Search results for: PadR family protein
4256 The Role of Glutamine-Rich Region of Candida Albicans Tec1p in Mediating Morphological Transition and Invasive Growth
Authors: W. Abu Rayyan, A. Singh, A. M. Al-Jaafreh, W. Abu Dayyih, M. Bustami, S. Salem, N. Seder, K. Schröppel
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Hyphal growth and the transcriptional regulation to the host environment are key issues during the pathogenesis of C. albicans. Tec1p is the C. albicans homolog of a TEA transcription factor family, which share a conserved DNA-binding TEA domain in their N-terminal. In order to define a structure-function relationship of the C. albicans Tec1p protein, we constructed several mutations on the N terminal, C terminal or in the TEA binding domain itself by homologous recombination technology. The modifications in the open reading frame of TEC1 were tested for reconstitution of the morphogenetic development of the tec1/tec1 mutant strain CaAS12. Mutation in the TEA consensus sequence did not confer transition to hyphae whereas the reconstitution of the full-length Tec1p has reconstituted hyphal development. A deletion in one of glutamine-rich regions either in the Tec1p N-terminal or the C-terminal in regions of 53-212 or 637–744 aa, respectively, did not restore morphological development in mutant CaAS12 strain. Whereas, the reconstitution with Tec1p mutants other than the glutamate-rich region has restored the morphogenetic switch. Additionally, the deletion of the glutamine-rich region has attenuated the invasive growth and the heat shock resistance of C. albicans. In conclusion, we show that a glutamine-rich region of Tec1p is essential for the hyphal development and mediating adaptation to the host environment of C. albicans.Keywords: Candida albicans, morphogenetic development, TEA domain, hyphal formation, TEC1
Procedia PDF Downloads 1454255 Airborne Pollutants and Lung Surfactant: Biophysical Impacts of Surface Oxidation Reactions
Authors: Sahana Selladurai, Christine DeWolf
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Lung surfactant comprises a lipid-protein film that coats the alveolar surface and serves to prevent alveolar collapse upon repeated breathing cycles. Exposure of lung surfactant to high concentrations of airborne pollutants, for example tropospheric ozone in smog, can chemically modify the lipid and protein components. These chemical changes can impact the film functionality by decreasing the film’s collapse pressure (minimum surface tension attainable), altering it is mechanical and flow properties and modifying lipid reservoir formation essential for re-spreading of the film during the inhalation process. In this study, we use Langmuir monolayers spread at the air-water interface as model membranes where the compression and expansion of the film mimics the breathing cycle. The impact of ozone exposure on model lung surfactant films is measured using a Langmuir film balance, Brewster angle microscopy and a pendant drop tensiometer as a function of film and sub-phase composition. The oxidized films are analyzed using mass spectrometry where lipid and protein oxidation products are observed. Oxidation is shown to reduce surface activity, alter line tension (and film morphology) and in some cases visibly reduce the viscoelastic properties of the film when compared to controls. These reductions in functionality of the films are highly dependent on film and sub-phase composition, where for example, the effect of oxidation is more pronounced when using a physiologically relevant buffer as opposed to water as the sub-phase. These findings can lead to a better understanding on the impact of continuous exposure to high levels of ozone on the mechanical process of breathing, as well as understanding the roles of certain lung surfactant components in this process.Keywords: lung surfactant, oxidation, ozone, viscoelasticity
Procedia PDF Downloads 3114254 Recovery of Food Waste: Production of Dog Food
Authors: K. Nazan Turhan, Tuğçe Ersan
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The population of the world is approximately 8 billion, and it increases uncontrollably and irrepressibly, leading to an increase in consumption. This situation causes crucial problems, and food waste is one of these. The Food and Agriculture Organization of the United Nations (FAO) defines food waste as the discarding or alternative utilization of food that is safe and nutritious for the consumption of humans along the entire food supply chain, from primary production to end household consumer level. In addition, according to the estimation of FAO, one-third of all food produced for human consumption is lost or wasted worldwide every year. Wasting food endangers natural resources and causes hunger. For instance, excessive amounts of food waste cause greenhouse gas emissions, contributing to global warming. Therefore, waste management has been gaining significance in the last few decades at both local and global levels due to the expected scarcity of resources for the increasing population of the world. There are several ways to recover food waste. According to the United States Environmental Protection Agency’s Food Recovery Hierarchy, food waste recovery ways are source reduction, feeding hungry people, feeding animals, industrial uses, composting, and landfill/incineration from the most preferred to the least preferred, respectively. Bioethanol, biodiesel, biogas, agricultural fertilizer and animal feed can be obtained from food waste that is generated by different food industries. In this project, feeding animals was selected as a food waste recovery method and food waste of a plant was used to provide ingredient uniformity. Grasshoppers were used as a protein source. In other words, the project was performed to develop a dog food product by recovery of the plant’s food waste after following some steps. The collected food waste and purchased grasshoppers were sterilized, dried and pulverized. Then, they were all mixed with 60 g agar-agar solution (4%w/v). 3 different aromas were added, separately to the samples to enhance flavour quality. Since there are differences in the required amounts of different species of dogs, fulfilling all nutritional needs is one of the problems. In other words, there is a wide range of nutritional needs in terms of carbohydrates, protein, fat, sodium, calcium, and so on. Furthermore, the requirements differ depending on age, gender, weight, height, and species. Therefore, the product that was developed contains average amounts of each substance so as not to cause any deficiency or surplus. On the other hand, it contains more protein than similar products in the market. The product was evaluated in terms of contamination and nutritional content. For contamination risk, detection of E. coli and Salmonella experiments were performed, and the results were negative. For the nutritional value test, protein content analysis was done. The protein contents of different samples vary between 33.68% and 26.07%. In addition, water activity analysis was performed, and the water activity (aw) values of different samples ranged between 0.2456 and 0.4145.Keywords: food waste, dog food, animal nutrition, food waste recovery
Procedia PDF Downloads 644253 Tailorability of Poly(Aspartic Acid)/BSA Complex by Self-Assembling in Aqueous Solutions
Authors: Loredana E. Nita, Aurica P. Chiriac, Elena Stoleru, Alina Diaconu, Tudorachi Nita
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Self-assembly processes are an attractive method to form new and complex structures between macromolecular compounds to be used for specific applications. In this context, intramolecular and intermolecular bonds play a key role during self-assembling processes in preparation of carrier systems of bioactive substances. Polyelectrolyte complexes (PECs) are formed through electrostatic interactions, and though they are significantly below of the covalent linkages in their strength, these complexes are sufficiently stable owing to the association processes. The relative ease way of PECs formation makes from them a versatile tool for preparation of various materials, with properties that can be tuned by adjusting several parameters, such as the chemical composition and structure of polyelectrolytes, pH and ionic strength of solutions, temperature and post-treatment procedures. For example, protein-polyelectrolyte complexes (PPCs) are playing an important role in various chemical and biological processes, such as protein separation, enzyme stabilization and polymer drug delivery systems. The present investigation is focused on evaluation of the PPC formation between a synthetic polypeptide (poly(aspartic acid) – PAS) and a natural protein (bovine serum albumin - BSA). The PPC obtained from PAS and BSA in different ratio was investigated by corroboration of various techniques of characterization as: spectroscopy, microscopy, thermo-gravimetric analysis, DLS and zeta potential determination, measurements which were performed in static and/or dynamic conditions. The static contact angle of the sample films was also determined in order to evaluate the changes brought upon surface free energy of the prepared PPCs in interdependence with the complexes composition. The evolution of hydrodynamic diameter and zeta potential of the PPC, recorded in situ, confirm changes of both co-partners conformation, a 1/1 ratio between protein and polyelectrolyte being benefit for the preparation of a stable PPC. Also, the study evidenced the dependence of PPC formation on the temperature of preparation. Thus, at low temperatures the PPC is formed with compact structure, small dimension and hydrodynamic diameter, close to those of BSA. The behavior at thermal treatment of the prepared PPCs is in agreement with the composition of the complexes. From the contact angle determination results the increase of the PPC films cohesion, which is higher than that of BSA films. Also, a higher hydrophobicity corresponds to the new PPC films denoting a good adhesion of the red blood cells onto the surface of PSA/BSA interpenetrated systems. The SEM investigation evidenced as well the specific internal structure of PPC concretized in phases with different size and shape in interdependence with the interpolymer mixture composition.Keywords: polyelectrolyte – protein complex, bovine serum albumin, poly(aspartic acid), self-assembly
Procedia PDF Downloads 2464252 A Systematic Review on Factors/Predictors and Outcomes of Parental Distress in Childhood Acute Lymphoblastic Leukemia
Authors: Ana Ferraz, Martim Santos, M. Graça Pereira
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Distress among parents of children with acute lymphoblastic leukemia (ALL) is common during treatment and can persist several years post-diagnosis, impacting the adjustment of children and parents themselves. Current evidence is needed to examine the scope and nature of parental distress in childhood ALL. This review focused on associated variables, predictors, and outcomes of parental distress following their ALL diagnosis of their child. PubMed, Web of Science, and PsycINFO databases were searched for English and Spanish papers published from 1983 to 2021. PRISMA statement was followed, and papers were evaluated through a standardized methodological quality assessment tool (NHLBI). Of the 28 papers included, 16 were evaluated as fair, eight as good, and four as poor. Regarding results, 11 papers reported subgroup differences, and 15 found potential predictors of parental distress, including sociodemographic, psychosocial, psychological, family, health, and ALL-specific variables. Significant correlations were found between parental distress, social support, illness cognitions, and resilience, as well as contradictory results regarding the impact of sociodemographic variables on parental distress. Family cohesion and caregiver burden were associated with distress, and the use of healthy coping strategies was associated with less anxiety. Caregiver strain contributed to distress, and the overall impact of illness positively predicted anxiety in mothers and somatization in fathers. Differences in parental distress were found regarding group risk, time since diagnosis, and treatment phases. Thirteen papers explored the outcomes of parental distress on psychological, family, health, and social/education outcomes. Parental distress was the most important predictor of family strain. Significant correlations were found between parental distress at diagnosis and further psychological adjustment of parents themselves and their children. Most papers reported correlations between parental distress on children’s adjustment and quality of life, although few studies reported no association. Correlations between maternal depression and child participation in education and social life were also found. Longitudinal studies are needed to better understand parental distress and its consequences on health outcomes, in particular. Future interventions should focus mainly on parents on distress reduction and psychological adjustment, both in parents and children over time.Keywords: childhood acute lymphoblastic leukemia, family, parental distress, psychological adjustment, quality of life
Procedia PDF Downloads 1084251 Computational Approach to Identify Novel Chemotherapeutic Agents against Multiple Sclerosis
Authors: Syed Asif Hassan, Tabrej Khan
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Multiple sclerosis (MS) is a chronic demyelinating autoimmune disorder, of the central nervous system (CNS). In the present scenario, the current therapies either do not halt the progression of the disease or have side effects which limit the usage of current Disease Modifying Therapies (DMTs) for a longer period of time. Therefore, keeping the current treatment failure schema, we are focusing on screening novel analogues of the available DMTs that specifically bind and inhibit the Sphingosine1-phosphate receptor1 (S1PR1) thereby hindering the lymphocyte propagation toward CNS. The novel drug-like analogs molecule will decrease the frequency of relapses (recurrence of the symptoms associated with MS) with higher efficacy and lower toxicity to human system. In this study, an integrated approach involving ligand-based virtual screening protocol (Ultrafast Shape Recognition with CREDO Atom Types (USRCAT)) to identify the non-toxic drug like analogs of the approved DMTs were employed. The potency of the drug-like analog molecules to cross the Blood Brain Barrier (BBB) was estimated. Besides, molecular docking and simulation using Auto Dock Vina 1.1.2 and GOLD 3.01 were performed using the X-ray crystal structure of Mtb LprG protein to calculate the affinity and specificity of the analogs with the given LprG protein. The docking results were further confirmed by DSX (DrugScore eXtented), a robust program to evaluate the binding energy of ligands bound to the ligand binding domain of the Mtb LprG lipoprotein. The ligand, which has a higher hypothetical affinity, also has greater negative value. Further, the non-specific ligands were screened out using the structural filter proposed by Baell and Holloway. Based on the USRCAT, Lipinski’s values, toxicity and BBB analysis, the drug-like analogs of fingolimod and BG-12 showed that RTL and CHEMBL1771640, respectively are non-toxic and permeable to BBB. The successful docking and DSX analysis showed that RTL and CHEMBL1771640 could bind to the binding pocket of S1PR1 receptor protein of human with greater affinity than as compared to their parent compound (Fingolimod). In this study, we also found that all the drug-like analogs of the standard MS drugs passed the Bell and Holloway filter.Keywords: antagonist, binding affinity, chemotherapeutics, drug-like, multiple sclerosis, S1PR1 receptor protein
Procedia PDF Downloads 2564250 The Effects of Fearing Cancer in Women
Authors: E. Kotrotsiou, A. S. Topsioti, S. Mantzoukas, E. Dragioti, M. Gouva
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Introduction: The literature has demonstrated that individual and psychological factors have a direct effect on the perceptions and attitudes of women with cancer. Objectives: To investigate the relationship between the fear of cancer and anxiety. Aim: To examine the impact of the fear of cancer in women with state and trait anxiety of women. Methods: A community sample of 286 women (mean age 39.6 years, SD = 9.5 ranged 20-60) participated in the current study. The women completed a) State - Trait Anxiety Inventory (STAI) and b) questionnaire concerning socio-demographic information and questions for fear of cancer. Results: The perception of fear in women with cancer is statistically independent from their age (t–test, p = 0.58), their family status (χ2, p = 0.519), their place of residency (χ2, p = 0.148), the manifestation of gynecological cancer (χ2, p = 0.979) or the manifestation of any type of cancer in the family (χ2, p = 0.277). In contrast, it was observed that there was a dependence in relation to a total of phobias (χ2, p = 0.003), the fear of illness (χ2, p< 0.001) and the fear of heights (χ2, p = 0.004). Furthermore, the participants that responded that they feared cancer displayed greater level of stress both as situation (t=-3.462; p=0.001) and as a trait of their personality (t=-4.377; p<0.001), and at the same time they displayed greater levels of depression in comparisons with the other participants. Furthermore, following multiple linear regression analysis it was observed that the participants that responded positively to the question if they feared cancer had 8, 3 units greater stress level as a personality trait in comparison to women that responded negatively to the question if they feared cancer (B=8.3; p=0.016; R2=0.506). Conclusion: Women’s fear of cancer is statistically independent from their age, family status, place of residency, the manifestation of gynaecological cancer and with the manifestation of cancer any type in the family. In contrast, there is a dependency with the total of phobias, fear of illness and fear of heights. Women that state that they have a fear of cancer manifest greater levels of stress from the rest of the participants both as situation and as a trait of their personality (p = 0.001 and p< 0.001 accordingly). In specific, the study demonstrated that the participants that positively to the question if they feared cancer had 8,3 units greater stress level as a personality trait in comparison to women that responded negatively.Keywords: fear, women health, anxiety, psychology, cancer
Procedia PDF Downloads 2624249 Identification of Common Indicators of Family Environment of Pupils of Alternative Schools
Authors: Yveta Pohnětalová, Veronika Nováková, Lucie Hrašová
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The paper presents the results of research in which we were looking for common characteristics of the family environment of students alternative and innovative education systems. Topicality comes from the fact that nowadays in the Czech Republic there are several civic and parental initiatives held with the aim to establish schools for their children. The goal of our research was to reveal key aspects of these families and to identify their common indicators. Among other things, we were interested what reasons lead parents to decide to enroll their child into different education than standard (common). The survey was qualitative and there were eighteen respondents of parents of alternative schools´ pupils. The reason to implement qualitative design was the opportunity to gain deeper insight into the essence of phenomena and to obtain detailed information, which would become the basis for subsequent quantitative research. There have been semi structured interviews done with the respondents which had been recorded and transcribed. By an analysis of gained data (categorization and by coding), we found out that common indicator of our respondents is higher education and higher economic level. This issue should be at the forefront of the researches because there is lack of analysis which would provide a comparison of common and alternative schools in the Czech Republic especially with regard to quality of education. Based on results, we consider questions whether approaches of these parents towards standard education come from their own experience or from the lack of knowledge of current goals and objectives of education policy of the Czech Republic.Keywords: alternative schools, family environment, quality of education, parents´ approach
Procedia PDF Downloads 3524248 Alternative Animal Feed Additive Obtain with Different Drying Methods from Carrot Unsuitable for Human Consumption
Authors: Rabia Göçmen, Gülşah Kanbur, Sinan Sefa Parlat
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This study was conducted to determine that carrot powder obtain by different drying methods (oven and vacuum-freeze dryer) of carrot unfit for human consumption that whether feed additives in animal nutrition or not. Carrots randomly divided 2 groups. First group was dried by using oven, second group was by using vacuum freeze dryer methods. Dried carrot prepared from fresh carrot was analysed nutrient matter (energy, crude protein, crude oil, crude ash, beta carotene, mineral concentration and colour). The differences between groups in terms of energy, crude protein, ash, Ca and Mg was not significant (P> 0,05). Crude oil, P, beta carotene content and colour values (L, a, b) with vacuum-freeze dryer group was greater than oven group (P<0,05). Consequently, carrot powder obtained by drying the vacuum-freeze dryer method can be used as a source of carotene.Keywords: carrot, vacuum freeze dryer, oven, beta carotene
Procedia PDF Downloads 3244247 Membrane Technologies for Obtaining Bioactive Fractions from Blood Main Protein: An Exploratory Study for Industrial Application
Authors: Fatima Arrutia, Francisco Amador Riera
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The meat industry generates large volumes of blood as a result of meat processing. Several industrial procedures have been implemented in order to treat this by-product, but are focused on the production of low-value products, and in many cases, blood is simply discarded as waste. Besides, in addition to economic interests, there is an environmental concern due to bloodborne pathogens and other chemical contaminants found in blood. Consequently, there is a dire need to find extensive uses for blood that can be both applicable to industrial scale and able to yield high value-added products. Blood has been recognized as an important source of protein. The main blood serum protein in mammals is serum albumin. One of the top trends in food market is functional foods. Among them, bioactive peptides can be obtained from protein sources by microbiological fermentation or enzymatic and chemical hydrolysis. Bioactive peptides are short amino acid sequences that can have a positive impact on health when administered. The main drawback for bioactive peptide production is the high cost of the isolation, purification and characterization techniques (such as chromatography and mass spectrometry) that make unaffordable the scale-up. On the other hand, membrane technologies are very suitable to apply to the industry because they offer a very easy scale-up and are low-cost technologies, compared to other traditional separation methods. In this work, the possibility of obtaining bioactive peptide fractions from serum albumin by means of a simple procedure of only 2 steps (hydrolysis and membrane filtration) was evaluated, as an exploratory study for possible industrial application. The methodology used in this work was, firstly, a tryptic hydrolysis of serum albumin in order to release the peptides from the protein. The protein was previously subjected to a thermal treatment in order to enhance the enzyme cleavage and thus the peptide yield. Then, the obtained hydrolysate was filtered through a nanofiltration/ultrafiltration flat rig at three different pH values with two different membrane materials, so as to compare membrane performance. The corresponding permeates were analyzed by liquid chromatography-tandem mass spectrometry technology in order to obtain the peptide sequences present in each permeate. Finally, different concentrations of every permeate were evaluated for their in vitro antihypertensive and antioxidant activities though ACE-inhibition and DPPH radical scavenging tests. The hydrolysis process with the previous thermal treatment allowed achieving a degree of hydrolysis of the 49.66% of the maximum possible. It was found that peptides were best transmitted to the permeate stream at pH values that corresponded to their isoelectric points. Best selectivity between peptide groups was achieved at basic pH values. Differences in peptide content were found between membranes and also between pH values for the same membrane. The antioxidant activity of all permeates was high compared with the control only for the highest dose. However, antihypertensive activity was best for intermediate concentrations, rather than higher or lower doses. Therefore, although differences between them, all permeates were promising regarding antihypertensive and antioxidant properties.Keywords: bioactive peptides, bovine serum albumin, hydrolysis, membrane filtration
Procedia PDF Downloads 2004246 Nursing System Development in Patients Undergoing Operation in 3C Ward
Authors: Darawan Augsornwan, Artitaya Sabangbal, Maneewan Srijan, Kanokarn Kongpitee, Lalida Petphai, Palakorn Surakunprapha
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Background: Srinagarind Hospital, Ward 3C, has patients with head and neck cancer, congenital urology anomalies such as hypospadis, cleft lip and cleft palate and congenital megacolon who need surgery. Undergoing surgery is a difficult time for patients/ family; they feel fear and anxiety. Nurses work closely with patients and family for 24 hours in the process of patients care, so should have the good nursing ability, innovation and an efficient nursing care system to promote patients self-care ability reducing suffering and preventing complications. From previous nursing outcomes we found patients did not receive appropriate information, could not take care of their wound, not early ambulation after the operation and lost follow-up. Objective: to develop the nursing system for patients who were undergoing an operation. Method: this is a participation action research. The sample population was 11 nurses and 60 patients. This study was divided into 3 phase: Phase 1. Situation review In this phase we review the clinical outcomes, the process of care from documents such as nurses note and interview nurses, patients and family about the process of care by nurses. Phase 2: focus group with 11 nurses, searching guideline for specific care, nursing care system then establish the protocol. This phase we have the protocol for giving information, teaching protocol and teaching record, leaflet for all of top five diseases, make video media to convey information, ambulation package and protocol for patients with head and neck cancer, patients zoning, primary nurse, improved job description for each staff level. Program to record number of patients, kind of medical procedures for showing nurses activity each day. Phase 3 implementation and evaluation. Result: patients/family receive appropriate information about deep breathing exercise, cough, early ambulation after the operation, information during the stay in the hospital. Patients family satisfaction is 95.04 percent, appropriate job description for a practical nurse, nurse aid, and worker. Nurses satisfaction is 95 percent. The complications can be prevented. Conclusion: the nursing system is the dynamic process using evidence to develop nursing care. The appropriate system depends on context and needs to keep an eye on every event.Keywords: development, nursing system, patients undergoing operation, 3C Ward
Procedia PDF Downloads 2654245 Factors of Successful Transition of Individuals with Intellectual Disabilities from School to Employment
Authors: Mubarak S. Aldosari
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Transition of adolescents with mild intellectual disabilities (ID) from secondary level to post-school employment level is a critical step for them and their families. Transition of adolescents with mild ID to post secondary levels faces serious difficulties and challenges. The current research highlighted the important factors related to the success of transition of students with mild ID to post-school employment such as vocational training, Self-determination skills, Social skills, and family involvement.Keywords: adolescents with mild intellectual disabilities, post-school employment, vocational training, self-determination skills, social skills, family involvement
Procedia PDF Downloads 2934244 Effect of Deer Antler Extract on Osteogenic Gene Expression and Longitudinal Bone Growth of Adolescent Male Rats
Authors: Kang-Hyun Leem, Myung-Gyou Kim, Hye Kyung Kim
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Deer antler, traditionally used as a tonic and valuable drug in oriental medicine, has been considered to possess bone-strengthening activity. The upper section, mid section, and base of the antler has been known to exhibit different biological properties. Present study was performed to examine the effects of different parts of deer antler extract (DH) on osteogenic gene expressions in MG-63 cells and longitudinal bone growth in adolescent male rats. The expressions of osteogenic genes, collagen, alkaline phosphatase, osteocalcin, and osteopontin, were measured by quantitative real-time PCR. Longitudinal bone growth was measured in 3-week-old male Sprague-Dawley rats using fluorescence microscopy. To examine the effects on the growth plate metabolism, the total height of growth plate and bone morphogenetic protein-2 (BMP-2) were measured. Collagen and osteocalcin mRNA expressions were increased by all three parts of the DH treatment while osteopontin gene expression was not affected by any of the DH treatment. Alkaline phosphatase gene expression was increased by upper and mid part of DH while base part of DH fails to affect alkaline phosphatase gene expression. The upper and mid parts of the DH treatment enhanced longitudinal bone growth and total height of growth plate. The induction of BMP-2 protein expression in growth plate assessed by immunostaining was also promoted by upper and mid parts of the DH treatment. These results suggest that DH, especially upper and mid parts, stimulate osteogenic gene expressions and have the effect on bone growth in adolescent rats and might be used for the growth delayed adolescent and inherent growth failure patient.Keywords: bone morphogenetic protein-2, deer antler, longitudinal bone growth, osteogenic genes
Procedia PDF Downloads 3794243 Development of Monoclonal Antibodies against the Acute Hepatopancreatic Necrosis Disease Toxins
Authors: Naveen Kumar B. T., Anuj Tyagi, Niraj Kumar Singh, Visanu Boonyawiwat, Shanthanagouda A. H., Orawan Boodde, Shankar K. M., Prakash Patil, Shubhkaramjeet Kaur
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Since 2009, Acute Hepatopancreatic Necrosis Disease (AHPND) outbreaks have increased rapidly, and these have led to the major economic losses to the global shrimp industry. In comparison to other treatments, passive immunity and monoclonal antibody (MAb) based farmer level kit have proved their importance in controlling and treating the diseases in the shrimp industry. In the present study, MAbs were produced against the recombinant PirB protein Vibrio parahaemolyticus strain causing AHPND. Briefly, Balb/C mice were immunized with rPirB at 15 days interval, and antibody titer was determined by ELISA. Spleen cells from mice showing high antibody titer were fused with SP2O myeloma cells for hybridoma production. Among 130 hybridomas, four showed high antibody titer and positive reactivity in an immunoblot assay. In Western blot assay, three out of four MAbs (4C4, 2C2 and 4G3) showed reactivity to rPirB protein. However, in the natural host, only Mab clone 4G3 show strong reactivity (with a strain of V. parahemolyticus causing EMS/AHPND). These clones also showed reactivity with less than 20 kDa proteins in AHPND free V. parahaemolyticus (Thailand stain). Further, on from MAb 4G3 clone, four panels of single cell MAbs clones (G3F5, G3B8, G3H2, and G3D6) were produced of which three showed strong positive reactivity to rPirB protein in the Western blot. These MAbs have potential for controlling and prevention of the AHPND through passive immunity and development of filed level rapid diagnostic kits.Keywords: shrimp, economic loss, AHPND, MAb
Procedia PDF Downloads 2534242 Gender Specific Nature of the Fiction Conflict in Modern Feminine Prose
Authors: Baglan Bazylova
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The purpose of our article is to consider the social and psychological conflicts in Lyudmila Petrushevskaya’s stories as an artistic presentation of gender structure of modern society; to reveal originality of the characters’ inner world, the models of their behavior expressing the gender specific nature of modern feminine prose. Gender conflicts have taken the leading place in the modern prose. L. Petrushevskaya represents different types of conflicts including those which are shown in the images of real contradictions in the stories "Narratrix", "Thanks to Life”, "Virgin's Case", "Father and Mother". In the prose of Petrushevskaya the gender conflicts come out in two dimensions: The first one is love relations between a man and a woman. Because of the financial indigence, neediness a woman can’t afford herself even to fall in love and arrange her family happiness. The second dimension is the family conflict because of the male adultery. Petrushevskaya fixed on the unmanifistated conflict in detail. In the real life such gender conflict can appear in different forms but for the writer is important to show it as a life basis, hidden behind the externally safe facade of “the family happiness”. In the stories of L. Petrushevskaya the conflicts reflect the common character of the social and historical situations in which her heroines find themselves, in situations where a woman feels her opposition to the customary mode of life. The types of gender conflicts of these stories differ in character of verbal images. They are presented by the verbal and event ranks creating the conflicts just in operation.Keywords: gender behavior of heroes, gender conflict, gender picture of the world, gender structure
Procedia PDF Downloads 5104241 Differentially Expressed Protein Biomarkers in Early and Advanced Stage Young Triple-Negative Breast Cancer Patients
Authors: Shamim Mushtaq, Moazzam Shahid
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Breast cancer (BC) claims the lives of half a million women every year and is the most common cause of death in the developing world. In 2019, it was estimated that BC alone accounts for 15% of all cancer deaths in younger women (aged < 45 years old) with advanced-stage lung metastasis. According to the World Health Organization & International Union against Cancer, in Asia, a high number of cancer-related deaths will be observed in 2020, whereas the burden will be reduced in Western countries due to awareness about the disease, better health facilities and advanced treatments. In the last 15 years, it has been reported that the incidence of BC has increased by 1.1% among Asian compared to the US population from 2003 to 2012. To date, several BC biological subtypes have been reported so far, which are associated with different treatment responses. The heterogeneity and diversity of BC reflected these different subtypes, including Luminal A (23.7% prevalence) and B (38.8% prevalence) that have pathological estrogen receptor (ER+)-positive tumors, the human epidermal growth factor receptor 2 (HER2) (11.2% prevalence) and triple-negative breast cancer (TNBC) (25% prevalence). According to Shaukat Khanum Memorial Cancer Hospital and Research Centre – Pakistan, ten years of data showed that among 636 BC patients, 30.5% had TNBC who were <40 years of age, which is an extremely alarming situation. Therefore, there is a dire need to explore and develop therapeutic targets for the treatment of early TNBC. Since the last decade, unfortunately, there has been little success in understanding the complexity of TNBC and in discovering new biological therapeutic targets. However, conventional chemotherapy is the only choice of treatment for TNBC patients. Many investigators revealed advances in multi-omics (multiple "omes", e.g., genome, proteome, transcriptome, epigenome, and microbiome) which were later identified as actionable targets and increased prevalence in TNBC patients. However, various drugs have been identified so far which are related to a particular diagnostic and prognostic biomarker. For example, Epidermal growth factor receptor ( EGFR or ErbB-1), HER-2/neu (ErbB-2), HER-3 (ErbB-3), and HER-4 (ErbB-4). Protein Transglin-2 (TAGLN 2 ) and Profilins-1 (Pfn-1 ) are the ubiquitously expressed large family of proteins present in all eukaryotes, enabling actin cytoskeletal reorganization. It is known that the oncogenic transformation of cells is accompanied by alteration in the actin cytoskeleton. There are causal connections between altered expression of actin cytoskeletal regulators and cancer progression. Our case-control study identified TAGLN-2 and Pfn-1 proteins in TNBC blood by mass spectrometry. Both TAGLN-2 and Pfn-1 proteins are differentially expressed in early and advanced stages of TNBS patients, which could be potential predictors or therapeutic targets for TNBC.Keywords: TNBC, blood biomarkers, mass spectrometry, qPCR, ELISA
Procedia PDF Downloads 434240 Interaction of Glycolipid S-TGA-1 with Bacteriorhodopsin and Its Functional Role
Authors: Masataka Inada, Masanao Kinoshita, Nobuaki Matsumori
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It has been demonstrated that lipid molecules in biological membranes are responsible for the functionalization and structuration of membrane proteins. However, it is still unclear how the interaction of lipid molecules with membrane proteins is correlated with the function of the membrane proteins. Here we first developed an evaluation method for the interaction between membrane proteins and lipid molecules via surface plasmon resonance (SPR) analysis. Bacteriorhodopsin (bR), which was obtained by the culture of halobacteria, was used as a membrane protein. We prepared SPR sensor chips covered with self-assembled monolayer containing mercaptocarboxylic acids, and immobilized bR onto them. Then, we evaluated the interactions with various lipids that have different structures. As a result, the halobacterium-specific glycolipid S-TGA-1 was found to have much higher affinity with bRs than other lipids. This is probably due to not only hydrophobic and electrostatic interactions but also hydrogen bonds with sugar moieties in the glycolipid. Next, we analyzed the roles of the lipid in the structuration and functionalization of bR. CD analysis showed that S-TGA-1 could promote trimerization of bR monomers more efficiently than any other lipids. Flash photolysis further indicated that bR trimers formed by S-TGA-1 reproduced the photocyclic activity of bR in purple membrane, halobacterium-membrane. These results suggest that S-TGA-1 promotes trimerization of bR through strong interactions and consequently fulfills the bR’s function efficiently.Keywords: membrane protein, lipid, interaction, bacteriorhodopsin, glycolipid
Procedia PDF Downloads 2534239 Analysing Representations of ‘Leftover’ Women in Chinese Media: Taking the Film ‘The Last Woman Standing’ and ‘I Do’ as Examples
Authors: Ting Li Liu
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‘Leftover woman’ or ‘3S’ woman is the term used to describe a well-educated, high income, independent woman who is single and never married around 30 years in Chinese society. With the naming of this demographic of ‘leftover women’, their family, dating culture, mate selection and marriage attract public concern. Massive media representations of ‘leftover women’ occur daily; the research aims to present several media representations of women’s anxiety about their singlehood and related marital issues around thirty. The research triangulates two areas of media representation of ‘leftover women’: films and audience reviews on ‘Douban Movie’ website. Drawing on traditional media studies, Fairclough’s critical discourse analysis combined with multimodal techniques is applied to the research to analyze the representations of ‘leftover women’ and their implications for marital culture in China, in conjunction with a feminist perspective. The conference paper will discuss two case studies: the film ‘The last woman standing’ and ‘I Do’. Paying attention to different aspects of ‘leftover women’, the research aims to re-examine the representations of ‘leftover women’ in selected scenes, such as their age anxiety, family, marriage, dating process, careers, etc. The paper also includes public beliefs about ‘leftover women’ from online audience reviews. In conclusion, the emergence of ‘leftover women’ is a reflection of Chinese tradition’s impact on people’s lives and new changes in Chinese families and their attitude to marriage.Keywords: leftover women, marriage, family, media culture, China
Procedia PDF Downloads 2544238 Effects of Anti-FGL2 Monoclonal Antibody SPF89 on Vascular Inflammation
Authors: Ying Sun, Biao Cheng, Qing Lu, Xuefei Tao, Xiaoyu Lai, Cheng Guo, Dan Wang
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Fibrinogen-like protein 2 (FGL2) has recently been identified to play an important role in inflammatory diseases such as atherosclerosis through a thrombin-dependent manner. Here, a murine monoclonal antibody was raised against the critical residue Ser(89) of FGL2, and the effects of the anti-FGL2 mAb (SPF89) were analyzed in human umbilical vein endothelial cells (HUVECs) and THP-1 cells. Firstly, it was proved that SPF89, which belongs to the IgG1 subtype with a KD value of 44.5 pM, could specifically show the expression levels of protein FGL2 in different cell lines of known target gene status. The lipopolysaccharide (LPS)-mediated endothelial cell proliferation was significantly inhibited with a decline of phosphorylation nuclear factor-κB (NF-κB) in a dose-dependent manner after SPF89 treatment. Furthermore, SPF89 reduced LPS-induced expression of adhesion molecules and inflammatory cytokines such as vascular cell adhesion molecule-1, tumor necrosis factor-α, Matrix metalloproteinase MMP-2, Integrin αvβ3, and interleukin-6 in HUVECs. In macrophage-like THP-1 cells, SPF89 effectively inhibited LPS and low-density lipoprotein-induced foam cell formation. However, these anti-inflammatory and anti-atherosclerotic effects of anti-FGL2 mAb in HUVECs and THP-1 cells were significantly reduced after treatment with an NF-κB inhibitor PDTC. All the above suggest, by efficiently inhibiting LPS-induced pro-inflammatory effects in vascular endothelial cells by attenuating NF-κB dependent pathway, the new anti-FGL2 mAb SPF89 could to be a potential therapeutic candidate for protecting the vascular endothelium against inflammatory diseases such as atherosclerosis. This work was supported by the Program of Sichuan Science and Technology Department (2017FZ0069) and Collaborative Innovation Program of Sichuan for Elderly Care and Health(YLZBZ1511).Keywords: monoclonal antibody, fibrinogen like protein 2, inflammation, endothelial cells
Procedia PDF Downloads 2714237 Production of Keratinase and Its Insilico Characterization
Authors: Akshita Bhardwaj
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Keratinase is an enzyme obtained from extracellular sources that is involved in biodegradation of keratin. It is a member of a group of proteases that can break down keratin into amino acids. Keratinases are produced only in the presence of substrate that contain keratin. It attacked the disulfide bond of substrate and involve in keratin degradation. Human hair, feathers, animal hard tissues, horns, claws, and hooves all contain keratin.. It exists in two form alpha keratin (found in soft tissues) and beta keratin (found in hard tissue). By taking part in the degradation of keratin, keratinases derived from microbial sources, often referred to as microbial keratinases, are important in the process of turning wastes containing keratin into products with added value. Chicken feathers contain high level of keratin protein content than other sources and became a suitable protein source. Keratinase production occurs at near alkaline pH and thermophilic temperatures. The bioprocessing of keratinous waste benefits greatly from the use of keratinases. Additionally, it lessens the issue caused by poultry excrement. The use of feather meal, along with keratinase, improves the digestion of proteins and amino acids.Keywords: mili litre (ml), micro litre (Ul), TCA - trichloroacetic acid, OD - optical density
Procedia PDF Downloads 774236 Nutritive Potential of Mealworm (Tenebrio molitor) in the Diet of Olive Flounder (Paralichthys olivaceus)
Authors: Joo-min Kim, Gi-wook Shin, Tae-ho Chung, Chul Park, Seong-hyun Kim, Namjung Kim
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Mealworm (Tenebrio molitor) was evaluated to investigate the effect of partial or total replacement of fish meal in diets for olive flounder, Paralichthys olivaceus. Experimental groups of fish with average initial body weight (287.5 ± 7.24 g) were fed each with 4 isonitrogeneous (52% crude protein) diets formulated to include 0, 7, 17 and 27% (diets 1 to 4, respectively) of fish meal substituted with mealworm. After six weeks of feeding trials, fish fed with diet 3 revealed the highest values for live weight gain(42.10), specific growth rates (0.445 ± 0.089) as well as better feed conversion ratio (12.08) compared to the other group with statistically significant manner (p<0.05). Hepatosomatic index was showed no significant difference in diet 3 compared to the control group. An increase in weight gain and other growth associated parameters was observed in diet 3. These results clearly indicate that 17% of fish meal protein in bastard halibut diet can be replaced by mealworm not only without any adverse effect but also the effect of promoting growth performance.Keywords: mealworm, olive flounder, Paralichthys olivaceus, Tenebrio molitor
Procedia PDF Downloads 4004235 From “Deafness” to “Aphasia”: A Study of Social Retreat of Families With Hearing-Impaired Children. Interviews with 12 Mothers of Hearing-Impaired Children in Beijing
Authors: Yang Jiayue, Liang Junwen
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Case interviews with 12 mothers of hearing-impaired children in Beijing found that families with hard-of-hearing children make all-out efforts for rescue and rehabilitation until they exhaust family resources, they travel to work places, homes, and rehabilitation institutions every day, and social relations gradually shrink inward. In the stigmatized social environment, children with hearing impairment find it difficult to integrate into regular communication and gradually return to a silent world; their parents also suffer from social discrimination and affiliate stigma, and they gradually reduce contact with others. Finally, families with hearing-impaired children become invisible people in society; the "deafness" of children leads families to "loss of voice."Keywords: family with hearing-impaired children, social retreat, social exclusion, stigma
Procedia PDF Downloads 744234 Effects of Novel Protease Enzyme From Bacillus subtilis on Low Protein and Low Energy Guar Meal (Cyamopsis tetragonoloba) Meal Based Diets on Performance and Nutrients Digestibility in Broilers
Authors: Aqeel Ahmed Shad, Tanveer Ahmad, Muhammad Farooq Iqbal, Muhammad Javaid Asad
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The supplemental effects of novel protease produced from Bacillus subtilis K-5 and beta-mannanase were evaluated on growth performance, carcass characteristics, nutrients digestibility, blood profile and intestinal morphometry of broilers fed guar meal (Cyamopsis tetragonoloba) based diets with reduced Crude Protein (CP), Essential Amino Acids (EAAs), and Metabolizable energy (ME) contents. One-day old Ross 308 broiler chicks (n=360) were randomly allotted to thirty six experimental units in a way that each of the nine dietary treatments received four replicates with ten birds per replicate. A control diet without guar meal (0GM) was formulated with standard nutrient specifications of Ross 308 for the starter and finisher phases. Two negative control diets, one with 5% (5GM) and second with 10% (10GM) guar meal, were formulated with reduction of 5% CP, 5% EAAs and 80 Kcal/kg ME. These three basal diets (no enzyme) were supplemented with novel protease enzyme (PROT) and commercial beta-mannanase (Beta-M) enzyme. The birds were reared up to 35d of age. The data on weekly body weight gain (BWG) and feed intake were recorded to compute feed:gain for the starter (0-21d) and finisher (22-35d) phases. At the end of 35d of experimental period, four birds per experimental unit were randomly selected for blood samples collection and later slaughtered for ileal digesta, intestinal tract and carcass trait sampling. The data on overall performance (1-35d) indicated improved (P<0.05) BWG and feed:gain in birds supplemented with PROT (1.41% and 1.67) and Beta-M (2.79% and 1.64) than non-supplemented groups. Improved (P<0.05) carcass yield, breast meat yield and thigh meat yield were noted with the supplementation of Beta-M. However, non-significant (P>0.05) effect on carcass traits was noted in broiler fed guar meal based PROT supplemented diets. Crude protein digestibility, nitrogen retention (Nret) and apparent digestibility coefficient for nitrogen (ADCN) were improved (P<0.05) only with PROT. The improvement in apparent metabolizable energy (AME) and apparent metabolizable energy corrected for nitrogen (AMEn) was noted (P<0.05) with both supplemented enzymes. However, no effect (P>0.05) of enzyme addition was noted on blood glucose, total protein and cholesterol. Improved villus height of duodenum, jejunum and ileum was noted (P<0.05) with the addition of both enzymes. The EAAs digestibility was improved (P<0.05) only with PROT. In conclusion, beta-mannanase and protease supplementation better improved the overall bird performance in low nutrient profile guar meal based diets than non-supplemented diets.Keywords: novel protease, guar meal, broilers, low protein diets, low metabolizable energy diets, nutrients digestibility
Procedia PDF Downloads 624233 Attachment and Self Esteem among Adolescents of Separated Parents
Authors: Aswathy Sampath
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The study examined the levels of self esteem and attachment among adolescents of divorced and non-divorced parents. Adolescent is a period which is most prodigious yet stressful period of development in a human’s life hence it is important to study the effects on them. The study was conducted on total 60 adolescents, 30 in each group, from the area of Trivandrum, Kerala as it is the top rated in the number of divorce cases in India. The data was collected using Rosenberg’s self esteem scale and IPPA (father, mother and peer) The results of this study were analyzed using t test and found that there is no significance difference in the level of self esteem and attachment (father, mother and peer). This is due to the cultural elements that give support to the individual and also the type of family as it is much different from the west. Although, there was no significant result, there were higher mean scores in the attachment towards peer for children who are from separated family background or in other words adolescents whose parents were divorced. This tells us the essence of social support.Keywords: adolescent, attachment, self esteem, separation
Procedia PDF Downloads 3864232 Critical Thinking Index of College Students
Authors: Helen Frialde-Dupale
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Critical thinking Index (CTI) of 150 third year college students from five State Colleges and Universities (SUCs) in Region I were determined. Only students with Grade Point Average (GPA) of at least 2.0 from four general classification of degree courses, namely: Education, Arts and Sciences, Engineering and Agriculture were included. Specific problem No.1 dealt with the profile variables, namely: age, sex, degree course, monthly family income, number of siblings, high school graduated from, grade point average, personality type, highest educational attainment of parents, and occupation of parents. Problem No. 2 determined the critical thinking index among the respondents. Problem No. 3 investigated whether or not there are significant differences in the critical thinking index among the respondents across the profile variables. While problem No.4 determined whether or not there are significant relationship between the critical thinking index and selected profile variables, namely: age, monthly family income, number of siblings, and grade point average of the respondents. Finally, on problem No. 5, the critical thinking instrument which obtained the lowest rates, were used as basis for outlining an intervention program for enhancing critical thinking index (CTI) of students. The following null hypotheses were tested at 0.05 level of significance: there are no significant differences in the critical thinking index of the third college students across the profile variables; there are no significant relationships between the critical thinking index of the respondents and selected variables, namely: age, monthly family income, number of siblings, and grade point average.Keywords: attitude as critical thinker, critical thinking applied, critical thinking index, self-perception as critical thinker
Procedia PDF Downloads 5174231 Revealing Potential Drug Targets against Proto-Oncogene Wnt10B by Comparative Molecular Docking
Authors: Shazia Mannan, Zunera Khalid, Hammad-Ul-Mubeen
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Wingless type Mouse mammary tumor virus (MMTV) Integration site-10B (Wnt10B) is an important member of the Wnt protein family that functions as cellular messenger in paracrine manner. Aberrant Wnt10B activity is the cause of several abnormalities including cancers of breast, cervix, liver, gastric tract, esophagus, pancreas as well as physiological problems like obesity, and osteoporosis. The objective of this study was to determine the possible inhibitors against aberrant expression of Wnt10B in order to prevent and treat the physiological disorders associated with it. Wnt10B3D structure was predicted by using comparative modeling and then analyzed by PROCHECK, Verify3D, and Errat. The model having 84.54% quality value was selected and acylated to satisfy the hydrophobic nature of Wnt10B. For search of inhibitors, virtual screening was performed on Natural Products (NP) database. The compounds were filtered and ligand-based screening was performed using the antagonist for mouse Wnt-3A. This resulted in a library of 272 unique compounds having most potent drug like activities for Wnt-4. Out of the 271 molecules analyzed three small molecules ZINC35442871, ZINC85876388, and ZINC00754234 having activity against Wnt4 abbarent expression were found common through docking experiment of Wnt10B. It is concluded that the three molecules ZINC35442871, ZINC85876388, and ZINC00754234 can be considered as lead compounds for performing further drug designing experiments against aberrant Wnt expressions.Keywords: Wnt10B inhibitors, comparative computational studies, proto-oncogene, molecular docking
Procedia PDF Downloads 1564230 Variation in Total Iron and Zinc Concentration, Protein Quality, and Quantity of Maize Hybrids Grown under Abiotic Stress and Optimal Conditions
Authors: Tesfaye Walle Mekonnen
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Maize is one of the most important staple food crops for most low-income households in the Sub-Saharan (SSA). Combined heat and drought stress is the major production threats that reduce the yield potential of biofortified maize and restrain various macro and micronutrient deficiencies highly prevalent in low-income people who rely solely on maize-based diets, SSA. This problem can be alleviated by crossing the biofortified inbred lines with different nutritional attributes, Fe, Zn, Protein, and Provitamin A, and developing agronomically superior and stable multi-nutrient maize of various genetic backgrounds. This aimed to understand the correlation between biofortified inbred lines per se and hybrid performance under combined heat and drought stress conditions (CSC). The experiment was conducted at CIMMYT, Zimbabwe, using α-lattice design with three replications. The hybrid effect was highly significant for zein fractions (α-, β-, γ- and δ-zein) zinc, (Zn), and iron (Fe) provitamin A, phytic acid, and grain yield. Under CSC, Fe, Zn concentration, provitamin A in grain and grain yield of hybrids were significantly decreased, however, the zein fraction content and phytic acid content increases in grain were increased under CSC. The phenotypic correlation between grain yield with Zn, Fe concentration, and Provitamin A in grain was strongly positive and higher under CSC than in well-watered conditions. The present investigation confirmed that under CSC, Fe, and Zn-enhanced hybrids could be forecasted to a certain scope based on the performance of and scientifically selected for desirable grain yield and related traits with CSC tolerance during hybrid development programs. In conclusion, the development of high-yielding and micronutrient-dense maize variety is possible under CSC, which could reduce the highly prevalent micronutrient in SSA.Keywords: drought, Fe, heat, maize, protein, zein fractions, Zn
Procedia PDF Downloads 664229 Assuming the Decision of Having One (More) Child: The New Dimensions of the Post Communist Romanian Family
Authors: Horea-Serban Raluca-Ioana, Istrate Marinela
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The first part of the paper analyzes the dynamics of the total fertility rate both at the national and regional level, pointing out the regional disparities in the distribution of this indicator. At the same time, we also focus on the collapse of the number of live births, on the changes in the fertility rate by birth rank, as well as on the failure of acquiring the desired number of children. The second part of the study centres upon a survey applied to urban families with 3 and more than 3 offspring. The preliminary analysis highlights the fact that an increased fertility (more than 3rd rank) is triggered by the parents’ above the average material condition and superior education. The current situation of Romania, which is still passing through a period of relatively rapid demographic changes, marked by numerous convulsions, requires a new approach, in compliance with the recent interpretations appropriate to a new post-transitional demographic regime.Keywords: fertility rate, family size intention, third birth rank, regional disparities
Procedia PDF Downloads 3274228 Disruption of MoNUC1 Gene Mediates Conidiation in Magnaporthe oryzae
Authors: Irshad Ali Khan, Jian-Ping Lu, Xiao-Hong Liu, Fu-Cheng Lin
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This study reports the functional analysis of a gene MoNUC1 in M. oryzae, which is homologous to the Saccharomyces cerevisiae NUC1 encoding a mitochondrial nuclease protein. The MoNUC1 having a gene locus MGG_05324 is 1002-bp in length and encodes an identical protein of 333 amino acids. We disrupted the gene through gene disruption strategy and isolated two mutants confirmed by southern blotting. The deleted mutants were then used for phenotypic studies and their phenotypes were compared to those of the Guy-11 strain. The mutants were first grown on CM medium to find the effect of MoNUC1 gene disruption on colony growth and the mutants were found to show normal culture colony growth similar to that of the Guy-11 strain. Conidial germination and appressorial formation were also similar in both the mutants and Guy-11 strains showing that this gene plays no significant role in these phenotypes. For pathogenicity, the mutants and Guy-11 mycelium blocks were inoculated on blast susceptible barley seedlings and it was found that both the strains exhibited full pathogenicity showing coalesced and necrotic blast lesions suggesting that this gene is not involved in pathogenicity. Mating of the mutants with 2539 strain formed numerous perithecia showing that MoNUC1 is not essential for sexual reproduction in M. oryzae. However, the mutants were found to form reduced conidia (1.06±8.03B and 1.08±9.80B) than those of the Guy-11 strain (1.46±10.61A) and we conclude that this protein is not required for the blast fungus to cause pathogenicity but plays significant role in conidiation. Proteins of signal transduction pathways that could be disrupted/ intervened genetically or chemically could lead to antifungal products of important fungal cereal diseases and reduce rice yield losses. Tipping the balance toward understanding the whole of pathogenesis, rather than simply conidiation will take some time, but clearly presents the most exciting challenge of all.Keywords: appressorium formation, conidiation, NUC1, Magnaporthe oryzae, pathogenicity
Procedia PDF Downloads 4994227 Gold-Mediated Modification of Apoferritin Surface with Targeting Antibodies
Authors: Simona Dostalova, Pavel Kopel, Marketa Vaculovicova, Vojtech Adam, Rene Kizek
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Protein apoferritin seems to be a very promising structure for use as a nanocarrier. It is prepared from intracellular ferritin protein naturally found in most organisms. The role of ferritin proteins is to store and transport ferrous ions. Apoferritin is a hollow protein cage without ferrous ions that can be prepared from ferritin by reduction with thioglycolic acid or dithionite. The structure of apoferritin is composed of 24 protein subunits, creating a sphere with 12 nm in diameter. The inner cavity has a diameter of 8 nm. The drug encapsulation process is based on the response of apoferritin structure to the pH changes of surrounding solution. In low pH, apoferritin is disassembled into individual subunits and its structure is “opened”. It can then be mixed with any desired cytotoxic drug and after adjustment of pH back to neutral the subunits are reconnected again and the drug is encapsulated within the apoferritin particles. Excess drug molecules can be removed by dialysis. The receptors for apoferritin, SCARA5 and TfR1 can be found in the membrane of both healthy and cancer cells. To enhance the specific targeting of apoferritin nanocarrier, it is possible to modify its surface with targeting moieties, such as antibodies. To ensure sterically correct complex, we used a a peptide linker based on a protein G with N-terminus affinity towards Fc region of antibodies. To connect the peptide to the surface of apoferritin, the C-terminus of peptide was made of cysteine with affinity to gold. The surface of apoferritin with encapsulated doxorubicin (ApoDox) was coated either with gold nanoparticles (ApoDox-Nano) or gold (III) chloride hydrate reduced with sodium borohydride (ApoDox-HAu). The applied amount of gold in form of gold (III) chloride hydrate was 10 times higher than in the case of gold nanoparticles. However, after removal of the excess unbound ions by electrophoretic separation, the concentration of gold on the surface of apoferritin was only 6 times higher for ApoDox-HAu in comparison with ApoDox-Nano. Moreover, the reduction with sodium borohydride caused a loss of doxorubicin fluorescent properties (excitation maximum at 480 nm with emission maximum at 600 nm) and thus its biological activity. Fluorescent properties of ApoDox-Nano were similar to the unmodified ApoDox, therefore it was more suited for the intended use. To evaluate the specificity of apoferritin modified with antibodies, we used ELISA-like method with the surface of microtitration plate wells coated by the antigen (goat anti-human IgG antibodies). To these wells, we applied ApoDox without targeting antibodies and ApoDox-Nano modified with targeting antibodies (human IgG antibodies). The amount of unmodified ApoDox on antigen after incubation and subsequent rinsing with water was 5 times lower than in the case of ApoDox-Nano modified with targeting antibodies. The modification of non-gold ApoDox with antibodies caused no change in its targeting properties. It can therefore be concluded that the demonstrated procedure allows us to create nanocarrier with enhanced targeting properties, suitable for nanomedicine.Keywords: apoferritin, doxorubicin, nanocarrier, targeting antibodies
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