Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1756

World Academy of Science, Engineering and Technology

[Bioengineering and Life Sciences]

Online ISSN : 1307-6892

1756 Applications of Copper Sensitive Fluorescent Dye to the Studies of the Role of Copper in Cisplatin Resistance in Human Cancer

Authors: Sumayah Mohammed Asiri A., Aviva Levina B., Elizabeth New C., Peter Lay D.


Pt compounds have been among the most successful anticancer drugs in the last 40 years, but the development of resistance to them is an increasing problem. Cellular homeostasis of an essential metal, Cu, is known to be involved in Pt resistance, but mechanisms of this process are poorly understood. We used a novel ratiometric Cu(I)-sensitive fluorescent probeInCCu1 dye to detect Cu(I) in the mitochondria. Total Cu and labile Cu pool measured using AAS and InCCu1 dye in A2780 cells and their corresponding resistant cells A2780-cis.R cells treated with Cu and cisplatin. The main difference between both cell lines in the presence and absence of Cu(II) is that resistant cells have lower total Cu content but higher labile Cu levels than cisplatin-sensitive cells. This means that resistant cells can metabolize and export excess Cu more efficiently. Furthermore, InCCu1 has emerged not only as an indicator of labile cellular Cu levels in the mitochondria but as a potentially versatile multi-organelle probe.

Keywords: AAS and ICPMS, A2780 and its resistant cells, ratiometric fluorescent sensors, inCCu1, and total and labile Cu

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1755 Bioinformatic Screening of Metagenomic Fosmid Libraries for Identification of Biosynthetic Pathways Derived from the Colombian Soils

Authors: María Fernanda Quiceno Vallejo, Patricia del Portillo, María Mercedes Zambrano, Jeisson Alejandro Triana, Dayana Calderon, Juan Manuel Anzola


Microorganisms from tropical ecosystems can be novel in terms of adaptations and conservation. Given the macrodiversity of Colombian ecosystems, it is possible that this diversity is also present in Colombian soils. Tropical soil bacteria could offer a potentially novel source of bioactive compounds. In this study we analyzed a metagenomic fosmid library constructed with tropical bacterial DNAs with the aim of understanding its underlying diversity and functional potential. 8640 clones from the fosmid library were sequenced by NANOPORE MiniOn technology, then analyzed with bioinformatic tools such as Prokka, AntiSMASH and Bagel4 in order to identify functional biosynthetic pathways in the sequences. The strains showed ample difference when it comes to biosynthetic pathways. In total we identified 4 pathways related to aryl polyene synthesis, 12 related to terpenes, 22 related to NRPs (Non ribosomal peptides), 11 related PKs (Polyketide synthases) and 7 related to RiPPs (bacteriocins). We designed primers for the metagenomic clones with the most BGCs (sample 6 and sample 2). Results show the biotechnological / pharmacological potential of tropical ecosystems. Overall, this work provides an overview of the genomic and functional potential of Colombian soil and sets the groundwork for additional exploration of tropical metagenomic sequencing.

Keywords: bioactives, biosyntethic pathways, bioinformatic, bacterial gene clusters, secondary metabolites

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1754 Seed Associated Microbial Communities of Holoparasitic Cistanche Species from Armenia and Portugal

Authors: Kristine Petrosyan, Renata Piwowarczyk, Karolina Ruraż, Jaco Vangronsveld, Wiesław Kaca


Holoparasitic plants are flowering heterotrophic angiosperms which, with the help of an absorbing organ - haustorium, attach to another plant, the so-called host. Due to the different hosts, unusual lifestyle, lack of roots, chlorophylls, and photosynthesis, these plants are interesting and unique study objects for global biodiversity. The seeds germination of the parasitic plants also is unique: they germinate only in response to germination stimulants, namely strigolactones produced by the root of an appropriate host. Resistance of the seeds on different environmental conditions allow them to stay viable in the soil for more than 20 years. Among the wide range of plant protection mechanisms, the endophytic communities have a specific role. In this way, they have the potential to mitigate the impacts of adverse conditions such as soil salinization. The major objective of our study was to compare the bacterial endo-microbiomes from seeds of two holoparasitic plants from Orobanchaceae family, Cistanche – C. armena (Armenia) and C. phelypaea (Portugal) – from saline habitats different in soil water status. The research aimed to perform how environmental conditions influence on the diversity of the bacterial communities of C. armena and C. phelypaea seeds. This was achieved by comparison of the endophytic microbiomes of two species and isolation of culturable bacteria. A combination of culture-dependent and molecular techniques was employed for the identification of the seed endomicrobiome (culturable and unculturable). Using the V3-V4 hypervariable region of the 16S rRNA gene, four main taxa were identified: Proteobacteria, Actinobacteria, Bacteroidetes, Firmicutes, but the relative proportion of the taxa was different in each type of seed. Generally, sixteen phyla, 323 genera, and 710 bacterial species were identified, mainly Gram negative, halotolerant bacteria with an environmental origin. However, also some unclassified and unexplored taxonomic groups were found in the seeds of both plants. 16S rRNA gene sequencing analysis from both species identified the gram positive, endospore forming, halotolerant, and alkaliphile Bacillus spp., which suggests that the endophytic bacteria of examined seeds possess traits that are correlated with the natural habitat of their hosts. The cultivable seed endophytes from C. armena and C. phelypaea were rather similar, notwithstanding the big distances between their growth habitats - Armenia and Portugal. Although the seed endophytic microbiomes of C. armena and C. phelypaea contain a high number of common bacterial taxa, also remarkable differences exist. We demonstrated that the environmental conditions or abiotic stresses influence the diversity of the bacterial communities of holoparasitic seeds. The research is the first report of endophytes from seeds of holoparasitic Cistanche armena and C. phelypaea plants.

Keywords: microbiome, parasitic plant, salinity, seeds

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1753 Characterizing Protein Conformational Space Using Robotics-Based Search and Topological Data Analysis

Authors: Ramin Dehghanpoor, Fatemeh Afrasiabi, Nurit Haspel


Investigating the conformational space of proteins is essential in order to associate their structures with their fundamental functions. Nonetheless, it is a challenging task, both experimentally and computationally. Because of the transient nature of these conformational changes and the fact that they are impermanent, empirical methods have failed to capture them. Computational methods, on the other hand, have shown great promise in exploring these conformational pathways. In this article, we provide an extensive evaluation of our algorithm called RRTMC that uses the Rapidly exploring Random Trees algorithm with Monte Carlo to explore the conformational space of proteins. This evaluation is conducted by identifying what intermediate conformations appear the most in RRTMC using Topological Data Analysis (TDA) methods and examining how close these conformations are to existing experimental data. We concluded that the intermediate conformations by RRTMC are, in fact, close to known PDB structures.

Keywords: conformational space of proteins, rapidly exploring random tree search, topological data analysis, principal component analysis, mapper algorithm

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1752 SUMOylation Enhances Nurr1/1a Mediated Transactivation in a Neuronal Cell Type

Authors: Jade Edey, Andrew Bennett, Gareth Hathway


Nuclear receptor-related 1 protein (also known as Nurr1 or NR4A2) is an orphan nuclear receptor which plays a vital role in the development, survival and maintenance of dopaminergic (DA) neurons particularly in the substantia nigra (SN). Increasing research has investigated Nurr1’s additional role within microglia and astrocytes where it has been suggested to act as a negative regulator of inflammation; potentially offering neuroprotection. Considering both DA neurodegeneration and neuroinflammation are commonly accepted constituents of Parkinson’s Disease (PD), understanding the mechanisms by which Nurr1 regulates inflammatory processes could provide an attractive therapeutic target. Nurr1 regulates inflammation via a transrepressive mechanism possibly dependent upon SUMOylation. In addition, Nurr1 can transactivate numerous genes involved in DA synthesis, such as Tyrosine Hydroxylase (TH). A C-terminal splice variant of Nurr1, Nurr-1a, has been reported in both neuronal and glial cells. However, research into its transcriptional activity is minimal. We employed in vitro methods such as SUMO-Pulldown experiments alongside Luciferase reporter assays to investigate the SUMOylation status and transactivation capabilities of Nurr1 and Nurr-1a respectively. The SUMO-Pulldown assay demonstrated Nurr-1a undergoes significantly more SUMO modification than its full-length variant. Consequently, despite having less transcriptional activation than Nurr1, Nurr1a may play a more prominent role in repression of microglial inflammation. Contrary to published literature we also identified that SUMOylation enhances transcriptional activation by Nurr1 and Nurr1a. SUMOylation-dependent increases in Nurr1 and Nurr1a transcriptional activation were only evident in neuronal SHSY5Y cells but not in HEK293 cells. This research provides novel insight into the regulation of Nurr-1a and indicates differential effects of SUMOylation dependent regulation in neuronal and inflammatory cells.

Keywords: nuclear receptors, Parkinson’s disease, inflammation, transcriptional regulation

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1751 New Experiences into Pancreatic Disease Science

Authors: Nadia Akbarpour


Pancreatic ductal adenocarcinoma is a forceful and obliterating illness, which is portrayed by intrusiveness, fast movement, and significant protection from treatment. Advances in neurotic arrangement and malignant growth hereditary qualities have worked on our illustrative comprehension of this infection; be that as it may, significant parts of pancreatic disease science remain ineffectively comprehended. A superior comprehension of pancreatic disease science should lead the way to more viable medicines. In the course of the most recent couple of years, there have been significant advances in the sub-atomic and organic comprehension of pancreatic malignancy. This included comprehension of the genomic intricacy of the illness, the job of pancreatic malignant growth undifferentiated organisms, the importance of the growth microenvironment, and the one-of-a-kind metabolic transformation of pancreas disease cells to acquire supplements under hypoxic climate. Endeavors have been made towards the advancement of the practical answer for its treatment with compelled achievement due to its complicated science. It is grounded that pancreatic malignancy undifferentiated cells (CSCs), yet present in a little count, contribute extraordinarily to PC inception, movement, and metastasis. Standard chemo and radiotherapeutic choices, notwithstanding, grow general endurance, the connected aftereffects are a huge concern. In the midst of the latest decade, our understanding with regards to atomic and cell pathways engaged with PC and the job of CSCs in its movement has expanded massively. By and by, the center is to target CSCs. The natural items have acquired a lot of thought as of late as they, generally, sharpen CSCs to chemotherapy and target atomic flagging engaged with different cancers, including PC. Some arranged investigations have demonstrated promising outcomes recommending that assessments in this course bring a ton to the table for the treatment of PC. Albeit preclinical investigations uncovered the significance of natural items in lessening pancreatic carcinoma, restricted examinations have been led to assess their part in centers. The current survey gives another knowledge to late advances in pancreatic malignancy science, treatment, and the current status of natural items in its expectation.

Keywords: pancreatic, genomic, organic, cancer

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1750 Design and in Slico Study of the Truncated Spike-M-N SARS-CoV-2 as a Novel Effective Vaccine Candidate

Authors: Aghasadeghi MR., Bahramali G., Sadat SM., Sadeghi SA., Yousefi M., Khodaei K., Ghorbani M., Sadat Larijani M.


Background:The emerging COVID-19 pandemic is a serious concernfor the public health worldwide. Despite the many mutations in the virus genome, it is important to find an effective vaccine against viral mutations. Therefore, in current study, we aimed at immunoinformatic evaluation of the virus proteins immunogenicity to design a preventive vaccine candidate, which could elicit humoral and cellular immune responses as well. Methods:Three antigenic regions are included;Spike, Membrane, and Nucleocapsid amino acid sequences were obtained, and possible fusion proteins were assessed andcompared by immunogenicity, structural features, and population coverage. The best fusion protein was also evaluated for MHC-I and MHC-II T-cell epitopes and the linear and conformational B-cell epitopes. Results: Among the four predicted models, the truncated Spike protein in fusion with M and N proteins is composed of 24 highly immunogenic human MHC class I and 29 MHC class II, along with 14 B-cell linear and 61 discontinues epitopes. Also, the selected protein has high antigenicity and acceptable population coverage of 82.95% in Iran and 92.51% in Europe. Conclusion: The data indicate that the truncated Spike-M-N SARS-CoV-2form which could be potential targets of neutralizing antibodies. The protein also has the ability to stimulate humoral and cellular immunity. The in silico study provided the fusion protein as a potential preventive vaccine candidate for further in vivo evaluation.

Keywords: SARS-CoV-2, immunoinformatic, protein, vaccine

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1749 Identification of the Target Genes to Increase the Immunotherapy Response in Bladder Cancer Patients using Computational and Experimental Approach

Authors: Sahar Nasr, Lin Li, Edwin Wang


Bladder cancer (BLCA) is known as the 13th cause of death among cancer patients worldwide, and ~575,000 new BLCA cases are diagnosed each year. Urothelial carcinoma (UC) is the most prevalent subtype among BLCA patients, which can be categorized into muscle-invasive bladder cancer (MIBC) and non-muscle-invasive bladder cancer (NMIBC). Currently, various therapeutic options are available for UC patients, including (1) transurethral resection followed by intravesical instillation of chemotherapeutics or Bacillus Calmette-Guérin for NMIBC patients, (2) neoadjuvant platinum-based chemotherapy (NAC) plus radical cystectomy is the standard of care for localized MIBC patients, and (3) systematic chemotherapy for metastatic UC. However, conventional treatments may lead to several challenges for treating patients. As an illustration, some patients may suffer from recurrence of the disease after the first line of treatment. Recently, immune checkpoint therapy (ICT) has been introduced as an alternative treatment strategy for the first or second line of treatment in advanced or metastatic BLCA patients. Although ICT showed lucrative results for a fraction of BLCA patients, ~80% of patients were not responsive to it. Therefore, novel treatment methods are required to augment the ICI response rate within BLCA patients. It has been shown that the infiltration of T-cells into the tumor microenvironment (TME) is positively correlated with the response to ICT within cancerous patients. Therefore, the goal of this study is to enhance the infiltration of cytotoxic T-cells into TME through the identification of target genes within the tumor that are responsible for the non-T-cell inflamed TME and their inhibition. BLCA bulk RNA-sequencing data from The Cancer Genome Atlas (TCGA) and immune score for TCGA samples were used to determine the Pearson correlation score between the expression of different genes and immune score for each sample. The genes with strong negative correlations were selected (r < -0.2). Thereafter, the correlation between the expression of each gene and survival in BLCA patients was calculated using the TCGA data and Cox regression method. The genes that are common in both selected gene lists were chosen for further analysis. Afterward, BLCA bulk and single-cell RNA-sequencing data were ranked based on the expression of each selected gene and the top and bottom 25% samples were used for pathway enrichment analysis. If the pathways related to the T-cell infiltration (e.g., antigen presentation, interferon, or chemokine pathways) were enriched within the low-expression group, the gene was included for downstream analysis. Finally, the selected genes will be used to calculate the correlation between their expression and the infiltration rate of the activated CD+8 T-cells, natural killer cells and the activated dendric cells. A list of potential target genes has been identified and ranked based on the above-mentioned analysis and criteria. SUN-1 got the highest score within the gene list and other identified genes in the literature as benchmarks. In conclusion, inhibition of SUN1 may increase the tumor-infiltrating lymphocytes and the efficacy of ICI in BLCA patients. BLCA tumor cells with and without SUN-1 CRISPR/Cas9 knockout will be injected into the syngeneic mouse model to validate the predicted SUN-1 effect on increasing tumor-infiltrating lymphocytes.

Keywords: data analysis, gene expression analysis, gene identification, immunoinformatic, functional genomics, transcriptomics

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1748 MARISTEM: A COST Action Focused on Stem Cells of Aquatic Invertebrates

Authors: Arzu Karahan, Loriano Ballarin, Baruch Rinkevich


Marine invertebrates, the highly diverse phyla of multicellular organisms, represent phenomena that are either not found or highly restricted in the vertebrates. These include phenomena like budding, fission, a fusion of ramets, and high regeneration power, such as the ability to create whole new organisms from either tiny parental fragment, many of which are controlled by totipotent, pluripotent, and multipotent stem cells. Thus, there is very much that can be learned from these organisms on the practical and evolutionary levels, further resembling Darwin's words, “It is not the strongest of the species that survives, nor the most intelligent, but the one most responsive to change”. The ‘stem cell’ notion highlights a cell that has the ability to continuously divide and differentiate into various progenitors and daughter cells. In vertebrates, adult stem cells are rare cells defined as lineage-restricted (multipotent at best) with tissue or organ-specific activities that are located in defined niches and further regulate the machinery of homeostasis, repair, and regeneration. They are usually categorized by their morphology, tissue of origin, plasticity, and potency. The above description not always holds when comparing the vertebrates with marine invertebrates’ stem cells that display wider ranges of plasticity and diversity at the taxonomic and the cellular levels. While marine/aquatic invertebrates stem cells (MISC) have recently raised more scientific interest, the know-how is still behind the attraction they deserve. MISC, not only are highly potent but, in many cases, are abundant (e.g., 1/3 of the entire animal cells), do not locate in permanent niches, participates in delayed-aging and whole-body regeneration phenomena, the knowledge of which can be clinically relevant. Moreover, they have massive hidden potential for the discovery of new bioactive molecules that can be used for human health (antitumor, antimicrobial) and biotechnology. The MARISTEM COST action (Stem Cells of Marine/Aquatic Invertebrates: From Basic Research to Innovative Applications) aims to connect the European fragmented MISC community. Under this scientific umbrella, the action conceptualizes the idea for adult stem cells that do not share many properties with the vertebrates’ stem cells, organizes meetings, summer schools, and workshops, stimulating young researchers, supplying technical and adviser support via short-term scientific studies, making new bridges between the MISC community and biomedical disciplines.

Keywords: aquatic/marine invertebrates, adult stem cell, regeneration, cell cultures, bioactive molecules

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1747 Improving Engagement: Dental Veneers, a Qualitative Analysis of Posts on Instagram

Authors: Matthew Sedgwick


Introduction: Social media continues to grow in popularity and Instagram is one of the largest platforms available. It provides an invaluable method of communication between health care professionals and patients. Both patients and dentists can benefit from seeing clinical cases posted by other members of the profession. It can prompt discussion about how the outcome was achieved and showcases what is possible with the right techniques and planning. This study aimed to identify what people were posting about the topic ‘veneers’ and inform health care professionals as to what content had the most engagement and make recommendations as to how to improve the quality of social media posts. Design: 150 consecutive posts for the search term ‘veneers’ were analyzed retrospectively between 21st October 2021 to 31st October 2021. Non-English language posts duplicated posts, and posts not about dental veneers were excluded. After exclusions were applied, 80 posts were included in the study for analysis. The content of the posts was analyzed and coded and the main themes were identified. The number of comments, likes and views were also recorded for each post. Results: The themes were: before and after treatment, cost, dental training courses, treatment process and trial smiles. Dentists were the most common posters of content (82.5%) and it was interesting to note that there were no patients who posted about treatment in this sample. The main type of media was photographs (93.75%) compared to video (6.25%). Videos had an average of 45,541 views and more comments and likes than the average for photographs. The average number of comments and likes per post were 20.88 and 761.58, respectively. Conclusion: Before and after photographs were the most common finding as this is how dentists showcase their work. The study showed that videos showing the treatment process had more engagement than photographs. Dentists should consider making video posts showing the patient journey, including before and after veneer treatment, as this can result in more potential patients and colleagues viewing the content. Video content could help dentists distinguish their posts from others as it can also be used across other platforms such as TikTok or Facebook reaching a wider audience. More informative posts about how the result has shown are achieved required, including potential costs. This will help increase transparency regarding this treatment method, including the financial and potential biological cost to teeth. As a result, this will improve patient understanding and become an invaluable adjunct in informed consent.

Keywords: content analysis, dental veneers, Instagram, social media

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1746 Palynological Investigation and Quality Determination of Honeys from Some Apiaries in Northern Nigeria

Authors: Alebiosu Olugbenga Shadrak, Victor Victoria


Honey bees exhibit preferences in their foraging behaviour on pollen and nectar for food and honey production, respectively. Melissopalynology is the study of pollen in honey and other honey products. Several work have been conducted on the palynological studies of honeys from the southern parts of Nigeria but with relatively scant records from the Northern region of the country. This present study aimed at revealing the favourably visited plants by honey bees, Apis melifera var. adansonii, at some apiaries in Northern Nigeria, as well as determining the quality of honeys produced. Honeys were harvested and collected from four apiaries of the region, namely: Sarkin Dawa missionary bee farm, Taraba State; Eleeshuwa Bee Farm, Keffi, Nassarawa State, Bulus Beekeeper Apiaries, Kagarko, Kaduna State and Mai Gwava Bee Farm, Kano State. These honeys were acetolysed for palynological microscopic analysis and subjected to standard treatment methods for the determination of their proximate composition and sugar profiling. Fresh anthers of two dominantly represented plants in the honeys were then collected for the quantification of their pollen protein contents, using the micro-kjeldhal procedure. A total of 30 pollen types were identified in the four honeys, and some of them were common to the honeys. A classification method for expressing pollen frequency class was employed: Senna cf. siamea, Terminalia cf. catappa, Mangifera indica, Parinari curatelifolia, Vitellaria paradoxa, Elaeis guineensis, Parkia biglobosa, Phyllantus muellerianus and Berlina Grandiflora, as “Frequent” (16-45%); while the others are either Rare (3-15%) or Sporadic (less than 3 %). Pollen protein levels of the two abundantly represented plants, Senna siamea (15.90mg/ml) and Terminalia catappa (17.33mg/ml) were found to be considerably lower. The biochemical analyses revealed varying amounts of proximate composition, non-reducing sugar and total sugar levels in the honeys. The results of this study indicate that pollen and nectar of the “Frequent” plants were preferentially foraged by honeybees in the apiaries. The estimated pollen protein contents of Senna same and Terminalia catappa were considerably lower and not likely to have influenced their favourable visitation by honeybees. However, a relatively higher representation of Senna cf. siamea in the pollen spectrum might have resulted from its characteristic brightly coloured and well scented flowers, aiding greater entomophily. Terminalia catappa, Mangifera indica, Elaeis guineensis, Vitellaria paradoxa, and Parkia biglobosa are typical food crops; hence they probably attracted the honeybees owing to the rich nutritional values of their fruits and seeds. Another possible reason for a greater entomophily of the favourably visited plants are certain nutritional constituents of their pollen and nectar, which were not investigated in this study. The nutritional composition of the honeys was observed to fall within the safe limits of international norms, as prescribed by Codex Alimentarius Commission, thus they are good honeys for human consumption. It is therefore imperative to adopt strategic conservation steps in ensuring that these favourably visited plants are protected from indiscriminate anthropogenic activities and also encourage apiarists in the country to establish their bee farms more proximally to the plants for optimal honey yield.

Keywords: honeybees, melissopalynology, preferentially foraged, nutritional, bee farms, proximally

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1745 Foamability and Foam Stability of Gelatine-Sodium Dodecyl Sulfate Solutions

Authors: Virginia Martin Torrejon, Song Hang


Gelatine foams are widely explored materials due to their biodegradability, biocompatibility, and availability. They exhibit outstanding properties and are currently subject to increasing scientific research due to their potential use in different applications, such as biocompatible cellular materials for biomedical products or biofoams as an alternative to fossil-fuel-derived packaging. Gelatine is a highly surface-active polymer, and its concentrated solutions usually do not require surfactants to achieve low surface tension. Still, anionic surfactants like sodium dodecyl sulfate (SDS) strongly interact with gelatine, impacting its viscosity and rheological properties and, in turn, their foaming behaviour. Foaming behaviour is a key parameter for cellular solids produced by mechanical foaming as it has a significant effect on the processing and properties of cellular materials. Foamability mainly impacts the density and the mechanical properties of the foams, while foam stability is crucial to achieving foams with low shrinkage and desirable pore morphology. This work aimed to investigate the influence of SDS on the foaming behaviour of concentrated gelatine foams by using a dynamic foam analyser. The study of maximum foam height created, foam formation behaviour, drainage behaviour, and foam structure with regard to bubble size and distribution were carried out in 10 wt% gelatine solutions prepared at different SDS/gelatine concentration ratios. Comparative rheological and viscometry measurements provided a good correlation with the data from the dynamic foam analyser measurements. SDS incorporation at optimum dosages and gelatine gelation led to highly stable foams at high expansion ratios. The viscosity increase of the hydrogel solution at SDS content increased was a key parameter for foam stabilization. In addition, the impact of SDS content on gelling time and gel strength also considerably impacted the foams' stability and pore structure.

Keywords: dynamic foam analyser, gelatine foams stability and foamability, gelatine-surfactant foams, gelatine-SDS rheology, gelatine-SDS viscosity

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1744 Comparison of Two Methods of Cryopreservation of Testicular Tissue from Prepubertal Lambs

Authors: Rensson Homero Celiz Ygnacio, Marco Aurélio Schiavo Novaes, Lucy Vanessa Sulca Ñaupas, Ana Paula Ribeiro Rodrigues


The cryopreservation of testicular tissue emerges as an alternative for the preservation of the reproductive potential of individuals who still cannot produce sperm; however, they will undergo treatments that may affect their fertility (e.g., chemotherapy). Therefore, the present work aims to compare two cryopreservation methods (slow freezing and vitrification) in testicular tissue of prepubertal lambs. For that, to obtain the testicular tissue, the animals were castrated and the testicles were collected immediately in a physiological solution supplemented with antibiotics. In the laboratory, the testis was split into small pieces. The total size of the testicular fragments was 3×3x1 mm³ and was placed in a dish contained in Minimum Essential Medium (MEM-HEPES). The fragments were distributed randomly into non-cryopreserved (fresh control), slow freezing (SF), and vitrified. To SF procedures, two fragments from a given male were then placed in a 2,0 mL cryogenic vial containing 1,0 mL MEM-HEPES supplemented with 20% fetal bovine serum (FBS) and 20% dimethylsulfoxide (DMSO). Tubes were placed into a Mr. Frosty™ Freezing container with isopropyl alcohol and transferred to a -80°C freezer for overnight storage. On the next day, each tube was plunged into liquid nitrogen (NL). For vitrification, the ovarian tissue cryosystem (OTC) device was used. Testicular fragments were placed in the OTC device and exposed to the first vitrification solution composed of MEM-HEPES supplemented with 10 mg/mL Bovine Serum Albumin (BSA), 0.25 M sucrose, 10% Ethylene glycol (EG), 10% DMSO and 150 μM alpha-lipoic acid for four min. The VS1 was discarded and then the fragments were submerged into a second vitrification solution (VS2) containing the same composition of VS1 but 20% EG and 20% DMSO. VS2 was then discarded and each OTC device containing up to four testicular fragments was closed and immersed in NL. After the storage period, the fragments were removed from the NL, kept at room temperature for one min and then immersed at 37 °C in a water bath for 30 s. Samples were warmed by sequentially immersing in solutions of MEM-HEPES supplemented with 3 mg/mL BSA and decreasing concentrations of sucrose. Hematoxylin-eosin staining to analyze the tissue architecture was used. The score scale used was from 0 to 3, classified with a score 0 representing normal morphologically, and 3 were considered a lot of alteration. The histomorphological evaluation of the testicular tissue shows that when evaluating the nuclear alteration (distinction of nucleoli and condensation of nuclei), there are no differences when using slow freezing with respect to the control. However, vitrification presents greater damage (p <0.05). On the other hand, when evaluating the epithelial alteration, we observed that the freezing showed scores statistically equal to the control in variables such as retraction of the basement membrane, formation of gaps and organization of the peritubular cells. The results of the study demonstrated that cryopreservation using the slow freezing method is an excellent tool for the preservation of pubertal testicular tissue.

Keywords: cryopreservation, slow freezing, vitrification, testicular tissue, lambs

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1743 The CDK Pho85 Inhibits Whi7 Repressor to Promote Cell Cycle Entry

Authors: Cristina Ros-Carrero, Mihai Spiridon-Bodi, Juan Carlos Igual, Mercè Gomar-Alba


Start (the G1/S transition) is the main decision point in the eukaryotic cell cycle at which cells irreversibly commit to a new round of cell division by activating the Start transcriptional program. In budding yeast, triggering Start involves the inactivation of the Start transcriptional repressors, Whi5 (Rb in mammals) and Whi7, which are inactivated by the CDK Cdc28 dependent phosphorylation. Pho85 is a CDK that regulates the cellular response to phosphate levels and diverse stresses. Pho85 is also linked to cell cycle control, and Start regulators have been proposed as Pho85 targets. Here we unravel a new mechanism by which Pho85 directly promotes Start. We saw that Pho85 specifically downregulates Whi7 but not Whi5 protein levels. We demonstrate that CDK Pho85-cyclins regulates Whi7 levels in two parallel ways: Pho85-Pho80 represses Whi7expression through the inactivation of the Pho4 transcription factor, and Pho85-Pcl1, Pcl2, Pcl9, Pho80 promotes its instability through the phosphorylation of Ser27 and Thr100 aminoacids. Strikingly, unlike in wild type cells, in the absence of Pho85, Whi7 is more potent than Whi5 repressing Start. First, only Whi7 causes G1 arrest in pho85 mutant cells when Whi5 or Whi7 are overexpressed to similar protein levels. Second, the G1 delay observed in pho85 mutant cells is restored by the deletion of Whi7, but not by Whi5 mutation. Furthermore, Pho85 inactivation causes an increased Whi7 association to G1/S promoters, which is dependent on the phosphorylation state of Ser27 and Thr100. Remarkably, Pho85 not only decreases Whi7 levels but also blocks Whi7 function as a Start repressor. Thus, this is a new mechanism that links the Pho85 pathway with the control of Start, unveiling a new role for the Whi7 transcriptional repressor under conditions of Pho85 inactivation.

Keywords: CDK Pho85, saccharomyces cerevisiae, start, whi7

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1742 Revisiting the Surgical Approaches to Decompression in Quadrangular Space Syndrome: A Cadaveric Study

Authors: Sundip Charmode, Simmi Mehra, Sudhir Kushwaha, Shalom Philip, Pratik Amrutiya, Ranjna Jangal


Introduction: Quadrangular space syndrome involves compression of the axillary nerve and posterior circumflex humeral artery and its management in few cases, requires surgical decompression. The current study reviews the surgical approaches used in the decompression of neurovascular structures and presents our reflections and recommendations. Methods: Four human cadavers, in the Department of Anatomy were used for dissection of the Axillae and the Scapular region by the senior residents of the Department of Anatomy and Department of Orthopedics, who dissected quadrangular space in the eight upper limbs, using anterior and posterior surgical approaches. Observations: Posterior approach to identify the quadrangular space and secure its contents was recognized as the easier and much quicker method by both the Anatomy and Orthopedic residents, but it may result in increased postoperative morbidity. Whereas the anterior (Delto-pectoral) approach involves more skill but reduces postoperative morbidity. Conclusions: Anterior (Delto-pectoral) approach with suggested modifications can prove as an effective method in surgical decompression of quadrangular space syndrome. The authors suggest more cadaveric studies to facilitate anatomists and surgeons with the opportunities to practice and evaluate older and newer surgical approaches.

Keywords: surgical approach, anatomical approach, decompression, axillary nerve, quadrangular space

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1741 De novo Transcriptome Assembly of Lumpfish (Cyclopterus lumpus L.) Brain Towards Understanding their Social and Cognitive Behavioural Traits

Authors: Likith Reddy Pinninti, Fredrik Ribsskog Staven, Leslie Robert Noble, Jorge Manuel de Oliveira Fernandes, Deepti Manjari Patel, Torstein Kristensen


Understanding fish behavior is essential to improve animal welfare in aquaculture research. Behavioral traits can have a strong influence on fish health and habituation. To identify the genes and biological pathways responsible for lumpfish behavior, we performed an experiment to understand the interspecies relationship (mutualism) between the lumpfish and salmon. Also, we tested the correlation between the gene expression data vs. observational/physiological data to know the essential genes that trigger stress and swimming behavior in lumpfish. After the de novo assembly of the brain transcriptome, all the samples were individually mapped to the available lumpfish (Cyclopterus lumpus L.) primary genome assembly (fCycLum1.pri, GCF_009769545.1). Out of ~16749 genes expressed in brain samples, we found 267 genes to be statistically significant (P > 0.05) found only in odor and control (1), model and control (41) and salmon and control (225) groups. However, genes with |LogFC| ≥0.5 were found to be only eight; these are considered as differentially expressed genes (DEG’s). Though, we are unable to find the differential genes related to the behavioral traits from RNA-Seq data analysis. From the correlation analysis, between the gene expression data vs. observational/physiological data (serotonin (5HT), dopamine (DA), 3,4-Dihydroxyphenylacetic acid (DOPAC), 5-hydroxy indole acetic acid (5-HIAA), Noradrenaline (NORAD)). We found 2495 genes found to be significant (P > 0.05) and among these, 1587 genes are positively correlated with the Noradrenaline (NORAD) hormone group. This suggests that Noradrenaline is triggering the change in pigmentation and skin color in lumpfish. Genes related to behavioral traits like rhythmic, locomotory, feeding, visual, pigmentation, stress, response to other organisms, taxis, dopamine synthesis and other neurotransmitter synthesis-related genes were obtained from the correlation analysis. In KEGG pathway enrichment analysis, we find important pathways, like the calcium signaling pathway and adrenergic signaling in cardiomyocytes, both involved in cell signaling, behavior, emotion, and stress. Calcium is an essential signaling molecule in the brain cells; it could affect the behavior of fish. Our results suggest that changes in calcium homeostasis and adrenergic receptor binding activity lead to changes in fish behavior during stress.

Keywords: behavior, De novo, lumpfish, salmon

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1740 Prevention of COVID-19 Using Herbs and Natural Products

Authors: Nada Alqadri, Omaima Nasir


Natural compounds are an important source of potential inhibitors; they have a lot of pharma potential with less adverse effects. The effective antiviral activities of natural products have been proved in different studies. The outbreak of COVID-19 in Wuhan, Hubei, in December 2019, coronavirus has had a significant impact on people's health and lives. Based on previous studies, natural products can be introduced as preventive and therapeutic agents in the fight against COVID-19; considering that no food or supplement has been authorized to prevent COVID-19, individuals continue to search for and consume specific herbs, foods, and commercial supplements for this purpose. This study will be aimed to estimate the uses of herbal and natural products during the COVID-19 infection to determine their usage reasons and evaluate their potential side effects. An online cross-sectional survey of different participants will be conducted and will be a focus on respondents’ chronic disease histories, socio-dmographic characteristics, and frequency and trends of using these products. Descriptive and univariate analyses will be performed to determine prevalence and associations between various products used and respondents’ socio-demographic data. Relationships will be tested using Pearson’s chi-square test or an exact probability test. Our main findings will give evidence of beneficial uses of natural products and herbal medicine as prophylactic and will be a vigorous approach to stop or at least slow down COVID-19 infection and transmission. This will be of great interest of public health, and the results of our study will lend health officials better control on the current pandemic.

Keywords: COVID-19, herbs, natural products, saudi arabia

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1739 Cascade Screening for Beta-Thalassemia in Pakistan: Relatives’ Experiences of a Decision Support Intervention in Routine Practice

Authors: Shenaz Ahmed, Hussain Jafri, Muhammed Faran, Wajeeha Naseer Ahmed, Yasmin Rashid, Yasmin Ehsan, Shabnam Bashir, Mushtaq Ahmed


Low uptake of cascade screening for βeta-Thalassaemia Major (β-TM) in the ‘Punjab Thalassaemia Prevention Project’ (PTPP) in Pakistan led to the development of a ‘decision support intervention for relatives’ (DeSIRe). This paper presents the experiences of relatives of children with β-TM of the DeSIRe following its use by PTPP field officers in routine clinical practice. Fifty-four semi-structured qualitative interviews were conducted (April to June 2021) with relatives in seven cities in the Punjab province (Lahore, Sheikhupura, Nankana Sahab, Kasur, Gujranwala, Multan, and Faisalabad). Thematic analysis shows that participants were satisfied with the content of the DeSIRe and its delivery by the field officers in a family meeting. They understood the main purpose of the DeSIRe was to improve their knowledge of β-TM and its inheritance, to enable them to make decisions about thalassemia carrier testing, particularly before marriage. While participants raised concerns about the stigma of testing positive, they believed the DeSIRe was an appropriate intervention, which supported relatives to make informed decisions. Our findings show the DeSIRe is appropriate for use by healthcare professionals in routine practice in a low-middle income country and has the potential to facilitate shared decision-making about cascade screening for thalassemia. Further research is needed to prove the efficacy of the DeSIRe.

Keywords: thalassemia, Pakistan, cascade screening, decision support

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1738 Evaluation of the Effectiveness of the Argon Plasma Jet on Healing Process of the Wagner Grade 2 Diabetic Foot Ulcer

Authors: M. Khaledi Pour, P. Akbartehrani, M. Amini, M. Khani, M. Mohajeri Tehrani, R. Radi, B. Shokri


Diabetic Foot Ulcer (DFU) is one of the costly severe complications of diabetes. Neuropathy and Peripheral Arterial Disease (PAD) due to diabetes are significant causes of this complication. In 10 years the patients with DFUs are twice as likely to die as patients without DFUs. Cold Atmospheric Plasma (CAP) is a promising tool for medical purposes. CAP generate reactive species at room temperature and are effective in killing bacteria and fibroblast proliferation. These CAP-based tools produce NO, which has bactericidal and angiogenesis properties. It also showed promising effects in the DFUs surface reduction and the time to wound closure. In this paper, we evaluated the effect of the Argon Plasma Jet (APJ) on the healing process of the Wagner Grade 2 DFUs in a randomized clinical trial. The 20 kHz sinusoidal voltage frequency derives the APJ. Patients (n=20) were randomly double-blinded assigned into two groups. These groups receive the standard care (SC, n=10) and the standard care with APJ treatment (SC+APJ, n=10) for five sessions in four weeks. The results showed that the APJ treatment along standard care could reduce the wound surface by 20 percent more than the standard care. Also, It showed a more influential role in controlling wound infection.

Keywords: argon plasma jet, cold atmospheric plasma, diabetes, diabetic foot ulcer

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1737 Investigation of Cold Atmospheric Plasma Exposure Protocol on Wound Healing in Diabetic Foot Ulcer

Authors: P. Akbartehrani, M. Khaledi Pour, M. Amini, M. Khani, M. Mohajeri Tehrani, E. Ghasemi, P. Charipoor, B. Shokri


A common problem between diabetic patients is foot ulcers which are chronic and require specialized treatment. Previous studies illustrate that Cold atmospheric plasma (CAP) has beneficial effects on wound healing and infection. Nevertheless, the comparison of different cap exposure protocols in diabetic ulcer wound healing remained to be studied. This study aims to determine the effect of two different exposure protocols on wound healing in diabetic ulcers. A prospective, randomized clinical trial was conducted at two clinics. Diabetic patients with G1 and G2 wanger classification diabetic foot ulcers were divided into two groups of study. One group was treated by the first protocol, which was treating wounds by argon-generated cold atmospheric plasma jet once a week for five weeks in a row. The other group was treated by the second protocol, which was treating wounds every three days for five weeks in a row. The wounds were treated for 40 seconds/cubic centimeter, while the nozzle tip was moved nonlocalized 1 cm above the wounds. A patient with one or more wounds could participate in different groups as wounds were separately randomized, which allow a participant to be treated several times during the study. The study's significant findings were two different reductions rate in wound size, microbial load, and two different healing speeds. This study concludes that CAP therapy by the second protocol yields more effective healing speeds, reduction in wound sizes, and microbial loads of foot ulcers in diabetic patients.

Keywords: wound healing, diabetic ulcers, cold atmospheric plasma, cold argon jet

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1736 FEDBD Plasma, A Promising Approach for Skin Rejuvenation

Authors: P. Charipoor, M. Khani, H. Mahmoudi, E. Ghasemi, P. Akbartehrani, B. Shokri


Cold air plasma could have a variety of effects on cells and living organisms and also shows good results in medical and cosmetic cases. Herein, plasma floating electrode dielectric barrier discharge (FEDBD) plasma was designed for mouse skin rejuvenation purposes. It is safe and easy to use in clinics, laboratories, and homes. The effects of this device were investigated on mouse skin. Vitamin C ointment in combination with plasma was also used as a new method to improve FEDBD results. In this study, 20 Wistar rats were evaluated in four groups. The first group received high-dose plasma, the second group received moderate-dose plasma (with vitamin C cream), the third group received low-dose plasma (with vitamin C cream) for 6 minutes, and the fourth group received only vitamin C cream. This process was done 3 times a week for 4 weeks. Skin temperature was monitored to evaluate the thermal effect of plasma. The presence of reactive species was also demonstrated using optical spectroscopy. Mechanical assays were performed to evaluate the effect of plasma and vitamin C on the mechanical strength of the tissue, which showed a positive effect of plasma on the treated tissue compared to the control group. Using pathological and biometric skin tests, an increase in collagen levels, epidermal thickness, and an increase in fibroblasts was observed in rat skin, as well as increased skin elasticity. This study showed the positive effect of using the FEDBD plasma device on the effective parameters in skin rejuvenation.

Keywords: plasma, skin rejuvenation, collagen, epidermal thickness

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1735 The Effects of Spark Plasma on Infectious Wound Healing

Authors: Erfan Ghasemi, Mohammadreza Khani, Hamidreza Mahmoudi, Mohammad Ali Nilforoushzadeh, Babak Shokri, Pouria Akbartehrani


Given the global significance of treating infectious wounds, the goal of this study is to use spark plasma as a new treatment for infectious wounds. To generate spark plasma, a high-voltage (7 kV) and high-frequency (75 kHz) source was used. Infectious wounds in the peritoneum of mice were divided into control and plasma-treated groups at random. The plasma-treated animals received plasma radiation every 4 days for 12 days, for 60 seconds each time. On the 15th day after the first session, the wound in the plasma-treated group had completely healed. The spectra of spark plasma emission and tissue properties were studied. The mechanical resistance of the wound healed in the plasma treatment group was considerably higher than in the control group (p<0.05), according to the findings. Furthermore, histological evidence suggests that wound re-epithelialization is faster in comparison to controls. Angiogenesis and fibrosis (collagen production) were also dramatically boosted in the plasma-treated group, whereas the stage of wound healing inflammation was significantly reduced. Plasma therapy accelerated wound healing by causing considerable wound constriction. The results of this investigation show that spark plasma has an influence on the treatment of infectious wounds.

Keywords: infectious wounds, mice, spark plasma, treatment

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1734 Discovery of Exoplanets in Kepler Data Using a Graphics Processing Unit Fast Folding Method and a Deep Learning Model

Authors: Kevin Wang, Jian Ge, Yinan Zhao, Kevin Willis


Kepler has discovered over 4000 exoplanets and candidates. However, current transit planet detection techniques based on the wavelet analysis and the Box Least Squares (BLS) algorithm have limited sensitivity in detecting minor planets with a low signal-to-noise ratio (SNR) and long periods with only 3-4 repeated signals over the mission lifetime of 4 years. This paper presents a novel precise-period transit signal detection methodology based on a new Graphics Processing Unit (GPU) Fast Folding algorithm in conjunction with a Convolutional Neural Network (CNN) to detect low SNR and/or long-period transit planet signals. A comparison with BLS is conducted on both simulated light curves and real data, demonstrating that the new method has higher speed, sensitivity, and reliability. For instance, the new system can detect transits with SNR as low as three while the performance of BLS drops off quickly around SNR of 7. Meanwhile, the GPU Fast Folding method folds light curves 25 times faster than BLS, a significant gain that allows exoplanet detection to occur at unprecedented period precision. This new method has been tested with all known transit signals with 100% confirmation. In addition, this new method has been successfully applied to the Kepler of Interest (KOI) data and identified a few new Earth-sized Ultra-short period (USP) exoplanet candidates and habitable planet candidates. The results highlight the promise for GPU Fast Folding as a replacement to the traditional BLS algorithm for finding small and/or long-period habitable and Earth-sized planet candidates in-transit data taken with Kepler and other space transit missions such as TESS(Transiting Exoplanet Survey Satellite) and PLATO(PLAnetary Transits and Oscillations of stars).

Keywords: algorithms, astronomy data analysis, deep learning, exoplanet detection methods, small planets, habitable planets, transit photometry

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1733 NanoCelle®: A Nano Delivery Platform to Enhance Medicine

Authors: Sean Hall


Nanosystems for drug delivery are not new; as medicines evolve, so too does the desire to deliver a more targeted, patient-compliant medicine. Though, historically the widespread use of nanosystems for drug delivery has been fouled by non-replicability, scalability, toxicity issues, and economics. Examples include steps of manufacture and thus cost to manufacture, toxicity for nanoparticle scaffolding, autoimmune response, and considerable technical expertise for small non-commercial yields. This, unfortunately, demonstrates the not-so-obvious chasm between science and drug formulation for regulatory approval. Regardless there is a general and global desire to improve the delivery of medicines, reduce potential side effect profiles, promote increased patient compliance, and increase and/or speed public access to medicine availability. In this paper, the author will discuss NanoCelle®, a nano-delivery platform that specifically addresses degradation and solubility issues that expands from fundamental micellar preparations. NanoCelle® has been deployed in several Australian listed medicines and is in use of several drug candidates across small molecules, with research endeavors now extending into large molecules. The author will discuss several research initiatives as they relate to NanoCelle® to demonstrate similarities seen in various drug substances; these examples will include both in vitro and in vivo work.

Keywords: NanoCelle®, micellar, degradation, solubility, toxicity

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1732 Night Shift Work as an Oxidative Stressor: A Systematic Review

Authors: Madeline Gibson


Night shift workers make up an essential part of the modern workforce. However, night shift workers have higher incidences of late in life diseases and earlier mortality. Night shift workers are exposed to constant light and experience circadian rhythm disruption. Sleep disruption is thought to increase oxidative stress, defined as an imbalance of excess pro-oxidative factors and reactive oxygen species over anti-oxidative activity. Oxidative stress can damage cells, proteins and DNA and can eventually lead to varied chronic diseases such as cancer, diabetes, cardiovascular disease, Alzheimer’s and dementia. This review aimed to understand whether night shift workers were at greater risk of oxidative stress and to contribute to a consensus on this relationship. Twelve studies published in 2001-2019 examining 2,081 workers were included in the review. Studies compared both the impact of working a single shift and in comparisons between those who regularly work night shifts and only day shifts. All studies had evidence to support this relationship across a range of oxidative stress indicators, including increased DNA damage, reduced DNA repair capacity, increased lipid peroxidation, higher levels of reactive oxygen species, and to a lesser extent, a reduction in antioxidant defense. This research supports the theory that melatonin and the sleep-wake cycle mediate the relationship between shift work and oxidative stress. It is concluded that night shift work increases the risk for oxidative stress and, therefore, future disease. Recommendations are made to promote the long-term health of shift workers considering these findings.

Keywords: night shift work, coxidative stress, circadian rhythm, melatonin, disease, circadian rhythm disruption

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1731 Mapping Structurally Significant Areas of G-CSF during Thermal Degradation with NMR

Authors: Mark-Adam Kellerman


Proteins are capable of exploring vast mutational spaces. This makes it difficult for protein engineers to devise rational methods to improve stability and function via mutagenesis. Deciding which residues to mutate requires knowledge of the characteristics they elicit. We probed the characteristics of residues in granulocyte-colony stimulating factor (G-CSF) using a thermal melt (from 295K to 323K) to denature it in a 700 MHz Bruker spectrometer. These characteristics included dynamics, micro-environmental changes experienced/ induced during denaturing and structure-function relationships. 15N-1H HSQC experiments were performed at 2K increments along with this thermal melt. We observed that dynamic residues that also undergo a lot of change in their microenvironment were predominantly in unstructured regions. Moreover, we were able to identify four residues (G4, A6, T133 and Q134) that we class as high priority targets for mutagenesis, given that they all appear in both the top 10% of measures for environmental changes and dynamics (∑Δ and ∆PI). We were also able to probe these NMR observables and combine them with molecular dynamics (MD) to elucidate what appears to be an opening motion of G-CSFs binding site III. V48 appears to be pivotal to this opening motion, which also seemingly distorts the loop region between helices A and B. This observation is in agreement with previous findings that the conformation of this loop region becomes altered in an aggregation-prone state of G-CSF. Hence, we present here an approach to profile the characteristics of residues in order to highlight their potential as rational mutagenesis targets and their roles in important conformational changes. These findings present not only an opportunity to effectively make biobetters, but also open up the possibility to further understand epistasis and machine learn residue behaviours.

Keywords: protein engineering, rational mutagenesis, NMR, molecular dynamics

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1730 The Effect of Different Cucumber (Cucumis sativus L.) Varieties on Growth and Development Time of Aphis gossypii Glover (Hemiptera: Aphididae)

Authors: Rochelyn Dona, Mohamed F. Nur, Serdar Satar


The biological response of Aphis gossypii Glover (Hom. Aphididae) was investigated on the effects of seven cucumber varieties (Cucumis sativus L.) such as Kitir, Muhika, Ayda, Beit, 14-F1, Ruzgar, and Ptk in the laboratory condition at 24±1°C, 65±5% relative humidity (RH) and a photoperiod of 16:8 (L:D) hour. The results were related that the developmental time of A. gossypii at the nymphal stages was presented a significant difference only on the first instar stage. From the lowest to the highest respectively, 0.98 days on ruzgar to 1.18 days on Kitir, the second nymphal stage 0.98 days to Beit alfa, 1.08 days on Muhika, the third from 0.94 days to Kitir, from 1.16 days to 14-F1, and the last instar 1.22 days on Ptk, 1.48 days on Kitir were investigated. The total development time was evaluated at 4.46 days Beit on alfa 4.72 days on Kitir. The offspring number was 60.42 aphids on ayda and 83.72 aphids on muhika, the significant differences between varieties were based on one-way ANOVA (Tukey test). The lifetime of A. gossypii was recorded 19.10 days on Kitir, 27.64 days on Ptk. The results showed that cucumber cultivars were affected by the biological life of A. gossypii. The combination of this study with the other methods of the IPM tactics can serve as the best strategy for controlling this pest on cucumber varieties into the greenhouse.

Keywords: cucumber cultivars, fecundity, intrinsic rate, mortality, resistance

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1729 Effect of Temperatures on Growth and Development Time of Aphis fabae Scopoli (Homoptera: Aphididae): On Bean (Phaseolus vulgaris L.)

Authors: Rochelyn Dona, Serdar Satar


The aim of this study was to evaluate the biological parameters of A. fabae Scopoli (Hemiptera: Aphididae). Developmental, survival, and reproductive data were collected for Aphis fabae reared on detached bean leaves (Phaseolus vulgaris L.) ‘pinto beans’ at five temperature regimes (12, 16, 20, 24, and 28 °C), 65% relative humidity (RH), relative and a photoperiod of 16:8 (LD) h. The developmental times of immature stages ranged from 16, 65 days at 12°C to 5.70 days at 24°C, but a slight increase again at 28°C (6.62 days). At 24°C from this study presented the developmental threshold for A. fabae slightly to 24°C. The average longevity of mature females significantly decreased from 42.32 days at 12°C to 16.12 days at 28°C. The reproduction rate per female was 62.27 at 16°C and 12.72 at 28°C. The mean generation period of the population ranged from 29.24 at 12°C to 11.50 at 28°C. The highest intrinsic rate of increase (rm = 0.41) were recorded at 24°C, the lowest at 12°C (rm = 0.15). It was evident that temperatures over 28°C augmented the development time, accelerated the death ratio of the nymphal stages, Shrunk Adult longevity, and reduced fecundity. The optimal range of temperature for the population growth of A. fabae on the bean was 16°C-24°C, according to this study.

Keywords: developmental time, intrinsic rate, reproduction period, temperature dependence

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1728 Polyhydroxybutyrate (PHB): Highly Porous Scaffold for Biomedicine

Authors: Neda Sinaei, Davood Zare, Mehrdad Azin


Polyhydroxyalkanoates (PHAs) are biocompatible and biodegradable polymers produced by a wide range of bacterial strains. These biopolymers are significantly studied for drug delivery and tissue engineering applications because of their fascinating physicochemical properties. Polyhydroxybutyrate (PHB) scaffold that has been extracted from a novel bacteria using oil wastewater was selected to study. Some physical parameters affecting scaffold properties such as PHB concentration, solvent evaporation speed, and ultrasonic time were investigated. Scanning electron microscopy was used to evaluate the porosity. Afterward, the biocompatibility of PHB scaffold was assessed. Initial results showed the highly porous PHB scaffold structure with a variety of pore sizes. Subsequent results indicated that more unique pore sizes can be obtained by optimizing physical factors. It would be noticed that the morphology of the pore structure was accordingly affected by ultrasonic time. Hence, In vitro cell viability tests on the PHB scaffold using human foreskin fibroblasts revealed strong cell attachment and proliferation supports. Therefore, it can be concluded that the cost-effective PHB scaffold has the potential using as a biomaterial cell adhesion substrate in therapeutic applications.

Keywords: Polyhydroxybutyrate, biocompatible, scaffold, porous, tissue engineering

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1727 Human Long-Term Deregulated Circadian Rhythm Alters Regenerative Properties of Skin and Hair Precursor Cells

Authors: Deshayes N, Genty G, Berthelot F, Paris M


Background: In mammals, desynchronized circadian rhythm leads to various biological symptoms. In skin and hair, human epidermal stem cell function in vitro is regulated by circadian oscillations and thus contributes to tissue aging when deregulated. In mice, circadian arrhythmia of hair follicle stem cells contributes to age-related hair follicle cycling defects. Despite the well-described impact of circadian oscillations through a feedback loop involving the clock pathway on hair and skin stem cell function in vitro, little is known about the change in characteristics or regenerative properties of hHF (human hair follicle keratinocytes), hEpi (human interfollicular epidermal keratinocytes), and hHFDP (hair follicle dermal papilla stem cells) after the long-term alteration of circadian rhythm in vivo. Objectives: The present study was designed to assess hHF, hEpi, and hHFDP precursors and stem cell properties in response to clock pathway alteration due to long-term deregulated circadian rhythm in vivo. A clinical study protocol involving two groups of women was designed: diurnal workers (control) and shift workers (deregulated). After informed consent, two 3-mm fresh punch biopsies were taken from the occipital region of each donor (10 donors/group). Cell culture characterization, measurement of colony area, culture medium analysis, and RT -qPCR analysis were carried out. Results: Long-term circadian rhythm deregulation affected clock pathway protein expression and correlated with alterations in hHF, hEpi, and hHFDP properties. Conclusion: This study provides, for the fi¬rst time in humans, evidence that in vivo deregulation of the clock pathway affects regenerative properties of human skin and hair precursor cells.

Keywords: circadian rhythm, clock pathway alteration, regenerative proprieties, human skin and hair cells

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