participants also struggled with in-store benefit redemption due to inconsistencies in shelf labelling, the WIC app, and low stock of WIC foods. These findings highlight the need to reexamine WIC operations and emergency food response policy in the United States during times of crisis to optimize public health and ensure federal nutrition assistance programs meeting the needs of low-income families with young children.

Keywords: benefit redemption, COVID-19 pandemic, infant formula shortage, inflation, shopping, WIC

Procedia PDF Downloads 58

Authors: Abhishek Pandey

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Capping and lamination are the most common tablet defects with poorly compressible drugs the common example of that Ibuprofen and Acetaminophen. Generally both these drugs are compressed by wet granulation method which is very time consuming process Ibuprofen and Acetaminophen is widely used as prescription & non-prescription medicine. Ibuprofen mainly used in the treatment of mild to moderate pain related to headache, migraine, postoperative condition and in the management of spondylitis, osteoarthritis Acetaminophen used as an analgesic and antipyretic drug. Ibuprofen having high tendency of sticking to punches of tablet punching machine while Acetaminophen is not ordinarily compressible to tablet formulation because Acetaminophen crystals are very hard and brittle in nature and fracture very easily when compressed producing capping and laminating tablet defects therefore wet granulation method is used to make them compressible. The aim of study was to prepare Ibuprofen and Acetaminophen tablets by direct compression technique and their evaluation. In this Investigation tablets were prepared by using directly compressible grade excipients. Dibasic calcium phosphate, lactose anhydrous (DCL21), microcrystalline cellulose (Avicel PH 101). In order to obtain best or optimize formulation nine different formulations were generated among them batch F5, F6, F7 shows good results and within the acceptable limit. Formulation (F7) selected as optimize product on the basis of evaluation parameters.

Keywords: capping, lamination, tablet defects, direct compression

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Authors: Sajad A. Rather, F. A. Masoodi, Rehana Akhter, S. M. Wani, Adil Gani

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Modern trend towards convenience foods has resulted in increased production and consumption of restructured meat products and are of great importance to the meat industry. In meat products fat plays an important role in cooking properties, texture & sensory scores, however, high fat contents in particular animal fats provide high amounts of saturated fatty acids and cholesterol and are associated with several types of non communicable diseases such as obesity, hypertension and coronary heart diseases. Thus, fat reduction has generally been seen as an important strategy to produce healthier meat products. This study examined the effects of reducing fat level from 20% to 10% and substituting mutton back fat with guar gum (0.5%, 1% & 1.5%) on cooking properties, proximate composition, lipid and protein oxidation, texture, microstructure and sensory characteristics of goshtaba- a traditional meat product of J & K, India were investigated and compared with high fat counterparts. Reduced- fat goshtaba samples containing guar gum had significantly (p ≤ 0.05) higher yield, less shrinkage, more moisture retention and more protein content than the control sample. TBARs and protein oxidation (carbonyl content) values of the control was significantly (p ≤ 0.05) higher than reduced fat goshtaba samples and showed a positive correlation between lipid and protein oxidation. Hardness, gumminess & chewiness of the control (20%) were significantly higher than reduced fat goshtaba samples. Microstructural differences were significant (p ≤ 0.05) between control and treated samples due to an increased moisture content in the reduced fat samples. Sensory evaluation showed significant (p ≤ 0.05) reduction in texture, flavour and overall acceptability scores of treatment products; however the scores for 0.5% and 1% treated samples were in the range of acceptability. Guar gum may also be used as a source of soluble dietary fibre in food products and a number of clinical studies have shown a reduction in postprandial glycemia and insulinemia on consumption of guar gum, with the mechanism being attributed to an increased transit time in the stomach and small intestine, which may have been due to the viscosity of the meal hindering the access of glucose to the epithelium.

Keywords: goshtaba, guar gum, traditional, fat reduction, acceptability

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Authors: Cheba Ben Amar

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Bacillus and Paenibacillus are Gram-positive aerobic endospore-forming bacteria (AEFB) and most abundant in the rhizosphere, they mediated plant growth promotion and disease protection by several complex and interrelated processes involving direct and indirect mechanisms that include nitrogen fixation, phosphate solubilization, siderophores production, phytohormones production and plant diseases control. In addition to their multiple PGPR properties, high secretory capacity, spore forming ability and spore resistance to unfavorable conditions enabling their extended commercial applications for long shelf-life. Due to these unique advantages, Bacillus species were the most an ideal candidate for developing efficient PGPR products such as biopesticides, fungicides and fertilizers. This review list all studied and reported plant growth promoting Bacillus species and strains, discuss their capacities to enhance plant growth and protection with special focusing on the most frequent species Bacillus subtilis, B. pumilus ,B. megaterium, B. amyloliquefaciens , B. licheniformis and B. sphaericus, furthermore we recapitulate the beneficial activities and mechanisms of several species and strains of the genus Paenibacillus involved in plant growth stimulation and plant disease control.

Keywords: bacillus, paenibacillus, PGPR, beneficial activities, mechanisms, growth promotion, disease control, agrobiotechnology

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Authors: A. Falenski, A. Kaesbohrer, M. Filter

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Introduction: Risk assessments can be performed in various ways and in different degrees of complexity. In order to assess risks associated with imported foods additional information needs to be taken into account compared to a risk assessment on regional products. The present review is an overview on currently available best practise approaches and data sources used for food import risk assessments (IRAs). Methods: A literature review has been performed. PubMed was searched for articles about food IRAs published in the years 2004 to 2014 (English and German texts only, search string “(English [la] OR German [la]) (2004:2014 [dp]) import [ti] risk”). Titles and abstracts were screened for import risks in the context of IRAs. The finally selected publications were analysed according to a predefined questionnaire extracting the following information: risk assessment guidelines followed, modelling methods used, data and software applied, existence of an analysis of uncertainty and variability. IRAs cited in these publications were also included in the analysis. Results: The PubMed search resulted in 49 publications, 17 of which contained information about import risks and risk assessments. Within these 19 cross references were identified to be of interest for the present study. These included original articles, reviews and guidelines. At least one of the guidelines of the World Organisation for Animal Health (OIE) and the Codex Alimentarius Commission were referenced in any of the IRAs, either for import of animals or for imports concerning foods, respectively. Interestingly, also a combination of both was used to assess the risk associated with the import of live animals serving as the source of food. Methods ranged from full quantitative IRAs using probabilistic models and dose-response models to qualitative IRA in which decision trees or severity tables were set up using parameter estimations based on expert opinions. Calculations were done using @Risk, R or Excel. Most heterogeneous was the type of data used, ranging from general information on imported goods (food, live animals) to pathogen prevalence in the country of origin. These data were either publicly available in databases or lists (e.g., OIE WAHID and Handystatus II, FAOSTAT, Eurostat, TRACES), accessible on a national level (e.g., herd information) or only open to a small group of people (flight passenger import data at national airport customs office). In the IRAs, an uncertainty analysis has been mentioned in some cases, but calculations have been performed only in a few cases. Conclusion: The current state-of-the-art in the assessment of risks of imported foods is characterized by a great heterogeneity in relation to general methodology and data used. Often information is gathered on a case-by-case basis and reformatted by hand in order to perform the IRA. This analysis therefore illustrates the need for a flexible, modular framework supporting the connection of existing data sources with data analysis and modelling tools. Such an infrastructure could pave the way to IRA workflows applicable ad-hoc, e.g. in case of a crisis situation.

Keywords: import risk assessment, review, tools, food import

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Authors: Mohamad Saab, Florent Real, Francois Virot, Laurent Cantrel, Valerie Vallet

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All spent nuclear fuel reprocessing plants use the PUREX process (Plutonium Uranium Refining by Extraction), which is a liquid-liquid extraction method. The organic extracting solvent is a mixture of tri-n-butyl phosphate (TBP) and hydrocarbon solvent such as hydrogenated tetra-propylene (TPH). By chemical complexation, uranium and plutonium (from spent fuel dissolved in nitric acid solution), are separated from fission products and minor actinides. During a normal extraction operation, uranium is extracted in the organic phase as the UO₂(NO₃)₂(TBP)₂ complex. The TBP solvent can form an explosive mixture called red oil when it comes in contact with nitric acid. The formation of this unstable organic phase originates from the reaction between TBP and its degradation products on the one hand, and nitric acid, its derivatives and heavy metal nitrate complexes on the other hand. The decomposition of the red oil can lead to violent explosive thermal runaway. These hazards are at the origin of several accidents such as the two in the United States in 1953 and 1975 (Savannah River) and, more recently, the one in Russia in 1993 (Tomsk). This raises the question of the exothermicity of reactions that involve TBP and all other degradation products, and calls for a better knowledge of the underlying chemical phenomena. A simulation tool (Alambic) is currently being developed at IRSN that integrates thermal and kinetic functions related to the deterioration of uranyl nitrates in organic and aqueous phases, but not of the n-butyl phosphate. To include them in the modeling scheme, there is an urgent need to obtain the thermodynamic and kinetic functions governing the deterioration processes in liquid phase. However, little is known about the thermodynamic properties, like standard enthalpies of formation, of the n-butyl phosphate molecules and of the UO₂(NO₃)₂(TBP)₂ UO₂(NO₃)₂(HDBP)(TBP) and UO₂(NO₃)₂(HDBP)₂ complexes. In this work, we propose to estimate the thermodynamic properties with Quantum Methods (QM). Thus, in the first part of our project, we focused on the mono, di, and tri-butyl complexes. Quantum chemical calculations have been performed to study several reactions leading to the formation of mono-(H₂MBP), di-(HDBP), and TBP in gas and liquid phases. In the gas phase, the optimal structures of all species were optimized using the B3LYP density functional. Triple-ζ def2-TZVP basis sets were used for all atoms. All geometries were optimized in the gas-phase, and the corresponding harmonic frequencies were used without scaling to compute the vibrational partition functions at 298.15 K and 0.1 Mpa. Accurate single point energies were calculated using the efficient localized LCCSD(T) method to the complete basis set limit. Whenever species in the liquid phase are considered, solvent effects are included with the COSMO-RS continuum model. The standard enthalpies of formation of TBP, HDBP, and H2MBP are finally predicted with an uncertainty of about 15 kJ mol⁻¹. In the second part of this project, we have investigated the fundamental properties of three organic species that mostly contribute to the thermal runaway: UO₂(NO₃)₂(TBP)₂, UO₂(NO₃)₂(HDBP)(TBP), and UO₂(NO₃)₂(HDBP)₂ using the same quantum chemical methods that were used for TBP and its derivatives in both the gas and the liquid phase. We will discuss the structures and thermodynamic properties of all these species.

Keywords: PUREX process, red oils, quantum chemical methods, hydrolysis

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Authors: Salwa Karboune, Amanda Waglay

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Recent studies indicate that health-promoting functional ingredients and nutraceuticals can help support and improve the overall public health, which is timely given the aging of the population and the increasing cost of health care. The development of novel ‘natural’ functional ingredients is increasingly challenging. Biocatalysis offers powerful approaches to achieve this goal. Our recent research has been focusing on the development of innovative biocatalytic approaches towards the isolation of protein isolates from potato by-products and the generation of peptides. Potato is a vegetable whose high-quality proteins are underestimated. In addition to their high proportion in the essential amino acids, potato proteins possess angiotensin-converting enzyme-inhibitory potency, an ability to reduce plasma triglycerides associated with a reduced risk of atherosclerosis, and stimulate the release of the appetite regulating hormone CCK. Potato proteins have long been considered not economically feasible due to the low protein content (27% dry matter) found in tuber (Solanum tuberosum). However, potatoes rank the second largest protein supplying crop grown per hectare following wheat. Potato proteins include patatin (40-45 kDa), protease inhibitors (5-25 kDa), and various high MW proteins. Non-destructive techniques for the extraction of proteins from potato pulp and for the generation of peptides are needed in order to minimize functional losses and enhance quality. A promising approach for isolating the potato proteins was developed, which involves the use of multi-enzymatic systems containing selected glycosyl hydrolase enzymes that synergistically work to open the plant cell wall network. This enzymatic approach is advantageous due to: (1) the use of milder reaction conditions, (2) the high selectivity and specificity of enzymes, (3) the low cost and (4) the ability to market natural ingredients. Another major benefit to this enzymatic approach is the elimination of a costly purification step; indeed, these multi-enzymatic systems have the ability to isolate proteins, while fractionating them due to their specificity and selectivity with minimal proteolytic activities. The isolated proteins were used for the enzymatic generation of active peptides. In addition, they were applied into a reduced gluten cookie formulation as consumers are putting a high demand for easy ready to eat snack foods, with high nutritional quality and limited to no gluten incorporation. The addition of potato protein significantly improved the textural hardness of reduced gluten cookies, more comparable to wheat flour alone. The presentation will focus on our recent ‘proof-of principle’ results illustrating the feasibility and the efficiency of new biocatalytic processes for the production of innovative functional food ingredients, from potato by-products, whose potential health benefits are increasingly being recognized.

Keywords: biocatalytic approaches, functional ingredients, potato proteins, peptides

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Authors: A. Albaridi Najla

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Flaxseed (Linum usitatissimum) is well known to have beneficial effect on health. The seeds are rich in protein, α-linolenic fatty acid and dietary fiber. Bakery products are important part of our daily meals. Functional food recently received considerable attention among consumers. The increase in bread daily consumption leads to the production of breads with functional ingredients such as flaxseed The aim of this Study was to improve the nutritional value of bread by adding flaxseed flour and assessing the effect of adding 0, 5, 10 and 15% flaxseed on whole wheat bread rheological and sensorial properties. The total consumer's acceptability of the flaxseed bread was assessed. Dough characteristics were determined using Farinograph (C.W. Brabender® Instruments, Inc). The result shows no change was observed in water absorption between the stander dough (without flaxseed) and the bread with flaxseed (67%). An Increase in the peak time and dough stickiness was observed with the increase in flaxseed level. Further, breads were evaluated for sensory parameters, colour and texture. High flaxseed level increased the bread crumb softness. Bread with 5% flaxseed was optimized for total sensory evaluation. Overall, flaxseed bread produced in this study was highly acceptable for daily consumption as a functional foods with a potentially health benefits.

Keywords: bread, flaxseed, rheological properties, whole-wheat bread

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Authors: Amir Amiri, Naghmeh Morakabati

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During the recent years the six-fold growth of cancer in Iran has led the production of healthy products to become a challenge in the food industry. Due to the young population in the country, the consumption of fast foods is growing. The chemical cancer-causing preservatives are used to produce these products more than the standard; so using an appropriate alternative seems to be important. On the one hand, the plant essential oils show the high antimicrobial potential against pathogenic and spoilage microorganisms and on the other hand they are highly volatile and decomposed under the processing conditions. The study aims to produce the loaded chitosan nanoparticles with different concentrations of savory essential oil to improve the anti-microbial property and increase the resistance of essential oil to oxygen and heat. The encapsulation efficiency was obtained in the range of 32.07% to 39.93% and the particle size distribution of the samples was observed in the range of 159 to 210 nm. The range of Zeta potential was obtained between -11.9 to -23.1 mV. The essential oil loaded in chitosan showed stronger antifungal activity against Rhizopus stolonifer. The results showed that the antioxidant property is directly related to the concentration of loaded essential oil so that the antioxidant property increases by increasing the concentration of essential oil. In general, it seems that the savory essential oil loaded in chitosan particles can be used as a food processor.

Keywords: chitosan, encapsulation, essential oil, nanogel

Procedia PDF Downloads 249

Authors: Nasser A. Al-Shabib

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Food allergens are most common non-native form when exposed to the immune system. Most food proteins undergo various treatments (e.g. thermal or proteolytic processing) during food manufacturing. Such treatments have the potential to impact the chemical structure of food allergens so as to convert them to more denatured or unfolded forms. The conformational changes in the proteins may affect the allergenicity of treated-allergens. However, most allergenic proteins possess high resistance against thermal modification or digestive enzymes. In the present study, ovomucoid (a major allergenic protein of egg white) was heated in phosphate-buffered saline (pH 7.4) at different temperatures, aqueous solutions and on different surfaces for various times. The results indicated that different antibody-based methods had different sensitivities in detecting the heated ovomucoid. When using one particular immunoassay‚ the immunoreactivity of ovomucoid increased rapidly after heating in water whereas immunoreactivity declined after heating in alkaline buffer (pH 10). Ovomucoid appeared more immunoreactive when dissolved in PBS (pH 7.4) and heated on a stainless steel surface. To the best of our knowledge‚ this is the first time that antibody-based methods have been applied for the detection of ovomucoid adsorbed onto different surfaces under various conditions. The results obtained suggest that use of antibodies to detect ovomucoid after food processing may be problematic. False assurance will be given with the use of inappropriate‚ non-validated immunoassays such as those available commercially as ‘Swab’ tests. A greater understanding of antibody-protein interaction after processing of a protein is required.

Keywords: ovomucoid, thermal treatment, solutions, surfaces

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Authors: A. Mohamed, A. El-Aziz

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The pH of the body plays a great role in the degradation kinetics of biodegradable Mg-Ca orthopedic implants. At the location of fracture, the pH of the body becomes no longer neutral which draws the attention towards studying a range of different pH values of the body fluid. In this study, the pH of Hank’s balanced salt solution (HBSS) was modified by phosphate buffers into an aggressive acidic pH 1.8, a slightly acidic pH 5.3 and an alkaline pH 8.1. The biodegradation of Mg-0.8Ca implant was tested in those three different media using immersion test and electrochemical polarization means. It was proposed that the degradation rate has increased with decreasing the pH of HBSS. The immersion test revealed weight gain for all the samples followed by weight loss as the immersion time increased. The highest weight gain was pronounced for the acidic pH 1.8 and the least weight gain was observed for the alkaline pH 8.1. This was in agreement with the electrochemical polarization test results where the degradation rate was found to be high (7.29 ± 2.2 mm/year) in the aggressive acidic solution of pH 1.8 and relatively minimum (0.31 ± 0.06 mm/year) in the alkaline medium of pH 8.1. Furthermore, it was confirmed that the pH of HBSS has reached a steady state of an alkaline pH (~pH 11) at the end of the two-month immersion period regardless of the initial pH of the solution. Finally, the corrosion products formed on the samples’ surface were investigated by SEM, EDX and XRD analyses that revealed the formation of magnesium and calcium phosphates with different morphologies according to the pH.

Keywords: biodegradable, electrochemical polarization means, orthopedics, immersion test, simulated body fluid

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Authors: Samira Melki, Moncef Gueddari

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In Tunisia, solid wastes storage continue to be uncontrolled. It is eliminated by land raising without any protection measurement against water table and soil contamination. Several industries are located in Sfax area, especially those of the Tunisian Chemical Group (TCG) for the enrichment and transformation of phosphate. The activity of the TCG focuses primarily on the production of chemical fertilizers and phosphoric acid, by transforming natural phosphates. This production generates gaseous emissions, liquid discharges and huge amounts of phosphogypsum (PG) stored directly on the soil surface. Groundwater samples were collected from Tunisian Chemical Group (TCG) site, to assess the effects of phosphogypsum leatchate on groundwater quality. The measurements of various physicochemical parameters including heavy metals (Al, Fe, Zn and F) and stable isotopes of the water molecule (¹⁸O, ²H) were determined in groundwater samples and are reported. The moderately high concentrations of SO₄⁼, Ortho-P, NH₄⁺ Al and F⁻ in groundwater particularly near to the phosphogypsum storage site, likely indicate that groundwater quality is being significantly affected by leachate percolation. The effect of distance of the piezometers from the pollution source was also investigated. The isotopic data of water molecule, showed that the waters of the Sfax-Agreb aquifer amount to recent-evaporation induced rainfall.

Keywords: phosphogypsum leatchate, groundwater quality, pollution, stable isotopes, Sfax-Agareb, Tunisia

Procedia PDF Downloads 169

Authors: B. Hamidu, A. Abdulhamid, S. Mohammed, S. Idi

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This study examined the household expenditure pattern on animal products in Bauchi metropolis. A cross-sectional data were collected from 157 households using systematic sampling technique. The data were analyzed using descriptive statistics, correlation and regression models. The results reveal that the mean age, mean household size, mean monthly income and mean total expenditure on animal products were found to be 39 years, 7 persons, N28,749 and N1,740 respectively. It was also found that household monthly income, number of children and educational level of the household heads (P<0.01) significantly influence the level of household expenditure on animal products. Similarly, income was found to be the most important factor determining the proportion of total expenditure on animal products (20.91%). Income elasticity was found to be 0.66 indicating that for every 1% increase in income, expenditure on animal products would increase by 0.66%. Furthermore, beef was found to be the most preferred (54.83%) and most regularly consumed (61.84%) animal products. However, it was discovered that the major constraints affecting the consumption of animal products were low-income level of the households (29.85%), high cost of animal products (15.82%) and increase in prices of necessities (15.82%). Therefore to improve household expenditure on animal products per capita real income of the households should be improved through creation of employment opportunities. Also stabilization of market prices of animal products and other foods items of necessities through increased production are recommended.

Keywords: animal products, economics, expenditure, households

Procedia PDF Downloads 220

Authors: J. Gabrielczyk, J. Kluitmann, T. Dammeyer, H. J. Jördening

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High-level expression of proteins in bacteria often causes production of insoluble protein aggregates, called inclusion bodies (IB). They contain mainly one type of protein and offer an easy and efficient way to get purified protein. On the other hand, proteins in IB are normally devoid of function and therefore need a special treatment to become active. Most refolding techniques aim at diluting the solubilizing chaotropic agents. Unfortunately, optimal refolding conditions have to be found empirically for every protein. For large-scale applications, a simple refolding process with high yields and high final enzyme concentrations is still missing. The constructed plasmid pASK-IBA63b containing the sequence of fructosyltransferase (FTF, EC 2.4.1.162) from Bacillus subtilis NCIMB 11871 was transformed into E. coli BL21 (DE3) Rosetta. The bacterium was cultivated in a fed-batch bioreactor. The produced FTF was obtained mainly as IB. For refolding experiments, five different amounts of IBs were solubilized in urea buffer with protein concentration of 0.2-8.5 g/L. Solubilizates were refolded with batch or continuous dialysis. The refolding yield was determined by measuring the protein concentration of the clear supernatant before and after the dialysis. Particle size was measured by dynamic light scattering. We tested the solubilization properties of fructosyltransferase IBs. The particle size measurements revealed that the solubilization of the aggregates is achieved at urea concentration of 5M or higher and confirmed by absorption spectroscopy. All results confirm previous investigations that refolding yields are dependent upon initial protein concentration. In batch dialysis, the yields dropped from 67% to 12% and 72% to 19% for continuous dialysis, in relation to initial concentrations from 0.2 to 8.5 g/L. Often used additives such as sucrose and glycerol had no effect on refolding yields. Buffer screening indicated a significant increase in activity but also temperature stability of FTF with citrate/phosphate buffer. By adding citrate to the dialysis buffer, we were able to increase the refolding yields to 82-47% in batch and 90-74% in the continuous process. Further experiments showed that in general, higher ionic strength of buffers had major impact on refolding yields; doubling the buffer concentration increased the yields up to threefold. Finally, we achieved corresponding high refolding yields by reducing the chamber volume by 75% and the amount of buffer needed. The refolded enzyme had an optimal activity of 12.5±0.3 x104 units/g. However, detailed experiments with native FTF revealed a reaggregation of the molecules and loss in specific activity depending on the enzyme concentration and particle size. For that reason, we actually focus on developing a process of simultaneous enzyme refolding and immobilization. The results of this study show a new approach in finding optimal refolding conditions for inclusion bodies at high concentrations. Straightforward buffer screening and increase of the ionic strength can optimize the refolding yield of the target protein by 400%. Gentle removal of chaotrope with continuous dialysis increases the yields by an additional 65%, independent of the refolding buffer applied. In general time is the crucial parameter for successful refolding of solubilized proteins.

Keywords: dialysis, inclusion body, refolding, solubilization

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Authors: Wayde Veldman, Ozlem T. Bishop, Igor Polikarpov

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In bacteria, mono and disaccharides are phosphorylated during uptake into the cell via the widely used phosphoenolpyruvate (PEP)-dependent phosphotransferase transport system. As an initial step in the phosphorylated disaccharide metabolism pathway, certain glycoside hydrolase family 1 (GH1) enzymes play a crucial role in releasing phosphorylated and non-phosphorylated monosaccharides. However, structural determinants for the specificity of these enzymes still need to be clarified. GH1 enzymes are known to have a wide array of functions. According to the CAZy database, there are twenty-one different enzymatic activities in the GH1 family. Here, the structure and substrate specificity of a GH1 enzyme from Bacillus licheniformis, hereafter known as BlBglH, was investigated. The sequence of the enzyme BlBglH was compared to the sequences of other characterized GH1 enzymes using sequence alignment, sequence identity calculations, phylogenetic analysis, and motif discovery. Through these various analyses, BlBglH was found to have sequence features characteristic of the 6-phospho-β-glucosidase activity enzymes. Additionally, motif and structure comparisons of the three most commonly studied GH1 enzyme-activities revealed a shared loop amongst the different structures that consist of different sequence motifs – this loop is thought to guide specific substrates (depending on activity) towards the active-site. To further affirm BlBglH enzyme activity, molecular docking and molecular dynamics simulations were performed. Docking was carried out using 6-phospho-β-glucosidase enzyme-activity positive (p-Nitrophenyl-beta-D-glucoside-6-phosphate) and negative (p-Nitrophenyl-beta-D-galactoside-6-phosphate) control ligands, followed by 400 ns molecular dynamics simulations. The positive-control ligand maintained favourable interactions within the active site until the end of the simulation. The negative-control ligand was observed exiting the enzyme at 287 ns. Binding free energy calculations showed that the positive-control complex had a substantially more favourable binding energy compared to the negative-control complex. Jointly, the findings of this study suggest that the BlBglH enzyme possesses 6-phospho-β-glucosidase enzymatic activity.

Keywords: 6-P-β-glucosidase, glycoside hydrolase 1, molecular dynamics, sequence analysis, substrate specificity

Procedia PDF Downloads 109