Abstracts | Bioengineering and Life Sciences
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2100

World Academy of Science, Engineering and Technology

[Bioengineering and Life Sciences]

Online ISSN : 1307-6892

1890 Reliability and Validity of a Portable Inertial Sensor and Pressure Mat System for Measuring Dynamic Balance Parameters during Stepping

Authors: Emily Rowe

Abstract:

Introduction: Balance assessments can be used to help evaluate a person’s risk of falls, determine causes of balance deficits and inform intervention decisions. It is widely accepted that instrumented quantitative analysis can be more reliable and specific than semi-qualitative ordinal scales or itemised scoring methods. However, the uptake of quantitative methods is hindered by expense, lack of portability, and set-up requirements. During stepping, foot placement is actively coordinated with the body centre of mass (COM) kinematics during pre-initiation. Based on this, the potential to use COM velocity just prior to foot off and foot placement error as an outcome measure of dynamic balance is currently being explored using complex 3D motion capture. Inertial sensors and pressure mats might be more practical technologies for measuring these parameters in clinical settings. Objective: The aim of this study was to test the criterion validity and test-retest reliability of a synchronised inertial sensor and pressure mat-based approach to measure foot placement error and COM velocity while stepping. Methods: Trials were held with 15 healthy participants who each attended for two sessions. The trial task was to step onto one of 4 targets (2 for each foot) multiple times in a random, unpredictable order. The stepping target was cued using an auditory prompt and electroluminescent panel illumination. Data was collected using 3D motion capture and a combined inertial sensor-pressure mat system simultaneously in both sessions. To assess the reliability of each system, ICC estimates and their 95% confident intervals were calculated based on a mean-rating (k = 2), absolute-agreement, 2-way mixed-effects model. To test the criterion validity of the combined inertial sensor-pressure mat system against the motion capture system multi-factorial two-way repeated measures ANOVAs were carried out. Results: It was found that foot placement error was not reliably measured between sessions by either system (ICC 95% CIs; motion capture: 0 to >0.87 and pressure mat: <0.53 to >0.90). This could be due to genuine within-subject variability given the nature of the stepping task and brings into question the suitability of average foot placement error as an outcome measure. Additionally, results suggest the pressure mat is not a valid measure of this parameter since it was statistically significantly different from and much less precise than the motion capture system (p=0.003). The inertial sensor was found to be a moderately reliable (ICC 95% CIs >0.46 to >0.95) but not valid measure for anteroposterior and mediolateral COM velocities (AP velocity: p=0.000, ML velocity target 1 to 4: p=0.734, 0.001, 0.000 & 0.376). However, it is thought that with further development, the COM velocity measure validity could be improved. Possible options which could be investigated include whether there is an effect of inertial sensor placement with respect to pelvic marker placement or implementing more complex methods of data processing to manage inherent accelerometer and gyroscope limitations. Conclusion: The pressure mat is not a suitable alternative for measuring foot placement errors. The inertial sensors have the potential for measuring COM velocity; however, further development work is needed.

Keywords: dynamic balance, inertial sensors, portable, pressure mat, reliability, stepping, validity, wearables

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1889 A Precursory Observation on Butterflies (Arthropoda, Insecta, Lepidoptera) of Umphang District, Tak Province, Western part of Thailand

Authors: Pisit Poolprasert, Auttpol Nakwa, Keerati Tanruean, Ezra Mongkolchaichana, Ezra Mongkolchaichana, Sinlapachai Senarat, Mark Tunmore

Abstract:

This present preliminary study aimed to observe the butterfly species diversity in two selected subdistricts i.e., Mae Klong and Umphang of Umphang district, Tak province, northern Thailand, during May to September 2018. A survey method using sweep net was employed along line transects between 10.00-12.00 a.m. and 13.00-15.00 p.m. A total of 337 butterflies representing 37 species, 26 genera in five families were encountered. The family Nymphalidae held the highest species richness (15 species), followed by Papilionidae (9 species) and Pieridae (6 speices), respectively. Herein, four uncommon species, namely Junonia iphita iphita, Tanaecia julii odilina, Penthema darlisa melema, and Papilio alcmenor alcmenor, were discovered in this time. The Shannon diversity (H’) for all samples obtained from the pooled data set of this observation valued 2.563 with relatively high values of Evenness (J’= 0.710) and Simpson index and (D = 0.829). For similarity index (Ss), the assemblage recorded of butterfly species between Mae Klang-Umphang shared about 0.629, implying that the environmental conditions in both surveyed zones were alike. Additionally, the accumulation curve in both locations of the butterfly was still increasing gradually when the collection ended, indicating that the lepidopteran species would be raised if we continue to survey more in next month. Nevertheless, to gain more butterfly taxa, observing different plant communities covering every season and using several survey techniques should be considered for further investigation.

Keywords: butterfly, biodiversity, tak province, thailand

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1888 Early Detection of Lymphedema in Post-Surgery Oncology Patients

Authors: Sneha Noble, Rahul Krishnan, Uma G., D. K. Vijaykumar

Abstract:

Breast-Cancer related Lymphedema is a major problem that affects many women. Lymphedema is the swelling that generally occurs in the arms or legs caused by the removal of or damage to lymph nodes as a part of cancer treatment. Treating it at the earliest possible stage is the best way to manage the condition and prevent it from leading to pain, recurrent infection, reduced mobility, and impaired function. So, this project aims to focus on the multi-modal approaches to identify the risks of Lymphedema in post-surgical oncology patients and prevent it at the earliest. The Kinect IR Sensor is utilized to capture the images of the body and after image processing techniques, the region of interest is obtained. Then, performing the voxelization method will provide volume measurements in pre-operative and post-operative periods in patients. The formation of a mathematical model will help in the comparison of values. Clinical pathological data of patients will be investigated to assess the factors responsible for the development of lymphedema and its risks.

Keywords: Kinect IR sensor, Lymphedema, voxelization, lymph nodes

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1887 Human Microbiome Hidden Association with Chronic and Autoimmune Diseases

Authors: Elmira Davasaz Tabrizi, Müşteba Sevil, Ercan Arican

Abstract:

In recent decades, there has been a sharp increase in the prevalence of several unrelated chronic diseases. The use of long-term antibiotics for chronic illnesses is increasing. The antibiotic resistance occurrence and its relationship with host microbiomes are still unclear. Properties of the identifying antibodies have been the focus of chronic disease research, such as prostatitis or autoimmune. The immune system is made up of a complicated but well-organized network of cell types that constantly monitor and maintain their surroundings. The regulated homeostatic interaction between immune system cells and their surrounding environment shapes the microbial flora. Researchers believe that the disappearance of special bacterial species from our ancestral microbiota might have altered the body flora that can cause a rise in disease during the human life span. This unpleasant pattern demonstrates the importance of focusing on discovering and revealing the root causes behind the disappearance or alteration of our microbiota. In this review, we gathered the results of some studies that reveal changes in the diversity and quantity of microorganisms that may affect chronic and autoimmune diseases. Additionally, a Ph.D. thesis that is still in process as Metagenomic studies in chronic prostatitis samples is mentioned.

Keywords: metagenomic, autoimmune, prostatitis, microbiome

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1886 John Cunningham Virus Interaction with Multiple Sclerosis Disease Progression

Authors: Sina Mahdavi

Abstract:

Background and Objective: Multiple sclerosis (MS) is the most common inflammatory autoimmune disease of the central nervous system (CNS) that affects the myelination process in the CNS. Complex interactions of various "environmental or infectious" factors may act as triggers in autoimmunity and disease progression. The association between viral infections, especially the John Cunningham virus (JCV) and MS is one potential cause that is not well understood. This study aims to summarize the available data on JCV infection in MS disease progression. Materials and Methods: For this study, the keywords "Multiple sclerosis", " John Cunningham virus ", and "central nervous system" in the databases PubMed, Google Scholar, Sid, and MagIran between 2019 and 2022 were searched, and 12 articles were chosen, studied, and analyzed. Results: MS patients are candidates for natalizumab therapy, which inhibits lymphocyte migration and increases the risk of progressive multifocal leukoencephalopathy (PML), a rare lytic infection of glial cells caused by JCV. Oligodendrocytes may be the target of JCV infection in the central nervous system (CNS). Conclusion: There is a high expression of JCV during the natalizumab treatment period for MS patients, suggesting that the virus may play a role in the development of MS by inducing an inflammatory state. Therefore, it is necessary to evaluate anti-JCV antibody serum as an important risk factor for the development of PML before deciding on the treatment course for these patients.

Keywords: multiple sclerosis, John Cunningham virus, central nervous system, autoimmunity

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1885 Fungal Profile and Antifungal Susceptibility Patterns among Symptomatic Pediatrics Patients Attending Aboozar Children’s Hospital, Ahvaz, Iran

Authors: Nasrin Amirrajab, Yasaman Razavi Ghahfarokhi, Zahra Tootak, Maryam Hadian, Fatemeh Abooali Shamshiri

Abstract:

Urinary tract infections (UTIs) have been reported in children with nephrotic syndrome. However, the only causes for the infection reported to date are bacteria, but not many prior reported occurrences of fungi or yeast as causative organisms. Hence, the present study aimed to describe the epidemiology of urinary tract fungal infections in a tertiary care pediatric. A single-center cross-sectional study was conducted at the nephrology ward of Aboozar Pediatric Hospital between March 21, 2021, and April 28, 2022. Urine was collected aseptically from children, inoculated onto culture media, and incubated at 37 °C for 18–48 hours. Yeast was identified following standard procedures. Antifungal susceptibility testing was determined by the disk diffusion method according to the CLSI guideline. Descriptive statistics and logistical regressions were used to estimate the crude ratio with a 95% confidence interval. P-value < 0.05 was considered significant. Among 68 individuals referred to the mycology lab, the result of direct examination and culture of all patients approved for C.albicans. Of these, 38 individuals (55.8%) were male, and 30 (44.2%) were female. The patients' age ranges were between one month and an 18-year-old. In the study of infection intensity, the patients were classified into three levels such as few (73.5%), moderate (20.6%), and many (5.9%). In the present study, all the patients were sensitive to Posaconazole. Also, the eagle effect was found in Amphotericin B, Voriconazole, and Fluconazole with frequencies of 91.7%, 91.7%, and 83%, respectively. In addition, just 8.3% of isolates were resistant to Itraconazole. It has not shown resistance in other mentioned medicine. The patients showed an intermediate response to Itraconazole (91.7%), Fluconazole (17%), Voriconazole (8.3%), and Amphotericin B (8.3%). There is a high prevalence of yeast infections in children with suspected UTIs. Also, boys are more likely to get yeast infections, and the severity of the infection is higher than girls. The present study demonstrated the importance of diagnosing and selecting the appropriate drug for urinary tract fungal infections in hospitalized children.

Keywords: urinary tract infections, children, fungal infections, yeast, antifungal susceptibility

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1884 Transcriptome and Metabolome Analysis of a Tomato Solanum Lycopersicum STAYGREEN1 Null Line Generated Using Clustered Regularly Interspaced Short Palindromic Repeats/Cas9 Technology

Authors: Jin Young Kim, Kwon Kyoo Kang

Abstract:

The SGR1 (STAYGREEN1) protein is a critical regulator of plant leaves in chlorophyll degradation and senescence. The functions and mechanisms of tomato SGR1 action are poorly understood and worthy of further investigation. To investigate the function of the SGR1 gene, we generated a SGR1-knockout (KO) null line via clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-mediated gene editing and conducted RNA sequencing and gas chromatography tandem mass spectrometry (GC-MS/MS) analysis to identify the differentially expressed genes. The SlSGR1 (Solanum lycopersicum SGR1) knockout null line clearly showed a turbid brown color with significantly higher chlorophyll and carotenoid content compared to wild-type (WT) fruit. Differential gene expression analysis revealed 728 differentially expressed genes (DEGs) between WT and sgr1 #1-6 line, including 263 and 465 downregulated and upregulated genes, respectively, for which fold change was >2, and the adjusted p-value was <0.05. Most of the DEGs were related to photosynthesis and chloroplast function. In addition, the pigment, carotenoid changes in sgr1 #1-6 line was accumulated of key primary metabolites such as sucrose and its derivatives (fructose, galactinol, raffinose), glycolytic intermediates (glucose, G6P, Fru6P) and tricarboxylic acid cycle (TCA) intermediates (malate and fumarate). Taken together, the transcriptome and metabolite profiles of SGR1-KO lines presented here provide evidence for the mechanisms underlying the effects of SGR1 and molecular pathways involved in chlorophyll degradation and carotenoid biosynthesis.

Keywords: tomato, CRISPR/Cas9, null line, RNA-sequencing, metabolite profiling

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1883 Fruiting Body Specific Sc4 Hydrophobin Gene Plays a Role in Schizophyllum Commune Hyphal Attachment to Structured Glass Surfaces

Authors: Evans Iyamu

Abstract:

Genes encoding hydrophobins play distinct roles at different stages of the life cycle of fungi, and they foster hyphal attachment to surfaces. The hydrophobin Sc4 is known to provide a hydrophobic membrane lining of the gas channels within Schizophyllum commune fruiting bodies. Here, we cultivated non-fruiting, monokaryotic S. commune 12-43 on glass surfaces that could be verified by micrography. Differential gene expression profiling of nine hydrophobin genes and the hydrophobin-like sc15 gene by quantitative PCR showed significant up-regulation of sc4 when S. commune was attached to glass surfaces, also confirmed with RNA-Seq data analysis. Another silicate, namely quartz sand, was investigated, and induction of sc4 was seen as well. The up-regulation of the hydrophobin gene sc4 may indicate involvement in S. commune hyphal attachment to glass as well as quartz surfaces. We propose that the covering of hyphae by Sc4 allows for direct interaction with the hydrophobic surfaces of silicates and that differential functions of specific hydrophobin genes depend on the surface interface involved. This study could help with the clarification of the biological functions of hydrophobins in natural surroundings, including hydrophobic surface attachment. Therefore, the analysis of growth on glass serves as a basis for understanding S. commune interaction with glass surfaces while providing the possibility to visualize the interaction microscopically.

Keywords: hydrophobin, structured glass surfaces, differential gene expression, quartz sand

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1882 In-Vitro Stability of Aspergillus terreus Phytases in Relation to Different Physico-Chemical Factors

Authors: Qaiser Akram, Ahsan Naeem, Hafiz Muhammad Rizwan, Waqas Ahmad, Rubeena Yasmeen

Abstract:

Aspergillus has good secretory potential for phytases. Morphologically and microscopically identified Aspergillus terreus (A. terreus) (n=20) were screened for phytase production and non-toxicity. Phytases produced by non-toxigenic A. terreus under optimum conditions were quantified. Phytases of highest producer A. terreus were evaluated for stability after exposure to temperature (35, 55, 75 and 95ºC) and pH (2, 4, 6 and 8). Effect of metal ions (Fe⁺³, Ba⁺², Ca⁺², Cu⁺², Mg⁺², Mn⁺², K⁺¹ and Na⁺¹) was assessed on phytase activity. Log reduction in phytase activity was calculated. The highest activity units of phytase produced by A. terreus were 271.49 ± 8.14 phytase unit / mL (FTU/ mL). The lowest reduction in phytase activity was 50.20 ± 7.36 (18.5%) and 68.22 ± 10.3 FTU/mL (25.13%) at 35ºC and pH 6, respectively for 15 minutes. The highest reduction 259 ± 0.84 (95.5%) and 211.99 ± 4.39 FTU/mL (78.1%) was recorded at 95ºC for 60 minutes and pH 2.0 for 45 minutes exposure, respectively. All metal ions negatively affected phytase activity. Phytase activity was inhibited minimum (45.32 ± 28.54 FTU/mL, 16.69%) by K⁺¹(1 mM) and maximum (231.48 ± 3.68 FTU/mL, 80.8%) by Cu⁺² (10 mM). It was concluded that A. terreus phytase stability and activity was dependent on physio-chemical factors.

Keywords: stability, phytase, aspergillus terreus, physio-chemical factors and metal ions

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1881 Coronin 1C and miR-128A as Potential Diagnostic Biomarkers for Glioblastoma Multiform

Authors: Denis Mustafov, Emmanouil Karteris, Maria Braoudaki

Abstract:

Glioblastoma multiform (GBM) is a heterogenous primary brain tumour that kills most affected patients. To the authors best knowledge, despite all research efforts there is no early diagnostic biomarker for GBM. MicroRNAs (miRNAs) are short non-coding RNA molecules which are deregulated in many cancers. The aim of this research was to determine miRNAs with a diagnostic impact and to potentially identify promising therapeutic targets for glioblastoma multiform. In silico analysis was performed to identify deregulated miRNAs with diagnostic relevance for glioblastoma. The expression profiles of the chosen miRNAs were then validated in vitro in the human glioblastoma cell lines A172 and U-87MG. Briefly, RNA extraction was carried out using the Trizol method, whilst miRNA extraction was performed using the mirVANA miRNA isolation kit. Quantitative Real-Time Polymerase Chain Reaction was performed to verify their expression. The presence of five target proteins within the A172 cell line was evaluated by Western blotting. The expression of the CORO1C protein within 32 GBM cases was examined via immunohistochemistry. The miRNAs identified in silico included miR-21-5p, miR-34a and miR-128a. These miRNAs were shown to target deregulated GBM genes, such as CDK6, E2F3, BMI1, JAG1, and CORO1C. miR-34a and miR-128a showed low expression profiles in comparison to a control miR-RNU-44 in both GBM cell lines suggesting tumour suppressor properties. Opposing, miR-21-5p demonstrated greater expression indicating that it could potentially function as an oncomiR. Western blotting revealed expression of all five proteins within the A172 cell line. In silico analysis also suggested that CORO1C is a target of miR-128a and miR-34a. Immunohistochemistry demonstrated that 75% of the GBM cases showed moderate to high expression of CORO1C protein. Greater understanding of the deregulated expression of miR-128a and the upregulation of CORO1C in GBM could potentially lead to the identification of a promising diagnostic biomarker signature for glioblastomas.

Keywords: non-coding RNAs, gene expression, brain tumours, immunohistochemistry

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1880 The Role of Sexual Satisfaction Sexual Satisfaction in Marital Satisfaction Married Men

Authors: Maghsoud Nader Pilehroud, Mohmmad Alizadeh, Soheila Golipour, Sedigeh Tajabadipour

Abstract:

Aim: in terms of importance, sexual issues are of the highest priority in married life issues and sexual compatibility is of the most important reasons of success in married life and consequently marital satisfaction.the present research was conducted with the aim of The role of sexual satisfaction sexual satisfaction in marital satisfaction married men. Study Design: this research is descriptive and is of correlation type.Method: The statistical population includes all the married men of Ardebil city out of which, 60 men were chosen using random sampling as the research samples. The research instruments were ENRICH couple scale and Hudson sexual satisfaction scale. The findings were analyzed using descriptive statistics method (mean and standard deviation) and inferential statistics (Pearson's correlation and regression) and SPSS-16 software. Results: the results showed that sexual satisfaction has a positive and significant relationship with marital satisfaction and all of its components, and that sexual satisfaction can predict marital satisfaction. The results also showed that sexual and marital satisfaction, are not significantly related to any of the variables of education level, duration of marriage and number of children. conclusion: according to the results, it can be claimed that sexual skills training for couples can be influential at increasing their martial satisfaction, and that also, sexual satisfaction has an important role in marital satisfaction.

Keywords: sexual satisfaction, marital satisfaction, married men, Iran

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1879 Altered Gene Expression: Induction/Suppression of some Pathogenesis Related Protein Genes in an Egyptian Isolate of Potato Leafroll Virus (PLRV)

Authors: Dalia G. Aseel

Abstract:

The potato (Solanum tubersum, L.) has become one of the major vegetable crops in Egypt and all over the world. Potato leafroll virus(PLRV) was observed on potato plants collected from different governorates in Egypt. Three cultivars, Spunta, Diamont, and Cara, infected with PLRV were collected; RNA was extracted and subjected to Real-Time PCR using the coat protein gene primers. The results showed that the expression of the coat protein was 39.6-fold, 12.45-fold, and 47.43-fold, respectively, for Spunta, Diamont, and Cara cultivars. Differential Display Polymerase Chain Reaction (DD-PCR) using pathogenesis-related protein 1 (PR-1), β-1,3-glucanases (PR-2), chitinase (PR-3), peroxidase (POD), and polyphenol oxidase (PPO) forward primers for pathogenesis-related proteins (PR). The obtained data revealed different banding patterns depending on the viral type and the region of infection. Regarding PLRV, 58 up-regulated and 19 down-regulated genes were detected. Sequence analysis of the up-and down-regulated genes revealed that infected plants were observed in comparison with the healthy control. Sequence analysis of the up-regulated gene was performed, and the encoding sequence analysis showed that the obtained genes include: induced stolen tip protein. On the other hand, two down-regulated genes were identified: disease resistance RPP-like protein and non-specific lipid-transfer protein. In this study, the expressions of PR-1, PR-2, PR-3, POD, and PPO genes in the infected leaves of three potato cultivars were estimated by quantitative real-time PCR. We can conclude that the PLRV-infection of potato plants inhibited the expression of the five PR genes. On the contrary, infected leaves by PLRV elevated the expression of some defense genes. This interaction may also induce and/or suppress the expression of some genes responsible for the plant's defense mechanisms.

Keywords: PLRV, pathogenesis-related proteins (PRs), DD-PCR, sequence, real-time PCR

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1878 Chikungunya Virus Detection Utilizing an Origami Based Electrochemical Paper Analytical Device

Authors: Pradakshina Sharma, Jagriti Narang

Abstract:

Due to the critical significance in the early identification of infectious diseases, electrochemical sensors have garnered considerable interest. Here, we develop a detection platform for the chikungunya virus by rationally implementing the extremely high charge-transfer efficiency of a ternary nanocomposite of graphene oxide, silver, and gold (G/Ag/Au) (CHIKV). Because paper is an inexpensive substrate and can be produced in large quantities, the use of electrochemical paper analytical device (EPAD) origami further enhances the sensor's appealing qualities. A cost-effective platform for point-of-care diagnostics is provided by paper-based testing. These types of sensors are referred to as eco-designed analytical tools due to their efficient production, usage of the eco-friendly substrate, and potential to reduce waste management after measuring by incinerating the sensor. In this research, the paper's foldability property has been used to develop and create 3D multifaceted biosensors that can specifically detect the CHIKVX-ray diffraction, scanning electron microscopy, UV-vis spectroscopy, and transmission electron microscopy (TEM) were used to characterize the produced nanoparticles. In this work, aptamers are used since they are thought to be a unique and sensitive tool for use in rapid diagnostic methods. Cyclic voltammetry (CV) and linear sweep voltammetry (LSV), which were both validated with a potentiostat, were used to measure the analytical response of the biosensor. The target CHIKV antigen was hybridized with using the aptamer-modified electrode as a signal modulation platform, and its presence was determined by a decline in the current produced by its interaction with an anionic mediator, Methylene Blue (MB). Additionally, a detection limit of 1ng/ml and a broad linear range of 1ng/ml-10µg/ml for the CHIKV antigen were reported.

Keywords: biosensors, ePAD, arboviral infections, point of care

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1877 Migration in Times of Uncertainty

Authors: Harman Jaggi, David Steinsaltz, Shripad Tuljapurkar

Abstract:

Understanding the effect of fluctuations on populations is crucial in the context of increasing habitat fragmentation, climate change, and biological invasions, among others. Migration in response to environmental disturbances enables populations to escape unfavorable conditions, benefit from new environments and thereby ride out fluctuations in variable environments. Would populations disperse if there is no uncertainty? Karlin showed in 1982 that when sub-populations experience distinct but fixed growth rates at different sites, greater mixing of populations will lower the overall growth rate relative to the most favorable site. Here we ask if and when environmental variability favors migration over no-migration. Specifically, in random environments, would a small amount of migration increase the overall long-run growth rate relative to the zero migration case? We use analysis and simulations to show how long-run growth rate changes with migration rate. Our results show that when fitness (dis)advantages fluctuate over time across sites, migration may allow populations to benefit from variability. When there is one best site with highest growth rate, the effect of migration on long-run growth rate depends on the difference in expected growth between sites, scaled by the variance of the difference. When variance is large, there is a substantial probability of an inferior site experiencing higher growth rate than its average. Thus, a high variance can compensate for a difference in average growth rates between sites. Positive correlations in growth rates across sites favor less migration. With multiple sites and large fluctuations, the length of shortest cycle (excursion) from the best site (on average) matters, and we explore the interplay between excursion length, average differences between sites and the size of fluctuations. Our findings have implications for conservation biology: even when there are superior sites in a sea of poor habitats, variability and habitat quality across space may be key to determining the importance of migration.

Keywords: migration, variable-environments, random, dispersal, fluctuations, habitat-quality

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1876 Classification of Potential Biomarkers in Breast Cancer Using Artificial Intelligence Algorithms and Anthropometric Datasets

Authors: Aref Aasi, Sahar Ebrahimi Bajgani, Erfan Aasi

Abstract:

Breast cancer (BC) continues to be the most frequent cancer in females and causes the highest number of cancer-related deaths in women worldwide. Inspired by recent advances in studying the relationship between different patient attributes and features and the disease, in this paper, we have tried to investigate the different classification methods for better diagnosis of BC in the early stages. In this regard, datasets from the University Hospital Centre of Coimbra were chosen, and different machine learning (ML)-based and neural network (NN) classifiers have been studied. For this purpose, we have selected favorable features among the nine provided attributes from the clinical dataset by using a random forest algorithm. This dataset consists of both healthy controls and BC patients, and it was noted that glucose, BMI, resistin, and age have the most importance, respectively. Moreover, we have analyzed these features with various ML-based classifier methods, including Decision Tree (DT), K-Nearest Neighbors (KNN), eXtreme Gradient Boosting (XGBoost), Logistic Regression (LR), Naive Bayes (NB), and Support Vector Machine (SVM) along with NN-based Multi-Layer Perceptron (MLP) classifier. The results revealed that among different techniques, the SVM and MLP classifiers have the most accuracy, with amounts of 96% and 92%, respectively. These results divulged that the adopted procedure could be used effectively for the classification of cancer cells, and also it encourages further experimental investigations with more collected data for other types of cancers.

Keywords: breast cancer, diagnosis, machine learning, biomarker classification, neural network

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1875 Relation between Biochemical Parameters and Bone Density in Postmenopausal Women with Osteoporosis

Authors: Shokouh Momeni, Mohammad Reza Salamat, Ali Asghar Rastegari

Abstract:

Background: Osteoporosis is the most prevalent metabolic bone disease in postmenopausal women associated with reduced bone mass and increased bone fracture. Measuring bone density in the lumbar spine and hip is a reliable measure of bone mass and can therefore specify the risk of fracture. Dual-energy X-ray absorptiometry(DXA) is an accurate non-invasive system measuring the bone density, with low margin of error and no complications. The present study aimed to investigate the relationship between biochemical parameters with bone density in postmenopausal women. Materials and methods: This cross-sectional study was conducted on 87 postmenopausal women referred to osteoporosis centers in Isfahan. Bone density was measured in the spine and hip area using DXA system. Serum levels of calcium, phosphorus, alkaline phosphatase and magnesium were measured by autoanalyzer and serum levels of vitamin D were measured by high-performance liquid chromatography(HPLC). Results: The mean parameters of calcium, phosphorus, alkaline phosphatase, vitamin D and magnesium did not show a significant difference between the two groups(P-value>0.05). In the control group, the relationship between alkaline phosphatase and BMC and BA in the spine was significant with a correlation coefficient of -0.402 and 0.258, respectively(P-value<0.05) and BMD and T-score in the femoral neck area showed a direct and significant relationship with phosphorus(Correlation=0.368; P-value=0.038). There was a significant relationship between the Z-score with calcium(Correlation=0.358; P-value=0.044). Conclusion: There was no significant relationship between the values ​​of calcium, phosphorus, alkaline phosphatase, vitamin D and magnesium parameters and bone density (spine and hip) in postmenopaus

Keywords: osteoporosis, menopause, bone mineral density, vitamin d, calcium, magnesium, alkaline phosphatase, phosphorus

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1874 Whole Coding Genome Inter-Clade Comparisons to Predict Global Cancer-Protecting Variants

Authors: Lamis Naddaf, Yuval Tabach

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We identified missense genetic variants with the potential to enhance resistance against cancer. Such a field has not been widely explored as researchers tend to investigate the mutations that cause diseases, in response to the suffering of patients, rather than those mutations that protect from them. In conjunction with the genomic revolution and the advances in genetic engineering and synthetic biology, identifying the protective variants will increase the power of genotype-phenotype predictions and have significant implications for improved risk estimation, diagnostics, prognosis, and even personalized therapy and drug discovery. To approach our goal, we systematically investigated the sites of the coding genomes and selected the alleles that showed a correlation with the species’ cancer resistance. Interestingly, we found several amino acids that are more generally preferred (like the Proline) or avoided (like the Cysteine) by the resistant species. Furthermore, Cancer resistance in mammals and reptiles is significantly predicted by the number of the predicted protecting variants (PVs) a species has. Moreover, PVs-enriched-genes are enriched in pathways relevant to tumor suppression. For example, they are enriched in the Hedgehog signaling and silencing pathways, which its improper activation is associated with the most common form of cancer malignancy. We also showed that the PVs are mostly more abundant in healthy people compared to cancer patients within different human races.

Keywords: cancer resistance, protecting variant, naked mole rat, comparative genomics

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1873 The Evolution of the Strategic Plasma Industry

Authors: Zahra Ghasemi, Fatemeh Babaei

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Plasma-derived medicinal products are vital categories of biological therapies. These products are used to treat rare, chronic, severe, and life-threatening conditions, such as bleeding disorders (Hemophilia A and B), hemolytic disease of the fetus and newborn, severe infections, burns and liver diseases, and other diseases caused by the absence or malfunction of certain proteins. In addition, they improve the patient’s quality of life. The process of producing plasma-derived medicinal products begins with the collection of human plasma from healthy donors. This initial stage is complex and is monitored with high precision and sensitivity by global authorities to maintain the quality and safety of the final products as well as the health of the donors. The amount of manufactured plasma-derived medicinal products depends on the availability of its raw material, human plasma, so collecting enough plasma for fractionation is essential. Therefore, adopting a suitable national policy regarding plasma donation, establishing collection centers, and increasing public awareness of the importance of plasma donation will improve any country’s conditions regarding the timely and sufficient supply of these medicines. In this study, we tried to briefly examine the importance of sustainability of the plasma industry and its situation in our beloved country of Iran.

Keywords: plasma, source plasma, plasma-derived medicinal products, fractionation

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1872 Last ca 2500 Yr History of the Harmful Algal Blooms in South China Reconstructed on Organic-Walled Dinoflagellate Cysts

Authors: Anastasia Poliakova

Abstract:

Harmful algal bloom (HAB) is a known negative phenomenon that is caused both by natural factors and anthropogenic influence. HABs can result in a series of deleterious effects, such as beach fouling, paralytic shellfish poisoning, mass mortality of marine species, and a threat to human health, especially if toxins pollute drinking water or occur nearby public resorts. In South China, the problem of HABs has an ultimately important meaning. For this study, we used a 1.5 m sediment core LAX-2018-2 collected in 2018 from the Zhanjiang Mangrove National Nature Reserve (109°03´E, 20°30´N), Guangdong Province, South China. High-resolution coastal environment reconstruction with a specific focus on the HABs history during the last ca 2500 yrs was attempted. Age control was performed with five radiocarbon dates obtained from benthic foraminifera. A total number of 71 dinoflagellate cyst types was recorded. The most common types found consistently throughout the sediment sequence were autotrophic Spiniferites spp., Spiniferites hyperacanthus and S. mirabilis, S. ramosus, Operculodinium centrocarpum sensu Wall and Dale 1966, Polysphaeridium zoharyi, and heterotrophic Brigantedinium ssp., cyst of Gymnodinium catenatum and cysts mixture of Protoperidinium. Three local dinoflagellate zones LAX-1 to LAX-3 were established based on the results of the constrained cluster analysis and data ordination; additionally, the middle zone LAX-2 was derived into two subzones, LAX-2a and LAX-2b based on the dynamics of toxic and heterotrophic cysts as well as on the significant changes (probability, P=0.89) in percentages of eutrophic indicators. The total cyst count varied from 106 to 410 dinocysts per slide, with 177 cyst types on average. Dinocyst assemblages are characterized by high values of the dost-depositional degradation index (kt) that varies between 3.6 and 7.6 (averaging 5.4), which is relatively high and is very typical for the areas with selective dinoflagellate cyst preservation that is related to bottom-water oxygen concentrations.

Keywords: reconstruction of palaeoenvironment, harmful algal blooms, anthropogenic influence on coastal zones, South China Sea

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1871 Evaluation of Total Antioxidant Activity (TAC) of Copper Oxide Decorated Reduced Graphene Oxide (CuO-rGO) at Different Stirring time

Authors: Aicha Bensouici, Assia Mili, Naouel Rdjem, Nacera Baali

Abstract:

Copper oxide decorated reduced graphene oxide (GO) was obtained successfully using two steps route synthesis was used. Firstly, graphene oxide was obtained using a modified Hummers method by excluding sodium nitrate from starting materials. After washing-centrifugation routine pristine GO was decorated by copper oxide using a refluxation technique at 120°C during 2h, and an equal amount of GO and copper acetate was used. Three CuO-rGO nanocomposite samples types were obtained at 30min, 24h, and 7 day stirring time. TAC results show dose dependent behavior of CuO-rGO and confirm no influence of stirring time on antioxidant properties, 30min is considered as an optimal stirring condition.

Keywords: copper oxide, reduced graphene oxide, TAC, GO

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1870 Predicting Open Chromatin Regions in Cell-Free DNA Whole Genome Sequencing Data by Correlation Clustering  

Authors: Fahimeh Palizban, Farshad Noravesh, Amir Hossein Saeidian, Mahya Mehrmohamadi

Abstract:

In the recent decade, the emergence of liquid biopsy has significantly improved cancer monitoring and detection. Dying cells, including those originating from tumors, shed their DNA into the blood and contribute to a pool of circulating fragments called cell-free DNA. Accordingly, identifying the tissue origin of these DNA fragments from the plasma can result in more accurate and fast disease diagnosis and precise treatment protocols. Open chromatin regions are important epigenetic features of DNA that reflect cell types of origin. Profiling these features by DNase-seq, ATAC-seq, and histone ChIP-seq provides insights into tissue-specific and disease-specific regulatory mechanisms. There have been several studies in the area of cancer liquid biopsy that integrate distinct genomic and epigenomic features for early cancer detection along with tissue of origin detection. However, multimodal analysis requires several types of experiments to cover the genomic and epigenomic aspects of a single sample, which will lead to a huge amount of cost and time. To overcome these limitations, the idea of predicting OCRs from WGS is of particular importance. In this regard, we proposed a computational approach to target the prediction of open chromatin regions as an important epigenetic feature from cell-free DNA whole genome sequence data. To fulfill this objective, local sequencing depth will be fed to our proposed algorithm and the prediction of the most probable open chromatin regions from whole genome sequencing data can be carried out. Our method integrates the signal processing method with sequencing depth data and includes count normalization, Discrete Fourie Transform conversion, graph construction, graph cut optimization by linear programming, and clustering. To validate the proposed method, we compared the output of the clustering (open chromatin region+, open chromatin region-) with previously validated open chromatin regions related to human blood samples of the ATAC-DB database. The percentage of overlap between predicted open chromatin regions and the experimentally validated regions obtained by ATAC-seq in ATAC-DB is greater than 67%, which indicates meaningful prediction. As it is evident, OCRs are mostly located in the transcription start sites (TSS) of the genes. In this regard, we compared the concordance between the predicted OCRs and the human genes TSS regions obtained from refTSS and it showed proper accordance around 52.04% and ~78% with all and the housekeeping genes, respectively. Accurately detecting open chromatin regions from plasma cell-free DNA-seq data is a very challenging computational problem due to the existence of several confounding factors, such as technical and biological variations. Although this approach is in its infancy, there has already been an attempt to apply it, which leads to a tool named OCRDetector with some restrictions like the need for highly depth cfDNA WGS data, prior information about OCRs distribution, and considering multiple features. However, we implemented a graph signal clustering based on a single depth feature in an unsupervised learning manner that resulted in faster performance and decent accuracy. Overall, we tried to investigate the epigenomic pattern of a cell-free DNA sample from a new computational perspective that can be used along with other tools to investigate genetic and epigenetic aspects of a single whole genome sequencing data for efficient liquid biopsy-related analysis.

Keywords: open chromatin regions, cancer, cell-free DNA, epigenomics, graph signal processing, correlation clustering

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1869 Effect of Removing Hub Domain on Human CaMKII Isoforms Sensitivity to Calcium/Calmodulin

Authors: Ravid Inbar

Abstract:

CaMKII (calcium-calmodulin dependent protein kinase II) makes up 2% of the protein in our brain and has a critical role in memory formation and long-term potentiation of neurons. Despite this, research has yet to uncover the role of one of the domains on the activation of this kinase. The following proposes to express the protein without the hub domain in E. coli, leaving only the kinase and regulatory segment of the protein. Next, a series of kinase assays will be conducted to elucidate the role the hub domain plays on CaMKII sensitivity to calcium/calmodulin activation. The hub domain may be important for activation; however, it may also be a variety of domains working together to influence protein activation and not the hub alone. Characterization of a protein is critical to the future understanding of the protein's function, as well as for producing pharmacological targets in cases of patients with diseases.

Keywords: CaMKII, hub domain, kinase assays, kinase + reg seg

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1868 Soil Enzyme Activity as Influenced by Post-emergence Herbicides Applied in Soybean [Glycine max (L.) Merrill]

Authors: Uditi Dhakad, Baldev Ram, Chaman K. Jadon, R. K. Yadav, D. L. Yadav, Pratap Singh, Shalini Meena

Abstract:

A field experiment was conducted during Kharif 2021 at Agricultural Research Station, Kota, to evaluate the effect of different post-emergence herbicides applied to soybean [Glycine max (L.) Merrill] on soil enzymes activity viz. dehydrogenase, phosphatase, and urease. The soil of the experimental site was clay loam (vertisols) in texture and slightly alkaline in reaction with 7.7 pH. The soil was low in organic carbon (0.49%), medium in available nitrogen (210 kg/ha), phosphorus (23.5 P2O5 kg/ha), and high in potassium (400 K2O kg/ha) status. The results elucidated that no significant adverse effect on soil dehydrogenase, urease, and phosphatase activity was determined with the application of post-emergence herbicides over the untreated control. Two hands weeding at 20 and 40 DAS registered maximum dehydrogenase enzyme activity (0.329 μgTPF/g soil/d) closely followed by herbicides mixtures and sole herbicide while pre-emergence application of pendimethalin + imazethapyr 960 g a.i./ha and pendimethalin 1.0 kg a.i./ha significantly reduced dehydrogenase enzyme activity compared to control. Urease enzyme activity was not much affected under different weed control treatments and weedy checks. The treatments were found statistically non-significant, and values ranged between 1.16-1.25 μgNH4N/g soil/d. Phosphatase enzyme activity was also not influenced significantly due to various weed control treatments. Though maximum phosphatase enzyme activity (30.17 μgpnp/g soil/hr) was observed under two-hand weeding, followed by fomesafen + fluazifop-p-butyl 220 g a.i./ha. Herbicidal weed control measures did not influence the total bacteria, fungi, and actinomycetes population.

Keywords: dehydrogenase, phosphatase, post-emergence, soil enzymes, urease.

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1867 Bioefficacy of Diclosulam for Controlling Weeds in Soybean [Glycine Max (L.) Merrill] and Its Carry Over Effect on Succeeding Wheat (Triticum Aestivum) Crop

Authors: Pratap Sing, Chaman. K. Jadon, H. P. Meena, D. L.yadav, S. L. Yadav, Uditi Dhakad

Abstract:

The experiment was conducted at Agricultural Research Station, Agriculture University, Kota, Rajasthan, India during kharif and rabi 2020-21 and 2021-22 to study the biofficacy of diclosulam and its residual effect on succeeding wheat crop. The treatments comprised of Diclosulam 84 % WDG viz. 6.25, 12.50, 25.00 and 37.50 g/ha as pre emergence (PE), Pendimethalin 30% EC 3.33 l/ha, Sulfentrazon 48% SC 750 g/ha, hand weeding at 30 and 45 DAS and weedy check, were evaluated in randomized block design in three replications. The experimental soil was clay in texture and non-calcareous. Experimental field was mainly dominated by grasses-Echinochloa colonum, E.crusgalli,Cynodon dactylon, Sedges-Cyperus rotundus and broad leaved weeds Celosia argentea and Digera arvensis.The result revealed that application of Diclosulam 84 % WDG 25 g/ha PE was found effective in controlling mostly weed species and registered higher weed control efficiency 81.2, 74.3, 69.6 per cent at 30, 45 days after sowing and at harvest. Diclosulam 84 % WDG (6.25-25.0 g/ha) was found selective to the soybean crop as no any phytotoxicity symptoms were observed. Among the herbicidal treatments, Diclosulam 84 % WDG 25 g/ha registered maximum and significantly higher soybean seed yield (1889 and 1431 kg/ha during kharif 2020 and 2021, respectively and was at par with Sulfentrazone 48% SC 750 g/ha and over weedy check( 1027 and 667 kg/ha).The wheat crop growth, yield attributes and seed yield were not influenced due to carry over effect of the Diclosulam 84 % WDG( 6.25-25.0 g/ha) and no any phytotoxicity symptoms were observed. Henceforth, the Diclosulam 84 % WDG 25.0 g/ha as pre emergence may be used in the soybean for effective weed control without carry over effect on succeeding wheat crop.

Keywords: Diclosulam, soybean, carry over effect, succeeding wheat

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1866 Functional Characterization of Transcriptional Regulator WhiB Proteins of Mycobacterium Tuberculosis

Authors: Sonam Kumari

Abstract:

Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis, possesses a remarkable feature of entering into and emerging from a persistent state. The mechanism by which Mtb switches from the dormant state to the replicative form is still poorly characterized. Proteome studies have given us an insight into the role of certain proteins in giving stupendous virulence to Mtb, but numerous dotsremain unconnected and unaccounted. The WhiB family of proteins is one such protein that is associated with developmental processes in actinomycetes.Mtb has seven such proteins (WhiB1 to WhiB7).WhiB proteins are transcriptional regulators; their conserved C-terminal HTH motif is involved in DNA binding. They regulate various essential genes of Mtbby binding to their promoter DNA. Biophysical Analysis of the effect of DNA binding on WhiB proteins has not yet been appropriately characterized. Interaction with DNA induces conformational changes in the WhiB proteins, confirmed by steady-state fluorescence and circular dichroism spectroscopy. ITC has deduced thermodynamic parameters and the binding affinity of the interaction. Since these transcription factors are highly unstable in vitro, their stability and solubility were enhanced by the co-expression of molecular chaperones. The present study findings help determine the conditions under which the WhiB proteins interact with their interacting partner and the factors that influence their binding affinity. This is crucial in understanding their role in regulating gene expression in Mtbandin targeting WhiB proteins as a drug target to cure TB.

Keywords: tuberculosis, WhiB proteins, mycobacterium tuberculosis, nucleic acid binding

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1865 Developing an Automated Protocol for the Wristband Extraction Process Using Opentrons

Authors: Tei Kim, Brooklynn McNeil, Kathryn Dunn, Douglas I. Walker

Abstract:

To better characterize the relationship between complex chemical exposures and disease, our laboratory uses an approach that combines low-cost, polydimethylsiloxane (silicone) wristband samplers that absorb many of the chemicals we are exposed to with untargeted high-resolution mass spectrometry (HRMS) to characterize 1000’s of chemicals at a time. In studies with human populations, these wristbands can provide an important measure of our environment: however, there is a need to use this approach in large cohorts to study exposures associated with the disease. To facilitate the use of silicone samplers in large scale population studies, the goal of this research project was to establish automated sample preparation methods that improve throughput, robustness, and scalability of analytical methods for silicone wristbands. Using the Opentron OT2 automated liquid platform, which provides a low-cost and opensource framework for automated pipetting, we created two separate workflows that translate the manual wristband preparation method to a fully automated protocol that requires minor intervention by the operator. These protocols include a sequence generation step, which defines the location of all plates and labware according to user-specified settings, and a transfer protocol that includes all necessary instrument parameters and instructions for automated solvent extraction of wristband samplers. These protocols were written in Python and uploaded to GitHub for use by others in the research community. Results from this project show it is possible to establish automated and open source methods for the preparation of silicone wristband samplers to support profiling of many environmental exposures. Ongoing studies include deployment in longitudinal cohort studies to investigate the relationship between personal chemical exposure and disease.

Keywords: bioinformatics, automation, opentrons, research

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1864 Haemoglobin Variants and Their Frequency Distribution in Human Population of Niger State, Nigeria

Authors: Akeem Akinboro, Bala Alhaj Kegun

Abstract:

Haemoglobinopathy is a genetic disorder that has the potentiality to cause death of individuals in whom both the alpha (α) and beta (β) globin chains of the haemoglobin molecule are defective due to mutations in their genes. The haemoglobin genotype variants among some residents of Niger state, Nigeria, were determined using the secondary data available at Bida, Minna and Kotangora general hospitals of the state. A total of 1,639 data, representing 434, 655 and 550, collected from the outside patients who visited the medical laboratory units of the three general hospitals, respectively, over five years period (2015-2020) were analyzed into gene frequency, sex and age to determine their haemoglobin genotypes status. More males (51.6 – 58.7%) than females (41.3 – 48.4%) visited the three hospitals during the period covered and most of the patients were between 11 - 20 years old. The frequency of HbA allele in the human population was 0.72, 0.65, 0.68 for Bida, Minna and Kotangora, respectively, while it was 0.25, 0.29 and 0.28 for HbS allele. The HbC allele was prevalent at 0.03, 0.06 and 0.05 among the human population in Bida, Minna and Kotangora cities of Niger state. In overall, the prevalence of HbA, HbS and HbC alleles in Niger state of Nigeria was 0.68, 0.28 and 0.05. Minna being the capital city of Niger state and the most populous among the three cities in the state seems to have influx of more people who are carriers of abnormal haemoglobin genotypes which has resulted to higher frequency of HbS and HbC than those of the other two cities in this study. These results show that the pattern of haemoglobin genotypes frequency of Kontagora could be a prediction for the whole of Niger state. It is therefore necessary and important to take screening of blood for haemoglobin genotype serious among intending couples to prevent and reduce the possibility of having increase in the number of people with abnormal haemoglobin genotypes in the state.

Keywords: haemoglobin, genotype, niger state, gene frequency, general hospitals

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1863 Genome Characterization and Phylogeny Analysis of Viruses Infected Invertebrates, Parvoviridae Family

Authors: Niloofar Fariborzi, Hamzeh Alipour, Kourosh Azizi, Neda Eskandarzade, Abozar Ghorbani

Abstract:

The family Parvoviridae consists of a large diversity of single-stranded DNA viruses, which cause mild to severe diseases in both vertebrates and invertebrates. The Parvoviridae are classified into three subfamilies: Parvovirinae infect vertebrates, Densovirinae infects invertebrates, while Hamaparovirinae infects both vertebrates and invertebrates. Except for the NS1 region, which is the prime criterion for phylogeny analysis, other parts of the parvoviruses genome, such as UTRs, are diverse even among closely related viruses or within the same genus. It is believed that host switching in parvoviruses may be related to genetic changes in regions other than NS1; therefore, whole-genome screening is valuable for studying parvoviruses' host-virus interactions. The aim of this study was to analyze genome organization and phylogeny of the complete genome sequence of the 132 Paroviridae family members, focusing on viruses that infect invertebrates. The maximum and minimum divergence within each subfamily belonged to Densovirinae and Parvovirinae, respectively. The greatest evolutionary divergence was between Hamaparovirinae and Parvovirinae. Unclassified viruses were mostly from Parovirinae and had the highest divergence to densoviruses and the lowest divergence to Parovirinae viruses. In a phylogenetic tree, all hamparoviruses were found in the center of densoviruses, with the exception of Syngnathid Ichthamaparvovirus 1 (NC_055527), which was positioned between two Parvovirinae members (NC _022089 and NC_038544). The proximity of hamparoviruses members to some densoviruses strengthens the possibility that densoviruses may be the ancestors of hamaparoviruses or vice versa. Therefore, examination and phylogeny analysis of the whole genome is necessary to understand Parvoviridae family host selection.

Keywords: densoviruses, parvoviridae, bioinformatics, phylogeny

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1862 New Recombinant Netrin-a Protein of Lucilia Sericata Larvae by Bac to Bac Expression Vector System in Sf9 Insect Cell

Authors: Hamzeh Alipour, Masoumeh Bagheri, Abbasali Raz, Javad Dadgar Pakdel, Kourosh Azizi, Aboozar Soltani, Mohammad Djaefar Moemenbellah-Fard

Abstract:

Background: Maggot debridement therapy is an appropriate, effective, and controlled method using sterilized larvae of Luciliasericata (L.sericata) to treat wounds. Netrin-A is an enzyme in the Laminins family which secreted from salivary gland of L.sericata with a central role in neural regeneration and angiogenesis. This study aimed to production of new recombinant Netrin-A protein of Luciliasericata larvae by baculovirus expression vector system (BEVS) in SF9. Material and methods: In the first step, gene structure was subjected to the in silico studies, which were include determination of Antibacterial activity, Prion formation risk, homology modeling, Molecular docking analysis, and Optimization of recombinant protein. In the second step, the Netrin-A gene was cloned and amplified in pTG19 vector. After digestion with BamH1 and EcoR1 restriction enzymes, it was cloned in pFastBac HTA vector. It was then transformed into DH10Bac competent cells, and the recombinant Bacmid was subsequently transfected into insect Sf9 cells. The expressed recombinant Netrin-A was thus purified in the Ni-NTA agarose. This protein evaluation was done using SDS-PAGE and western blot, respectively. Finally, its concentration was calculated with the Bradford assay method. Results: The Bacmid vector structure with Netrin-A was successfully constructed and then expressed as Netrin-A protein in the Sf9 cell lane. The molecular weight of this protein was 52 kDa with 404 amino acids. In the in silico studies, fortunately, we predicted that recombinant LSNetrin-A have Antibacterial activity and without any prion formation risk.This molecule hasa high binding affinity to the Neogenin and a lower affinity to the DCC-specific receptors. Signal peptide located between amino acids 24 and 25. The concentration of Netrin-A recombinant protein was calculated to be 48.8 μg/ml. it was confirmed that the characterized gene in our previous study codes L. sericata Netrin-A enzyme. Conclusions: Successful generation of the recombinant Netrin-A, a secreted protein in L.sericata salivary glands, and because Luciliasericata larvae are used in larval therapy. Therefore, the findings of the present study could be useful to researchers in future studies on wound healing.

Keywords: blowfly, BEVS, gene, immature insect, recombinant protein, Sf9

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1861 Lucilia Sericata Netrin-A: Secreted by Salivary Gland Larvae as a Potential to Neuroregeneration

Authors: Hamzeh Alipour, Masoumeh Bagheri, Tahereh Karamzadeh, Abbasali Raz, Kourosh Azizi

Abstract:

Netrin-A, a protein identified for conducting commissural axons, has a similar role in angiogenesis. In addition, studies have shown that one of the netrin-A receptors is expressed in the growing cells of small capillaries. It will be interesting to study this new group of molecules because their role in wound healing will become clearer in the future due to angiogenesis. The greenbottle blowfly Luciliasericata (L. sericata) larvae are increasingly used in maggot therapy of chronic wounds. This aim of this was the identification of moleculareatures of Netrin-A in L. sericata larvae. Larvae were reared under standard maggotarium conditions. The nucleic acid sequence of L. sericataNetrin-A (LSN-A) was then identified using Rapid Amplification of cDNA Ends (RACE) and Rapid Amplification of Genomic Ends (RAGE). Parts of the Netrin-A gene, including the middle, 3′-, and 5′-ends were identified, TA cloned in pTG19 plasmid, and transferred into DH5ɑ Escherichia coli. Each part was sequenced and assembled using SeqMan software. This gene structure was further subjected to in silico analysis. The DNA of LSN-A was identified to be 2407 bp, while its mRNA sequence was recognized as 2115 bp by Oligo0.7 software. It translated the Netrin-A protein with 704 amino acid residues. Its molecular weight is estimated to be 78.6 kDa. The 3-D structure ofNetrin-A drawn by SWISS-MODEL revealed its similarity to the Netrin-1 of humans with 66.8% identity. The LSN-A protein conduces to repair the myelin membrane in neuronal cells. Ultimately, it can be an effective candidate in neural regeneration and wound healing. Furthermore, our next attempt is to deplore recombinant proteins for use in medical sciences.

Keywords: maggot therapy, netrin-A, RACE, RAGE, lucilia sericata

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