Search results for: shoot initiation

Authors: Andrej Golowin, Viktor Denk, Axel Riepe

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Solid fan blades and discs in aero engines are subjected to high combined low and high cycle fatigue loads especially around the contact areas between blade and disc. Therefore, special coatings (e.g. dry film lubricant) and surface treatments (e.g. shot peening or laser shock peening) are applied to increase the strength with respect to combined cyclic fatigue and fretting fatigue, but also to improve damage tolerance capability. The traditional deterministic damage tolerance assessment based on fracture mechanics analysis, which treats service damage as an initial crack, often gives overly conservative results especially in the presence of vibratory stresses. A probabilistic damage tolerance methodology using crack initiation data has been developed for fan discs exposed to relatively high vibratory stresses in cross- and tail-wind conditions at certain resonance speeds for limited time periods. This Monte-Carlo based method uses a damage databank from similar designs, measured vibration levels at typical aircraft operations and wind conditions and experimental crack initiation data derived from testing of artificially damaged specimens with representative surface treatment under combined fatigue conditions. The proposed methodology leads to a more realistic prediction of the minimum damage tolerance life for the most critical locations applicable to modern fan disc designs.

Keywords: combined fatigue, damage tolerance, engine, surface treatment

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Authors: A. Majeed, M.K. Abbasi, S. Hameed, A. Imran, T. Naqqash, M. K. Hanif

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Plant growth-promoting rhizobacteria (PGPR) are free-living soil bacteria that aggressively colonize the rhizosphere/plant roots, and enhance the growth and yield of plants when applied to seed or crops. Root associated (endophytic and rhizospheric) PGPR were isolated from Sunflower (Helianthus anus) grown in soils collected from 16 different sites of sub division Dhirkot, Poonch, Azad Jammu & Kashmir, Pakistan. A total of 150 bacterial isolates were isolated, purified, screened in vitro for their plant growth promoting (PGP) characteristics. 11 most effective isolates were selected on the basis of biochemical assays (nitrogen fixation, phosphate solubilization, growth hormone production, biocontrol assay, and carbon substrates utilization assay through gas chromatography (GCMS), spectrophotometry, high performance liquid chromatography HPLC, fungal and bacterial dual plate assay and BIOLOG GN2/GP2 microplate assay respectively) and were tested on the crop under controlled and field conditions. From the inoculation assay, the most promising 4 strains (on the basis of increased root/shoot weight, root/shoot length, seed oil content, and seed yield) were than selected for colonization studies through confocal laser scanning and transmission electron microscope. 16Sr RNA gene analysis showed that these bacterial isolates belong to Pseudononas, Enterobacter, Azospirrilum, and Citobacter genera. This study is the clear evident that such isolates have the potential for application as inoculants adapted to poor soils and local crops to minimize the chemical fertilizers harmful for soil and environment

Keywords: PGPR, nitrogen fixation, phosphate solubilization, colonization

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Authors: Ireneusz Majsterek, Dariusz Pytel, J. Alan Diehl

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PKR-like ER kinase (PERK) is serine/threonie endoplasmic reticulum (ER) transmembrane kinase activated during ER-stress. PERK can activate signaling pathways known as unfolded protein response (UPR). Attenuation of translation is mediated by PERK via phosphorylation of eukaryotic initiation factor 2α (eIF2α), which is necessary for translation initiation. PERK activation also directly contributes to activation of Nrf2 which regulates expression of anti-oxidant enzymes. An increased phosphorylation of eIF2α has been reported in Alzheimer disease (AD) patient hippocampus, indicating that PERK is activated in this disease. Recent data have revealed activation of PERK signaling in non-Hodgkins lymphomas. Results also revealed that loss of PERK limits mammary tumor cell growth in vitro and in vivo. Consistent with these observations, activation of UPR in vitro increases levels of the amyloid precursor protein (APP), the peptide from which beta-amyloid plaques (AB) fragments are derived. Finally, proteolytic processing of APP, including the cleavages that produce AB, largely occurs in the ER, and localization coincident with PERK activity. Thus, we expect that PERK-dependent signaling is critical for progression of many types of diseases (human cancer, neurodegenerative disease and other). Therefore, modulation of PERK activity may be a useful therapeutic target in the treatment of different diseases that fail to respond to traditional chemotherapeutic strategies, including Alzheimer’s disease. Our goal will be to developed therapeutic modalities targeting PERK activity.

Keywords: PERK kinase, small molecule inhibitor, neurodegenerative disease, Alzheimer’s disease

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Authors: Maria Manzoor, Iram Gul, Muhammad Arshad, Jean Kallerhoff

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The potential of fungal isolates to be used in phytoremediation of widespread lead contaminated soil has been evaluated in this study. Five different fungal isolates (Trichoderma harzianum, Penicillium simplicissimum, Aspergillus flavus, Aspergillus niger and Mucor spp.) were obtained and tested for their tolerance to increasing concentration of lead (Pb) i.e. 100, 200, 300, 400 and 500 mgL-1 on PDA and PDB culture experiment. All strains were tolerant up to 500 mgL-1 following sequence; A. flavus > A. niger > Mucor spp. > P. simplicissimum > T. harzianum. Further the isolates were then monitored for possible effect on Pb solubility/mobility through soil incubation experiments and characterized for essays including pathogenicity, germination and root elongation and plant growth promoting activities including IAA (indole acetic acid), phosphorus solubilization and gibberellic acid (GA3) production. Results revealed that fungal isolates have positive effect on Pb mobility in soil and plant biomass production. Pb solubility was significantly (P> 0.05) increased in soil upon application of Mucor spp. P. simplicissimum and T. harzianum. when compared to control. Among different strains three isolates (Mucor spp., P. simplicissimum and T. harzianum) were nonpathogenic because no inhibitory effect of fungus was observed to plant growth when exposed to these strains in root shoot elongation essay. Particularly T. harzianum and P. simplicissimum showed great ability to increase root length by 1.1 and 1.3 folds and shoot length by 1.47 and 1.5 folds respectively under Pb stress (500 mgL-1). Significantly high production of IAA was observed in A. niger (26.7 μg/ml), Phosphorus solubilization was observed in T. harzianum (9.15 μg/ml) and GA3 production was observed in P. simplicissimum (11.02 μg/ml). From results it is concluded that Mucor spp., P. simplicissimum and T. harzianum have potential to increase Pb mobility and improving plant growth under highy Pb contamination, therefore can be used in microbially assisted phytoremediation of Pb contaminated soil.

Keywords: Pb tolerant fungus, Pb mobility, plant growth promoting activities, indole acetic acid (IAA)

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Authors: Ajmal Khan Kassi, Humayun Javed, Muhammad Asif Aziz

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Response of different aubergine cultivars against Brinjal shoot and fruit borer (Leucinodes orbonalis Guenee.) was evaluated at research farm of PMAS, Arid Agriculture University, Rawalpindi, during 2013. Field trials were conducted in randomized completed block design with four replications for the screening of five cultivars of Brinjal (Solanum melongena L) (Short Purpal, Singhnath 666, Brinjal long 6275, Round Brinjal 86602, Round Egg Plant White). Cultivar Round White Brinjal showed maximum fruit infestation (54.44%) followed by Singhnath 666 (53.19%), while minimum fruit infestation was observed in Round Brinjal 86602 (42.39%). Cultivar Short Purpal showed maximum larval population (0.43) followed by Round White Brinjal (0.39), while the minimum larval population was observed in Round Brinjal 86602 with (0.27). It was observed that Round Brinjal 86602 cultivar showed comparatively minimum (L. orbonalis) larval population per leaf. The correlation of Brinjal fruit infestation and larval population of (L. orbonalis) with the different environmental factors showed that, the average relative humidity was positively and significantly correlated with fruit infestation on cultivars average precipitation showed positive but non- significant correlation on all the cultivars except Singhnath 666 with the value of (0.79) which was positive and significant. The average temperature showed non-significant and negative correlation with Brinjal long 6275, Round Brinjal 86602 and Singhnath 666, but significant negative correlation with Short Purpal and Round White Brinjal. Maximum temperature also showed the significant and negative correlation on all the five Brinjal cultivars which were significant and highly significant. Minimum temperature showed negative correlation and not significant correlation with all the cultivars. Consequently, based on the (L. orbonalis) larval density and Brinjal fruit infestation, the Round Brinjal 86602 proved least susceptible and Short Purpal highly susceptible cultivar.

Keywords: evaluation, Brinjal (Solanum melongena L), Cultivars, L. orbonalis

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Authors: Pankaj Singh Rawat, Rajeew Kumar, Pradeep Ram, Priyanka Pandey

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Wheat is an important cereal crop for food security. Boosting the wheat production and productivity is the major challenge across the nation. Good quality of seed is required for maintaining optimum plant stand which ultimately increases grain yield. Ensuring a good germination is one of the key steps to ensure proper plant stand and moisture assurance during seed germination may help to speed up the germination. The tiny size of nanoparticles may help in entry of water into seed without disturbing their internal structure. Considering above, a laboratory experiment was conducted during 2012-13 at G.B. Pant University of Agriculture and Technology, Pantnagar, India. The completely randomized design was used for statistical analysis. The experiment was conducted in two phases. In the first phase, the appropriate concentration of nanoparticles for seed treatment was screened. In second phase seed soaking hours of nanoparticles for better seed germination were standardized. Wheat variety UP2526 was taken as test crop. Four nanoparticles (TiO₂, ZnO, nickel and chitosan) were taken for study. The crop germination studies were done in petri dishes and standard package and practices were used to raise the seedlings. The germination studies were done by following standard procedure. In first phase of the experiment, seeds were treated with 50 and 300 ppm of nanoparticles and control was also maintained for comparison. In the second phase of experiment, seeds were soaked for 4 hours, 6 hours and 8 hours with 50 ppm nanoparticles of TiO₂, ZnO, nickel and chitosan along with control treatment to identify the soaking time for better seed germination. Experiment revealed that the application of nanoparticles help to enhance seed germination. The study revealed that seed treatment with  nanoparticles at 50 ppm concentration increases root length, shoot length, seedling length, shoot dry weight, seedling dry weight, seedling vigour index I and seedling vigour index II as compared to seed soaking at 300 ppm concentration. This experiment showed that seed soaking up to 4 hr was better as compared to 6 and 8 hrs. Seed soaking with nanoparticles specially TiO₂, ZnO, and chitosan proved to enhance germination and seedling growth indices of wheat crop.

Keywords: nanoparticles, seed germination, seed soaking, wheat

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Authors: Md Abdullah Al Mamun

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Background: The current situation of Ensuring WHO recommended feeding practices for infant and young children which is becoming a challenge nowadays in many developing countries, especially in areas where household food security is at risk. Because many households of the developing countries often encounter severe food insecurity hence provision of adequate child nutrition is threatened. Aim: The study aimed to assess the child feeding practices of 0-24 months childbearing mothers and explore their household food insecurity in a coastal region of Bangladesh. Methods: This study was conducted in Suborno Char (one of the coastal suburbs in Noakhali District in Bangladesh) from October 2019 to April 2020. A total of 400 mothers were selected with their children of 0-24 months following a cross sectional study sampling procedure of the population. Data were collected through a standard questionnaire and analyzed using statistical tests in SPSS version 20.0.0. Results: The frequency of exclusive breastfeeding, timely initiation of complementary feeding, and giving foods from four food groups to the children were 53.5%, 75.5%, and 22.2%, respectively. Mother's level of education showed a strong association with the child feeding practices of the mothers. Mothers of severely food insecure households showed lower odds in exclusive breastfeeding practice (COR 0.233 at 95% CI 0.083, 0.655; and AOR 0.478 at 95% CI 0.133, 1.713) than mothers of food secured households. Similar results have also been found in case of timely initiation of complementary feeding and minimum dietary diversity of the children.

Keywords: household food insecurity, exclusive breastfeeding, complementary feeding, maternal education, mothers age, household income

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Authors: Rumana Keyani, Haleema Sadia, Asia Nosheen, Rabia Naz, Humaira Yasmin, Sidra Zahoor

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Tomato is a significant crop of the world and is one of the staple foods of Pakistan. A vast number of plant pathogens from simple viruses to complex parasites cause diseases in tomatoes but fungal infection in our country is quite high. Sometimes the symptoms are too harsh destroying the crop altogether. Countries like our own with continuously increasing massive population and limited resources cannot afford such an economic loss. There is an array of morphological, genetic, biochemical and molecular processes involved in plant resistance mechanisms to biotic stress. The study of different metabolic pathways like Jasmonic acid (JA) pathways and most importantly signaling molecules like ROS/RNS and their redoxin enzymes i.e. TRX and NRX is crucial to disease management, contributing to healthy plant growth. So, improving tolerance in crop plants against biotic stresses is a dire need of our country and world as whole. In the current study, fungal pathogenic strains Alternaria solani and Rhizoctonia solani were used to inoculate tomatoes to check the defense responses of tomato plant against these pathogens at molecular as well as phenotypic level with jasmonic acid and spermidine pretreatment. All the growth parameters (root and shoot length, dry and weight root, shoot weight measured 7 days post-inoculation, exhibited that infection drastically declined the growth of the plant whereas jasmonic acid and spermidine assisted the plants to cope up with the infection. Thus, JA and Spermidine treatments maintained comparatively better growth factors. Antioxidant assays and expression analysis through real time quantitative PCR following time course experiment at 24, 48 and 72 hours intervals also exhibited that activation of JA defense genes and a polyamine Spermidine helps in mediating tomato responses against fungal infection when used alone but the two treatments combined mask the effect of each other.

Keywords: fungal infection, jasmonic acid defence, tomato, spermidine

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Authors: S. K. Thind, Aparjot Kaur

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Heat stress is one of the major environmental factors drastically reducing wheat production. Crop heat tolerance can be enhanced by preconditioning of plants by exogenous application of osmoprotectants. Presently, the effect of trehalose pretreatment (at 1 mM, and 1.5 nM) under heat stress of 35±2˚C (moderate) and 40±2˚ (severe) for four and eight hour was conducted in wheat (Tricticum aestivum L.) genotypes viz. HD2967, PBW 175, PBW 343, PBW 621, and PBW 590. Heat stress affects wide spectrum of physiological processes within plants that are irreversibly damaged by stress. Membrane thermal stability (MTS) and cell viability was significantly decreased under heat stress for eight hours. Pretreatment with trehalose improved MTS and cell viability under stress and this effect was more promotory with higher concentration. Thermal stability of photosynthetic apparatus differed markedly between genotypes and Hill reaction activity was recorded more in PBW621 followed by C306 as compared with others. In all genotypes photolysis of water showed decline with increase in temperature stress. Trehalose pretreatment helped in sustaining Hill reaction activity probably by stabilizing the photosynthetic apparatus against heat-induced photo inhibition. Both plant growth and development were affected by temperature in both shoot and root under heat stress. The reduction was compensated partially by trehalose (1.5 mM) application. Adaption to heat stress is associated with the metabolic adjustment which led to accumulation of soluble sugars including non-reducing and reducing for their role in adaptive mechanism. Higher acid invertase activity in shoot of tolerant genotypes appeared to be a characteristic for stress tolerance. As sucrose synthase play central role in sink strength and in studied wheat genotype was positively related to dry matter accumulation. The duration of heat stress for eight hours had more severe effect on these parameters and trehalose application at 1.5 mM ameliorated it to certain extent.

Keywords: heat stress, Triticum aestivum, trehalose, membrane thermal stability, triphenyl tetrazolium chloride, reduction test, growth, sugar metabolism

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Authors: Radheshyam Yadav, Ramakrishna Wusirika

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Plant Growth Promoting Bacteria (PGPB) and Arbuscular Mycorrhizal (AM) Fungi are ubiquitous in soil and often very critical for crop yield and agriculture sustainability, and this has motivated the agricultural practices to support and promote PGPB and AM Fungi in agriculture. PGPB can be involved in a range of processes that affect Nitrogen (N) and Phosphorus (P) transformations in soil and thus influence nutrient availability and uptake to the plants. A field study with two wheat cultivars, HD-3086, and HD-2967 was performed in Malwa region, Bathinda of Punjab, India, to evaluate the effect of native and non-native PGPB alone and in combination with AM fungi as an inoculant on wheat grain yield, nutrient uptake and soil health parameters (dehydrogenase, urease, β‐glucosidase). Our results showed that despite an early insignificant increase in shoot length, plants treated with PGPB (Bacillus sp.) and AM Fungi led to a significant increase in shoot growth at maturity, aboveground biomass, nitrogen (45% - 40%) and phosphorus (40% - 34%) content in wheat grains relative to untreated control plants. Similarly, enhanced grain yield and nutrients uptake i.e. copper (27.15% - 36.25%) iron (43% - 53%) and zinc (44% - 47%) was recorded in PGPB and AM Fungi treated plants relative to untreated control. Overall, inoculation with native PGPB alone and in combination with AM Fungi provided benefits to enhance grain yield, wheat biofortification, and improved soil fertility, despite this effect varied depending on different PGPB isolates and wheat cultivars. These field study results provide evidence of the benefits of agricultural practices involving native PGPB and AM Fungi to the plants. These native strains and AM Fungi increased accumulations of copper, iron, and zinc in wheat grains, enhanced grain yield, and soil fertility.

Keywords: AM Fungi, biofortification, PGPB, soil microbial enzymes

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Authors: Cristina Fernandez, Felix Amparano, John Tumberger, Stephani Stancil, Sarah Hampl, Brooke Sweeney, Amy R. Beck, Helena H Laroche, Jared Tucker, Eileen Chaves, Sara Gould, Matthew Lindquist, Lora Edwards, Renee Arensberg, Meredith Dreyer, Jazmine Cedeno, Alleen Cummins, Jennifer Lisondra, Katie Cox, Kelsey Dean, Rachel Perera, Nicholas A. Clark

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Background: Adolescents with obesity are at higher risk of disordered eating than the general population. Detection of eating disorders (ED) is difficult. Screening questionnaires may aid in early detection of ED. Our team’s prior efforts focused on increasing ED screening rates to ≥90% using a validated 10-question adolescent binge eating disorder screening questionnaire (ADO-BED). This aim was achieved. We then aimed to improve treatment plan initiation of patients ≥12 years of age who screen positive for BED within our WMC from 33% to 70% within 12 months. Methods: Our WMC is within a tertiary-care, free-standing children’s hospital. A3, an improvement framework, was used. A multidisciplinary team (physicians, nurses, registered dietitians, psychologists, and exercise physiologists) was created. The outcome measure was documentation of treatment plan initiation of those who screen positive (goal 70%). The process measure was ADO-BED screening rate of WMC patients (goal ≥90%). Plan-Do-Study-Act (PDSA) cycle 1 included provider education on current literature and treatment plan initiation based upon ADO-BED responses. PDSA 2 involved increasing documentation of treatment plan and retrain process to providers. Pre-defined treatment plans were: 1) repeat screen in 3-6 months, 2) resources provided only, or 3) comprehensive multidisciplinary weight management team evaluation. Run charts monitored impact over time. Results: Within 9 months, 166 patients were seen in WMC. Process measure showed sustained performance above goal (mean 98%). Outcome measure showed special cause improvement from mean of 33% to 100% (n=31). Of treatment plans provided, 45% received Plan 1, 4% Plan 2, and 46% Plan 3. Conclusion: Through a multidisciplinary improvement team approach, we maintained sustained ADO-BED screening performance, and, prior to our 12-month timeline, achieved our project aim. Our efforts may serve as a model for other multidisciplinary WMCs. Next steps may include expanding project scope to other WM programs.

Keywords: obesity, pediatrics, clinic, eating disorder

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Authors: Jose G. Tshikuka, Goabaone Rankgoane-Pono, Mgaywa G. M. D. Magafu, Julius C. Mwita, Tiny Masupe, Fortunat M. Kandanda, Shimeles G. Hamda, Roy Tapera, Mooketsi Molefi, John T. Tlhakanelo

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Background: Factors associated with overweight and obesity among antiretroviral therapy (ART) recipients have not been sufficiently studied in Botswana. We aimed to study (i) the prevalence and trends in overweight/obesity by duration of exposure to ART among recipients, (ii) changes in body mass index (BMI) categories among recipients before ART initiation (BMI-1) and after ART initiation (BMI-2), (iii) associations between ART and overweight/obesity and (iv) factors associated with BMI changes among ART recipients. Methods: A 12 years retrospective record-based review was conducted. Factors potentially associated with BMI change among patients after at least three years of ART exposure were examined using multiple regression model. Adjusted odds ratios (AOR) and their 95% confidence intervals (CIs) were computed. ART regimens, duration of exposure to ART, and recipients’ demographic and biomedical characteristics including the presence or absence of diabetes mellitus related comorbidities (DRC) were investigated as potential factors associated with overweight/obesity. Results: Twenty-nine percent of recipients were overweight, 16.6% had obesity of whom 2.4% were morbidly-obese at the last clinic visit. Overweight/obesity recipients were more likely to be female, to have DRC and less likely to have nadir CD4 count or CD4 count between 201 – 249 cells/mm³. Neither the first-line nor the second-, third-line ART regimens predicted overweight/obesity more than the other and neither did the duration of exposure to ART. No significant linear trends were observed in the prevalence of overweight/obesity by the duration of exposure to ART. Conclusions: These results indicate that overweight/obesity seen among ART recipients is not directly induced by ART. ART used CD4 and/or DRC pathway to induce overweight/obesity seen among recipients; suggesting that, weight gain documented herein is likely a reflection of improved health status that mirrors trends in the general population or a DRC related effect. Weight management programs may be important components of HIV care.

Keywords: overweight/obesity, recipients of antiretroviral therapy, HIV/AIDS, Botswana

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Authors: B. Kenny

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The complications associated with transfusions are well documented, with the serious hazards of transfusion (SHOT) reporting system continuing to report deaths and serious morbidity due to the transfusion of allogenic blood. Many different sources including the TRICC trial, NHMRC and Cochrane recommending similar transfusion triggers/guidelines. Recent studies found the rate of infection (deep infection, wound infection, chest infection, urinary tract infection, and others) were purely a dose response relationship, increasing the Relative Risk by 3.44. It was also noted that each transfused patient stayed in hospital for one additional day. We hypothesise that providing an approved/standardised, guideline with a graphical summary of decision pathways for anaemic patients will reduce unnecessary transfusions. We retrospectively assessed patients undergoing primary knee or hip arthroplasties over a 4 year period, 1459 patients. Of these, 339 (23.24%) patients received allogenic blood transfusions and 858 units of blood were transfused, 9.14% of patients transfused had haemoglobin levels above 100 g/L, 7.67% of patients were transfused without knowing the haemoglobin level, 24 hours prior to transfusion initiation and 4.5% had possible transfusion reactions. Overall, 17% of allogenic transfusions topatients admitted to the Orthopaedic department within a 4 year period were outside NHMRC and Cochrane guidelines/recommendations. If our transfusion frequency is compared with that of other authors/hospitals, transfusion rates are consistently being high. We subsequently implemented a simple guideline for transfusion initiation. This guideline was then assessed. We found the transfusion rate post transfusion implementation to be significantly lower, without increase in patient morbidity or mortalitiy, p <0.001). Transfusion rates and patient outcome can be optimized by a simple graphical aid for decision making.

Keywords: transfusion, morbidity, mortality, neck of femur, fracture, arthroplasty, rehabilitation

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Authors: Juhi Pandya, Elekes Zsuzsanna

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Health is related to every aspect of human being. Even a quintal change leads to ill-health of an individual. Gender plays an eminent role in determining an individual health exposure. Political implications on health have implicit effects on the individual, societal and economical. The inclusion of gender perspective into policies have plunged enormous attention globally, nationally and locally to detract inequalities and achieve economic growth. Simultaneously, there is an initiation of policies with gender perspective which are named differently but hold similar meaning or objective. They are named gender mainstreaming policies or gender sensitization policies. Gender budgeting acts as a tool for the application of gender mainstreaming policies. It incorporates gender perspective into the budgetary process by restricting the revenues and expenditures at all level of the budget. The current study takes into account the analysis of Gender Budgeting reports in terms of healthcare from the 2014-16 year of Gujarat State, India. The expenditures and literature under the heading of gender budgeting reports named “Health and Family Welfare Department” are discussed in the paper. The data analytics is done with the help of reports published by the Gujarat government on Gender Budgeting. The results discuss upon the expenditure and initiation of new policies as a roadmap for the promotion of gender equality from the path of gender budgeting. It states with the escalation of the budgetary numbers for the health expenditure. Additionally, the paper raises the questions on the hypothetical loopholes pertaining to the gender budgeting in Gujarat. The budget reports do not show a specify explanation to the expenditure use of budget for the schemes mentioned in healthcare. It also does not clarify that how many beneficiaries are benefited through gender budget. The explanation just provides an overlook of theory for healthcare Schemes/Yojana or Abhiyan.

Keywords: gender, gender budgeting, gender equality, healthcare

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Authors: S. Padma, D. H. Tejavathi

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Somatic embryogenesis is a remarkable illustration of the dictum of plant totipotency where developmental reconstruction of somatic cells takes place towards the embryogenic pathway. It recapitulates the morphological and developmental process that occurs in zygotic embryogenesis. S. androgynous commonly called as multivitamin plant. The leaves are consumed as green leafy vegetable by the Southeast Asian communities due to their rich nutritional profile. Despite being a good nutritional vegetable with proteins, vitamins, minerals, amino acids, it is warned for excessive intake due to the presence of alkoloid called papaverine. Papaverine at higher concentrations is toxic and leads to a syndrome called Bronchiolitis Obliterans. In the present study, morphogenetic potential of shoot tip, leaf and nodal explants of Sauropus androgynous was investigated to develop and enhance the reliable plant regeneration protocol via somatic embryogenesis. Somatic embryos were derived directly from the embryogenic callus derived from shoot tip, node and leaf cultures on Phillips and Collins (L2) medium supplemented with NAA at various concentrations ranging from 5.3 µM/l to 26.85 µM/l within two months of inoculation. Thus obtained embryos were sub cultured to modified L2 media supplemented with increased vitamin level for the further growth. Somatic embryos with well-developed cotyledons were transferred to normal and modified L2 basal medium for conversion. The plantlets thus obtained were subjected to brief acclimatization before transferring them to land. About 95% of survival rate was recorded. The augmentation process of culturing various explants through somatic embryogenesis using synthetic medium with various plant growth regulators under controlled conditions have aggrandized the commercial production of Sauropus making it easily available over the conventional propagation methods. In addition, regeneration process through somatic embryogenesis has ameliorated the development of desired character in Sauropus with low papaverine content thereby providing a valuable resource to the food and pharmaceutical industry. Based on this research, plant tissue culture techniques have shown promise for economical and convenient application in Sauropus androgynous breeding.

Keywords: L2 medium, multivitamin plant, NAA, papaverine

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Authors: Muhammad Naveed, Sohail Yousaf, Zahir Ahmad Zahir, Birgit Mitter, Angela Sessitsch

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Most bacterial endophytes originate from the soil and enter plants via the roots followed by further spread through the inner tissues. The mechanisms allowing bacteria to colonize plants endophytically are still poorly understood for most bacterial and plant species. Specific bacterial functions are required for plant colonization, but also the plant itself is a determining factor as bacterial ability to establish endophytic populations is very often dependent on the plant genotype (cultivar) and inoculums density. The effect of inoculum density (107, 108, 109 CFU mL-1) of Burkholderia phytofirmans strain PsJN was evaluated on growth and endophytic colonization of different maize and potato cultivars under axenic and natural soil conditions. PsJN inoculation significantly increased maize seedling growth and tuber yield of potato at all inoculum density compared to uninoculated control. Under axenic condition, PsJN inoculation (108 CFU mL-1) significantly improved the germination, root/shoot length and biomass up to 62, 115, 98 and 135% of maize seedling compared to uninoculated control. In case of potato, PsJN inoculation (109 CFU mL-1) showed maximum response and significantly increased root/shoot biomass and tuber yield under natural soil condition. We confirmed that PsJN is able to colonize the rhizosphere, roots and shoots of maize and potato cultivars. The endophytic colonization increased linearly with increasing inoculum density (within a range of 8 x 104 – 3 x 107 CFU mL-1) and were highest for maize (Morignon) and potato (Romina) as compared to other cultivars. Efficient colonization of cv. Morignon and Romina by strain PsJN indicates the specific cultivar colonizing capacity of the bacteria. The findings of the study indicate the non-significant relationship between colonization and plant growth promotion in maize under axenic conditions. However, the inoculum level (109 CFU mL-1) that promoted colonization of rhizosphere and plant interior (endophytic) also best promoted growth and tuber yield of potato under natural soil conditions.

Keywords: crop genotype, inoculum density, Burkholderia phytofirmans PsJN, colonization, growth, potato

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Authors: Semayat Fanta, P.M. Mohite, C.S. Upadhyay

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Damage meso-model for laminates is one of the most widely applicable approaches for the analysis of damage induced in laminated fiber-reinforced polymeric composites. Damage meso-model for laminates has been developed over the last three decades by many researchers in experimental, theoretical, and analytical methods that have been carried out in micromechanics as well as meso-mechanics analysis approaches. It has been fundamentally developed based on the micromechanical description that aims to predict the damage initiation and evolution until the failure of structure in various loading conditions. The current damage meso-model for laminates aimed to act as a bridge between micromechanics and macro-mechanics of the laminated composite structure. This model considers two meso-constituents for the analysis of damage in ply and interface that imparted from low-velocity impact. The damages considered in this study include fiber breakage, matrix cracking, and diffused damage of the lamina, and delamination of the interface. The damage initiation and evolution in laminae can be modeled in terms of damaged strain energy density using damage parameters and the thermodynamic irreversible forces. Interface damage can be modeled with a new concept of spherical micro-void in the resin-rich zone of interface material. The damage evolution is controlled by the damage parameter (d) and the radius of micro-void (r) from the point of damage nucleation to its saturation. The constitutive martial model for meso-constituents is defined in a user material subroutine VUMAT and implemented in ABAQUS/Explicit finite element modeling tool. The model predicts the damages in the meso-constituents level very accurately and is considered the most effective technique of modeling low-velocity impact simulation for laminated composite structures.

Keywords: mesomodel, laminate, low-energy impact, micromechanics

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Authors: Jacques M. Berner, Monja Gerber, Gillian L. Maggs-Kolling, Stuart J. Piketh

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The enigmatic melon species, Acanthosicyos horridus Welw. ex Hook. f., locally known as !nara, is endemic to the hyper-arid Namib Desert, where it thrives in sandy dune areas and dry river banks. The Namib Desert is characterized by extreme weather conditions which include high temperatures, very low rainfall, and extremely dry air. Plant and animals that have made the Namib Dessert their home are dependent on non-rainfall water inputs, like fog, dew and water vapor, for survival. Fog is believed to be the most important non-rainfall water input for most of the coastal Namib Desert and is a life line to many Namib plants and animals. It is commonly assumed that the !nara plant is adapted and dependent upon coastal fog events. The !nara plant shares many comparable adaptive features with other organisms that are known to exploit fog as a source of moisture. These include groove-like structures on the stems and the cone-like structures of thorns. These structures are believed to be the driving forces behind directional water flow that allow plants to take advantage of fog events. The !nara-fog interaction was investigated in this study to determine the dependence of !nara on these fog events, as it would illustrate strategies to benefit from non-rainfall water inputs. The direct water uptake capacity of !nara shoots was investigated through absorption tests. Furthermore, the movement and behavior of fluorescent water droplets on a !nara stem were investigated through time-lapse macrophotography. The shoot water potential was measured to investigate the effect of fog on the water status of !nara stems. These tests were used to determine whether the morphology of !nara has evolved to exploit fog as a non-rainfall water input and whether the !nara plant has adapted physiologically in response to fog. Chlorophyll a fluorescence was used to compare the photochemical efficiency of !nara plants on days with fog events to that on non-foggy days. The results indicate that !nara plants do have the ability to take advantage of fog events as commonly believed. However, the !nara plant did not exhibit visible signs of drought stress and this, together with the strong shoot water potential, indicates that these plants are reliant on permanent underground water sources. Chlorophyll a fluorescence data indicated that temperature stress and wind were some of the main abiotic factors influencing the plants’ overall vitality.

Keywords: Acanthosicyos horridus, chlorophyll a fluorescence, fog, foliar absorption, !nara

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Authors: Dawit Thomas

Abstract:

Feeding and weaning practices vary from culture to culture and depend on different child-rearing values. The socio-cultural dimensions that influence the acceptable infant feeding practices are varied and complex. Understanding cultural differences in beliefs and practices relating to infant feeding is important to enhance designing programs for delivering successful psychological, social, physiological and economic well-being of mothers and infants. The main purpose of this study was exploring mothers infant feeding practices in the context of Hadiyya culture. After purposively selecting Harche Huyaya Uyaya Kebele eight infant feeding mothers were selected using snowball sampling technique. The study employed interviews and focus group discussion. The study found out early initiation and prolonged breastfeeding and early complementary feeding in some instances immediately after birth. In addition, infants were not forced to wean unless the mothers encounter pressing issues like pregnancy and health related problems. Furthermore, the main weaning techniques were putting unpleasant materials on the tip of nipples and sending infants to grandparents home. The study also found out gender difference in weaning, i.e., early initiation of weaning for girls. This can be indicative of gender-based bias on weaning practice. Finally, health extension workers, office of women and children affairs and Hadiyya Zone Tourism office should organize awareness raising programs to preserve vital infant feeding practices like prolonged breastfeeding and length of weaning. In addition, the offices should raise awareness among communities on negative side effects of sending infant to grandparents home that may weaken infant-mothers attachment and create favorable ground for the development of phobia.

Keywords: feeding, infant, practices, weaning

Procedia PDF Downloads 277

Authors: Zhiguo Ju

Abstract:

Both ancient written documents and archaeological studies showed that the ancient Chinese was people who worshiped the Sun God. There is a legend of Dayi in China, however, told a story of Dayi, an ancient hero who sacrificed his life to shoot nine suns out of the ten in the sky, restored the order of the universe, and saved the people’s life. By investigating its oral inheritance and folk customs, we found that the story was originated from Rizhao, the east coralline in east China. This could provide valuable cultural resources and could be used in local tourism. How the Sun-worshipping-people developed such a contradictory legend, how was it be carried by generations, and how to use it as a cultural source to promote the local tourism was discussed.

Keywords: Legend of Dayi, sun worship, ancient Chinese, China

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Authors: M. A. Barros, M. V. Barros

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The aim of this work is to analyze the discursive dynamics of preservice teachers during the implementation of a didactic sequence on topics of Quantum Mechanics for High School. Our research methodology was qualitative, case study type, in which we selected two prospective teachers on the Physics Teacher Training Course of the Sao Carlos Institute of Physics, at the University of Sao Paulo/Brazil. The set of modes of communication analyzed were the intentions and interventions of the teachers, the established communicative approach, the patterns and the contents of the interactions between teachers and students. Data were collected through video recording, interviews and questionnaires conducted before and after an 8 hour mini-course, which was offered to a group of 20 secondary students. As teaching strategy we used an active learning methodology, called: Peer Instruction. The episodes pointed out that both future teachers used interactive dialogic and authoritative communicative approaches to mediate the discussion between peers. In the interactive dialogic dimension the communication pattern was predominantly I-R-F (initiation-response-feedback), in which the future teachers assisted the students in the discussion by providing feedback to their initiations and contributing to the progress of the discussions between peers. Although the interactive dialogic dimension has been preferential during the use of the Peer Instruction method the authoritative communicative approach was also employed. In the authoritative dimension, future teachers used predominantly the type I-R-E (initiation-response-evaluation) communication pattern by asking the students several questions and leading them to the correct answer. Among the main implications the work contributes to the improvement of the practices of future teachers involved in applying active learning methodologies in classroom by identifying the types of communicative approaches and communication patterns used, as well as researches on curriculum innovation in physics in high school.

Keywords: curricular innovation, high school, physics teaching, discursive dynamics

Procedia PDF Downloads 157

Authors: Anthony J. Kondracki, Bonzo Reddick, Jennifer L. Barkin

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Background: Many pregnancies in the United States are affected each year with the most common sexually transmitted infections (STIs), including Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Treponema pallidum (TP, syphilis), and the rate of congenital syphilis has reached a 20-year high. We sought to estimate the prevalence of CT, NG, and TP in pregnancy and the risk of preterm birth (PTB) (<37 weeks gestation) and low birthweight (LBW) (<2500g) deliveries according to utilization of prenatal care (PNC) during the COVID-19 pandemic. Methods: This study was based on the 2020 National Center for Health Statistics (NCHS) Natality File restricted to singleton births (N=3,512,858). We estimated the prevalence of CT, NG, TP, PTBand LBW across timing and the number of prenatal care (PNC) visits attended. In multivariable logistic regression models, adjusted odds ratios of PTB and LBW were assessed according to STIs and PNC status. E-values, based on effect size estimates and the lower bound of the 95% confidence intervals (CIs) of the association, examined the potential impact of unmeasured confounding. Results: CT (1.8%) was most prevalent in pregnancy, followed by NG (0.3%) and TP (0.1%). The strongest predictors of PTB and LBW were maternal NG (12.2% and 12.1%, respectively), late initiation/no PNC (8.5% and 7.6%, respectively), and ≤10 prenatal visits (13.1% and 10.3%, respectively). The odds of PTB and LBW were 2.5- to 3-fold greater for each STI in women who received ≤10 compared to >10 prenatal visits. E-values demonstrated the minimum strength of potential unmeasured confounding necessary to explain away observed associations. Conclusions: Timely initiation and receipt of recommended number of prenatal visits benefits screening and treatment of all women for STIs, including NG to substantially reduce infant morbidity and mortality related to PTB and LBW among infants born during the COVID-19 pandemic.

Keywords: COVID-19 pandemic, sexually transmitted infections, preterm birth, low birthweight, prenatal care

Procedia PDF Downloads 127

Authors: Alfathony Krisnabudhi, Syahli Reza Ananda, M. Edo Marshal, M. Maaruf Mukti

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This study investigates the kinematics and basin development of pull-apart basin Sunda Strait based on palinspastic reconstructions of new acquired seismic reflection data to unravel hydrocarbon exploration opportunity in frontier area, fore-arc basin western Indonesia. We use more than 780 km seismic reflection data that cover whole basin. Structural patterns in Sunda Strait are dominated by northwest-southeast trending planar and listric-normal faults which appear to be graben and half-graben system. The main depocentre of this basin is East Semangko graben and West Semangko graben that are formed by overstepping of Sumatra Fault Zone and Ujungkulon Fault Zone. In father east, another depocentre is recognized as the Krakatau graben. The kinematic evolution started in Middle Miocene, characterized by the initiation of basement faulting with 0% to 7.00% extension. Deposition stratigraphic unit 1 and unit 2 started at 7.00% to 10.00% extension in Late Miocene and recognized as pre-transtensional deposit. The Plio-Pleistocene unit 3 and 4 were deposited as syn-transtensional deposit with 10.00% to 17.00% extension contemporaneously with the initiation of uplift NW-SE trending ridges due to the evolution of cross-basin fault in central basin and the development of en-echelon basin margin in a transtensional system. The control of sedimentation rate and basin subsidence cause the Neogene sediment to be very thick. We suggest that both controls allow thermal and pressure to generate hydrocarbon habitats in the pre-transtensional deposits. It is reinforced by stable kinematic evolution and interpretation of the deposition environment of pre-transtensional deposits that are deposited in the marine environment.

Keywords: kinematics, palinspastic, Sunda Strait, hydrocarbon exploration, fore-arc basin

Procedia PDF Downloads 155

Authors: Abobaker Abrahem M. Saad, Asma Abudasalam

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The stem node segments were cultured on Murashige and Skoog (MS) medium. In the case of using MS+ Zeatin (1 mg/l), small shoot buds were formed directly in 70% of explants after 15 days, their length range between 0.1 to 0.3 cm after two weeks and reached 0.3 cm in length and three shoots in numbers after 4 weeks. When those small shoots were sub cultured on the same medium, they increased in length, number and reached 0.4 cm with 4 shoots, 0.4 cm with 5 shoots after six, eight and ten weeks respectively. In the case of using MS free hormones, MS+IAA (0.2mg/l) +BA (0.5mg/l), MS + kin(0.5mg/l), MS + kin (3mg/l) and MS +NAA (3mg/l) +BA (1mg/l), no sign of responses were noticed and only change in color in some cases. Different types of parenchyma cells and many layers of thick wall sclerenchyma cells were observed on MS+BA (1mg/l).

Keywords: Maerua, stem node, shoots, buds, In vitro

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Authors: Carl-Philipp Valentin Beichert, Marcel Seger

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Digital transformation and climate change require small and medium-sized enterprises (SME) to rethink their way of doing business. Inter-firm collaboration is recognized as helpful means of promoting innovation and competitiveness. In this context, collaborations with start-ups offer valuable opportunities through their innovative products, services, and business models. SMEs, and in particular German craft enterprises, play an important role in the country’s society and economy. Companies in this heterogeneous economic sector have unique characteristics and are limited in their ability to innovate due to their small size and lack of resources. Collaborating with start-ups could help to overcome these shortcomings. To investigate how collaborations emerge and what factors are decisive to successfully drive collaboration, we apply an explorative, qualitative research design. A sample of ten case studies was selected, with the collaboration between a start-up and a craft enterprise forming the unit of analysis. Semi-structured interviews with 20 company representatives allow for a two-sided perspective on the respective collaboration. The interview data is enriched by publicly available data and three expert interviews. As a result, objectives, initiation practices, applied collaboration types, barriers, as well as key success factors could be identified. The results indicate a three-phase collaboration process comprising an initiation, concept, and partner phase (ICP). The ICP framework proposed accordingly highlights the success factors (personal fit, communication, expertise, structure, network) for craft enterprises and start-ups for each collaboration phase. The role of a mediator in the start-up company, with strong expertise in the respective craft sector, is considered an important lever for overcoming barriers such as cultural and communication differences. The ICP framework thus provides promising directions for further research and can help practitioners establish successful collaborations.

Keywords: open innovation, SME, craft businesses, startup collaboration, qualitative research

Procedia PDF Downloads 68

Authors: Ira V. Stancheva, Ely G. Zayova, Maria P. Geneva, Marieta G. Hristozkova, Lyudmila I. Dimitrova, Maria I. Petrova

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The study describes a highly efficient and low-cost protocol for rapid and mass in vitro propagation of medicinal and aromatic plant species (Hyssopus officinalis L., Lamiaceae). Hyssop is an important aromatic herb used for its medicinal values because of its antioxidant, anti-inflammatory and antimicrobial properties. The protocol for large-scale multiplication of this aromatic plant was developed using young stem tips explants. The explants were sterilized with 0.04% mercuric chloride (HgCl₂) solution for 20 minutes and washing three times with sterile distilled water in 15 minutes. The cultural media was full and half strength Murashige and Skoog medium containing indole-3-butyric acid. Full and ½ Murashige and Skoog media without auxin were used as controls. For each variant 20 glass tubes with two plants were used. In each tube two tip and nodal explants were inoculated. Maximum shoot and root number were obtained on ½ Murashige and Skoog medium supplemented with 0.1 mg L-1 indole-3-butyric acid at the same time after four weeks of culture. The number of shoots per explant and shoot height were considered. The data on rooting percentage, the number of roots per plant and root length were collected after the same cultural period. The highest percentage of survival 85% for this medicinal plant was recorded in mixture of soil, sand and perlite (2:1:1 v/v/v). This mixture was most suitable for acclimatization of all propagated plants. Ex vitro acclimatization was carried out at 24±1 °C and 70% relative humidity under 16 h illuminations (50 μmol m⁻²s⁻¹). After adaptation period, the all plants were transferred to the field. The plants flowered within three months after transplantation. Phenotypic variations in the acclimatized plants were not observed. An average of 90% of the acclimatized plants survived after transferring into the field. All the in vitro propagated plants displayed normal development under the field conditions. Developed in vitro techniques could provide a promising alternative tool for large-scale propagation that increases the number of homologous plants for field cultivation. Acknowledgments: This study was conducted with financial support from National Science Fund at the Bulgarian Ministry of Education and Science, Project DN06/7 17.12.16.

Keywords: Hyssopus officinalis L., in vitro culture, micro propagation, acclimatization

Procedia PDF Downloads 287

Authors: Osama M. Ahmed, Walaa G. Hozayen, Haidy Tamer Abo Sree

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The effect of doxorubicin (4 mg/kg b.w.week) without or with oral administration of ethanolic purslane (Portulaca oleracea) shoot (leaves and stems) extract (50 mg/kg b.w.day) or ethanolic purslane seeds extract (50 mg/kg b.w.day) co-treatments for 6 weeks was evaluated in adult male rats. There was an increase in serum levels of ALT, AST, ALP, GGT and total bilirubin. In addition, hepatic glutathine, glutathione transferase, peroxidase, SOD, CAT activities were decreased while lipid peroxidation in the liver was increased. Co-administration of ethanolic purslane and seed extracts successfully improved the adverse changes in the liver functions with an increase in antioxidants activities and reduction of lipid peroxidation.

Keywords: antioxidants, doxorubicin, hepatotoxicity, purslane

Procedia PDF Downloads 381

Authors: Kinda AlSamara

Abstract:

The nineteenth century was characterized by major educational reforms in the Arab World. One of the unintended outcomes of colonization in Arab countries was the initiation of women liberation as well as the introduction of modern education and its application in sensitizing people on the rights of women and their liberation. The reforms were often attributed to various undercurrents that took place at different levels within the Ottoman Empire, and particularly the arrival and influence of the Christian missionaries were supported by the American and European governments. These trends were also significantly attributed to the increase in the presence of Europeans in the region, as well as the introduction of secular ideas and approaches related to the meaning of modernity. Using literary analysis as a method, this paper examines the role of an important male figure like the political activist and writer Qāsim Amīn and the religious reformer Muḥammad ʻAbduh in starting this discourse and shows their impact on the emancipation of women movement (Taḥrīr), and how later women led the movement with their published work. This paper explores Arab Salons and the initiation of women’s literary circles. Women from wealthy families in Egypt and Syria who had studied in Europe or interacted with European counterparts began these circles. These salons acted as central locations where people could meet and hold discussions on political, social, and literary trends as they happened each day. The paper concludes with a discussion of current debates between the Islamist and the secularist branches of the movement today. While the Islamists believe that adhering to the core of Islam with some of its contested position on women is a modern ideology of liberation that fits the current culture of modern time Egypt; the secularists argue that the influence that Islam has on the women’s liberation movement in Egypt has been a threat to the natural success and progress of the movement, which was initiated in the early nineteenth century independent of the more recent trends towards religiosity in the country.

Keywords: educational model, crisis of terminologies, Arab awakening, nineteenth century

Procedia PDF Downloads 187

Authors: Gudakesh Yadav

Abstract:

Utter Pradesh, which is one of the largest states of India with unequal distribution of resources and different socioeconomic and cultural characteristics, level of different new born health care indicators varies a lot from one district to another district. State shared more than 21 percent of total live births of India; whereas, it accounts for 28 percent of total infant deaths of the country, with the 53 per thousand infant mortality rate. The present paper attempts to examine tempo-spatial changes in new born care practices during NFHS-1 to NFHS-3 and DLHS-2 to DLHS-3 in Uttar Pradesh and different regions. Descriptive statistics, rate-ratios, concentration index, multivariate and decomposition analysis has been used for the study. Findings of the study reveal that new born care practices have improved over the time in the state and across all the regions because of giving more emphasis on venerable groups like poor, rural, less educated mothers and scheduled caste & tribes but still it did not achieve the desired successes. Regional analysis of third rounds of DLHS shows that, coverage of intuitional delivery was the lowest in the central region. Performance of the southern region was the lowest in terms of initiation of breastfeeding, keeping baby warm and dry after the birth. The study calls for proper follow up of new born children to accelerate new born and child health care service and prioritises increasing antenatal check-ups and institutional delivery, which helps to improve level of other new born care services. At the policy level there is need to reach venerable groups like scheduled caste and tribes, poor and uneducated, and new mother especially in rural areas. High focused district should be allocated for better implementation of new born care promotion programme in low performing districts. Partnership with the private sector health professional is necessary to reach the every part of population.

Keywords: decomposition, inequality, initiation of breastfeeding, institutional delivery

Procedia PDF Downloads 214

Authors: S. Najafi, E. Bertini, M. Pezzotti, G.B. Tornielli, S. Zenoni

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Grapevine is an important agricultural fruit crop plant consumed worldwide and with a key role in the global economy. Grapevine is strongly affected by both biotic and abiotic stresses, which impact grape growth at different stages, such as during plant and berry development and pre- and post-harvest, consequently causing significant economic losses. Recently global warming has propelled the anticipation of the onset of berry ripening, determining the reduction of a grape color and increased volatilization of aroma compounds. Climate change could negatively alter the physiological characteristics of the grape and affect the berry and wine quality. Modern plant breeding can provide tools such as genome editing for improving grape resilience traits while maintaining intact the viticultural and oenological quality characteristics of the genotype. This study aims at developing a platform for genome editing application in grapevine plants with the final goal to improve berry quality, biotic, and abiotic resilience traits. We chose to directly deliver ribonucleoproteins (RNP, preassembled Cas protein and guide RNA) into plant protoplasts, and, from these cell structures, regenerate grapevine plants edited in specific selected genes controlling traits of interest. Edited plants regenerated by somatic embryogenesis from protoplasts will then be sequenced and molecularly characterized. Embryogenic calli of Sultana and Shiraz cultivars were initiated from unopened leaves of in-vitro shoot tip cultures and from stamens, respectively. Leaves were placed on NB2 medium while stamens on callus initiation medium (PIV) medium and incubated in the dark at 28 °C for three months. Viable protoplasts, tested by FDA staining, isolated from embryogenic calli were cultured by disc method at 1*105 protoplasts/ml. Mature well-shaped somatic embryos developed directly in the protoplast culture medium two months later and were transferred in the light into to shooting medium for further growth. Regenerated plants were then transferred to the greenhouse; no phenotypic alterations were observed when compared to non in-vitro cultured plants. The performed experiments allowed to established an efficient protocol of embryogenic calli production, protoplast isolation, and regeneration of the whole plant through somatic embryogenesis in both Sultana and Shiraz. Regenerated plants, through direct somatic embryogenesis deriving from a single cell, avoid the risk of chimerism during the regeneration process, therefore improving the genome editing process. As pre-requisite of genome editing, an efficient method for transfection of protoplast by yellow fluorescent protein (YFP) marker genes was also established and experiments of direct delivery of CRISPR–Cas9 ribonucleoproteins (RNPs) in protoplasts to achieve efficient DNA-free targeted mutations are in progress.

Keywords: CRISPR-cas9, plant regeneration, protoplast isolation, Vitis vinifera

Procedia PDF Downloads 121