Search results for: mature embryo
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 441

Search results for: mature embryo

381 Clinical Outcome after in Vitro Fertilization in Women Aged 40 Years and Above: Reasonable Cut-Off Age for Successful Pregnancy

Authors: Eun Jeong Yu, Inn Soo Kang, Tae Ki Yoon, Mi Kyoung Koong

Abstract:

Advanced female age is associated with higher cycle cancelation rates, lower clinical pregnancy rate, increased miscarriage and aneuploidy rates in IVF (In Vitro Fertilization) cycles. This retrospective cohort study was conducted at a Cha Fertility Center, Seoul Station. All fresh non-donor IVF cycles performed in women aged 40 years and above from January 2016 to December 2016 were reviewed. Donor/recipient treatment, PGD/PGS (Preimplantation Genetic Diagnosis/ Preimplantation Genetic Screening) were excluded from analysis. Of the 1,166 cycles from 753 women who completed ovulation induction, 1,047 were appropriate for the evaluation according to inclusion and exclusion criterion. IVF cycles were categorized according to age and grouped into the following 1-year age groups: 40, 41, 42, 43, 44, 45 and > 46. The mean age of patients was 42.4 ± 1.8 years. The median AMH (Anti-Mullerian Hormone) level was 1.2 ± 1.5 ng/mL. The mean number of retrieved oocytes was 4.9 ± 4.3. The clinical pregnancy rate and live birth rate in women > 40 years significantly decreased with each year of advancing age (p < 0.001). The clinical pregnancy rate decreased from 21% at the age of 40 years to 0% at ages above 45 years. Live birth rate decreased from 12.3% to 0%, respectively. There were no clinical pregnancy outcomes among 95 patients aged above 45 years of age. The overall miscarriage rate was 40.7% (range, 36.7%-70%). The transfer of at least one good quality embryo was associated with about 4-9% increased chance of a clinical pregnancy rate. Therefore, IVF in old age women less than 46 had a reasonable chance for successful pregnancy outcomes especially when good quality embryo is transferred.

Keywords: advanced maternal age, in vitro fertilization, pregnancy rate, live birth rate

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380 Noninvasive Technique for Measurement of Heartbeat in Zebrafish Embryos Exposed to Electromagnetic Fields at 27 GHz

Authors: Sara Ignoto, Elena M. Scalisi, Carmen Sica, Martina Contino, Greta Ferruggia, Antonio Salvaggio, Santi C. Pavone, Gino Sorbello, Loreto Di Donato, Roberta Pecoraro, Maria V. Brundo

Abstract:

The new fifth generation technology (5G), which should favor high data-rate connections (1Gbps) and latency times lower than the current ones (<1ms), has the characteristic of working on different frequency bands of the radio wave spectrum (700 MHz, 3.6-3.8 GHz and 26.5-27.5 GHz), thus also exploiting higher frequencies than previous mobile radio generations (1G-4G). The higher frequency waves, however, have a lower capacity to propagate in free space and therefore, in order to guarantee the capillary coverage of the territory for high reliability applications, it will be necessary to install a large number of repeaters. Following the introduction of this new technology, there has been growing concern in recent years about the possible harmful effects on human health and several studies were published using several animal models. This study aimed to observe the possible short-term effects induced by 5G-millimeter waves on heartbeat of early life stages of Danio rerio using DanioScope software (Noldus). DanioScope is the complete toolbox for measurements on zebrafish embryos and larvae. The effect of substances can be measured on the developing zebrafish embryo by a range of parameters: earliest activity of the embryo’s tail, activity of the developing heart, speed of blood flowing through the vein, length and diameters of body parts. Activity measurements, cardiovascular data, blood flow data and morphometric parameters can be combined in one single tool. Obtained data are elaborate and provided by the software both numerical as well as graphical. The experiments were performed at 27 GHz by a no commercial high gain pyramidal horn antenna. According to OECD guidelines, exposure to 5G-millimeter waves was tested by fish embryo toxicity test within 96 hours post fertilization, Observations were recorded every 24h, until the end of the short-term test (96h). The results have showed an increase of heartbeat rate on exposed embryos at 48h hpf than control group, but this increase has not been shown at 72-96 h hpf. Nowadays, there is a scant of literature data about this topic, so these results could be useful to approach new studies and also to evaluate potential cardiotoxic effects of mobile radiofrequency.

Keywords: Danio rerio, DanioScope, cardiotoxicity, millimeter waves.

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379 Defense Mechanism Maturity and the Severity of Mood Disorder Symptoms

Authors: Maja Pandža, Sanjin Lovrić, Iva Čolak, Josipa Mandarić, Miro Klarić

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This study explores the role of symptoms related to mood disorders salience on different types of defense mechanisms (mature, neurotic, immature) predominance. Total of 177 both clinical and non-clinical participants in Mostar, Bosnia & Herzegovina, completed a battery of questionnaires associated with defense mechanisms and self-reported depression and anxiety symptoms. The sample was additionally divided into four groups, given the level of symptoms experienced: 1. minimal, 2. mild, 3. moderate, 4. severe depression/anxiety. Participants with minimal anxiety and depression symptoms use mature defense mechanisms more often than other three groups. Immature mechanisms are most commonly used by the group with severe depression/anxiety levels in comparison with other groups. These differences are discussed on the dynamic level of analysis to have a better understanding of the relationship between defense mechanisms' maturity and degree of mood disorders' symptom severity. Also, results given could serve as an implication for the psychotherapeutic treatment plans.

Keywords: anxiety/depression symptoms, clinical/non-clinical sample, defense mechanism maturity, dynamic approach

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378 The Status of BIM Adoption in Six Continents

Authors: Wooyoung Jung, Ghang Lee

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This paper paper reports the worldwide status of building information modeling (BIM) adoption from the perspectives of the engagement level, the Hype Cycle model, the technology diffusion model, and BIM-uses. An online survey was distributed, and 156 experts from six continents responded. Overall, North America was the most advanced continent, followed by Oceania and Europe. Countries in Asia perceived their phase mainly as slope of enlightenment (mature) in the Hype Cycle model. In the technology diffusion model, the main BIM-users worldwide were “early majority” (third phase), but those in the Middle East/Africa and South America were “early adopters” (second phase). In addition, the more advanced the country, the more number of BIM services employed in general. In summary, North America, Europe, Oceania, and Asia were advancing rapidly toward the mature stage of BIM, whereas the Middle East/Africa and South America were still in the early phase. The simple indexes used in this study may be used to track the worldwide status of BIM adoption in long-term surveys.

Keywords: BIM adoption, BIM services, hype cycle model, technology diffusion model

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377 In Ovo Injection of N-Carbamylglutamate Improves Growth Performance, Muscle Fiber Development, and Meat Quality in Broiler Chickens

Authors: Wang Yuan-hao, Habtamu Ayalew, Jing Wang, Shugeng Wu, Kai Qiu, Guanghai Qi, Haijun Zhang

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N-carbamylglutamate (NCG) has emerged as a promising candidate for regulating endogenous arginine synthesis, thereby promoting desirable growth, carcass traits, and muscle development in broilers. Thus, this study aimed to investigate the effects of NCG in ovo feeding on the growth performance, growth hormones, and meat quality of Ross 308 breeder broilers. A total of 1680 embryo eggs were equally allocated into three treatment groups: non punctured control (NC), saline-injected control (SC; 100μL/egg), and N-carbamylglutamate injected group (NCG; 2 mg/egg). The treatment solution was injected into the amniotic cavity of the embryo at 17.5 days of incubation (DOI). For the subsequent 42 days of post hatch experimental sampling, a total of 360 broiler chicks with 6 replications per treatment and 15 chicks per replication were used. Chickens in the NCG group showed significantly higher (P<0.05) body weight gain (BWG) and final body weight (FBW) at both 21 and 42 days after hatch (DAH), while feed conversion efficiency (FCE) was significantly improved (P<0.05) at 42 DAH. The weight and percentage of drums at 21 DAH and the weight and percentage of breast muscle at 42 DAH were significantly higher (P<0.05) in the NCG group. In addition, insulin (INS), growth hormone (GH), and testosterone (T) levels were significantly higher (P<0.05) in the NCG groups at 21 and 42 DAH. Furthermore, triiodothyronine (T3) and tetraiodothyronine (T4) levels were significantly higher (P<0.05) in the NCG treatment group. Interestingly, meat color values were also significantly higher (P<0.05) in the NCG group at 24 hrs postmortem. Collectively, these findings show that 2 mg NCG in ovo injection improves the growth performance and meat quality of broilers; even the effects extend into the market age of the chickens.

Keywords: N-carbamylglutamate, broiler, in ovo injection, growth performance, meat quality

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376 The Life-Cycle Theory of Dividends: Evidence from Indonesia

Authors: Vashti Carissa

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The main objective of this study is to examine whether the life-cycle theory of dividends could explain the determinant of an optimal dividend policy in Indonesia. The sample that was used consists of 1,420 non-financial and non-trade, services, investment firms listed in Indonesian Stock Exchange during the period of 2005-2014. According to this finding using logistic regression, firm life-cycle measured by retained earnings as a proportion of total equity (RETE) significantly has a positive effect on the propensity of a firm pays dividend. The higher company’s earned surplus portion in its capital structure could reflect firm maturity level which will increase the likelihood of dividend payment in mature firms. This result provides an additional empirical evidence about the existence of life-cycle theory of dividends for dividend payout phenomenon in Indonesia. It can be known that dividends tend to be paid by mature firms while retention is more dominating in growth firms. From the testing results, it can also be known that majority of sample firms are being in the growth phase which proves the fact about infrequent dividend distribution in Indonesia during the ten years observation period.

Keywords: dividend, dividend policy, life-cycle theory of dividends, mix of earned and contributed capital

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375 Alpha Lipoic Acid: An Antioxidant for Infertility

Authors: Chiara Di Tucci, Giulia Galati, Giulia Mattei, Valentina Bonanni, Oriana Capri, Renzo D'Amelio, Ludovico Muzii, Pierluigi Benedetti Panici

Abstract:

Objective: Infertility is an increasingly frequent health condition, which may depend on female or male factors. Oxidative stress (OS), resulting from a disrupted balance between reactive oxygen species (ROS) and protective antioxidants, affects the reproductive lifespan of men and women. In this review, we examine if alpha lipoic acid (ALA), among the oral supplements currently in use, has an evidence-based beneficial role in the context of female and male infertility. Methods: We performed a search from English literature using the PubMed database with the following keywords: 'female infertility', 'male infertility', 'semen', 'sperm', 'sub-fertile man', 'alpha-lipoic acid', ' alpha lipoic acid', 'lipoid acid', 'endometriosis', 'chronic pelvic pain', 'follicular fluid' and 'oocytes'. We included clinical trials, multicentric studies, and reviews. The total number of references found after automatically and manually excluding duplicates was 180. After the primary and secondary screening, 28 articles were selected. Results: The available literature demonstrates the positive effects of ALA in multiple processes, from oocyte maturation (0.87 ± 0.9% of oocyte in MII vs 0.81 ± 3.9%; p < .05) to fertilization, embryo development (57.7% vs 75.7% grade 1 embryo; p < .05) and reproductive outcomes. Its regular administration both in sub-fertile women and men has been shown to reduce pelvic pain in endometriosis (p < .05), regularize menstrual flow and metabolic disorders (p < .01), and improve sperm quality (p < .001). Conclusions: ALA represents a promising new molecule in the field of couple infertility. More clinical studies are needed in order to enhance its use in clinical practice.

Keywords: alpha lipoic acid, endometriosis, infertility, male factor, polycystic ovary syndrome

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374 The Effect of TiO₂ Nanoparticles on Zebrafish Embryos

Authors: Elena Maria Scalisi

Abstract:

Currently, photodegradation by nanoparticles (NPs) is a common solution for wastewater treatment. Nanoparticles are efficient for removing organic and inorganic pollutants, heavy metals from wastewater and killing microorganisms through environmentally friendly. In this context, the major representative of photocatalytic technology for industrial wastewater treatment are TiO₂ nanoparticles (TiO₂-NPs). TiO₂-NPs have a strong catalytic activity that depends to their physicochemical properties. Thanks to their small size (between 1-100 nm), nanoparticles occupy less volume, then their surface area increases. The increase in the surface-to-volume ratio results in the increase of the particle surface energy, which improve their reactivity potential. However, these unique properties represent risks to the ecosystems and organisms when unintentionally TiO₂-NPs are release into the environment and absorbed by living organisms. Several studies confirm that there is a high level of interest concerning the safety of TiO₂-NPs in the aquatic environment, furthermore, ecotoxicological tools are useful to correctly evaluate their toxicity. In the current study, we aimed to characterize potential toxic effects of TiO₂-NP suspension to zebrafish during embryo-larval stages to evaluate parameters such as survival rates, malformation, hatching, the overall length of the larvae heartbeat, and biochemical biomarkers that reflect the acute toxicity and sublethal effects of TiO₂-NPs. Zebrafish embryos were exposed to titanium dioxide nanoparticles (TiO₂-NPs at 1mg/L, 2mg/L, and 4mg/L) from fertilization to the free swimming stage (144hpf). Every day, we recorded the toxicological endpoints, moreover, immunohistochemical analysis has been performed at the end of the exposure. In particular, we have evaluate the expression of the following biomarkers: Heat Shock Protein 70 (HSP70), Poly ADP-Ribose Polymerase-1 (PARP-1), Metallothioneins (MTs). Our results have shown that hatch ability, survival, and malformation rate were not affected by TiO₂ NPs at these exposure levels. However, TiO₂-NPs caused an increase of heartbeat and reduction of body length; at the same time, TiO₂-NPs have inducted the production of ROS and the expression of oxidative stress biomarkers HSP70 and PARP-1. Hight positivity for PARP-1 at all concentration tested was observed. As regards MT, positivity was found in the expression of this biomarker in the whole body of the embryo, with the exception of the end of the tail. Metallothioneins (MT) are biomarkers widely used in environmental monitoring programs for aquatic creatures. At the light of our results i.e. no death until the end of the experiment (144hpf), no malformation and expression of the biomarkers mentioned, it is evident that zebrafish larvae with their natural detoxification pathways are able to resist the presence of toxic substances and then they can tolerate the presence of metal concentrations. However, an excessive oxidative state can compromise cell function, therefore the uncontrolled release of nanoparticles into the environment is severe and must be constantly monitored.

Keywords: nanoparticles, embryo zebrafish, HSP70, PARP-1

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373 Teratogenic Effect of Bisphenol A in Development of Balb/C Mouse

Authors: Nazihe Sedighi, Mohsen Nokhbatolphoghaei

Abstract:

Bisphenol A (BPA) is a monomer used in the manufacture of polycarbonate plastics. Due to having properties such as transparency, heat and impact resistance, it is used widely in medicine, sorts, electronic components, and food containers. It is also used in the production of resins which is applied for lining cans. BPA releases from resins and polycarbonate when it is heated or continuously used the containers from which BPA can enter the body. There are several reports indicating the presence of BPA in the placenta, amniotic fluid, and the embryo itself. While researchers investigated the teratogenic effect of BPA on embryos, very limited work has been done on the effects of BPA when applied from early stages of development. In this study, The teratogenic effect of BPA was investigated at earliest preimplantation (day zero) through day 15.5 of the development of Balb/C mouse embryos. After ensuring the pregnancy via observing vaginal plug, Pregnant mice were divided into five groups. For the three experimental groups, the amount of 500, 750, and 1000 mg/kg/d Bisphenol A was given orally according to body weight. The sham group that was treated with sesame oil, which was used as vehicle and control group remained intact. On day 18.5 of gestation, embryos were removed from the uterus. Randomly half of the embryo were fixed in Bouin for tissue analysis. The other half were prepared for skeletal system staining using Alizarin Red and alcian blue dies. The results showed that the embryonic weight and the crown-rump length of embryos decreased significantly (P < 0.05) in all experimental groups compared to the control group and the sham. In this study, skeletal abnormalities such as delay in ossification of skull and limbs as well as the deviation in the backbone were seen. This research suggests that pregnant mothers need to be aware of possible teratogenic effects of BPA at any stage of pregnancy especially from early to mid stages. In this case, pregnant mothers may need to stop using any manufacture of polycarbonate plastics, as a container for food or drinking.

Keywords: bisphenol A, development, polycarbonate plastic, skeletal system, teratogenicity

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372 Chemical Composition and Antibacterial Activity of Ceratonia siliqua L. Growing in Boumerdes, Algeria

Authors: N. Meziou-Chebouti, A. Merabet, Y. Chebouti N. Behidj

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This work is a contribution to the knowledge of physicochemical characteristics of mature carob followed by evaluation of the activity, antimicrobial phenolics leaves and green pods of Ceratonia siliqua L. physicochemical study shows that mature carob it has a considerable content of sugar (50.90%), but poor in proteins (7%), fat (8%) and also has a high mineral content. The results obtained from phenolic extracts of leaves and green pods of Ceratonia siliqua L. show a wealth leaf phenolic extract especially flavonoids (0,545 mg EqQ/g) relative to the extract of green pods (0,226 mgEqQ/g). Polyphenols leaves have a slightly inhibitory effect on the growth of strains: Staphylococcus aureus, Escherichia coli, Klebsiella pneumoiae, Streptococcus sp and Sanmonella enteritidis, a strong inhibitory effect on the growth of Pseudomonas strain aerogenosa. Moreover, polyphenols pod have a slightly inhibitory effect on the growth of Streptococcus sp strains, Pseudomonas and aerogenosa Sanmonella enteritidis, a slightly inhibitory effect on the growth of Klebsiella pneumoniae strains, E. coli and Staphylococcus aureus.

Keywords: antimicrobial activity, bacteria, clove, Ceratonia siliqua, polyphenols

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371 Effects of Supplementation with Annatto (Bixa Orellana)-Derived δ-Tocotrienol on the Nicotine-Induced Reduction in Body Weight and 8-Cell Preimplantation Embryonic Development in Mice

Authors: M. H. Rajikin, S. M. M. Syairah, A. R. Sharaniza

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Effects of nicotine on pre-partum body weight and preimplantation embryonic development has been reported previously. Present study was conducted to determine the effects of annatto (Bixa orellana)-derived delta-tocotrienol (TCT) (with presence of 10% gamma-TCT isomer) on the nicotine-induced reduction in body weight and 8-cell embryonic growth in mice. Twenty four 6-8 weeks old (23-25g) female balb/c mice were randomly divided into four groups (G1-G4; n=6). Those groups were subjected to the following treatments for 7 consecutive days: G1 (control) were gavaged with 0.1 ml tocopherol stripped corn oil, G2 was subcutaneously (s.c.) injected with 3 mg/kg/day of nicotine, G3 received concurrent treatment of nicotine (3 mg/kg/day) and 60 mg/kg/day of δ-TCT mixture (contains 90% delta & 10% gamma isomers) and G4 was given 60 mg/kg/day of δ-TCT mixture alone. Body weights were recorded daily during the treatment. On Day 8, females were superovulated with 5 IU Pregnant Mare’s Serum Gonadotropin (PMSG) for 48 hours followed with 5 IU human Chorionic Gonadotropin (hCG) before mated with males at the ratio of 1:1. Females were sacrificed by cervical dislocation for embryo collection 48 hours post-coitum. Collected embryos were cultured in vitro. Results showed that throughout Day 1 to Day 7, the body weight of nicotine treated group (G2) was significantly lower (p<0.05) than that of G1, G3 and G4. Intervention with δ-TCT mixture (G3) managed to increase the body weight close to the control group. This is also observed in the group treated with δ-TCT mixture alone (G4). The development of 8-cell embryos following in vitro culture (IVC) was totally inhibited in G2. Intervention with δ-TCT mixture (G3) resulted in the production of 8-cell embryos, although it was not up to that of the control group. Treatment with δ-TCT mixture alone (G4) caused significant increase in the average number of produced 8-cell embryo compared to G1. Present data indicated that δ-TCT mixture was able to reverse the body weight loss in nicotine treated mice and the development of 8-cell embryos was also improved.

Keywords: δ-tocotrienol, body weight, nicotine, preimplantation embryonic development

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370 Influence of IL-1β on Hamster Blastocyst Hatching via Regulation of Hatching Associated Proteases

Authors: Madhulika Pathak, Polani Seshagiri, Vani Venkatappa

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Blastocyst hatching is an indispensable process for successful implantation. One of the major reasons for implantation failure in IVF clinic is poor quality of embryo, which are not development/hatching-competent. Therefore, attempts are required to develop or enhance the culture system with a molecule recapitulating the autocrine/paracrine factors containing the environment of in-vivo endometrial milieu. We have tried to explore the functional molecules involved in the hamster hatching phenomenon. Blastocyst hatching is governed by several molecules that are entwined and regulate this process, among which cytokines are known to be expressed and are still least explored. Two of such cytokines we have used for our study are IL-1β and its natural antagonist IL-1ra to understand the functional dynamics of cytokines involved in the hatching process. Using hamster, an intriguing experimental model for hatching behavior, we have shown the mRNA (qPCR) and protein (ICC) expression of IL-1β, IL-1ra and IL-1 receptor type 1 throughout all the stages of morula, blastocyst and hatched blastocyst. Post-asserting the expression, the functional role is shown by supplementation studies, where IL-1β supplementation showed enhancement in hatching level (IL-1β treated: 84.1 ± 4.2% vs control: 63.7 ± 3.1 %, N=11), further confirmed by the diminishing effect of IL-1ra on hatching rate (IL-1ra treated: 27.5 ± 11.1% vs control: 67.9 ± 3.1%). The exogenous supplementation of IL-1ra decreased the survival rate of embryos and affected the viability in dose-dependent manner, establishing the importance of IL-1β in blastocyst cell survival. Previously, the cathepsin L and B were established as the proteases that were involved in the hamster hatching process. The inducing effect of IL-1β was correlated with enhanced mRNA level, as analyzed by qPCR, for both CAT L (by 1.9 ± 0.5 fold) and CAT B (by 3.5 ± 0.1) fold which was diminished in presence of IL-1ra (Cat L by 88 percent and Cat B by 94 percent. Moreover, using a specific fluorescent substrate-based assay kit, the enzymatic activity of these proteases was found to be increased in presence of IL-1β (Cat L by 2.1 ± 0.1 fold and CAT B by 2.3 ± 0.7 fold) and was curtailed in presence of IL-1ra. These observations provide functional insights with respect to the involvement of cytokines in the hatching process. This has implications in understanding the hatching biology and improving the embryo development quality in IVF clinics.

Keywords: Blastocyst, Cytokines, Hatching, Interleukin

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369 Seasonal Variation in Free Radical Scavenging Properties of Indian Moringa (Moringa Oleifera)

Authors: Awadhesh Kishore, Tushar Sharma

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The goal of this study was to compare the free radical-scavenging (FRS) characteristics of four Indian moringa (Moringa oleifera) plant components: flowers, tender and mature leaves, and seeds that were collected from three Indian districts: Jaipur, Dehra Dun, and Gwalior; in every month of 2021–2022. The samples were collected from three randomly selected agroforest locations from each district. The samples were extracted, and antioxidant properties were determined following the DPPH method with minor modifications. The FRS properties were calculated as the non-absorbance values of the sample in percentage. The factorial ANOVA statistical analysis technique was implemented for comparing FRS properties, and an MS Office Excel 2016 analysis pack was used to compare data. The flowers from Dehra Dun had superior FRS properties (27.06±1.03%), while the seeds from the same location were inferior (8.64±0.17%). The FRS properties of flowers (26.27±0.61%) were not statistically different (P > 0.05) compared to those of tender (27.30±0.63%) and mature leaves (28.37±0.59%), but significantly higher (P < 0.05) than those of seeds (9.31±0.16%). However, the FRS properties in Indian moringa were significantly higher during the winter (Jan 28.67±1.48%) compared to that in the summer (Jun 14.03±0.79%) season, but collected from three locations, viz. Gwalior (22.35±0.70%), Jaipur (23.06±0.73%), and Dehra Dun (23.10±0.76%), were not significantly different (P > 0.05). Based on this study, it can be concluded that the FRS value of flowers during the winter season is superior.

Keywords: flowers, free radical-scavenging, leaves, moringa oleifera, seeds

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368 Nitrification and Denitrification Kinetic Parameters of a Mature Sanitary Landfill Leachate

Authors: Tânia F. C. V. Silva, Eloísa S. S. Vieira, João Pinto da Costa, Rui A. R. Boaventura, Vitor J. P. Vilar

Abstract:

Sanitary landfill leachates are characterized as a complex mixture of diverse organic and inorganic contaminants, which are usually removed by combining different treatment processes. Due to its simplicity, reliability, high cost-effectiveness and high nitrogen content (mostly under the ammonium form) inherent in this type of effluent, the activated sludge biological process is almost always applied in leachate treatment plants (LTPs). The purpose of this work is to assess the effect of the main nitrification and denitrification variables on the nitrogen's biological removal, from mature leachates. The leachate samples were collected after an aerated lagoon, at a LTP nearby Porto, presenting a high amount of dissolved organic carbon (1.0-1.3 g DOC/L) and ammonium nitrogen (1.1-1.7 g NH4+-N/L). The experiments were carried out in a 1-L lab-scale batch reactor, equipped with a pH, temperature and dissolved oxygen (DO) control system, in order to determine the reaction kinetic constants at unchanging conditions. The nitrification reaction rate was evaluated while varying the (i) operating temperature (15, 20, 25 and 30ºC), (ii) DO concentration interval (0.5-1.0, 1.0-2.0 and 2.0-4.0 mg/L) and (iii) solution pH (not controlled, 7.5-8.5 and 6.5-7.5). At the beginning of most assays, it was verified that the ammonium stripping occurred simultaneously to the nitrification, reaching up to 37% removal of total dissolved nitrogen. The denitrification kinetic constants and the methanol consumptions were calculated for different values of (i) volatile suspended solids (VSS) content (25, 50 and 100 mL of centrifuged sludge in 1 L solution), (ii) pH interval (6.5-7.0, 7.5-8.0 and 8.5-9.0) and (iii) temperature (15, 20, 25 and 30ºC), using effluent previously nitrified. The maximum nitrification rate obtained was 38±2 mg NH4+-N/h/g VSS (25ºC, 0.5-1.0 mg O2/L, pH not controlled), consuming 4.4±0.3 mg CaCO3/mg NH4+-N. The highest denitrification rate achieved was 19±1 mg (NO2--N+NO3--N)/h/g VSS (30ºC, 50 mL of sludge and pH between 7.5 and 8.0), with a C/N consumption ratio of 1.1±0.1 mg CH3OH/mg (NO2--N+NO3--N) and an overall alkalinity production of 3.7±0.3 mg CaCO3/mg (NO2--N+NO3--N). The denitrification process showed to be sensitive to all studied parameters, while the nitrification reaction did not suffered significant change when DO content was changed.

Keywords: mature sanitary landfill leachate, nitrogen removal, nitrification and denitrification parameters, lab-scale activated sludge biological reactor

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367 Rheometer Enabled Study of Tissue/biomaterial Frequency-Dependent Properties

Authors: Polina Prokopovich

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Despite the well-established dependence of cartilage mechanical properties on the frequency of the applied load, most research in the field is carried out in either load-free or constant load conditions because of the complexity of the equipment required for the determination of time-dependent properties. These simpler analyses provide a limited representation of cartilage properties thus greatly reducing the impact of the information gathered hindering the understanding of the mechanisms involved in this tissue replacement, development and pathology. More complex techniques could represent better investigative methods, but their uptake in cartilage research is limited by the highly specialised training required and cost of the equipment. There is, therefore, a clear need for alternative experimental approaches to cartilage testing to be deployed in research and clinical settings using more user-friendly and financial accessible devices. Frequency dependent material properties can be determined through rheometry that is an easy to use requiring a relatively inexpensive device; we present how a commercial rheometer can be adapted to determine the viscoelastic properties of articular cartilage. Frequency-sweep tests were run at various applied normal loads on immature, mature and trypsinased (as model of osteoarthritis) cartilage samples to determine the dynamic shear moduli (G*, G′ G″) of the tissues. Moduli increased with increasing frequency and applied load; mature cartilage had generally the highest moduli and GAG depleted samples the lowest. Hydraulic permeability (KH) was estimated from the rheological data and decreased with applied load; GAG depleted cartilage exhibited higher hydraulic permeability than either immature or mature tissues. The rheometer-based methodology developed was validated by the close comparison of the rheometer-obtained cartilage characteristics (G*, G′, G″, KH) with results obtained with more complex testing techniques available in literature. Rheometry is relatively simpler and does not require highly capital intensive machinery and staff training is more accessible; thus the use of a rheometer would represent a cost-effective approach for the determination of frequency-dependent properties of cartilage for more comprehensive and impactful results for both healthcare professional and R&D.

Keywords: tissue, rheometer, biomaterial, cartilage

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366 Shale Gas Accumulation of Over-Mature Cambrian Niutitang Formation Shale in Structure-Complicated Area, Southeastern Margin of Upper Yangtze, China

Authors: Chao Yang, Jinchuan Zhang, Yongqiang Xiong

Abstract:

The Lower Cambrian Niutitang Formation shale (NFS) deposited in the marine deep-shelf environment in Southeast Upper Yangtze (SUY), possess excellent source rock basis for shale gas generation, however, it is currently challenged by being over-mature with strong tectonic deformations, leading to much uncertainty of gas-bearing potential. With emphasis on the shale gas enrichment of the NFS, analyses were made based on the regional gas-bearing differences obtained from field gas-desorption testing of 18 geological survey wells across the study area. Results show that the NFS bears low gas content of 0.2-2.5 m³/t, and the eastern region of SUY is higher than the western region in gas content. Moreover, the methane fraction also presents the similar regional differentiation with the western region less than 10 vol.% while the eastern region generally more than 70 vol.%. Through the analysis of geological theory, the following conclusions are drawn: Depositional environment determines the gas-enriching zones. In the western region, the Dengying Formation underlying the NFS in unconformity contact was mainly plateau facies dolomite with caves and thereby bears poor gas-sealing ability. Whereas the Laobao Formation underling the NFS in eastern region was a set of siliceous rocks of shelf-slope facies, which can effectively prevent the shale gas from escaping away from the NFS. The tectonic conditions control the gas-enriching bands in the SUY, which is located in the fold zones formed by the thrust of the Southern China plate towards to the Sichuan Basin. Compared with the western region located in the trough-like folds, the eastern region at the fold-thrust belts was uplifted early and deformed weakly, resulting in the relatively less mature level and relatively slight tectonic deformation of the NFS. Faults determine whether shale gas can be accumulated in large scale. Four deep and large normal faults in the study area cut through the Niutitang Formation to the Sinian strata, directly causing a large spillover of natural gas in the adjacent areas. For the secondary faults developed within the shale formation, the reverse faults generally have a positive influence on the shale accumulation while the normal faults perform the opposite influence. Overall, shale gas enrichment targets of the NFS, are the areas with certain thickness of siliceous rocks at the basement of the Niutitang Formation, and near the margin of the paleouplift with less developed faults. These findings provide direction for shale gas exploration in South China, and also provide references for the areas with similar geological conditions all over the world.

Keywords: over-mature marine shale, shale gas accumulation, structure-complicated area, Southeast Upper Yangtze

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365 Epigenetics Regulation Play Role in the Pathogenesis of Adipose Tissue Disorder, Lipedema

Authors: Musarat Ishaq, Tara Karnezis, Ramin Shayan

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Lipedema, a poorly understood chronic disease of adipose hyper-deposition, is often mistaken for obesity and causes significant impairment to mobility and quality-of-life. To identify molecular mechanisms underpinning lipedema, we employed comprehensive omics-based comparative analyses of whole tissue, adipocyte precursors (adipose-derived stem cells (ADSCs)), and adipocytes from patients with or without lipedema. Transcriptional profiling revealed significant differences in lipedema tissue, adipocytes, and ADSCs, with altered levels of mRNAs involved inproliferation and cell adhesion. One highly up-regulated gene in lipedema adipose tissue, adipocytes and ADSCs, ZIC4, encodes Zinc Finger Protein ZIC 4, a class of transcription factor which may be involved in regulating metabolism and adipogenesis. ZIC4 inhibition impaired the adipogenesis of ADSCs into mature adipocytes. Epigenetic regulation study revealed overexpression of ZIC4 is involved in decreased promoter DNA methylation and subsequent decrease in adipogenesis. These epigenetic modifications can alter adipocytes microenvironment and adipocytes differentiation. Our study show that epigenetic events regulate the ability of ADSCs to commit and differentiate into mature adipocytes by modulating ZIC4.

Keywords: lipedema, adipose-derived stem cells, adipose tisue, adipocytes, zinc finger protein, epigenetic

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364 Evaluation of Toxicity of Cerium Oxide on Zebrafish Developmental Stages

Authors: Roberta Pecoraro, Elena Maria Scalisi

Abstract:

Engineered Nanoparticles (ENPs) and Nanomaterials (ENMs) concern an active research area and a sector in full expansion. They have physical-chemical characteristics and small size that improve their performance compared to common materials. Due to the increase in their production and their subsequent release into the environment, new strategies are emerging to assess risk of nanomaterials. NPs can be released into the environment through aquatic systems by human activities and exert toxicity on living organisms. We evaluated the potential toxic effect of cerium oxide (CeO2) nanoparticles because it’s used in different fields due to its peculiar properties. In order to assess nanoparticles toxicity, Fish Embryo Toxicity (FET) test was performed. Powders of CeO2 NPs supplied by the CNR-IMM of Catania are indicated as CeO2 type 1 (as-prepared) and CeO2 type 2 (modified), while CeO2 type 3 (commercial) is supplied by Sigma-Aldrich. Starting from a stock solution (0.001g/10 ml dilution water) of each type of CeO2 NPs, the other concentration solutions were obtained adding 1 ml of the stock solution to 9 ml of dilution water, leading to three different solutions of concentration (10-4, 10-5, 10-6 g/ml). All the solutions have been sonicated to avoid natural tendency of NPs to aggregate and sediment. FET test was performed according to the OECD guidelines for testing chemicals using our internal protocol procedure. A number of eight selected fertilized eggs were placed in each becher filled with 5 ml of each concentration of the three types of CeO2 NPs; control samples were incubated only with dilution water. Replication was performed for each concentration. During the exposure period, we observed four endpoints (embryo coagulation, lack of formation of somites, failure to lift the yolk bag, no heartbeat) by a stereomicroscope every 24 hours. Immunohistochemical analysis on treated larvae was performed to evaluate the expression of metallothioneins (MTs), Heat Shock Proteins 70 (HSP70) and 7-ethoxyresorufin-O-diethylase (EROD). Our results have not shown evident alterations on embryonic development because all embryos completed the development and the hatching of the eggs, started around the 48th hour after exposure, took place within the last observation at 72 hours. A good reactivity, both in the embryos and in the newly hatched larvae, was found. The presence of heartbeat has also been observed in embryos with reduced mobility confirming their viability. A higher expression of EROD biomarker was observed in the larvae exposed to the three types of CeO2, showing a clear difference with the control. A weak positivity was found for MTs biomarker in treated larvae as well as in the control. HSP70 are expressed homogeneously in all the type of nanoparticles tested but not too much greater than control. Our results are in agreement with other studies in the literature, in which the exposure of Danio rerio larvae to other metal oxide nanoparticles does not show adverse effects on survival and hatching time. Further studies are necessary to clarify the role of these NPs and also to solve conflicting opinions.

Keywords: Danio rerio, endpoints, fish embryo toxicity test, metallic nanoparticles

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363 Carbon Capture and Storage in Geological Formation, its Legal, Regulatory Imperatives and Opportunities in India

Authors: Kalbende Krunal Ramesh

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The Carbon Capture and Storage Technology (CCS) provides a veritable platform to bridge the gap between the seemingly irreconcilable twin global challenges of ensuring a secure, reliable and diversified energy supply and mitigating climate change by reducing atmospheric emissions of carbon dioxide. Making its proper regulatory policy and making it flexible for the government and private company by law to regulate, also exploring the opportunity in this sector is the main aim of this paper. India's total annual emissions was 1725 Mt CO2 in 2011, which comprises of 6% of total global emission. It is very important to control the greenhouse gas emission for the environment protection. This paper discusses the various regulatory policy and technology adopted by some of the countries for successful using CCS technology. The brief geology of sedimentary basins in India is studied, ranging from the category I to category IV and deep water and potential for mature technology in CCS is reviewed. Areas not suitable for CO2 storage using presently mature technologies were over viewed. CSS and Clean development mechanism was developed for India, considering the various aspects from research and development, project appraisal, approval and validation, implementation, monitoring and verification, carbon credit issued, cap and trade system and its storage potential. The opportunities in oil and gas operations, power sector, transport sector is discussed briefly.

Keywords: carbon credit issued, cap and trade system, carbon capture and storage technology, greenhouse gas

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362 Use of Didactic Bibliographic Resources to Improve the Teaching and Learning Processes of Animal Reproduction in Veterinary Science

Authors: Yasser Y. Lenis, Amy Jo Montgomery, Diego F. Carrillo-Gonzalez

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Introduction: The use of didactic instruments in different learning environments plays a pivotal role in enhancing the level of knowledge in veterinary science students. The direct instruction of basic animal reproduction concepts in students enrolled in veterinary medicine programs allows them to elucidate the biological and molecular mechanisms that perpetuate the animal species in an ecosystem. Therefore, universities must implement didactic strategies that facilitate the teaching and learning processes for students and, in turn, enrich learning environments. Objective: to evaluate the effect of the use of a didactic textbook on the level of theoretical knowledge in embryo-maternal recognition for veterinary medicine students. Methods: the participants (n=24) were divided into two experimental groups: control (Ctrl) and treatment (Treat). Both groups received 4 hours of theoretical training regarding the basic concepts in bovine embryo-maternal recognition. However, the Treat group was also exposed to a guided lecture and the activity play-to-learn from a cow reproduction didactic textbook. A pre-test and a post-test were applied to assess the prior and subsequent knowledge in the participants. Descriptive statistics were applied to identify the success rates for each of the tests. Afterwards, a repeated measures model was applied where the effect of the intervention was considered. Results: no significant difference (p>0,05) was observed in the number of right answers for groups Ctrl (54,2%±12,7) and Treat (40,8%±16,8) in the pre-test. There was no difference (p>0,05) compering the number of right answers in Ctrl pre-test (54,2%±12,7) and post-test (60,8±18,8). However, the Treat group showed a significant (p>0,05) difference in the number of right answers when comparing pre-test (40,8%±16,8) and post-test (71,7%±14,7). Finally, after the theoretical training and the didactic activity in the Treat group, an increase of 10.9% (p<0,05) in the number of right answers was found when compared with the Ctrl group. Conclusion: the use of didactic tools that include guided lectures and activities like play-to-learn from a didactic textbook enhances the level of knowledge in an animal reproduction course for veterinary medicine students.

Keywords: animal reproduction, pedagogic, level of knowledge, learning environment

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361 Significance of Water Saving through Subsurface Drip Irrigation for Date Palm Trees

Authors: Ahmed I. Al-Amoud

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A laboratory and field study were conducted on subsurface drip irrigation systems. In the first laboratory study, eight subsurface drip irrigation lines available locally, were selected and a number of experiments were made to evaluate line hydraulic characteristics to insure it's suitability for drip irrigation design requirements and high performance to select the best for field experiments. The second study involves field trials on mature date palm trees to study the effect of subsurface drip irrigation system on the yield and water consumption of date palms, and to compare that with the traditional surface drip irrigation system. Experiments were conducted in Alwatania Agricultural Project, on 50 mature palm trees (17 years old) of Helwa type with 10 meters spacing between rows and between trees. A high efficiency subsurface line (Techline) was used based on the results of the first study. Irrigation scheduling was made through a soil moisture sensing device to ensure enough soil water levels in the soil. Experiment layouts were installed during 2001 season, measurements continued till end of 2008 season. Results have indicated that there is an increase in the yield and a considerable saving in water compared to the conventional drip irrigation method. In addition there were high increases in water use efficiency using the subsurface system. The subsurface system proves to be durable and highly efficient for irrigating date palm trees.

Keywords: drip irrigation, subsurface drip irrigation, date palm trees, date palm water use, date palm yield

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360 Potential Serological Biomarker for Early Detection of Pregnancy in Cows

Authors: Shveta Bathla, Preeti Rawat, Sudarshan Kumar, Rubina Baithalu, Jogender Singh Rana, Tushar Kumar Mohanty, Ashok Kumar Mohanty

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Pregnancy is a complex process which includes series of events such as fertilization, formation of blastocyst, implantation of embryo, placental formation and development of fetus. The success of these events depends on various interactions which are synchronized by endocrine interaction between a receptive dam and competent embryo. These interactions lead to change in expression of hormones and proteins. But till date no protein biomarker is available which can be used to detect successful completion of these events. We employed quantitative proteomics approach to develop putative serological biomarker which has diagnostic applicability for early detection of pregnancy in cows. For this study, sera were collected from control (non-pregnant, n=6) and pregnant animals on successive days of pregnancy (7, 19, 45, n=6). The sera were subjected to depletion for removal of albumin using Norgen depletion kit. The tryptic peptides were labeled with iTRAQ. The peptides were pooled and fractionated using bRPLC over 80 min gradient. Then 12 fractions were injected to nLC for identification and quantitation in DDA mode using ESI. Identification using Mascot search revealed 2056 proteins out of which 352 proteins were differentially expressed. Twenty proteins were upregulated and twelve proteins were down-regulated with fold change > 1.5 and < 0.6 respectively (p < 0.05). The gene ontology studies of DEPs using Panther software revealed that majority of proteins are actively involved in catalytic activities, binding and enzyme regulatory activities. The DEP'S such as NF2, MAPK, GRIPI, UGT1A1, PARP, CD68 were further subjected to pathway analysis using KEGG and Cytoscape plugin Cluego that showed involvement of proteins in successful implantation, maintenance of pluripotency, regulation of luteal function, differentiation of endometrial macrophages, protection from oxidative stress and developmental pathways such as Hippo. Further efforts are continuing for targeted proteomics, western blot to validate potential biomarkers and development of diagnostic kit for early pregnancy diagnosis in cows.

Keywords: bRPLC, Cluego, ESI, iTRAQ, KEGG, Panther

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359 Design of an Improved Distributed Framework for Intrusion Detection System Based on Artificial Immune System and Neural Network

Authors: Yulin Rao, Zhixuan Li, Burra Venkata Durga Kumar

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Intrusion detection refers to monitoring the actions of internal and external intruders on the system and detecting the behaviours that violate security policies in real-time. In intrusion detection, there has been much discussion about the application of neural network technology and artificial immune system (AIS). However, many solutions use static methods (signature-based and stateful protocol analysis) or centralized intrusion detection systems (CIDS), which are unsuitable for real-time intrusion detection systems that need to process large amounts of data and detect unknown intrusions. This article proposes a framework for a distributed intrusion detection system (DIDS) with multi-agents based on the concept of AIS and neural network technology to detect anomalies and intrusions. In this framework, multiple agents are assigned to each host and work together, improving the system's detection efficiency and robustness. The trainer agent in the central server of the framework uses the artificial neural network (ANN) rather than the negative selection algorithm of AIS to generate mature detectors. Mature detectors can distinguish between self-files and non-self-files after learning. Our analyzer agents use genetic algorithms to generate memory cell detectors. This kind of detector will effectively reduce false positive and false negative errors and act quickly on known intrusions.

Keywords: artificial immune system, distributed artificial intelligence, multi-agent, intrusion detection system, neural network

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358 Depositional Environment and Source Potential of Devonian Source Rock, Ghadames Basin, Southern Tunisia

Authors: S. Mahmoudi, A. Belhaj Mohamed, M. Saidi, F. Rezgui

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Depositional environment and source potential of the different organic rich levels of Devonian age (up to 990m thick) from the onshore EC-1 well (Southern Tunisia) were investigated using different geochemical techniques (Rock-Eval pyrolysis, GC-MS) of over than 130 cutting samples. The obtained results including Rock Eval Pyrolysis data and biomarker distribution (terpanes, steranes and aromatics) have been used to describe the depositional environment and to assess the thermal maturity of the Devonian organic matter. These results show that the Emsian deposits exhibit poor to fair TOC contents. The associated organic matter is composed of mixed kerogen (type II/III), as indicated by the predominance of C29 steranes over C27 and C28 homologous, that was deposited in a slightly reduced environment favoring organic matter preservation. Thermal maturity assessed from Tmax, TNR and MPI-1 values shows a mature stage of organic matter. The Middle Devonian (Eifelian) shales are rich in type II organic matter that was deposited in an open marine depositional environment. The TOC values are high and vary between 2 and 7 % indicating good to excellent source rock. The relatively high IH values (reaching 547 mg HC/g TOC) and the low values of t19/t23 ratio (down to 0.2) confirm the marine origin of the organic matter (type II). During the Upper Devonian, the organic matter was deposited under variable redox conditions, oxic to suboxic which is clearly indicated by the low C35/C34 hopanes ratio, immature to marginally mature with the vitrinite reflectance ranging from 0.5 to 0.7 Ro and Tmax value of 426°C-436 °C and the TOC values range between 0.8% to 4%.

Keywords: biomarker, depositional environment, devonian, source rock

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357 Plantlet Regeneration from Zygotic Embryos of Securidaca longepedunculata Fresen

Authors: Uche C. Okafor, Nwanneka M. Okpokwu, Felix Nwafor, Carl E. A. Okezie

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Securidaca longepedunculata Fresen (Violet tree) belongs to the family Polygalaceae characterised by papillionaceous purplish flowers. This medicinally valued plant disappears at an alarming rate due to intensified anthropopressure particularly the unregulated manner of subterranean plant parts' collection from natural stands. Some indiscriminately harvested plants bear seeds containing both mature and immature zygotic embryos that are often discarded. Here, such seeds are collected for this experiment. Seeds were collected, washed, de-coated, and dipped in 70 % (v/v) ethanol for 30 s followed by rising in 5 % solution sodium hypochlorite, containing two drops of tween 20, for another 25 min. Mature zygotic embryos (MZEs) were excised from seeds and cultured in two basal media (MS and B5), three carbon sources (sucrose, glucose and fructose) at five concentrations (0-40 g/L) while immature zygotic embryos (iMZEs) were composed on similar basal media and carbon source supplemented with 0-2 mg/L Benzylaminopurine (BAP) and 0-2 mg/L Indole acetic acid (IAA). MZEs cultured on MS + 30g/L sucrose differed significantly from other treatments at p≤0.05 with maximum percent sprouting (85.24± 5.67 %) and shoot length (7.53±0.67 cm). MZEs culture had the maximum percent sprouting (85.24± 5.67 %) and shoot length (7.53±0.67 cm) in medium containing MS+ 30g L-1 sucrose. iMZEs on the other hand had maximum growth on MS + 40g/L sucrose supplemented with 1.5 mg/L IAA+ 1.0 mg/L BAP. This study is a geared towards creating an alternative path for the maximum production of plants in vitro, thereby, preventing the plants from disappearing.

Keywords: Gamborg's medium, Murashige and Skoog medium, Securidaca longepedunculata, zygotic embryos

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356 Over Expression of Mapk8ip3 Patient Variants in Zebrafish to Establish a Spectrum of Phenotypes in a Rare-Neurodevelopmental Disorder

Authors: Kinnsley Travis, Camerron M. Crowder

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Mapk8ip3 (Mitogen-Activated Protein Kinase 8 Interacting Protein 3) is a gene that codes for the JIP3 protein, which is a part of the JIP scaffolding protein family. This protein is involved in axonal vesicle transport, elongation and regeneration. Variants in the Mapk8ip3 gene are associated with a rare-genetic condition that results in a neurodevelopmental disorder that can cause a range of phenotypes including global developmental delay and intellectual disability. Currently, there are 18 known individuals diagnosed to have sequenced confirmed Mapk8ip3 genetic disorders. This project focuses on examining the impact of a subset of missense patient variants on the Jip3 protein function by overexpressing the mRNA of these variants in a zebrafish knockout model for Jip3. Plasmids containing cDNA with individual missense variants were reverse transcribed, purified, and injected into single-cell zebrafish embryos (Wild Type, Jip3 -/+, and Jip3 -/-). At 6-days post mRNA microinjection, morphological, behavioral, and microscopic phenotypes were examined in zebrafish larvae. Morphologically, we compared the size and shape of the zebrafish during their development over a 5-day period. Total locomotive activity was assessed using the Microtracker assay and patterns of movement over time were examined using the DanioVision assay. Lastly, we used confocal microscopy to examine sensory axons for swelling and shortened length, which are phenotypes observed in the loss-of-function knockout Jip3 zebrafish model. Using these assays during embryonic development, we determined the impact of various missense variants on Jip3 protein function, compared to knockout and wild-type zebrafish embryo models. Variants in the gene Mapk8ip3 cause rare-neurodevelopmental disorders due to an essential role in axonal vesicle transport, elongation and regeneration. A subset of missense variants was examined by overexpressing the mRNA of these variants in a Jip3 knock-out zebrafish. Morphological, behavioral, and microscopic phenotypes were examined in zebrafish larvae. Using these assays, the spectrum of disorders can be phenotypically determined and the impact of variant location can be compared to knockout and wild-type zebrafish embryo models.

Keywords: rare disease, neurodevelopmental disorders, mrna overexpression, zebrafish research

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355 Effects of Achillea millefolium L. Extract on Rat Spermatogenesis

Authors: Nasrin Takzaree, Gholamreza Hassanzadeh, Abbas Hadjiakhoondi, Mohammadreza Rouini

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Introduction: Today herbal medicine are extensively used for various therapeutic reasons. Whereas Achillea millefolium L. comprises different chemical compounds it is used in classic and modern medicine for different purposes. Concerning the family planning as a principle matter, the idea of using specific herbal medicine is of great importance. Purpose: To investigate the effects of Achillea millefolium L. extract on fertility power and spermatogenesis process in male mature Wistar rats and the anti-fertility effects of this extract in male genital system. Material and methods: In this study 32 male mature Wistar rats were randomly divided in to 4 experimental groups. 1st experimental group included 8 rats receiving Achillea millefolium extract at the dose of 200 mg/kg intraperitoneally. Second and third groups received the extract the same at the doses of 400 and 800 mg/kg respectively. 4th group was considered as control group in which the parenteral distilled water was administered. after 20 days, rats were sacrificed and the spermatogenesis process was histologically examined. Results: In experimental groups receiving high doses of extract comparing with control group, thickness in seminiferous tubules basal membrane, decrease in germinal epithelium cells, congestion in testicular tissue, disarrangement in germinal epithelium cells as well as decrease in cellular condense were observed (p<0.001). Conclusion: Findings suggest that alcoholic extract of Achillea millefolium at high concentrations lead to the structural alterations and changes in spermatogenesis in testicular tissue.

Keywords: spermatogenesis, alcoholic extract of Achillea millefolium L., testis, Wistar rat

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354 Prediction of Solanum Lycopersicum Genome Encoded microRNAs Targeting Tomato Spotted Wilt Virus

Authors: Muhammad Shahzad Iqbal, Zobia Sarwar, Salah-ud-Din

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Tomato spotted wilt virus (TSWV) belongs to the genus Tospoviruses (family Bunyaviridae). It is one of the most devastating pathogens of tomato (Solanum Lycopersicum) and heavily damages the crop yield each year around the globe. In this study, we retrieved 329 mature miRNA sequences from two microRNA databases (miRBase and miRSoldb) and checked the putative target sites in the downloaded-genome sequence of TSWV. A consensus of three miRNA target prediction tools (RNA22, miRanda and psRNATarget) was used to screen the false-positive microRNAs targeting sites in the TSWV genome. These tools calculated different target sites by calculating minimum free energy (mfe), site-complementarity, minimum folding energy and other microRNA-mRNA binding factors. R language was used to plot the predicted target-site data. All the genes having possible target sites for different miRNAs were screened by building a consensus table. Out of these 329 mature miRNAs predicted by three algorithms, only eight miRNAs met all the criteria/threshold specifications. MC-Fold and MC-Sym were used to predict three-dimensional structures of miRNAs and further analyzed in USCF chimera to visualize the structural and conformational changes before and after microRNA-mRNA interactions. The results of the current study show that the predicted eight miRNAs could further be evaluated by in vitro experiments to develop TSWV-resistant transgenic tomato plants in the future.

Keywords: tomato spotted wild virus (TSWV), Solanum lycopersicum, plant virus, miRNAs, microRNA target prediction, mRNA

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353 Mature Field Rejuvenation Using Hydraulic Fracturing: A Case Study of Tight Mature Oilfield with Reveal Simulator

Authors: Amir Gharavi, Mohamed Hassan, Amjad Shah

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The main characteristics of unconventional reservoirs include low-to ultra low permeability and low-to-moderate porosity. As a result, hydrocarbon production from these reservoirs requires different extraction technologies than from conventional resources. An unconventional reservoir must be stimulated to produce hydrocarbons at an acceptable flow rate to recover commercial quantities of hydrocarbons. Permeability for unconventional reservoirs is mostly below 0.1 mD, and reservoirs with permeability above 0.1 mD are generally considered to be conventional. The hydrocarbon held in these formations naturally will not move towards producing wells at economic rates without aid from hydraulic fracturing which is the only technique to assess these tight reservoir productions. Horizontal well with multi-stage fracking is the key technique to maximize stimulated reservoir volume and achieve commercial production. The main objective of this research paper is to investigate development options for a tight mature oilfield. This includes multistage hydraulic fracturing and spacing by building of reservoir models in the Reveal simulator to model potential development options based on sidetracking the existing vertical well. To simulate potential options, reservoir models have been built in the Reveal. An existing Petrel geological model was used to build the static parts of these models. A FBHP limit of 40bars was assumed to take into account pump operating limits and to maintain the reservoir pressure above the bubble point. 300m, 600m and 900m lateral length wells were modelled, in conjunction with 4, 6 and 8 stages of fracs. Simulation results indicate that higher initial recoveries and peak oil rates are obtained with longer well lengths and also with more fracs and spacing. For a 25year forecast, the ultimate recovery ranging from 0.4% to 2.56% for 300m and 1000m laterals respectively. The 900m lateral with 8 fracs 100m spacing gave the highest peak rate of 120m3/day, with the 600m and 300m cases giving initial peak rates of 110m3/day. Similarly, recovery factor for the 900m lateral with 8 fracs and 100m spacing was the highest at 2.65% after 25 years. The corresponding values for the 300m and 600m laterals were 2.37% and 2.42%. Therefore, the study suggests that longer laterals with 8 fracs and 100m spacing provided the optimal recovery, and this design is recommended as the basis for further study.

Keywords: unconventional, resource, hydraulic, fracturing

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352 Variation in Wood Anatomical Properties of Acacia seyal var. seyal Tree Species Growing in Different Zones in Sudan

Authors: Hanadi Mohamed Shawgi Gamal, Ashraf Mohamed Ahmed Abdalla

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Sudan is endowed by a great diversity of tree species; nevertheless, the utilization of wood resources has traditionally concentrated on a few number of species. With the great variation in the climatic zones of Sudan, great variations are expected in the anatomical properties between and within species. This variation needs to be fully explored in order to suggest the best uses for the species. Modern research on wood has substantiated that the climatic condition where the species grow has significant effect on wood properties. Understanding the extent of variability of wood is important because the uses for each kind of wood are related to its characteristics; furthermore, the suitability or quality of wood for a particular purpose is determined by the variability of one or more of these characteristics. The present study demonstrates the effect of rainfall zones in some anatomical properties of Acacia seyal var. seyal growing in Sudan. For this purpose, twenty healthy trees were collected randomly from two zones (ten trees per zone). One zone with relatively low rainfall (273mm annually) which represented by North Kordofan state and White Nile state and the second with relatively high rainfall (701 mm annually) represented by Blue Nile state and South Kordofan state. From each sampled tree, a stem disc (3 cm thick) was cut at 10% from stem height. One radius was obtained in central stem dices. Two representative samples were taken from each disc, one at 10% distance from pith to bark, the second at 90% in order to represent the juvenile and mature wood. The investigated anatomical properties were fibers length, fibers and vessels diameter, lumen diameter, and wall thickness as well as cell proportions. The result of the current study reveals significant differences between zones in mature wood vessels diameter and wall thickness, as well as juvenile wood vessels, wall thickness. The higher values were detected in the drier zone. Significant differences were also observed in juvenile wood fiber length, diameter as well as wall thickness. Contrary to vessels diameter and wall thickness, the fiber length, diameter as well as wall thickness were decreased in the drier zone. No significant differences have been detected in cell proportions of juvenile and mature wood. The significant differences in some fiber and vessels dimension lead to expect significant differences in wood density. From these results, Acacia seyal var. seyal seems to be well adapted with the change in rainfall and may survive in any rainfall zone.

Keywords: Acacia seyal var. seyal, anatomical properties, rainfall zones, variation

Procedia PDF Downloads 115