Search results for: protein purification

Authors: Ting Zou, Chengfei Zhang

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Objectives: The purpose of this study was to investigate the role of Semaphorin 4D (Sema4D)/Plexin-B1 signaling on osteo/odontogenic differentiation of human dental pulp stem cells (DPSCs) and uncover its molecular mechanism. Methods: DPSCs were cultured in osteo/odontogenic medium. After treatment with Sema4D (10μg/mL), osteo/odontogenic differentiation and mineralization was evaluated by measuring alkaline phosphatase (ALP) activity and alizarin red S staining respectively. The expression of osteo/odontogenic genes (ALP, Col1A1, BSP, and Runx2) was determined by real-time polymerase chain reaction. p-Plexin-B1, Plexin-B1, Col1A1, RhoA, and ErbB2 were analyzed by western. Results: ALP activity and mineralization formation of DPSCs were significantly decreased after treatment with Sema4D (P<0.05). Sema4D significantly down-regulated osteo/odontogenic-related genes expression (ALP, Col1A1, BSP, and Runx2). p-Plexin-B1, Plexin-B1 and RhoA protein expression levels increased after stimulated with Sema4D, while the expression of Col1A1 decreased. Pretreatment with Plexin-B1 antibody blocked Sema4D induced p-Plexin-B1 expression. Conclusion: Sema4D suppressed osteo/odontogenic differentiation of DPSCs via RhoA-mediated pathways.

Keywords: Sema4D/Plexin-B1, dental pulp stem cells, osteo/odontogenic differentiation, alkaline phosphatase (ALP)

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Authors: N. Cetinkaya, Y. S. Kuleyin

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The aim of this study was to estimate the digestibility of the fruit internal skin of different varieties of hazelnuts to propose hazelnut fruit skin as an alternative feed source as roughage in ruminant nutrition. In 2015, the fruit internal skins of three different varieties of round hazelnuts (RH), pointed hazelnuts (PH) and almond hazelnuts (AH) were obtained from hazelnut processing factory then their crude nutrients analysis were carried out. Organic matter digestibility (OMD) and metabolisable energy (ME) values of hazelnut fruit skins were estimated from gas measured by in vitro gas production method. Their antioxidant activities were determined by spectrophotometric method. Crude nutrient values of three different varieties were; organic matter (OM): 87.83, 87.81 and 87.78%), crude protein (CP): 5.97, 5.93 and 5.89%, neutral detergent fiber (NDF): 30.30, 30.29 and 30.29%, acid detergent fiber (ADF): 48.68, 48.67 and 48.66% and acid detergent lignin (ADL): 25.43, 25.43 and 25.39% respectively. OMD from 24 h incubation time of RH, PH and AH were 22.04, 22.46 and 22.74%; ME_GP values were 3.69, 3.75 and 3.79 MJ/kg DM; and antioxidant activity values were 94.60, 94.54 and 94.52 IC 50 mg/mL respectively. The fruit internal skin of different varieties of hazelnuts may be considered as an alternative roughage for ruminant nutrition regarding to their crude and digestible nutritive values. Moreover, hazelnut fruit skin has a rich antioxidant content so it may be used as a feed additive for both ruminant and non-ruminant animals.

Keywords: antioxidant activity, hazelnut fruit skin, metabolizable energy, organic matter digestibility

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Authors: Pushpa Debnath

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Buddhism implies the ultimate methodology to obtain the acknowledgment to get out from cycling existence applied by the sutras. The Buddha’s natural methodology is the highest way of cessation from suffering existence. To be out of it, one must know the suffering before having tentativeness. According to the Buddha’s methodology, one can observe every being suffer from chronologically grasping craving. It is because lack of knowledge that the Buddha finds the four noble truths which are the basic states. These are suffering, the origin of suffering, cessation of suffering, and the path leading to the cessation of suffering. The Buddha describes that birth is suffering, aging is suffering, sickness is suffering, death is suffering, association with the unexpected is suffering, separation from the pleasant is suffering, and not receiving what one desires is suffering, In brief, the five aggregates of clinging are suffering. As the five aggregates are form, feeling, perception, mental formation, and consciousness. These are known as the matter that we identify with “You, Me” or “He.” The second truth cause of suffering is craving which has three types: craving for sense pleasures, craving for existence, and craving for non-existence. The third truth is the obliteration of craving, suffering can be eliminated to attain the Nibbana. The fourth truth is the path of liberation is the noble eight-fold path consisting of the right view, right intention, right speech, right action, right livelihood, right effort, right mindfulness, and right concentration. The six senses are the media of the eye, ear, nose, tongue, body, and mind sense faculties relating with the five aggregates and the six senses objects visual objects, sounds, smells, tastes, touch, and mind-objects that are contained by every visible being. The first five internal sense bases are material while the mind is a non-material phenomenon. Contact with the external world maintains by receiving through the six senses; visual objects through the eye, sounds through the ear, smells through the nose, tastes through the tongue, touch through the body, and mind-objects through sense faculties. These are the six senses a living being experiences by craving. Everything is conglomerated with all senses faculties through the natural phenomenon which are earth, water, fire, and air element. In this analysis, it is believed that beings are well adapted to the natural phenomenon. Everybody has fear of life because we have hatred, delusion, and anger which are the primary resources of falling into (Samsara) continuously that is the continuity of the natural way. These are the reasons for the suffering that chronically self-diluting through the threefold way. These are the roots of the entire beings suffering so the Buddha finds the enlightenment to uproot from cycling existence and the understanding of the natural consequence. When one could uproot ignorance, one could able to realize the ultimate happiness of Nirvana. From the craving of ignorance, everything starts to be present to the future which gives us mental agonies in existence.

Keywords: purification, morality, natural phenomenon, analysis, development of mind, observatory, Nirvana

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Authors: Cristina Lavilla, Andreas Heise

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The synthesis of sequence-controlled polymers has received increasing attention in the last years. Well-defined polyacrylates, polyacrylamides and styrene-maleimide copolymers have been synthesized by sequential or kinetic addition of comonomers. However this approach has not yet been introduced to the synthesis of polypeptides, which are in fact polymers developed by nature in a sequence-controlled way. Polypeptides are natural materials that possess the ability to self-assemble into complex and highly ordered structures. Their folding and properties arise from precisely controlled sequences and compositions in their constituent amino acid monomers. So far, solid-phase peptide synthesis is the only technique that allows preparing short peptide sequences with excellent sequence control, but also requires extensive protection/deprotection steps and it is a difficult technique to scale-up. A new strategy towards sequence control in the synthesis of polypeptides is introduced, based on the sequential addition of α-amino acid-N-carboxyanhydrides (NCAs). The living ring-opening process is conducted to full conversion and no purification or deprotection is needed before addition of a new amino acid. The length of every block is predefined by the NCA:initiator ratio in every step. This method yields polypeptides with a specific sequence and controlled molecular weights. A series of polypeptides with varying block sequences have been synthesized with the aim to identify structure-property relationships. All of them are able to adopt secondary structures similar to natural polypeptides, and display properties in the solid state and in solution that are characteristic of the primary structure. By design the prepared polypeptides allow selective modification of individual block sequences, which has been exploited to introduce functionalities in defined positions along the polypeptide chain. Poly(ethylene glycol)(PEG) was the functionality chosen, as it is known to favor hydrophilicity and also yield thermoresponsive materials. After PEGylation, hydrophilicity of the polypeptides is enhanced, and their thermal response in H2O has been studied. Noteworthy differences in the behavior of the polypeptides having different sequences have been found. Circular dichroism measurements confirmed that the α-helical conformation is stable over the examined temperature range (5-90 °C). It is concluded that PEG units are the main responsible of the changes in H-bonding interactions with H2O upon variation of temperature, and the position of these functional units along the backbone is a factor of utmost importance in the resulting properties of the α-helical polypeptides.

Keywords: α-amino acid N-carboxyanhydrides, multiblock copolymers, poly(ethylene glycol), polypeptides, ring-opening polymerization, sequence control

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Authors: Katerina Zaharieva, Vicente Rives, Martin Tsvetkov, Raquel Trujillano, Boris Kunev, Ivan Mitov, Maria Milanova, Zara Cherkezova-Zheleva

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The nanostructured cobalt ferrite-type materials Sample A - Co0.25Fe2.75O4, Sample B - Co0.5Fe2.5O4, and Sample C - CoFe2O4 were prepared by co-precipitation in our previous investigations. The co-precipitated Sample B and Sample C were mechanochemically activated in order to produce Sample D - Co0.5Fe2.5O4 and Sample E- CoFe2O4. The PXRD, Moessbauer and FTIR spectroscopies, specific surface area determination by the BET method, thermal analysis, element chemical analysis and temperature-programmed reduction were used to investigate the prepared nano-sized samples. The changes of the Malachite green dye concentration during reaction of the photocatalytic decolorization using nanostructured cobalt ferrite-type catalysts with different chemical composition are included. The photocatalytic results show that the increase in the degree of incorporation of cobalt ions in the magnetite host structure for co-precipitated cobalt ferrite-type samples results in an increase of the photocatalytic activity: Sample A (4 х10-3 min-1) < Sample B (5 х10-3 min-1) < Sample C (7 х10-3 min-1). Mechanochemically activated photocatalysts showed a higher activity than the co-precipitated ferrite materials: Sample D (16 х10-3 min-1) > Sample E (14 х10-3 min-1) > Sample C (7 х10-3 min-1) > Sample B (5 х10-3 min-1) > Sample A (4 х10-3 min-1). On decreasing the degree of substitution of iron ions by cobalt ones a higher sorption ability of the dye after the dark period for the co-precipitated cobalt ferrite materials was observed: Sample C (72 %) < Sample B (78 %) < Sample A (80 %). Mechanochemically treated ferrite catalysts and co-precipitated Sample B possess similar sorption capacities, Sample D (78 %) ~ Sample E (78 %) ~ Sample B (78 %). The prepared nano-sized cobalt ferrite-type materials demonstrate good photocatalytic and sorption properties. Mechanochemically activated Sample D - Co0.5Fe2.5O4 (16х10-3 min-1) and Sample E-CoFe2O4 (14х10-3 min-1) possess higher photocatalytic activity than that of the most common used UV-light catalyst Degussa P25 (12х10-3 min-1). The dependence of the photo-catalytic activity and sorption properties on the preparation method and different degree of substitution of iron ions by cobalt ions in synthesized cobalt ferrite samples is established. The mechanochemical activation leads to formation of nano-structured cobalt ferrite-type catalysts (Sample D and Sample E) with higher rate constants than those of the ferrite materials (Sample A, Sample B, and Sample C) prepared by the co-precipitation procedure. The increase in the degree of substitution of iron ions by cobalt ones leads to improved photocatalytic properties and lower sorption capacities of the co-precipitated ferrite samples. The good sorption properties between 72 and 80% of the prepared ferrite-type materials show that they could be used as potential cheap absorbents for purification of polluted waters.

Keywords: nanodimensional cobalt ferrites, photocatalyst, synthesis, mechanochemical activation

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Authors: Mohsen Rabbani, Trista McLaughlin, Ehsan Vahidi

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the remediation of polluted sites with heavy metals, such as rare earth elements (REEs), has been a primary concern of researchers to decontaminate the soil. Among all developed methods to address this concern, phytoremediation has been established as efficient, cost-effective, easy-to-use, and environmentally friendly way, providing a long-term solution for addressing this global concern. Furthermore, this technology has another great potential application in the metals production sector through returning metals buried in soil via metals cropping. Considering the significant metal concentration in hyper-accumulators, the utilization of bioaccumulated metals to extract metals from plant matter has been proposed as a sub-economic area called phytomining. As a recent, more advanced technology to eliminate such pollutants from the soil and produce critical metals, bioharvesting (phytomining/agromining) has been considered another compromising way to produce metals and meet the global demand for critical/target metals. The bio-ore obtained from phytomining can be safely disposed of or introduced to metal production pathways to obtain the most demanded metals, such as REEs. It is well-known that some hyperaccumulators, e.g., fern Dicranopteris linearis, can be used to absorb REE metals from the polluted soils and accumulate them in plant organs, such as leaves and stems. After soil remediation, the plant species can be harvested and introduced to the downstream steps, namely crushing/grinding, leaching, and purification processes, to extract REEs from plant matter. This novel interdisciplinary field can fill the gap between agriculture, mining, metallurgy, and the environment. Despite the advantages of agromining for the REEs production industry, key issues related to the environmental sustainability of the entire life cycle of this new concept have not been assessed yet. Hence, a comparative life cycle assessment (LCA) study was conducted to quantify the environmental footprints of REEs phytomining. The current LCA study aims to estimate and calculate environmental effects associated with phytomining by considering critical factors, such as climate change, land use, and ozone depletion. The results revealed that phytomining is an easy-to-use and environmentally sustainable approach to either eliminate REEs from polluted sites or produce REEs, offering a new source of such metals production. This LCA research provides guidelines for researchers active in developing a reliable relationship between agriculture, mining, metallurgy, and the environment to encounter soil pollution and keep the earth green and clean.

Keywords: phytoremediation, phytomining, life cycle assessment, environmental impacts, rare earth elements, hyperaccumulator

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Authors: Kazi Muhammad Rezaul Karim, Tasmia Tasnim

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Introduction: Pregnancy is a condition of higher nutrient requirement but in developing countries like Bangladesh most of the pregnant women can not meet their nutrient requirement and sometimes they are neglected in the family. The purpose of the study was to assess the nutritional status and dietary nutrient intake by the pregnant women, in Dhaka city, Bangladesh. Methods: The study population comprised of pregnant women from urban or semi-urban, aged between 18 to 35 and free of pregnancy related complication and other diseases. Under a cross-sectional design, 30 healthy non-pregnant as well as 130 pregnant women, at 3 different trimesters of pregnancy were assessed. A questionnaire was developed to obtain demographic, socio-economic, anthropometric, drug and medical history. Three day consecutive 24-hour food recalls were used to assess food intake and then converted to nutrient intake. Results: The average BMI of the nonpregnant women was 22.89 ± 3.4 kg/m2 and that of pregnant women was 23.52 ± 3.71 kg/m2. The mean dietary nutrient intake of dietary fiber, calorie, protein, fat, carbohydrate, calcium, iron, thiamine, riboflavin, vitamin C, Vitamin A, folate, vitamin B6 and Vitamin B12 of the pregnant mothers were 4.38 g, 1619 kcal, 60.05 g, 30.38 g, 268.79 g, 537.21 mg, 21.53 mg, 1.15 mg, 0.94 mg, 97.36 mg, 647.6 µg, 153.93 µg, 1.41 mg and 4.09 µg respectively. Most of pregnant women (more than 90%) can not meet their energy, calcium and folate requirements. Conclusion: Most of the pregnant mother in Bangladesh can not meet their dietary requirements during pregnancy.

Keywords: pregnancy, dietary nutrient, nutritional status, BMI

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Authors: Ghazi Chabchoub, Mouna Feki, Mohamed Abid, Hammadi Ayadi

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Autoimmune thyroid diseases (AITDs), including Graves’ disease (GD) and Hashimoto’s thyroiditis (HT), are inherited as complex traits. Genetic factors associated with AITDs have been tentatively identified by candidate gene and genome scanning approaches. We analysed three intragenic microsatellite markers in the thyroid hormone receptor associated protein 2 gene (THRAP2), mapped near D12S79 marker, which have a potential role in immune function and inflammation [THRAP2-1(TG)n, THRAP2-2 (AC)n and THRAP2-3 (AC)n]. Our study population concerned 12 patients affected with AITDs belonging to a multiplex Tunisian family with high prevalence of AITDs. Fluorescent genotyping was carried out on ABI 3100 sequencers (Applied Biosystems USA) with the use of GENESCAN for semi-automated fragment sizing and GENOTYPER peak-calling software. Statistical analysis was performed using the non parametric Lod score (NPL) by Merlin software. Merlin outputs non-parametric NPLall (Z) and LOD scores and their corresponding asymptotic P values. The analysis for three intragenic markers in the THRAP2 gene revealed strong evidence for linkage (NPL=3.68, P=0.00012). Our results suggested the possible role of THRAP2 gene in AITDs susceptibility in this family.

Keywords: autoimmunity, autoimmune disease, genetic, linkage analysis

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Authors: C. O. Agu, C. C. Okafor

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Maize and soybean flours were produced using different methods of processing which include fermentation (FWF), roasting (RWF) and malting (MWF). Products from the different methods were mixed in the ratio 60:40 maize/soybean, respectively. These composites mixed with other ingredients such as sugar, vegetable oil, vanilla flavour and vitamin mix were analyzed for proximate composition, physical/functional, sensory and nutritional properties. The results for the protein content ranged between 6.25% and 16.65% with sample RWF having the highest value. Crude fibre values ranged from 3.72 to 10.0%, carbohydrate from 58.98% to 64.2%, ash from 1.27 to 2.45%. Physical and functional properties such as bulk density, wettability, gelation capacity have values between 0.74 and 0.76g/ml, 20.33 and 46.33 min and 0.73 to 0.93g/ml, respectively. On the sensory quality colour, flavour, taste, texture and general acceptability were determined. In terms of colour and flavour there was no significant difference (P < 0.05) while the values for taste ranged between 4.89 and 7.1 l, texture 5.50 to 8.38 and general acceptability 6.09 and 7.89. Nutritionally there is no significant difference (P < 0.05) between sample RWF and the control in all parameters considered. Samples FWF and MWF showed significantly (P < 0.5) lower values in all parameters determined. In the light of the above findings, roasting method is highly recommend in the production of weaning foods.

Keywords: fermentation, malting, ratio, roasting, wettability

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Authors: Petr Konvalina, Jan Moudry

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As organic farmers are searching foregoing crops for horticultural crops, there is possible to choice neglected wheat species and also have a new market and sale opportunities. Concerning wheat, there are landraces so called hulled wheat species (einkorn, emmer wheat, spelt) comprising parts of collections of the world gene banks. The advantage of this wheat species are small demands on growing conditions and also droughtiness in conditions of changing climate. Our paper aims at presenting the results of the study and the assessment of spring wheat forms, four einkorn cultivars, eight emmer wheat cultivars, seven spelt wheat cultivars in particular, as compared to modern bread wheat variety. Small-plot trials were established at two different localities within the Czech Republic and Austria in 2009 and 2012. The results of the trials show that some varieties were inclined to lodging. On the other hand, they were resistant to common wheat diseases (mildew, brown rust). Hulls served as barriers and obstacles against the DON grain contamination. The yield rate was lower. The grains were characterized by a high proportion of protein in grain (up to 18.1 %). However, they may be difficult to use for common baking. Moreover, new food products demonstrating a different technological quality of the hulled wheat species have to be launched on the market. They will be suitable for regional marketing.

Keywords: organic farming, hulled wheat species, einkorn, emmer, spelt

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Authors: Khaled S. Al Salhen

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Aldehyde oxidase is molybdo-flavoenzyme involved in the oxidation of hundreds of endogenous and exogenous and N-heterocyclic compounds and environmental pollutants. Uncharged N-heterocyclic aromatic compounds such phenanthridine are commonly distributed pollutants in soil, air, sediments, surface water and groundwater, and in animal and plant tissues. Phenanthridine as uncharged N-heterocyclic aromatic compound was incubated with partially purified aldehyde oxidase from rainbow trout fish liver. Reversed-phase HLPC method was used to separate the oxidation products from phenanthridine and the metabolite was identified. The 6(5H)-phenanthridinone was identified the major metabolite by partially purified aldehyde oxidase from fish liver. Kinetic constant for the oxidation reactions were determined spectrophotometrically and showed that this substrate has a good affinity (Km = 78 ± 7.6 µM) for hepatic aldehyde oxidase, coupled with a relatively high oxidation rate (0.77± 0.03 nmol/min/mg protein). In addition, the kinetic parameters of hepatic fish aldehyde oxidase towards the phenanthridine substrate indicate that in vitro biotransformation by hepatic fish aldehyde oxidase will be a significant pathway. This study confirms that partially purified aldehyde oxidase from fish liver is indeed the enzyme responsible for the in vitro production 6(5H)-phenanthridinone metabolite as it is a major metabolite by mammalian aldehyde oxidase.

Keywords: aldehyde oxidase, fish, phenanthridine, specificity

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Authors: Abul B. M. M. K. Islam, Nuria Lopez-Bigas, Elizaveta V. Benevolenskaya

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RBP2 has shown to be important for cell differentiation control through epigenetic mechanism. The main aim of the present study is genome-wide location analysis of human RBP2 isoforms that differ in a histone-binding domain by ChIPseq. It is conceivable that the larger isoform (LI) of RBP2, which contains a specific H3K4me3 interacting domain, differs from the smaller isoform (SI) in genomic location, may account for the observed diversity in RBP2 function. To distinguish the two RBP2 isoforms, we used the fact that the SI lacks the C-terminal PHD domain and hence used the antibodies detecting both RBP2 isoforms (AI) through a common central domain, and the antibodies detecting only LI but not SI, through a C-terminal PHD domain. Overall our analysis suggests that RBP2 occupies about 77 nucleotides and binds GC rich motifs of active genes, does not bind to centromere, telomere, or enhancer regions, and binding sites are conserved compare to random. A striking difference between the only-SI and only-LI is that a large number of only-SI peaks are located in CpG islands and close to TSS compared to only-LI peaks. Enrichment analysis of the related genes indicates that several oncogenic pathways and metabolic pathways/processes are significantly enriched among only-SI/AI targets, but not LI/only-LI peak’s targets.

Keywords: bioinformatics, cancer, ChIP-seq, KDM5A

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Authors: M. Hetmańczyk, R. Polaniak, K. Brukało, E. Grochowska-Niedworok

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Eating behaviours of people are determined by knowledge gained at different stages of life. Children’s diet is especially important. They have to eat meals regularly. Meals should consist of protein, carbohydrates and fat, and drinking the right amount of water. Mistakes in children’s diets affect their health and may lead to health issues such as diabetes, overweight, obesity or malnutrition. The aim of the study was to assess the eating habits among 10-15-year-old children. To achieve this aim, the study included children aged 10-15 years living in Silesia Province, Poland; the participants consisted of 52.08% girls and 47.92% boys. Authorial questionnaire contains 28 questions about eating habits. The results of 192 students were subjected to analysis. The results show that half of the surveyed students participated in physical activity every day. Most children ate 4-5 meals every day, but the breaks between them were too long (four and more hours). Children generally ate cooked meals. Most children ate first breakfast every day, but only one third of studied children ate a second breakfast daily, while 93.75% ate vegetables at least once a day, 94.79% ate fruit at least once a day, and 79.17% drink a daily glass of milk or more. The study found that the eating behaviours of the surveyed children were unsatisfying. While the children did not participate in physical activity often enough, girls took part slightly more often. Children eat second breakfast not often enough. Younger children (10-12 years old) are doing it more often than the older children (13-15 years old). Gender is not a determinant of the frequency of second breakfast consumption.

Keywords: eating habits, children, diet, nutrition status

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Authors: Zuoyong Zhang, Min Yu, Jinlong Zhao, Shudong He

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The Maillard reaction can produce the flavor enhancing substance through the chemical crosslinking between free amino group of the protein or polypeptide with the carbonyl of the reducing sugar. In this research, solutions of rapeseed meal peptide and D-xylose with or without L-cysteine (RXC or RX) were heated over a range of temperatures (80-140 °C) for 2 h. It was observed that RXs had a severe browning,while RXCs accompanied by more pH decrement with the temperature increasing. Then the correlation among data of quantitative sensory descriptive analysis, free amino acid (FAA) and GC–MS of RXCs and RXs were analyzed using the partial least square regression method. Results suggested that the Maillard reaction product (MRPs) with cysteine formed at 120 °C (RXC-120) had greater sensory properties especially meat-like flavor compared to other MRPs. Meanwhile, it revealed that glutamic and glycine not only had a positive contribution to meaty aroma but also showed a significant and positive influence on umami taste of RXs based on the FAA data. Moreover, the sulfur-containing compounds showed a significant positive correlation with the meat-like flavor of RXCs, while RXs depended on furans and nitrogenous-containing compounds with more caramel-like flavor. Therefore, a MRP with strong meaty flavor could be obtained at 120 °C by addition of cysteine.

Keywords: rapeseed meal, Maillard reaction, sensory characteristics, FAA, GC–MS, partial least square regression

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Authors: K. O. Igwe, E. N. Onyeike, A. A. Uwakwe

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Comparative proximate composition of selected edible vegetables (Abelmoschus esculentus, Telfairia occidentalis, vernonia amygdalina and Talinum triangulare ) harvested in farmland nearby oil impacted sites in Rivers State was investigated. The result obtained was compared to the proximate composition of similar edible vegetables harvested from non oil producing areas in Imo State. Values from the study areas indicated vegetables with high moisture levels (highest values in Abelmoschus esculentus and Telfairia occidentalis; 6.07±0.11% and 4.10±0.12%) low ash contents (lowest value in Telfairia occidentalis 9.80±0.08%) low crude protein (lowest value in Talinum triangulare 8.15±0.06 %) high lipid content (highest value in Talinum triangulare 4.12± 0.20%) lower crude fibre (lowest value in Abelmoschus esculentus 9.58±0.08 %) and higher Carbohydrate contents (highest value in vernonia amygdalina 62.18± 0.67%). Values obtained indicated vegetables with lower quality in the study areas.

Keywords: edible vegetables, proximate composition, oil spill, farmland

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Authors: J. Zekri, N. Souilah, W. Abdelaziz, D. Alatou

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Our study shall focus on the ability of trees cork oak that showed vis-à-vis sensitivity to climate change, including late spring frosts. The combination of these factors resulted in damage alarmed, therefore forest ecosystems weakened trees that can affect their ability to support other abiotic and biotic stresses, For this we tested its tolerance to thermal variations and cold weather conditions by estimating some stress markers (quantification of proteins, RNA, soluble sugars) that are quantified to evaluate the cold tolerance of seedlings. Sowing of cork oak (Quercus suber L.) is grown in controlled conditions at 25° C ± 2° C in long days 16h. These seedlings are transferred at low temperatures between 5° C and -6° C for a period of 3 hours. Biochemical analyzes were performed in the various organs of the cork oak seedlings. Cool temperatures induced a significant accumulation of proline in different organs of seedlings and the optimum concentrations were observed in the roots with very high concentrations (4 times larger than those of the control). The accumulation of soluble sugars is significantly in stems and roots at 0° C. Protein concentrations are very high in leaves of both growth and high waves in rod at -4° C to -2° C. Tolerance cork oak seems to be at the thermal limit of -2°C. The concentration of these metabolites in the various organs showed the ability oak cork hardening during the winter.

Keywords: climate change, thermal change, semi-aride, biochemical markers, heat stress

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Authors: Ravi R. Patel, Vasudev R. Thakkar

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Use of plant growth-promoting rhizobacteria (PGPR) to increase the production and decrees disease occurrence is a recent method in agriculture. An RA2 rhizospheric culture was isolated from peanut rhizosphere from Junagadh region of Gujarat, India and showed different direct and indirect plant growth promoting activity like indole acetic acid, gibberellic acid, siderophore, hydrogen cyanide, Ammonia and (1-Aminocyclopropane-1-Carboxylate) deaminase production, N2 fixation, phosphate and potassium solubilization in vitro. RA2 was able to protect peanut germinating seedling from A. niger infection and reduce collar rot disease incidence 60-35% to 72-41% and increase germination percentage from 70-82% to 75-97% in two varieties GG20 and GG2 of peanut. RA2 was found to induce resistance in A. hypogaea L. seedlings via induction of different defense-related enzymes like phenylalanine ammonia lyase, peroxidase, polyphenol oxidase, lipoxygenase and pathogenesis related protein like chitinase, ß – 1,3- glucanase. Jasmonic acid one of the major signaling molecules of inducing systemic resistance was also found to induced due to RA2 treatments. RA2 bacterium was also promoting peanut growth and reduce A. niger infection in pot studies. 16S rDNA sequence of RA2 showed 99 % homology to Azotobacter species.

Keywords: plant growth promoting rhizobacteria, peanut, aspergillus niger, induce systemic resistance

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Authors: Chengwei Wang, Shuqing Xu, Philipp M. Schlüter

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The genus Ophrys is remarkable for its mimicry, flower-lip closely resembling pollinator females in a species-specific manner. Therefore, floral traits associated with pollinator attraction, especially scent, are suitable models for investigating the molecular basis of adaption, speciation, and evolution. Within the two Ophrys species groups: O. sphegodes (S) and O. fusca (F), pollinator shifts among the same insect species have taken place. Preliminary data suggest that they involve a comparable hydrocarbon profile in their scent, which is mainly composed of alkanes and alkenes. Genes encoding stearoyl-acyl carrier protein desaturases (SAD) involved in alkene biosynthesis have been identified in the S group. This study aims to investigate the control and parallel evolution of ecologically significant alkene production in Ophrys. Owing to the central role those SAD genes play in determining positioning of the alkene double-bonds, a detailed understanding of their functional mechanism and of regulatory aspects is of utmost importance. We have identified 5 transcription factors potentially related to SAD expression in O. sphegodes which belong to the MYB, GTE, WRKY, and MADS families. Ultimately, our results will contribute to understanding genes important in the regulatory control of floral scent synthesis.

Keywords: floral traits, transcription factors, biosynthesis, parallel evolution

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Authors: Maesoomeh Khorshidi Mehr, Mohammad Sajadian, Shadi Alipour

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The purpose of the present study was to compare the effects of eight weeks of continuous and periodic aerobic exercises on serum levels of CRP in overweight woman. 36 woman aged between 20 and 35 years from the city of Ahwaz were randomly selected as the sample of the study. This sample was further divided into three groups (n= 12) of continuous aerobic exercise, periodic aerobic exercise, and control. Subjects of the groups of continuous and periodic aerobic exercise participated in 8 weeks of specialized exercises while the control group subjects did not take part in any regular physical activity program. Blood samples were collected from subjects in 24 hours prior to and 48 hours past to the intervention period. Afterwards, the serum level of CRP was measured for each blood sample. Results showed that BMI and serum level of CRP both significantly reduced as a result of aerobic exercises. However, no statistically significant difference was recorded between the extent of effects of the former and latter aerobic exercise types. Eight weeks of aerobic exercise will probably result in reduced inflammation and cardiovascular diseases risk in overweight women. The reason for lack of difference between effects of continuous and periodic aerobic exercise may lie in the similarity of average intensity and length of physical administered activities.

Keywords: heart diseases, aerobic exercise, inflammation, CRP, overweight

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Authors: Christina Roman, Deepak Koirala, Joseph A. Piccirilli

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Phage displaying synthetic Fab libraries have been used to obtain Fabs that bind to specific RNA targets with high affinity and specificity. These Fabs have been demonstrated to facilitate RNA crystallization. However, the antibody framework used in the construction of these phage display libraries contains numerous bulky, flexible, and charged residues, which facilitate solubility and hinder aggregation. These residues can interfere with crystallization due to the entropic cost associated with burying them within crystal contacts. To systematically reduce the surface entropy of the Fabs and improve their crystallization properties, a protein engineering strategy termed surface entropy reduction (SER) is being applied to the Fab framework. In this approach, high entropy residues are mutated to smaller ones such as alanine or serine. Focusing initially on Fab BL3-6, which binds an RNA AAACA pentaloop with 20nM affinity, the SER P server (http://services.mbi.ucla.edu/SER/) was used and analysis was performed on existing RNA-Fab BL3-6 co-crystal structures. From this analysis twelve surface entropy reduced mutants were designed. These SER mutants were expressed and are now being measured for their crystallization and diffraction performance with various RNA targets. So far, one mutant has generated 3.02 angstrom diffraction with the yjdF riboswitch RNA. Ultimately, the most productive mutations will be combined into a new Fab framework to be used in a optimized phage displayed Fab library.

Keywords: antibody fragment, crystallography, RNA, surface entropy reduction

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Authors: Roudabeh Vakil Monfared, Farhad Mashayekhi

Abstract:

Breast cancer is the most common malignancy type among women with about 1 million new cases each year. The immune system plays an important role in the breast cancer development. OX40L (also known as TNFSF4), a membrane protein, which is a member of the tumor necrosis factor super family binds to its receptor OX40 and this co-stimulation has a crucial role in T-cell proliferation, survival and cytokine release. Due to the importance of the T-cells in anti-tumor activities of OX40L we studied the association of rs3850641 (T→C) polymorphism of OX40L gene with breast cancer. The study included 123 women with breast cancer and 126 healthy volunteers with no signs of cancer. Genomic DNA was extracted from blood leucocytes. Genotype and allele frequencies were determined in patients and control cases with the method of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and the analysis was performed by Med Calc. The prevalence of genotype frequencies of TT, CT and CC were 60.9%, 30.08% and 8.9 % in patients with breast cancer and 74.6 %, 18.25 % and 7.14 % in healthy volunteers while the T and C allelic frequency was 76.01% and 23.98 % in patients and 83.73% and 16.26% in healthy controls. Respectively Statistical analysis has shown no significant difference from the comparison of either genotype (P=0.06). According to these results, the rs3850641 SNP has no association with the susceptibility of breast cancer in a population in northern Iran. However, further studies in larger populations including other genetic and environmental factors are required to achieve conclusion.

Keywords: OX40L, gene, polymorphism, breast cancer

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Authors: Debasish Pradhan, Shaktiprasad Pradhan, Rakesh Kumar Pradhan, Gitanjali Tripathy

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Suppressors of cytokine signalling (SOCS1), a newly indentified antiapoptotic molecule is a downstream effector of the receptor tyrosine kinase-Ras signalling pathway. The current study has uncovered that SOCS1 may have wide and imperative capacities, particularly because of its close correlation with malignant tumors. To investigate the impact of SOCS1 on MDR, we analyzed the expression of P-gp and SOCS1 by immunohistochemistry and found there was a positive correlation between them. At that point, we effectively interfered with RNA translation by the contamination of siRNA of SOCS1 into MCF7/ADM breast cancer cell lines through a lentivirus, and the expression of the target gene was significantly inhibited. After RNAi, the drug resistance was reduced altogether and the expression of MDR1 mRNA and P-gp in MCF7/ADM cell lines demonstrated a significant decrease. Likewise, the expression of P53 protein increased in a statistically significant manner (p ≤ 0.01) after RNAi exposure. Moreover, flow cytometry analysis uncovers that cell cycle and anti-apoptotic enhancing capacity of cells changed after RNAi treatment. These outcomes proposed SOCS1 may take part in breast cancer MDR by managing MDR1 and P53 expression, changing cell cycle and enhancing the anti-apoptotic ability.

Keywords: breast cancer, multidrug resistance, SOCS1 gene, MDR1 gene, RNA interference

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Authors: Laila Ozola, Evita Straumite

Abstract:

Celiac disease is a permanent enteropathy caused by the ingestion of gluten, a protein occurring in wheat, rye and barley. The only way of the effective daily treatment is a strict gluten-free diet. From the investigation of products available in the local market, it was found that Latvian producers do not offer gluten-free products. The aim of this research was to study and analyze changes of celiac patient’s attitude to gluten-free product quality and availability in the Latvian market and purchasing habits. The survey was designed using website www.visidati.lv, and a questionnaire was sent to people suffering from celiac disease. The first time the respondents were asked to fill in the questionnaire in 2011, but now repeatedly from the beginning of September 2013 till the end of January 2014. The questionnaire was performed with 75 celiac patients, respondents were from all Latvian regions and they answered 16 questions. One of the most important questions was aimed to find out consumers’ opinion about quality of gluten-free products, consumption patterns of gluten-free products, and, moreover, their interest in products made in Latvia. Respondents were asked to name gluten-free products they mainly buy and give specific purchase locations, evaluate the quality of products and necessity for products produced in Latvia. The results of questionnaire show that the consumers are satisfied with the quality of gluten-free flour, flour blends, sweets and pasta, but are not satisfied with the quality of bread and confectionery available in the Latvian markets.

Keywords: consumers, gluten-free products, quality, survey

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Authors: C. Minguez, J. Bueso-Rodenas, C. Ibanez, A. Calvo

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Mannan oligosaccharides (MOS) is one of the prebiotic most used in livestock nutrition. In this research, the effect of MOS dietary supplementation on growth performance and mortality in meat guinea pigs were studied. Three different experimental groups were compared: Control group (no additives); MOS 1 (1.5 g kg−1); MOS 2 (2 g kg−1). Guinea pigs were housed in 15 collective cages (n = 50 animals in each trial; 10 animals per cage). The young guinea pigs were weaning at day 28 and individually identified by a little ear tag. The fattening period was 49 days. Guinea pigs in both groups were fed ad libitum, with a standard commercial pellet diet (10 MJ of digestible energy/kg, 17% crude protein, 11% crude fiber, and 4.5% crude fat) and alfalfa (Medicago sativa) as forage. Growth traits, including body weight (BW), average daily gain (ADG), feed intake (FI), and feed conversion ratio (FCR), were measured weekly. On day 74, the animals were slaughtered. Contrasts between groups were obtained by calculated generalized least squares values. Mortality were evaluated by Fisher's exact test. Between MOS groups no significant differences were observed for growth traits and mortality. However, significant differences against the control group were observed for traits studied (pvalue < 0.05). In conclusion, the use of MOS could be a good prebiotic supplement to raise guinea pigs because it MOS has shown positive effects in growth traits and immune response in animals.

Keywords: guinea pig, growth, mannan oligosaccharides, mortality

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Authors: Shan-Ying Wu, Sheng-Hui Lan, Xi-Zhang Lin, Ih-Jen Su, Ting-Fen Tsai, Chia-Jui Yen, Tsung-Hsueh Lu, Fu-Wen Liang, Huey-Jen Su, Chun-Li Su, Hsiao-Sheng Liu

Abstract:

In hepatocelluar carcinoma (HCC), dysregulated expression of cyclin D1 and impaired autophagy has been reported separately. However, the relationship between them has not been explored. In this study, we demonstrated that autophagy was inversely correlated with cyclin D1 expression in 147 paired HCC patient specimens. HCC specimen with highly expression of cyclin D1 shows correlation with poor overall survival rate. Furthermore, induction of autophagy by amiodarone (antiarrhythmic drug) in Hep 3B cells, cyclin D1 was recruited into autophagosomes demonstrated by immune-gold labeling of cyclin D1 after extraction of autophagosomes. We further demonstrated that autophagy suppresses Hep 3B cell proliferation, and further analysis revealed that cell cycle was arrested at G1 phase. The interaction between LC3 (maker of autophagy) and cyclin D1 was increased after autophagy induction. In addition, ubiquitinated-cyclin D1 was also increased after autophagy induction, which is selectively degraded by autophagosome through binding with SQSTM1/p62 (an adaptor protein). In vivo study showed that amiodarone induced autophagy suppresses liver tumor formation in xenograft mouse and orthotopic rat model through decreasing cyclin D1 expression and inhibition of cell proliferation. Altogether, we reveal a novel mechanism that ubiquitinated cyclin D1 degraded by autophagic pathway by p62 and amiodarone is a promising drug for targeting cyclin D1 in liver cancer therapy.

Keywords: autophagy, cyclin D1, hepatocellular carcinoma, amiodarone

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Authors: Giang Tran Thi Huong, Hieu Dong Van, Tung Dao Duy, Saadanov Iskender, Isakeev Mairambek, Tsutomu Omatsu, Yukie Katayama, Tetsuya Mizutani, Yuki Ozeki, Yohei Takeda, Haruko Ogawa, Kunitoshi Imai

Abstract:

Newcastle disease virus (NDV) is a contagious viral disease of the poultry industry and other birds throughout the world. At present, very little is known about molecular epidemiological data regarding the causes of ND outbreak in commercial poultry farms in Kyrgyzstan. In the current study, the NDV isolated from the one out of three samples from the unvaccinated flock was confirmed as NDV. Phylogenetic analysis indicated that this NDV strain is clustered in the Class II subgenotype VIId, and closely related to the Chinese NDV isolate. Phylogenetic analyses revealed that the isolated NDV strain has an origin different from the 4 NDV strains previously identified in Kyrgyzstan. According to the mean death time (MDT: 61.1 h) and a multibasic amino acid (aa) sequence at the F0 proteolytic cleavage site (¹¹²R-R-Q-K-R-F¹¹⁷), the NDV isolate was determined as mesogenic strain. Several mutations in the neutralizing epitopes (notably, ³⁴⁷E→K) and the global head were observed in the hemagglutinin-neuraminidase (HN) protein of the current isolate. The present study represents the molecular characterization of the coding gene region of NDV in Kyrgyzstan. Additionally, further study will be investigated on the antigenic characterization using monoclonal antibody.

Keywords: Kyrgyzstan, Newcastle disease, genotype, genome characterization

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Authors: Muhammad Mazhar, Yong Zhu, Likang Qin

Abstract:

Foods contain endogenous components known as dietary fibers, which are classified into soluble and insoluble forms. Dietary fibers are resistant to gut digestive enzymes, modulating anaerobic intestinal microbiota (AIM) and fabricating short-chain fatty acids (SCFAs). Acetate, butyrate, and propionate dominate in the gut, and different pathways, including Wood-Ljungdahl and acrylate pathways, generate these SCFAs. In pancreatic dysfunction, the release of insulin/glucagon is impaired, which leads to hyperglycemia. SCFAs enhance insulin sensitivity or secretion, beta-cell functions, leptin release, mitochondrial functions, and intestinal gluconeogenesis in human organs, which positively affect type 2 diabetes (T2D). Research models presented that SCFAs either enhance the release of peptide YY (PYY) and glucagon-like peptide-1 (GLP-1) from L-cells (entero-endocrine) or promote the release of leptin hormone satiation in adipose tissues through G-protein receptors, i.e., GPR-41/GPR-43. Dietary fibers are the components of foods that influence AIM and produce SCFAs, which may be offering beneficial effects on T2D. This review addresses the effectiveness of SCFAs in modulating gut AIM in the fermentation of dietary fiber and their worth against T2D.

Keywords: dietary fibers, intestinal microbiota, short-chain fatty acids, fermentation, type 2 diabetes

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Authors: Maha Deif, Alaa Eldin Hassan, Eman Shaat, Nesrine Elazhary, Eman Magdy

Abstract:

Background: Erythropoietin (EPO) is a hematopoietic factor with multiple protective effects. The aim of the present study was to investigate the potential effect of EPO administration on renal functions and hypoxia inducible factor 1-alpha (HIF-1a) in diabetic rat kidneys. Methodology: The current study was carried out on 40 male albino rats divided into four groups (n= 10 in each). Group I served as normal control, group II was the diabetic control, group III rats received EPO on the same day of diagnosis of diabetes mellitus (DM), while group IV received the first dose of EPO 2 weeks after the diagnosis of DM. Results: The results showed that EPO supplementation leads to a significant decrease in serum urea, urinary protein and creatinine clearance as well as a significant increase in renal HIF-1a in group III and IV rats compared to the diabetic control group (group II). However, fasting blood glucose was significantly decreased in group III as compared to the diabetic control group in the third week, but no significant difference was reported in the fourth week among groups II, III and IV. Conclusion: EPO administration leads to the improvement of renal functions and increased levels of HIF-1a in diabetic rats.

Keywords: erythropoietin, diabetic nephropathy, hypoxia-inducible factor1-alpha, renal functions

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Authors: Uzma Saqib, Mirza S. Baig

Abstract:

Myeloid differentiation primary response protein 88 (MYD88) has long been considered a central player in the inflammatory pathway. Recent studies clearly suggest that it is an important therapeutic target in inflammation. On the other hand, a recent study on the interaction between the orphan nuclear receptor (Nur77) and p38α, leading to increased lipopolysaccharide-induced hyperinflammatory response, suggests this binary complex as a therapeutic target. In this study, we have designed inhibitors that can inhibit both MYD88 and Nur77 at the same time. Since both MYD88 and Nur77 are an integral part of the pathways involving lipopolysaccharide-induced activation of NF-κB-mediated inflammation, we tried to target both proteins with the same library in order to retrieve compounds having dual inhibitory properties. To perform this, we developed a homodimeric model of MYD88 and, along with the crystal structure of Nur77, screened a virtual library of compounds from the traditional Chinese medicine database containing ~61,000 compounds. We analyzed the resulting hits for their efficacy for dual binding and probed them for developing a common pharmacophore model that could be used as a prototype to screen compound libraries as well as to guide combinatorial library design to search for ideal dual-target inhibitors. Thus, our study explores the identification of novel leads having dual inhibiting effects due to binding to both MYD88 and Nur77 targets.

Keywords: drug design, Nur77, MYD88, inflammation

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Authors: Derya Yucel, Nigar Angın, Dürdane Mart, Meltem Turkeri, Volkan Catalkaya, Celal Yucel

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Chickpea (Cicer arietinum L.) with high protein content is a vital food, especially in under-developed and developing countries for the people who do not consume enough meat due to low-income level. The objective of the proposed study is to evaluate growing, yield and yield components of chickpea genotypes under Mediterranean condition so determine tolerance of chickpea genotypes against drought and heat stress. For this purpose, a total of 34 chickpea genotypes were used as material. The experiment was conducted according to factorial randomized complete block design with 3 reps at the Eastern Mediterranean Research Institute, Adana, TURKEY for 2014-15 growing season under three different growing conditions (Winter sowing, irrigated-late sowing and non-irrigated- late sowing). According to results of this experiment, vegetative period, flowering time, poding time, maturity time, plant height, height of first pod, seed yield and 100 seed weight were ranged between 68.33 to 78.77 days, 94.22 to 85.00 days, 94.11 to 106.44 days, 198.56 to 214.44 days, 37.18 to 64.89 cm, 18.33 to 34.83 cm, 417.1 to 1746.4 kg/ha and 14.02 to 45.02 g, respectively. Among the chickpea genotypes, the Aksu, Arda, Çakır, F4 09 (X 05 TH 21-16189), FLIP 03-108 were least affected by drought and heat stress. Therefore, these genotypes can be used as sources of drought and heat tolerance in further breeding programme for evolving the drought and heat tolerant genotypes in chickpea.

Keywords: chickpea, drought stress, heat stress, yield

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