Search results for: maturation

Authors: Ibrahim A. H. Barakat, Ahmed R. Al-Himaidi

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The present study was conducted to determine the role and optimum concentration of fenugreek seed extract during in-vitro maturation on in-vitro maturation and developmental competence of Neaimi sheep oocytes following in-vitro fertilization. The Cumulus Oocyte Complexes (COCs) collected from sheep slaughterhouse ovaries were randomly divided into three groups, and they were matured for 24 hrs. in maturation medium containing fenugreek seed extract (0, 1 and 10 µg ml-1). Oocytes of a control group were matured in a medium containing 1 µg ml-1 estradiol 17β. After maturation, half of oocytes were fixed and stained for evaluation of nuclear maturation. The rest of oocytes were fertilized in vitro with fresh semen, then cultured for 9 days for the assessment of the developmental capacity of the oocytes. The results showed that the mean values of oocytes with expanded cumulus cells percentage were not significantly different among all groups (P < 0.05). But nuclear maturation rate of oocytes matured with 10 µg ml-1 fenugreek seed extract was significantly higher than that of the control group. The maturation rate and development to morula and blastocyst stage for oocytes matured at 10 µg ml-1 fenugreek seed extract was significantly higher than those matured at 1µg ml-1 of fenugreek seed extract and the control group. In conclusion, better maturation and developmental capacity rate to morula and blastocyst stage were obtained by the addition of 10 µg ml-1 fenugreek seed extract to maturation medium than addition of 1 µg ml-1 estradiol-17β (P < 0.05).

Keywords: fenugreek seed extract, in vitro maturation, sheep oocytes, in vitro fertilization, embryo development

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Authors: Babak Qasemi-Panahi, Gholamali Moghaddam, Seyed-Abbas Rafat, Hossein Daghigh Kia, Mansoureh Movahedin, Reza Hadavi

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The objective of this study was to determine the effects of IGF-I on ovine oocytes maturation and subsequent development of embryos derived from in vitro fertilization (IVF). In vitro maturation (IVM) of oocytes and in vitro culture (IVC) of embryos was conducted with or without 100 ng/mL IGF-1. In the IGF-I treated group, mean percentage of oocyte maturation was significantly higher than the control group (57.67 ± 3.04 versus 49.81 ± 3.04%, respectively, P < 0.05). However, in comparison with control group, there was no significant effect of IGF-1 on rates of cleavage, morula, and blastocyst formation (85% versus 84%; 63% versus 65%, and 40% to 39%, respectively). These data demonstrate that IGF-I has a positive effect on ovine oocyte maturation rate, but it has not the significant outcome on embryo development.

Keywords: ovine, IGF-I, IVM, ICSI

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Authors: Debora P. Arce, Flavia J. Krsticevic, Marco R. Bertolaccini, Joaquín Ezpeleta, Estela M. Valle, Sergio D. Ponce, Elizabeth Tapia

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Small Heat Shock Proteins (sHSPs) are low molecular weight chaperones that play an important role during stress response and development in all living organisms. Fruit maturation and oxidative stress can induce sHSP synthesis both in Arabidopsis and tomato plants. RNA-Seq technology is becoming widely used in various transcriptomics studies; however, analyzing and interpreting the RNA-Seq data face serious challenges. In the present work, we de novo assembled the Solanum lycopersicum transcriptome for three different maturation stages (mature green, breaker and red ripe). Differential gene expression analysis was carried out during tomato fruit development. We identified 12 sHSPs differentially expressed that might be involved in breaker and red ripe fruit maturation. Interestingly, these sHSPs have different subcellular localization and suggest a complex regulation of the fruit maturation network process.

Keywords: sHSPs, maturation, tomato, RNA-Seq, assembly

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Authors: Al-Mutary M., Alhimaidi A., Al-Ghadi M. Iwamoto D., Javed Ahmad. Abdulaziz A. Al-Khedhairy

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The aim of the present study was to evaluate the effect of ovine follicular fluid supplementation during IVM of sheep oocytes on the resumption of meiosis, glutathione (GSH) content and expression of Bax, Bcl-2, and HSPB1 genes. Sheep ovaries were collected from Riyadh slaughterhouse, KSA. Oocytes were aspirated from 3-6 mm follicles. Ovine oocytes were cultured in maturation medium with 0% (control), 10%, 20%, 40% of ovine follicular fluid for 24 h. Results indicated that the rate of oocyte maturation was significantly (P≤0.05) decreased in 40% OFF (36.87%) versus the control (61.3%), 10% OFF (63.95%) and 20% OFF (64.08%). Supplementation of 10% OFF to IVM medium induced an intra-oocyte GSH concentration significantly higher than that found in ovine oocytes cultured with 20% OFF and 40% OFF and similar to the GSH content in oocytes cultured without FF. Real time polymerase chain reaction analysis for gene expression revealed no differences in Bax, Bcl-2, HSPB1 genes between control and 10% OFF group, whereas they were strongly expressed in 20% OFF and 40% OFF (P < 0.05) when compared to the control and 10% OFF. In conclusion the addition of 10% OFF to the IVM culture of sheep oocytes is recommended to support cytoplasmic maturation and increase oocytes competence.

Keywords: IVM, oocyte maturation, gene expression, follicular fluid

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Authors: Goreti Botelho, Filomena Gomes, Fernanda M. Ferreira, Ilda Caldeira

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The strawberry tree (Arbutus unedo L.) is a small tree or shrub from botanical Ericaceae family that grows spontaneously nearby the Mediterranean basin and produce edible red fruits. A traditional processed fruit application, in Mediterranean countries, is the production of a spirit (known as aguardente de medronho, in Portugal) obtained from the fermented fruit. The main objective of our study was to contribute to the knowledge about the influence of the degree of maturation of fruits in the volatile composition and quality of arbutus spirit. The major volatiles in the three distillates fractions (head, heart and tail) obtained from fermentation of two different fruit maturation levels were quantified by GC-FID analysis and ANOVA one-way was performed. Additionally, the total antioxidant capacity and total phenolic compounds of both arbutus fruit spirits were determined, by ABTS and Folin-Ciocalteau method, respectively. The methanol concentration is superior (1022.39 g/hL a.a.) in the spirit made from fruits with highest total soluble solids, which is a value above the legal limit (1000 g/hL a.a.). Overall, our study emphasizes, for the first time, the influence of maturation degree of arbutus fruits in the spirit volatile composition and quality.

Keywords: arbutus fruit, maturation, quality, spirit

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Authors: D. C. Soares, J. T. S. Santos, D. G. Costa, A. K. S. Abud, T. P. Nunes, A. V. D. Figueiredo, A. M. de Oliveira Junior

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Mangabeira (Hancornia speciosa Gomes), a native plant in Brazil, is found growing spontaneously in various regions of the country. The high perishability of tropical fruits such as mangaba, causes it to be necessary to use technologies that promote conservation, aiming to increase the shelf life of this fruit and add value. The objective of this study was to compare the mangabas lyophilisation curves behaviours with different sizes and maturation stages. The fruits were freeze-dried for a period of approximately 45 hours at lyophilizer Liotop brand, model L -108. It has been considered large the fruits between 38 and 58 mm diameter and small, between 23 and 28 mm diameter and the two states of maturation, intermediate and mature. Large size mangabas drying curves in both states of maturation were linear behaviour at all process, while the kinetic drying curves related to small fruits, independent of maturation state, had a typical behaviour of drying, with all the well-defined steps. With these results it was noted that the time of lyophilisation was suitable for small mangabas, a fact that did not happen with the larger one. This may indicate that the large mangabas require a longer time to freeze until reaches the equilibrium level, as it happens with the small fruits, going to have constant moisture at the end of the process. For both types of fruit were analysed water activity, acidity, protein, lipid, and vitamin C before and after the process.

Keywords: freeze dryer, mangaba, conservation, chemical characteristics

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Authors: Shifu Hu, Qiong Yu, Wei Xia, Changhong Zhu

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Background: P38α MAPK, which is a member of the canonical MAPK family, is activated in response to various extracellular stresses and plays a role in multiple cellular processes. It is well known that p38α MAPK play vital roles in oocyte maturation, but the localization and functional roles of p38α MAPK during the meiotic maturation of rat oocytes remain unknown. Study Design: In this study, western-blot and immunofluorescent staining were used to investigate the expression and subcellular localization of p38α MAPK during the meiotic maturation of rat oocytes. SB203580, a specific inhibitor of p38α MAPK, was used to study the roles of p38α MAPK in the meiotic cell cycle of rat oocytes. Results: The results found that p38α MAPK phosphorylation (p-p38α MAPK, indicative of p38α MAPK activation) was low at the germinal vesicle (GV) stage, increased 3 h after germinal vesicle breakdown (GVBD), and maintained its maximum at MI (metaphase I) or M II (metaphase II). The p-p38α MAPK mainly accumulated in the germinal vesicle and had no obvious expression in the nucleus. From GVBD to M II, p-p38α MAPK was distributed in the cytoplasm around either the chromosomes or the spindle. We used SB203580, an inhibitor of p38α MAPK, to investigate the possible functional role of p38α MAPK during rat oocyte meiotic maturation. Treatment of GV stage oocytes with 20 μM SB203580 blocked p-p38α MAPK activity, and the spindles appeared abnormal. Additionally, the rate of GVBD after 3h of culture with 20 μM SB203580 (58.8%) was significantly inhibited compared with the control (82.5%, p < 0.05), and the polar body extrusion rate after 12 h of culture with SB203580 was also significantly decreased compared with the control (40.1 vs. 73.3%, p < 0.05). Conclusions: These data indicate that p38α MAPK may play a vital role in rat oocyte meiotic maturation.

Keywords: meiotic maturation, oocyte, p38α MAPK, spindle

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Authors: Thanyaporn Senarai, Rapeepun Vanichviriyakit, Shinji Miyata, Chihiro Sato, Prapee Sretarugsa, Wattana Weerachatyanukul, Ken Kitajima

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In this study, we characterized a region-specific 250 kDa protein that was secreted of MSD fluid, which is believed to play dual functions in forming a spermatophoric wall for sperm physical protection, and in sperm membrane modification as part of sperm maturation process. The partial amino acid sequence and N-terminal sequencing revealed that the MSD-specific 250 kDa protein showed a high similarity with a plasma-rich protein, α-2 macroglobulin (α2M), so termed pp-α2M. This protein was a large glycoprotein contained predominantly mannose and GlcNAc. The expression of pp-α2M mRNA was detected in spermatic duct (SD), androgenic gland (AG) and hematopoietic tissue, while the protein expression was rather specific to the apical cytoplasm of MSD epithelium. The secretory pp-α2M in MSD fluid was acquired onto the MSD sperm membrane and was also found within the matrix of the acrosome. Distally, pp-α2M was removed from spermathecal sperm membrane, while its level kept constant in the sperm AC. Together the results indicate that pp-α2M is a 250 kDa region-specific secretory protein which plays roles in sperm physical protection and also acts as maturation factor in the P. pelagicus sperm.

Keywords: alpha-2 macroglobulin, blue swimming crab, sperm maturation, spermatic duct

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Authors: Maimuna Nontji

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The life cycle of rice plant has three phases of growth; they are the vegetative, reproductive and maturation phase. They greatly affect the life of dynamics metanotrof bacterial as reducer methane emissions in the rice field, both of population and on the rate of growth. The aim of this study was to analyze the population and growth rate of methanotrof isolates which has been isolated in previous studies. Isolates were taken at all the life cycle of rice plant. Population of analysis was conducted by standard plate count method and growth rate was analysed by logarithmic calculation. The results showed that each isolate varied in population and growth rate. The highest population was obtained in the isolates Gowa Methanotrof Reproductive (GMR 8) about 7.06 x 10 11 cfu / ml on 3 days of incubation and the lowest population was obtained in the Gowa Methanotrof Maturation (GMP 5) about 0.27 x 10 11 cfu / ml on 7 day of incubation. Some isolate were demonstrated in long growth rate about 5 days of incubation and another are 3 days.

Keywords: emission, methanotrof, methane, population

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Authors: Sri Wahjuningsih, Nur Ihsan, Hadiah

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In the embryo transfer program, to address the limited production of embryos in vivo, in vitro embryo production has become an alternative approach that is relatively inexpensive. One potential source of embryos that can be developed is to use immature oocytes then conducted in vitro maturation and in vitro fertilization. However, obstacles encountered were oocyte viability mammals have very limited that it cannot be stored for a long time, so we need oocyte cryopreservation. The research was conducted to know the optimal concentration use of ethylene glycol as a cryoprotectant on oocytes freezing.Material use in this research was immature oocytes; taken from abbatoir which was aspirated from follicle with diameter 2-6 mm. Concentration ethylen glycol used were 0,5 M, I M, 1,5 M and 2M. The freezing method used was conventional method combined with a five-step protocol washing oocytes from cryoprotectant after thawing. The result showed that concentration ethylen glycol have the significant effect (P<0.05) on oocytes quality after thawing and in vitro maturation. It was concluded that concentration 1,5 M was the best concentration for freezing oocytes using conventional method.

Keywords: bovine, conventional freezing, ethylen glycol, oocytes

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Authors: Ihtisham Bukhari

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In the current study, we enrolled a 46, XY phenotypically female patient bearing testes in her inguinal canal. DNA sequencing of the AR gene detected a missense mutation C.1715A > G (p. Y572C) in exon 2 which is already known to cause Complete androgen insensitivity syndrome (CAIS). We further studied the effects of this mutation on the testicular histopathology of the patient. No spermatocytes were seen in the surface spreading of testicular tissues while H&E staining showed that seminiferous tubules predominantly have only Sertoli cells. To confirm this meiotic failure is likely due to the current AR mutation we performed mRNA expression of genes associated with AR pathway, expression and location of the associated proteins in testicular tissues. Western blot and real-time PCR data showed that the patient had high levels of expression of AMH, SOX9, and INNB in testis. Tubules were stained with SOX9 and AMH which revealed Sertoli cell maturation arrest. Therefore, we suggest that AR mutation enhances AMH expression which ultimately leads to failure in the maturation of Sertoli cells and failure in spermatogenesis.

Keywords: androgen receptor, spermatogenesis, infertility, Sertoli cell only syndrome

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Authors: Saha Saradindu, Das Payel, Somdeb BoseDasgupta

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Tuberculosis caused by Mycobacteria tuberculosis is a dreadful disease and more so with the advent of extreme and total drug-resistant species. Mycobacterial pathogenesis is an ever-changing paradigm from phagosome maturation block to phagosomal escape into macrophage cytosol and finally acid tolerance and survival inside the lysosome. Mycobacteria are adept at subverting the host immune response by highjacking host cell signaling and secreting virulence factors. One such virulence factor is a ser/thr kinase; Protein kinase G (PknG), which is known to prevent phagosome maturation. The host substrates of PknG, allowing successful pathogenesis still remain an enigma. Hence we carried out a comparative phosphoproteomic screen and identified a number of substrates phosphorylated by PknG. We characterized some of these substrates in vivo and in vitro and observed that PknG mediated phosphorylation of these substrates leads to reduced TNFa production as well as decreased response to TNFa induced macrophage necroptosis, thus enabling mycobacterial survival and proliferation.

Keywords: mycobacteria, Protein kinase G, phosphoproteomics, necroptosis

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Authors: T. Topór, A. Derkowski, P. Ziemiański

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Formation of organic matter (OM)-hosted nanopores upon thermal maturation are one of the key factor controlling methane storage potential in unconventional shale-gas reservoirs. In this study, the subcritical CO₂ and N₂ gas adsorption measurements combined with scanning electron microscopy and supercritical methane adsorption have been used to characterize pore system and methane storage potential in black shales from the Baltic Basin (Poland). The samples were collected from a virtually equivalent Llandovery strata across the basin and represent a complete digenetic sequence, from thermally immature to overmature. The results demonstrate that the thermal maturation is a dominant mechanism controlling the formation of OM micro- and mesopores in the Baltic Basin shales. The formation of micro- and mesopores occurs in the oil window (vitrinite reflectance; leavedVR; ~0.5-0.9%) as a result of oil expulsion from kerogenleft OM highly porous. The generated hydrocarbons then turn into solid bitumen causing pore blocking and substantial decrease in micro- and mesopore volume in late-mature shales (VR ~0.9-1.2%). Both micro- and mesopores are regenerated in a middle of the catagenesis range (VR 1.4-1.9%) due to secondary cracking of OM and gas formation. The micropore volume in investigated shales is almost exclusively controlled by the OM content. The contribution of clay minerals to micropore volume is insignificant and masked by a strong contribution from OM. Methane adsorption capacity in the Baltic Basin shales is predominantly controlled by microporous OM with pores < 1.5 nm. The mesopore volume (2-50 nm) and mesopore surface area have no effect on methane sorption behavior. The adsorbed methane density equivalent, calculated as absolute methane adsorption divided by micropore volume, reviled a decrease of the methane loading potential in micropores with increasing maturity. The highest methane loading potential in micropores is observed for OM before metagenesis (VR < 2%), where the adsorbed methane density equivalent is greater than the density of liquid methane. This implies that, in addition to physical adsorption, absorption of methane in OM may occur before metagenesis. After OM content reduction using NaOCl solution methane adoption capacity substantially decreases, suggesting significantly greater adsorption potential for OM microstructure than for the clay minerals matrix.

Keywords: maturation, methane sorption, organic matter, porosity, shales

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Authors: Ratmawati Malaka, Effendi Abustam, Kusumandari Indah Prahesti, Sudirman Baco

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Dangke is natural cheese from Enrekang South Sulawesi, Indonesia produced through aglutination buffalo milk, cow, goat or sheep using the sap of papaya (Carica papaya). Dangke has been widely known in South Sulawesi but this soft cheese product diversification by using passion fruit juice as milk clotting agents has not been used. Passion fruit juice has a high acidity with a pH of around 4 - 4.5 and has a proteolytic enzyme, so that it can be used to agglutinate milk. The purpose of this study was to investigate the nature Dangke using passion fruit juice as coagulate milk. Dangke made by 10 lt of raw milk by heating at a temperature of 73oC with coagulant passion fruit juice (7.5% and 10%), and added 1% salt. Curd clot and then be formed using a coconut shell, is then pressed until the cheese is compact. The cheese is then observed for 28 days ripening at a temperature of about 5 ° C. Dangke then studied to violence, pH, fat levels and microstructure. Hardness is determined using CD-shear Force, pH is measured using a pH meter Hanna, and fat concentrations were analyzed with methods of proximate. Microstructure viewed using a light microscope with magnification 1000 x. The results showed that the levels of clotting material very significant influence on hardness, pH, and lipid levels. Maturation increase the hardness but lower the pH, the level of fat soft cheese with an average Dangke respectively 21.4% and 30.5% on 7.5% addition of passion fruit juice and 10%. Dangke violence is increasing with the increasing maturation time (1.38 to 3.73 kg / cm), but Dangke pH was decreased by the increase in storage maturation (5.34 to 4.1). Microktrukture cheeses coagulated with 10% of the passion fruit are very firmer and compact with a full globular fat of 7.5%. But the sensory properties of the soft cheese similar in both treatment. The manufacturing process with the addition of coagulant passion fruit juice on making Dangke affect hardness, pH, fat content and microstructure during storage at 5 ° C for 1 d - 28 d.

Keywords: dangke, passion fruits, microstructure, cheese

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Authors: Udoaka A. I., Didia B. C.

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Background: The maturation of a child’s bones as it grows to adulthood can be viewed radiologically. The skeletal age (bone age) is the average age at which a particular stage of bone maturation is achieved. The Greulich and Pyle standard is the commonest method used to assess the skeletal age using the hand and wrist radiograph throughout the world. This atlas was compiled solely from Caucasian children and made use of the orderly sequence of carpal ossification to determine the skeletal age. Several authors have faulted this atlas for not being suitable for other races. Aim: The aim of this study is to determine if the Greulich and Pyle Atlas is applicable to Nigerian children when compared to their chronological ages. Methods: The total number of 78 normal radiographs of the hand and wrist of Nigerian children obtained from several hospitals were used for this study . These radiographs were compared with the atlas and their skeletal ages noted form the atlas. The child’s chronological age in each case was also recorded. Results: The result shows a mean increase of two months in the skeletal ages of the Nigerian children compared to the atlas. This difference, however, was not significant. The skeletal age (in months) was greater in 77% of the children than the expected age in the atlas. Conclusion: The mean skeletal age of Nigerian children, though more than the standard in the atlas, is not statistically significant; as a result the study finds the radiographic atlas of Greulich and Pyle atlas applicable to Nigerian children.

Keywords: Greulich and Pyle Atlas, radiograph, skeletal age

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Authors: Hatairuk Tungkasen, Somrudee Phetchrid, Suwapat Jaidee, Supinya Yoomak, Chantana Kankamol, Mayuree Pumipaiboon, Mayuva Areekijseree

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Ovaries of reproductive female pigs were obtained from local slaughterhouses in Nakorn Pathom Province, Thailand. Follicular fluid of medium follicle (5-6 diameters) and large follicles (7-8 mm and 10 mm in diameter) were aspirated and collected by sterile technique and analyzed protein pattern. The follicular fluid protein bands were found by SDS-PAGE which has no protein band in difference compared to standard protein band. So we chose protein band molecular weight 50, 62-65, 75-80, 90, 120-160, and >220 kDa were analyzed by LC/MS/MS. The result was found immunoglobulin gamma chain, keratin, transferrin, heat shock protein, and plasminogen precursor, ceruloplasmin, and hemopexin, and protease, respectively. All proteins play important roles in promotion and regulation on growth and development of reproductive cells. The result of this study found many proteins which were useful and important for in vitro oocyte maturation and embryonic development of cell technology in animals. The further study will be use porcine follicular fluid protein of medium and large follicles as feeder cells in in vitro condition to promote oocyte and embryo maturation.

Keywords: follicular fluid protein, LC/MS/MS, porcine oocyte, SDS-PAGE

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Authors: Ali. Marjani, M. Farsi, M. Rahimizadeh

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Chickpea (Cicer arietinum L.) is one of the important grain legume crops in the world. However, drought stress is a serious threat to chickpea production, and development of drought-resistant varieties is a necessity. Field experiments were conducted to evaluate the response of 8 chickpea genotypes (MCC* 696, 537, 80, 283, 392, 361, 252, 397) and drought stress (S1: non-stress, S2: stress at vegetative growth stage, S3: stress at early bloom, S4: stress at early pod visible) at different growth stages. Experiment was arranged in split plot design with four replications. Difference among the drought stress time was found to be significant for investigated traits except biological yield. Differences were observed for genotypes in flowering time, pod information time, physiological maturation time and yield. Plant height reduced due to drought stress in vegetative growth stage. Stem dry weight reduced due to drought stress in pod visibly. Flowering time, maturation time, pod number, number of seed per plant and yield cause of drought stress in flowering was also reduced. The correlation between yield and number of seed per plant and biological yield was positive. The MCC283 and MCC696 were the high-tolerance genotypes. These results demonstrated that drought stress delayed phonological growth in chickpea and that flowering stage is sensitive.

Keywords: chickpea, drought stress, growth stage, tolerance

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Authors: K. Ö. Kartal, L. Maul, M. Hägele

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With the development of new technologies come additional opportunities for the founding of companies and new markets to be created. The barriers to entry are lowered and technology makes old business models obsolete. Incumbent companies have to be adaptable to this quickly changing environment. They have to start the process of digital maturation and they have to be able to adapt quickly to new and drastic changes that might arise. One of the biggest barriers for organizations in order to do so is their culture. This paper shows the core elements of a corporate culture that supports the process of digital maturation in incumbent organizations. Furthermore, it is explored how ideation and innovation can be used in a strategy in order to facilitate these core elements of culture that promote digital maturity. Focus areas are identified for the design of ideation strategies, with the aim to make the facilitation and incitation process more effective, short to long term. Therefore, one in-depth case study is conducted with data collection from interviews, observation, document review and surveys. The findings indicate that digital maturity is connected to cultural shift and 11 relevant elements of digital culture are identified which have to be considered. Based on these 11 core elements, five focus areas that need to be regarded in the design of a strategy that uses ideation and innovation to facilitate the cultural shift are identified. These are: Focus topics, rewards and communication, structure and frequency, regions and new online formats.

Keywords: digital transformation, innovation management, ideation strategy, creativity culture, change

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Authors: Jose A. Aznar-Moreno, Xi Jiang, Stefan Burén, Luis M. Rubio

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Integration of prokaryotic nitrogen fixation (nif) genes into the plastid genome for expression of functional nitrogenase components could render plants capable of assimilating atmospheric N2 making their crops less dependent of nitrogen fertilizers. The nitrogenase Fe protein component (NifH) has been used as proxy for expression and targeting of Nif proteins within plant and yeast cells. Here we use tobacco plants with the Azotobacter vinelandii nifH and nifM genes integrated into the plastid genome. NifH and its maturase NifM were constitutively produced in leaves, but not roots, during light and dark periods. Nif protein expression in transplastomic plants was stable throughout development. Chloroplast NifH was soluble, but it only showed in vitro activity when isolated from leaves collected at the end of the dark period. Exposing the plant extracts to elevated temperatures precipitated NifM and apo-NifH protein devoid of [Fe4S4] clusters, dramatically increasing the specific activity of remaining NifH protein. Our data indicate that the chloroplast endogenous [Fe-S] cluster biosynthesis was insufficient for complete NifH maturation, albeit a negative effect on NifH maturation due to excess NifM in the chloroplast cannot be excluded. NifH and NifM constitutive expression in transplastomic plants did not affect any of the following traits: seed size, germination time, germination ratio, seedling growth, emergence of the cotyledon and first leaves, chlorophyll content and plant height throughout development.

Keywords: NifH, chloroplast, nitrogen fixation, crop improvement, transplastomic plants, fertilizer, biotechnology

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Authors: Clorinda Hogan, Shaun Abbott, Mark Halaki, Marcela Torres Catiglioni, Goshi Yamauchi, Lachlan Mitchell, James Salter, Michael Romann, Stephen Cobley

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Introduction: Common annual age-group competition structures unintentionally introduce participation inequalities, performance (dis)advantages and selection biases due to the effect of maturational variation between youth swimmers. On this basis, there are implications for improving performance evaluation strategies. Therefore the aim was to: (1) To determine maturity timing distributions in female youth swimming; (2) quantify the relationship between maturation status and 100-m FC performance; (3) apply Maturational-based Corrective Adjustment Procedures (Mat-CAPs) for removal of maturational status performance influences. Methods: (1) Cross-sectional analysis of 663 female (10-15 years) swimmers who underwent assessment of anthropometrics (mass, height and sitting height) and estimations of maturity timing and offset. (2) 100-m front-crawl performance (seconds) was assessed at Australian regional, state, and national-level competitions between 2016-2020. To determine the relationship between maturation status and 100-m front-crawl performance, MO was plotted against 100-m FC performance time. The expected maturity status - performance relationship for females aged 10-15 years of age was obtained through a quadratic function (y = ax2 + bx + c) from unstandardized coefficients. The regression equation was subsequently used for Mat-CAPs. (3) Participants aged 10-13 years were categorised into maturity-offset categories. Maturity offset distributions for Raw (‘All’, ‘Top 50%’ & ‘Top 25%’) and Correctively Adjusted swim times were examined. Chi-square, Cramer’s V and ORs determined the occurrence of maturation biases for each age group and selection level. Results—: (1) Maturity timing distributions illustrated overrepresentation of ‘normative’ maturing swimmers (11.82 ± 0.40 years), with a descriptive shift toward the early maturing relative to the normative population. (2) A curvilinear relationship between maturity-offset and swim performance was identified (R2 = 0.53, P < 0.001) and subsequently utilised for Mat-CAPs. (3) Raw maturity offset categories identified partial maturation status skewing towards biologically older swimmers at 10/11 and 12 years, with effect magnitudes increasing in the ‘Top 50%’ and ‘25%’ of performance times. Following Mat-CAPs application, maturity offset biases were removed in similar age groups and selection levels. When adjusting performance times for maturity offset, Mat-CAPs was successful in mitigating against maturational biases until approximately 1-year post Peak Height Velocity. The overrepresentation of ‘normative’ maturing female swimmers contrasted with the substantial overrepresentation of ‘early’ maturing male swimmers found previously in 100-m front-crawl. These findings suggest early maturational timing is not advantageous in females, but findings associated with Aim 2, highlight how advanced maturational status remained beneficial to performance. Observed differences between female and male maturational biases may relate to the differential impact of physiological development during pubertal years. Females experience greater increases of fat mass and potentially differing changes in body shape which can negatively affect swim performance. Conclusions: Transient maturation status-based participation and performance advantages were apparent within a large sample of Australian female youth 100-m FC swimmers. By removing maturity status performance biases within female youth swimming, Mat-CAPs could help improve participation experiences and the accuracy of identifying genuinely skilled female youth swimmers.

Keywords: athlete development, long-term sport participation, performance evaluation, talent identification, youth competition

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Authors: Ntalamagka N., Sanchis-Benlloch P. J., Mayoral-Serrano R., Tena-Medialdea J., García-March J. R.

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The pen shell Pinna nobilis population has declined dramatically since 2016 due to die-off events observed in the whole extent of the Mediterranean Sea associated with the protozoan Haplosporidium pinnae. As of 2019, it is considered a critically endangered species. Due to its ecological importance and its endangered status, several initiatives have been developed for its salvation and recovery. This research is an effort to understand and control its reproduction under captivity. As a limited number of Pinna nobilis individuals could be used for experimentation, the possibility of using the Pinna rudis as a model animal was explored. The molecular mechanism that regulates the reproduction of both species is unknown; consequently, transcriptomic analysis was performed to identify neuropeptides that are expressed in the key regulatory tissues of the visceral ganglia and gonads of both species. Neuropeptides form an important group of signaling peptides that regulate reproductive, behavioral and physiological functions in molluscs. In total, 17 neuropeptide precursors were identified in P. nobilis and 14 in P. rudis transcriptomes; 14 of them were identical in both species. This affinity verified the genetic similarity of these species at the reproduction level. APGWamide, buccalin, ELH and GnRH were tested in P. rudis and demonstrated their capacity to advance gonadal maturation and trigger spawning while spawning was recorded in P. nobilis after the usage of APGWamide and buccalin. The neuropeptides were administered using intramuscular injection and cholesterol implants following relative literature as well as a new method was developed for external administration without the use of anesthesia using a mathematical model. The know-how of this research will not only lead to the survival of the species but also will narrow the horizons of broodstock conditioning of other similar species.

Keywords: neuropeptides, Pinna nobilis, reproduction, transcriptomics

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Authors: Zakir Hossain, Saddam Hossain

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A nutritionally balanced diet and selection of appropriate species are important criteria in aquaculture. The present study was conducted to evaluate the effects of polyunsaturated fatty acids (PUFAs) and beta glucan-containing diets on growth performance, feed utilization, maturation, immunity, early embryonic and larval development of endangered Pabdah catfish, Ompok pabda. In this study, squid extracted lipids and mushroom powder were used as the source of PUFAs and beta-glucan, respectively, and formulated two isonitrogenous diets such as a basal or control (CON) diet and a treated (PBG) diet with maintaining 30% protein levels. During the study period, similar physicochemical conditions of water such as temperature, pH, and dissolved oxygen (DO) were 26.5±2 °C, 7.4±0.2, and 6.7±0.5 ppm, respectively, in each cistern. The results showed that final mean body weight, final mean length gain, food conversion ratio (FCR), specific growth rate (SGR), food conversion efficiency (%), hepato somatic index (HSI), kidney index (KI), and viscerosomatic index (VSI) were significantly (P<0.01 and P<0.05) higher in fish fed the PBG diet than that of fish fed the CON diet. The length-weight relationship and relative condition factor (K) of O. pabda were significantly (P<0.05) affected by the PBG diet. The gonadosomatic index (GSI), sperm viability, blood serum calcium ion concentrations (Ca²⁺), and vitellogenin level were significantly (P<0.05) higher in fish fed the PBG diet than that of fish fed the CON diet; which was used to the indication of fish maturation. During the spawning season, lipid granules and normal morphological structure were observed in the treated fish liver, whereas fewer lipid granules of liver were observed in the control group. Based on the immunity and stress resistance-related parameters such as hematological indices, antioxidant activity, lysozyme level, respiratory burst activity, blood reactive oxygen species (ROS), complement activity (ACH50 assay), specific IgM, brain AChE, plasma PGOT, and PGPT enzyme activity were significantly (P<0.01 and P<0.05) higher in fish fed the PBG diet than that of fish fed the CON diet. The fecundity, fertilization rate (92.23±2.69%), hatching rate (87.43±2.17 %), and survival (76.62±0.82%) of offspring were significantly higher (P˂0.05) in the PBG diet than in control. Consequently, early embryonic and larval development was better in PBG treated group than in control. Therefore, the present study showed that the polyunsaturated fatty acids (PUFAs) and beta-glucan enriched experimental diet were more effective and achieved better growth, feed utilization, maturation, immunity, and spawning performances of O. pabda.

Keywords: lipids, beta-glucan, fish maturity, fish immunity

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Authors: Elisabeth Louise Ndjengue Mindang, Charline Florence Awounfack, Derek Tantoh Ndinteh, Rui W. M. Krause, Dieudonne Njamen

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The evolution of human fertility over the last 50 years has shown considerable problems due to a growing number of couples that find it difficult to procreate without medical assistance. In Africa, this inability to conceive affect 30 to 40% of couples. A number of contaminants in the environment are thought to contribute significantly to the observed infertility epidemic. Methods: On this basis, the impact of 40-days unique oral administration (between 9 and 10 am) of potassium sorbate (at of 12.5, 45, and 78 mg/kg BW doses) was evaluated on sexual maturation and hematologic parameters on immature Wistar rats (21-22 days of age). At the end of the treatment, animals were sacrificed. Vaginal opening was evaluated before the sacrifice. After the sacrifice, relative weight of reproductive organs, pituitary gonadotrophin level (LH and FSH), and sexuals steroids (estrogen and progesterone), cholesterol level in ovaries, folliculogenesis, and some hematological parameters were evaluated. Results: Compared to the control group, no significant variation was observed in the body weight of the animals treated with patassium sorbate. On the other hand, potassium sorbate, a significantly lower percentage (25%) of vaginal-opening in these rats, was observed from day 46 of age (p <0.01); likewise, a significant decrease was observed on the relative weight of the ovaries (p <0.01), number of primary follicles (p <0.01), and a significant increase of follicle number (p <0.001) at 78 mg/kg BW have been obseved. Potassium sorbate always decreased the number of white blood cells (p <0.05). Taken together, these results confirm the disturbing effects on the endocrine system, causing a decrease in fertility by increasing the number of follicles in atresia. A deleterious effect on the immune system was also observed. Overall, these results validate at least in part the global observations on the growing decline in fertility in populationsfeeding increasingly on industrial processed foods.

Keywords: potassium sorbate, early puberty, folliculogenesis, endocrine disruptor, immatur wistar rat

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Authors: M. Areekijseree, W. Pongsawat, M. Pumipaiboon, C. Thepsithar, S. Sengsai, T. Chuen-Im

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Morphological interaction of porcine cumulus-oocyte complexes (pCOCs) was investigated on in vitro condition using electron microscope (SEM and TEM). The totals of 1,923 oocytes were round in shape, surrounded by zona pellucida with layer of cumulus cells ranging between 59.29-202.14 µm in size. They were classified into intact-, multi-, partial cumulus cell layer oocyte, and completely denuded oocyte, at the percentage composition of 22.80% 32.70%, 18.60%, and 25.90 % respectively. The pCOCs classified as intact- and multi cumulus cell layer oocytes were further culturing at 37°C with 5% CO2, 95% air atmosphere and high humidity for 44 h in M199 with Earle’s salts supplemented with 10% HTFCS, 2.2 mg/mL NaHCO3, 1 M Hepes, 0.25 mM pyruvate, 15 µg/mL porcine follicle-stimulating hormone, 1 µg/mL LH, 1µg/mL estradiol with ethanol, and 50 µg/mL gentamycin sulfate. On electron microscope study, cumulus cells were found to stick their processes to secrete substance from the sac-shape end into zona pellucida of the oocyte and also communicated with the neighboring cells through their microvilli on the beginning of incubation period. It is believed that the cumulus cells communicate with the oocyte by inserting the microvilli through this gap and embedded in the oocyte cytoplasm before secreting substance, through the sac-shape end of the microvilli, to inhibit primary oocyte development at the prophase I. Morphological changes of the complexes were observed after culturing for 24-44 h. One hundred percentages of the cumulus layers were expanded and cumulus cells were peeling off from the oocyte surface. In addition, the round-shape cumulus cells transformed themselves into either an elongate shape or a columnar shape, and no communication between cumulus neighboring cells. After 44 h of incubation time, diameter of oocytes surrounded by cumulus cells was larger than 0 h incubation. The effect of hormones in culture medium is exerted by their receptors present in porcine oocyte. It is likely that all morphological changes of the complexes after hormone treatment were to allow maturation of the oocyte. This study demonstrated that the association of hormones in M199 could promote porcine follicle activation in 44 h in vitro condition. This culture system should be useful for studying the regulation of early follicular growth and development, especially because these follicles represent a large source of oocytes that could be used in vitro for cell technology.

Keywords: cumulus cells, electron microscopy, in vitro, porcine oocyte

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Authors: Akbarova Gulnozakhon

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Relevance. Physical exercises can prolong the function of the growth zones of long tubular bones, delay the fusion of the epiphyses and diaphyses of bones and, thus, increase the growth of the body. At the same time, intensive strength exercises can accelerate the process of ossification of bone growth zones and slow down their growth in length. The influence of physical exercises on the process of biological maturation is noted. Gymnastics, which requires intense speed and strength loads, delays puberty. On the other hand, it is indicated that the relatively slow puberty of gymnasts is associated with the selection of girls with a special somatotype in this sport. It was found that the later onset of menstruation in female athletes does not have a negative effect on the maturation process and fertility (the ability to procreate). Observations are made about the normalizing influence of sports on the puberty of girls. The purpose of the study. Our goal is to study physical activity of varying intensity on the formation of secondary sexual characteristics and hormonal status of girls in adolescence. Each biological process peculiar to a given organism is not in a stationary state, but fluctuates with a certain frequency. According to the duration, there are, for example, circadian cycles, and infradian cycles, a typical example of which is the menstrual cycle. Materials and methods, results. Violations of menstrual function in athletes were detected by applying a questionnaire survey that contains several paragraphs and sub-paragraphs where passport data, anthropometric indicators, taking into account anthropometric indices, information about the menstrual cycle are indicated. Of 135 female athletes aged 1-3 to 16 years engaged in various sports - gymnasts, menstrual function disorders were noted in 86.7% (primary or secondary amenorrhea, irregular MC), in swimming-in 57.1%. The general condition also changes during the menstrual cycle. In a large percentage of cases, athletes indicate an increase in irritability in the premenstrual (45%) and menstrual (36%) phases. During these phases, girls note an increase in fatigue of 46.5% and 58% (respectively). In girls, secondary sexual characteristics continue to form during puberty and the clearest indicator of the onset of puberty is the age of the onset of the first menstruation - menarche. Conclusions. 1. Physical exercise has a positive effect on all major systems of the body and thus promotes health.2. Along with a beneficial effect on human health, physical exercise, if the requirements of sports are not observed, can be harmful.

Keywords: girls health, anthropometric, physical development, reproductive health

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Authors: Gredson Keif Souza, Nehemias Curvelo Pereira

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Kernel oil from Macaúba is an important source of essential fatty acids. Thus, a new knowledge of the oil of this species could be used in new applications, such as pharmaceutical drugs based in the manufacture of cosmetics, and in various industrial processes. The aim of this study was to characterize the kernel oil of macaúba (Acrocomia Totai) at different times of their maturation. The physico-chemical characteristics were determined in accordance with the official analytical methods of oils and fats. It was determined the content of water and lipids in kernel, saponification value, acid value, water content in the oil, viscosity, density, composition in fatty acids by gas chromatography and molar mass. The results submitted to Tukey test for significant value to 5%. Found for the unripe fruits values superior to unsaturated fatty acids.

Keywords: extraction, characterization, kernel oil, acrocomia totai

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Authors: El Hadji Mamadou Sonko, Mbaye Mbéguéré, Cheikh Diop, Linda Strande

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In new approaches that are being developed for the treatment of sludge, the valorization of by-product is increasingly encouraged. In this perspective, Echinochloa pyramidalis has been successfully tested in Cameroon. Echinochloa pyramidalis is an efficient forage plant in the treatment of faecal sludge. It provides high removal rates and biosolids of high agronomic value. Thus in order to advise the use of this plant in planted drying beds in Senegal its comparison with the plants long been used in the field deserves to be carried out. That is the aim of this study showing the influence of the nature of the plants on the drainage, the purifying performances and the quality of the biosolids. Echinochloa pyramidalis, Typha australis, and Phragmites australis are the three macrophytes used in this study. The drainage properties of the beds were monitored through the frequency of clogging, the percentage of recovered leachate and the dryness of the accumulated sludge. The development of plants was followed through the measurement of the density. The purification performances were evaluated from the incoming raw sludge flows and the outflows of leachate for parameters such as Total Solids (TS), Total Suspended Solids (TSS), Total Volatile Solids (TVS), Chemical Oxygen Demand (COD), Total Kjeldahl Nitrogen (TKN), Ammonia (NH₄⁺), Nitrate (NO₃⁻), Total Phosphorus (TP), Orthophosphorus (PO₄³⁻) and Ascaris eggs. The quality of the biosolids accumulated on the beds was measured after 3 months of maturation for parameters such as dryness, C/N ratio NH₄⁺/NO₃⁻ ratio, ammonia, Ascaris eggs. The results have shown that the recovered leachate volume is about 40.4%; 45.6% and 47.3%; the dryness about 41.7%; 38.7% and 28.7%, and clogging frequencies about 6.7%; 8.2% and 14.2% on average for the beds planted with Echinochloa pyramidalis, Typha australis and Phragmites australis respectively. The plants of Echinochloa pyramidalis (198.6 plants/m²) and Phragmites australis (138 plants/m²) have higher densities than Typha australis (90.3 plants/m²). The nature of the plants has no influence on the purification performance with reduction percentages around 80% or more for all the parameters followed whatever the nature of the plants. However, the concentrations of these various leachate pollutants are above the limit values of the Senegalese standard NS 05-061 for the release into the environment. The biosolids harvested after 3 months of maturation are all mature with C/N ratios around 10 for all the macrophytes. The NH₄⁺/NO₃⁻ ratio is lower than 1 except for the biosolids originating from the Echinochloa pyramidalis beds. The ammonia is also less than 0.4 g/kg except for biosolids from Typha australis beds. Biosolids are also rich in mineral elements. Their concentrations of Ascaris eggs are higher than the WHO recommendations despite a percentage of inactivation around 80%. These biosolids must be stored for an additional time or composted. From these results, the use of Echinochloa pyramidalis as the main macrophyte can be recommended in the various drying beds planted in sub-Saharan climate conditions.

Keywords: faecal sludge, nature of plants, quality of biosolids, treatment performances

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Authors: Chien-Chun Huang, P. W. Yuan

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Banana is one of the important fruits which constitute a valuable source of energy, vitamins and minerals and an important food component throughout the world. The fruit ripening and maturity standards vary from country to country depending on the expected shelf life of market. During ripening there are changes in appearance, texture and chemical composition of banana. The changes of component of banana during ethylene-induced ripening are categorized as nutritive values and commercial utilization. The objectives of this study were to investigate the changes of chemical composition and physicochemical properties of banana during ethylene-induced ripening. Green bananas were harvested and ripened by ethylene gas at low temperature (15℃) for seven stages. At each stage, banana was sliced and freeze-dried for banana flour preparation. The changes of total starch, resistant starch, chemical compositions, physicochemical properties, activity of amylase, polyphenolic oxidase (PPO) and phenylalanine ammonia lyase (PAL) of banana were analyzed each stage during ripening. The banana starch was isolated and analyzed for gelatinization properties, pasting properties and microscopic appearance each stage of ripening. The results indicated that the highest total starch and resistant starch content of green banana were 76.2% and 34.6%, respectively at the harvest stage. Both total starch and resistant starch content were significantly declined to 25.3% and 8.8%, respectively at the seventh stage. Soluble sugars content of banana increased from 1.21% at harvest stage to 37.72% at seventh stage during ethylene-induced ripening. Swelling power of banana flour decreased with the progress of ripening stage, but solubility increased. These results strongly related with the decreases of starch content of banana flour during ethylene-induced ripening. Both water insoluble and alcohol insoluble solids of banana flour decreased with the progress of ripening stage. Both activity of PPO and PAL increased, but the total free phenolics content decreased, with the increases of ripening stages. As ripening stage extended, the gelatinization enthalpy of banana starch significantly decreased from 15.31 J/g at the harvest stage to 10.55 J/g at the seventh stage. The peak viscosity and setback increased with the progress of ripening stages in the pasting properties of banana starch. The highest final viscosity, 5701 RVU, of banana starch slurry was found at the seventh stage. The scanning electron micrograph of banana starch showed the shapes of banana starch appeared to be round and elongated forms, ranging in 10-50 μm at the harvest stage. As the banana closed to ripe status, some parallel striations were observed on the surface of banana starch granular which could be caused by enzyme reaction during ripening. These results inferred that the highest resistant starch was found in the green banana could be considered as a potential application of healthy foods. The changes of chemical composition and physicochemical properties of banana could be caused by the hydrolysis of enzymes during the ethylene-induced ripening treatment.

Keywords: maturation of banana, appearance, texture, soluble sugars, resistant starch, enzyme activities, physicochemical properties of banana starch

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Authors: Mehdi Abbasi, Masoumeh Majidi Zolbin

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Varicocele is one of the common causes of infertility in 30-50% of married men which occurs within the spermatic cord. It can be considered as an abnormal dilatation and stasis of veins of the pampiniform plexus that drain the testis. It occurs in 15-20% of the male population. Inducible nitric oxide synthase (NOS) activity has been frequently reported in varicose veins. Several studies have considered the relationship between varicocele and semen NO concentrations. NOS isoforms have been shown to regulate a number of functions, e.g., sperm motility and maturation and germ cell apoptosis in the testes. In adult patients with varicocele, the amount of NO levels in the varicose veins are 25 times higher than in serum of peripheral veins. The aim of this study was to review the effect of different antioxidant that we applied so far on sperm parameters as well as sperm DNA fragmentation. The findings of this study suggest that antioxidants improve sperm parameters which are associated with infertility in varicocelized rats, and treatment can reduce damage to sperm DNA and increase the chance of fertility.

Keywords: antioxidant, rat, sperm parameter, varicocele

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Authors: Joao Almeida, Ana Azevedo, Myriam Kanoun-Boule, Maria Ines Santos, Antonio Tadeu

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The effects of the increasing expansion of cities and climate changes have encouraged European countries and regions to adopt nature-based solutions with ability to mitigate environmental issues and improve life in cities. Among these strategies, green roofs and urban gardens have been considered ingenious solutions, since they have the desirable potential to improve air quality, prevent floods, reduce the heat island effect and restore biodiversity in cities. However, an additional consumption of fresh water and mineral nutrients is necessary to sustain larger green urban areas. This communication discusses the main technical features of a new system to manage urine recovery at the building level and its application in green roofs. The depletion of critical nutrients like phosphorus constitutes an emergency. In turn, their elimination through urine is one of the principal causes for their loss. Thus, urine recovery in buildings may offer numerous advantages, constituting a valuable fertilizer abundantly available in cities and reducing the load on wastewater treatment plants. Although several urine-diverting toilets have been developed for this purpose and some experiments using urine directly in agriculture have already been carried out in Europe, several challenges have emerged with this practice concerning collection, sanitization, storage and application of urine in buildings. To our best knowledge, current buildings are not designed to receive these systems and integrated solutions with ability to self-manage the whole process of urine recovery, including separation, maturation and storage phases, are not known. Additionally, if from a hygiene point of view human urine may be considered a relatively safe fertilizer, the risk of disease transmission needs to be carefully analysed. A reduction in microorganisms can be achieved by storing the urine in closed tanks. However, several factors may affect this process, which may result in a higher survival rate for some pathogens. In this work, urine effluent was collected under real conditions, stored in closed containers and kept in climatic chambers under variable conditions simulating cold, temperate and tropical climates. These samples were subjected to a first physicochemical and microbiological control, which was repeated over time. The results obtained so far suggest that maturation conditions were reached for all the three temperatures and that a storage period of less than three months is required to achieve a strong depletion of microorganisms. The authors are grateful for the Project WashOne (POCI-01-0247-FEDER-017461) funded by the Operational Program for Competitiveness and Internationalization (POCI) of Portugal 2020, with the support of the European Regional Development Fund (FEDER).

Keywords: sustainable green roofs and urban gardens, urban nutrient cycle, urine-based fertilizers, urine recovery in buildings

Procedia PDF Downloads 130