Search results for: gene expression datasets

Authors: Alok Kumar, Hari Ram, Lebin Thomas, Ved Pal Singh

Abstract:

Organic solvent tolerant amylases and proteases of microbial origin are in great demand for their application in transglycosylation of water-insoluble flavanoids and in peptide synthesizing reaction in organic media. Most of the amylases and proteases are unstable in presence of organic solvent. In the present work two different bacterial strains M-11 and VP-07 were isolated from the soil sample of a dairy farm in Delhi, India, for the efficient production of extracellular amylase and protease through their screening on starch agar (SA) and skimmed milk agar (SMA) plates, respectively. Both the strains (M-11 and VP-07) were identified based on morphological, biochemical and 16S rRNA gene sequencing methods. After analysis through Ez-Taxon software, the strains M-11 and VP-07 were found to have maximum pairwise similarity of 98.63% and 100% with Bacillus subtilis subsp. inaquosorum BGSC 3A28 and Bacillus anthracis ATCC 14578 and were therefore identified as Bacillus sp. UKS1 and Bacillus sp. UKS2, respectively. Time course study of enzyme activity and bacterial growth has shown that both strains exhibited typical sigmoid growth behavior and maximum production of amylase (180 U/ml) and protease (78 U/ml) by these strains (UKS1 and UKS2) was commenced during stationary phase of growth at 24 and 20 h, respectively. Thereafter, both amylase and protease were tested for their tolerance towards organic solvents and were found to be active as well stable in p-xylene (130% and 115%), chloroform (110% and 112%), isooctane (119% and 107%), benzene (121% and 104%), n-hexane (116% and 103%) and toluene (112% and 101%, respectively). Owing to such properties, these enzymes can be exploited for their potential application in industries for organic synthesis.

Keywords: amylase, enzyme activity, industrial applications, organic solvent tolerant, protease

Procedia PDF Downloads 328

Authors: Pinel Sebastien, Bourrin François, De Madron Du Rieu Xavier, Ludwig Wolfgang, Arnau Pedro

Abstract:

Heavy precipitation constitutes a major meteorological threat in the western Mediterranean. Research has investigated the relationship between the states of the Mediterranean Sea and the atmosphere with the precipitation for short temporal windows. However, at a larger temporal scale, the precursor signals of heavy rainfall in the sea and atmosphere have drawn little attention. Moreover, despite ongoing improvements in numerical weather prediction, the medium-term forecasting of rainfall events remains a difficult task. Here, we aim to investigate the influence of early-spring environmental parameters on the following autumnal heavy precipitations. Hence, we develop a machine learning model to predict extreme autumnal rainfall with a 6-month lead time over the Spanish Catalan coastal area, based on i) the sea pattern (main current-LPC and Sea Surface Temperature-SST) at the mesoscale scale, ii) 4 European weather teleconnection patterns (NAO, WeMo, SCAND, MO) at synoptic scale, and iii) the hydrological regime of the main local river (Rhône River). The accuracy of the developed model classifier is evaluated via statistical analysis based on classification accuracy, logarithmic and confusion matrix by comparing with rainfall estimates from rain gauges and satellite observations (CHIRPS-2.0). Sensitivity tests are carried out by changing the model configuration, such as sea SST, sea LPC, river regime, and synoptic atmosphere configuration. The sensitivity analysis suggests a negligible influence from the hydrological regime, unlike SST, LPC, and specific teleconnection weather patterns. At last, this study illustrates how public datasets can be integrated into a machine learning model for heavy rainfall prediction and can interest local policies for management purposes.

Keywords: extreme hazards, sensitivity analysis, heavy rainfall, machine learning, sea-atmosphere modeling, precipitation forecasting

Procedia PDF Downloads 115

Authors: Hae-Jun Lee, Ji-Sun Shin, Kyung-Tae Lee

Abstract:

Potentilla species (Rosasease) have been used in traditional medicine to treat different ailment, disease or malady. In this study, we investigated the anti-inflammatory effects of ethanol extracts of NUTT (EPP) in lipopolysaccharide (LPS)-induced Raw 264.7 macrophages and septic mice. EPP suppressed LPS-induced nitric oxide (NO) and prostaglandin E2 (PGE2) production in LPS-induced Raw 264.7 macrophages. Consistent with these observations, EPP reduced the expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) by downregulation of their promoter activities. EPP inhibited tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) at production and mRNA levels. Molecularly, EPP attenuated the LPS-induced transcriptional activity, and DNA-binding activity of nuclear factor-κB (NF-κB), and this was associated with a decrease of translocation and phosphorylation of p65 NF-κB by inhibiting the inhibitory κB-α (IκB-α) degradation and IκB kinase-α/β (IKK-α/β) phosphorylation. Furthermore, EPP suppressed the LPS-induced activation of activator protein-1 (AP-1) by reducing the expression of c-Fos and c-Jun in nuclear. EPP also reduced the phosphorylation of mitogen-activated protein kinase (MAPK), such as p38 MAPK and c-Jun N-terminal kinase/stress-activated protein kinase (JNK). In a sepsis model, pretreatment with EPP reduced the LPS-induced lethality. Collectively, these results suggest that the anti-inflammatory effects of EPP were associated with the suppression of NF-κB and AP-1 activation, and support its possible therapeutic role for the treatment of sepsis.

Keywords: anti-inflammation, activator protein-1, nuclear factor κB, Potentilla paradoxa Nutt

Procedia PDF Downloads 315

Authors: Sumeeta Khurana, Kirti Megha, Amit Gupta, Rakesh Sehgal

Abstract:

Background: Amoebic keratitis is a painful vision threatening infection caused by a free living pathogenic amoeba Acanthamoeba. It can be misdiagnosed and very difficult to treat if not suspected early. The epidemiology of Acanthamoeba genotypes causing infection in our geographical area is not yet known to the best of our knowledge. Objective: To characterize Acanthamoeba isolates from amoebic keratitis patients. Methods: A total of 19 isolates obtained from patients with amoebic keratitis presenting to the Advanced Eye Centre at Postgraduate Institute of Medical Education and Research, a tertiary care centre of North India over a period of last 10 years were included. Their corneal scrapings, lens solution and lens case (in case of lens wearer) were collected for microscopic examination, culture and molecular diagnosis. All the isolates were maintained in the Non Nutrient agar culture medium overlaid with E.coli and 13 strains were axenised and maintained in modified Peptone Yeast Dextrose Agar. Identification of Acanthamoeba genotypes was based on amplification of diagnostic fragment 3 (DF3) region of the 18srRNA gene followed by sequencing. Nucleotide similarity search was performed by BLAST search of sequenced amplicons in GenBank database (http//www.ncbi.nlm.nih.gov/blast). Multiple Sequence alignments were determined by using CLUSTAL X. Results: Nine out of 19 Acanthamoeba isolates were found to belong to Genotype T4 followed by 6 isolates of genotype T11, 3 T5 and 1 T3 genotype. Conclusion: T4 is the predominant Acanthamoeba genotype in our geographical area. Further studies should focus on differences in pathogenicity of these genotypes and their clinical significance.

Keywords: Acanthamoeba, free living amoeba, keratitis, genotype, ocular

Procedia PDF Downloads 224

Authors: Hela Gargouri, Nizar Moalla, Hassen Hadj Kacem

Abstract:

Meat adulteration affecting the safety and quality of food is becoming one of the main concerns of public interest across the world. The drastic consequences on the meat industry highlighted the urgent necessity to control the products' quality and to point out the complexity of both supply and processing circuits. Due to the expansion of this problem, the authentic testing of foods, particularly meat and its products, is deemed crucial to avoid unfair market competition and to protect consumers from fraudulent practices of meat adulteration. The adoption of authentication methods by the food quality-control laboratories is becoming a priority issue. However, in some developing countries, the number of food tests is still insignificant, although a variety of processed and traditional meat products are widely consumed. Little attention has been paid to provide an easy, fast, reproducible, and low-cost molecular test, which could be conducted in a basic laboratory. In the current study, the 359 bp fragment of the cytochrome-b gene was mapped by PCR-RFLP using firstly fresh biological supports (DNA and meat) and then turkey salami as an example of commercial processed meat. This technique has been established through several optimizations, namely: the selection of restriction enzymes. The digestion with BsmAI, SspI, and TaaI succeed to identify the seven included animal species when meat is formed by individual species and when the meat is a mixture of different origin. In this study, the PCR-RFLP technique using universal primer succeed to meet our needs by providing an indirect sequencing method identifying by restriction enzymes the specificities characterizing different species on the same amplicon reducing the number of potential tests.

Keywords: adulteration, animal species, authentication, meat, mtDNA, PCR-RFLP

Procedia PDF Downloads 98

Authors: Anthony Proschka, Deepak Mishra, Merlyn Ramanan, Zurab Baratashvili

Abstract:

Small and medium-sized businesses receive over 160 billion invoices every year. Since these documents exhibit many subtle differences in layout and text, extracting structured fields such as sender name, amount, and VAT rate from them automatically is an open research question. In this paper, existing work in template-based document extraction is extended, and a system is devised that is able to reliably extract all required fields for up to 70% of all documents in the data set, more than any other previously reported method. The approaches are described for 1) detecting through visual features which template a given document belongs to, 2) automatically generating extraction rules for a given new template by composing regular expressions from multiple components, and 3) computing confidence scores that indicate the accuracy of the automatic extractions. The system can generate templates with as little as one training sample and only requires the ground truth field values instead of detailed annotations such as bounding boxes that are hard to obtain. The system is deployed and used inside a commercial accounting software.

Keywords: data mining, information retrieval, business, feature extraction, layout, business data processing, document handling, end-user trained information extraction, document archiving, scanned business documents, automated document processing, F1-measure, commercial accounting software

Procedia PDF Downloads 110

Authors: Nayra Martin-Key, Erich Graf, Wendy Adams, Graeme Fairchild

Abstract:

Children and adolescents with Conduct Disorder (CD) have been shown to demonstrate impairments in emotion recognition, but it is currently unclear whether this deficit is related to specific emotions or whether it represents a global deficit in emotion recognition. An emotion recognition task with concurrent eye-tracking was employed to further explore this relationship in a sample of male and female adolescents with CD. Participants made emotion categorization judgements for presented dynamic and morphed static facial expressions. The results demonstrated that males with CD, and to a lesser extent, females with CD, displayed impaired facial expression recognition in general, whereas callous-unemotional (CU) traits were linked to specific problems in sadness recognition in females with CD. A region-of-interest analysis of the eye-tracking data indicated that males with CD exhibited reduced fixation times for the eye-region of the face compared to typically-developing (TD) females, but not TD males. Females with CD did not show reduced fixation to the eye-region of the face relative to TD females. In addition, CU traits did not influence CD subjects’ attention to the eye-region of the face. These findings suggest that the emotion recognition deficits found in CD males, the worst performing group in the behavioural tasks, are partly driven by reduced attention to the eyes.

Keywords: attention, callous-unemotional traits, conduct disorder, emotion recognition, eye-region, eye-tracking, sex differences

Procedia PDF Downloads 291

Authors: Lian Zeng

Abstract:

Double-Spear 1-H2-1 (DS1-H2-1) is an oncolytic virus and an innovative biological drug candidate. The chemical composition of the drug product is a live attenuated West Nile virus (WNV) containing the human T cell costimulator (CD86) gene. After intratumoral injection, the virus can rapidly self-replicate in the injected site and lyse/kill the tumor by repeated infection among tumor cells. We also established xenograft tumor models in mice to evaluate the drug candidate's efficacy on those tumors. The results from preclinical studies on transplanted tumors in immunodeficient mice showed that DS1-H2-1 had significant oncolytic effects on human-origin cancers: it completely (100%) shrieked human glioma; limited human neuroblastoma growth reached as high as 95% growth inhibition rate (%TGITW). The safety data of preclinical animal experiments confirmed that DS1-H2-1 is safe as a biological drug for clinical use. In the preclinical drug efficacy experiment, virus-drug administration with different doses did not show abnormal signs and disease symptoms in more than 300 tested mice, and no side effects or death occurred through various administration routes. Intravenous administration did not cause acute infectious disease or other side effects. However, the replication capacity of the virus in tumor tissue via intravenous administration is only 1% of that of direct intratumoral administration. The direct intratumoral administration of DS1-H2-1 had a higher rate of viral replication. Therefore, choosing direct intratumoral injection can ensure both efficacy and safety.

Keywords: oncolytic virus, WNV-CD86, immunotherapy drugs, glioma, neuroblastoma

Procedia PDF Downloads 89

Authors: Joshua W. Edefo, Shafiul Azam, Murray D. Smith

Abstract:

Introduction: In April 2022, the Welsh Government introduced the six goals for urgent and emergency care programs. One of these goals was to provide access to clinically safe alternatives, leading to the establishment of the Same Day Emergency Care (SDEC) program. The SDEC initiative aims to offer viable options that maintain patient safety while avoiding unnecessary hospital stays. The aim of the study is to determine the duration of patient stay in SDEC and compare it with that of Emergency department (ED) stay to ascertain if one of the objectives of SDEC is achieved. Methods: Patient stays and attendance datasets were constructed from Withybush SDEC and ED patient records. These records were provided by Hywel Dda University Health Board Informatics. Some hypothetical pathways were identified, notably SDEC visits involving a single attendance and ED visits then immediately transferred to SDEC. Descriptive statistics were used to summarise the data, and hypothesis tests for mean differences used the student t-test. Propensity scoring was employed to match a set of ED patient stays to SDEC patient stays which were then used to determine the average treatment effect (ATE) to compare durations of stay in SDEC with ED. Regression methods were used to model the natural logarithm of the duration of SDEC attendance, and the level of statistical significance was set to 0.05. Results: SDEC visits involving a single attendance (170 of 384; 44.3%) is the most frequently observed pathway with patient length of stay at 256 minutes (95%CI 237.4 - 275.1). The next most frequently observed pathway of patient stay was SDEC attendance after presenting to ED (80 of 384; 20.8%) and gave the length of stay of 440 minutes (95%CI 351.6 - 529.2). Time spent in this pathway significantly increased by 184 minutes (95%CI 118.0 - 250.2, support for no difference p<0.001) compared to the most seen pathway. When SDEC data were compared with ED, the estimate for the ATE from SDEC single attendance was -272 minutes (95%CI -334.1 - -210.5; p<0.001), while that of ED then SDEC pathway was -50.6 min (95%CI -182.7-81.5; p=0.453). Conclusion: When patients are admitted to SDEC and successfully discharged, their stays are significantly shorter, approximately 4.5 hours, compared to patients who spend their entire stay in the Emergency Department. That difference vanishes when the patient stay includes a period spent previously in ED before transfer to SDEC.

Keywords: attendance, emergency-department, patient-stay, same-day-emergency-care

Procedia PDF Downloads 34

Authors: Mingxi Zhou, Jiří Kubásek, Iva Mozgová

Abstract:

In Arabidopsis thaliana, histone 3 lysine 27 tri-methylation catalysed byPRC2 is playing essential functions in the regulation of plant development, growth, and reproduction[1-2]. Despite numerous studies related to the role of PRC2 in developmental control, how PRC2 works in the operational control in plants is unknown. In the present, the evidence that PRC2 probably participates in the regulation of retrograde singalling pathway in Arabidopsisis found. Firstly, we observed that the rosette size and biomass in PRC2-depletion mutants (clf-29 and swn-3) is significantly higher than WTunder medium light condition (ML: 125 µmol m⁻² s⁻²), while under medium high light condition (MHL: 300 µmol m⁻² s-2), the increase was reverse. Under ML condition, the photosynthesis related parameters determined by fluorCam did not show significant differences between WT and mutants, while the pigments concentration increased in the leaf of PRC2-depletion mutants, especially in swn. The dynamic of light-responsive genes and circadian clock genes expression by RT-qPCRwithin 24 hours in the mutants were comparable to WT. However, we observed upregulation of photosynthesis-associated nuclear genes in the PRC2-depletion mutants under chloroplast damaging condition (treated by lincomycin), corresponding to the so-called genome uncoupled (gun) phenotype. Here, we will present our results describing these phenotypes and our suggestion and outlook for studying the involvement of PRC2 in chloroplast-to-nucleus retrograde signalling.

Keywords: PRC2, retrograde signalling, light acclimation, photosyntheis

Procedia PDF Downloads 89

Authors: Shahzad Bhatti, M. Aslamkhan, Sana Abbas, Marcella Attimonelli, Hikmet Hakan Aydin, Erica Martinha Silva de Souza,

Abstract:

Despite being situated at the crossroad of Asia, Pakistan has gained crucial importance because of its pivotal role in subsequent migratory events. To highlight the genetic footprints and to contribute an enigmatic picture of the relative population expansion pattern among four major Pashtun tribes in Khyber Pakhtunkhwa viz., Bangash, Khattak, Mahsuds and Orakzai, the complete mitochondrial control region of 100 Pashtun were analyzed. All Pashtun tribes studied here revealed high genetic diversity; that was comparable to the other Central Asian, Southeast Asian and European populations. The configuration of genetic variation and heterogeneity further unveiled through Multidimensional Scaling, Principal Component Analysis, and phylogenetic analysis. The results revealed that the Pashtun is a composite mosaic of West Eurasian ancestry of numerous geographic origin. They received substantial gene flow during different invasions and have a high element of the Western provenance. The most common haplogroups reported in this study are: South Asian haplogroup M (28%) and R (8%); whereas, West Asians haplogroups are present, albeit in high frequencies (67%) and widespread over all; HV (15%), U (17%), H (9%), J (8%), K (8%), W (4%), N (3%) and T (3%). Herein we linked the unexplored genetic connection between Ashkenazi Jews and Pashtun. The presence of specific haplotypes J1b (4%) and K1a1b1a (5%) point to a genetic connection of Jewish conglomeration with Khattak tribe. This was a result of an ancient genetic influx in the early Neolithic period that led to the formation of a diverse genetic substratum in present day Pashtun.

Keywords: mtDNA haplogroups, control region, Pakistan, KPK, ethnicity

Procedia PDF Downloads 467

Authors: Piedad Lucia Bolivar Goez, Daniel Ignacio Garzon Luna, Maria Camila Balcero Angel, Sara Carolina Martinez Roman, Daniela Natalia Polo Rivas, Sandra Liliana Rocha Guitierrez

Abstract:

This article provides a critical analysis on the conception of disorders of sexual development (DSDs) within the bioethics framework. By means of analytical thought, the objective is to approach topics such as the rediscovery of the body, the reinvention of sexuality and link them to the liability that health personnel have to inform people about the options they have to decide over their health and body. The medicalization of sexed bodies in both psychosocial and anatomo-morpho-physiological dimensions from a legal standpoint were analyzed. Its also explored the gender stereotypes established by society and the role of laws in guaranteeing the right of autonomy that takes on greater relevance in DSD. Through this analysis, it was concluded that despite intersexuality having been analyzed by Colombia’s Constitutional Court, that it is stated as a fair entity, the stigmatization by society has not allowed these individuals to belong to an egalitarian context in which everyone has the same opportunities of access to the goods and services that they need. This leads individuals to hide their identity and expression of genre in order to be accepted in a set of contexts. Thus creating a vulnerability that the health system must be able to identify and in which it is necessary to intervene at a biopsychosocial level, in order to guarantee the protection of the individual within an unquestionable frame of equality and solidarity.

Keywords: disorders of sex development, gender identity, sexuality, transgender persons

Procedia PDF Downloads 176

Authors: Khalda Amr, Noha Eltaweel, Sherif Ismail, Hala Raslan

Abstract:

Introduction: Osteoarthritis is the most common form of arthritis that involves the wearing away of the cartilage that caps the bones in the joints. While rheumatoid arthritis is an autoimmune disease in which the immune system attacks the joints, beginning with the lining of joints. In this study, we aimed to study the top deregulated miRNAs that might be the cause of pathogenesis in both diseases. Methods: Eight cases were recruited in this study: 4 rheumatoid arthritis (RA), 2 osteoarthritis (OA) patients, as well as 2 healthy controls. Total RNA was isolated from plasma to be subjected to miRNA profiling by NGS. Sequencing libraries were constructed and generated using the NEBNextR UltraTM small RNA Sample Prep Kit for Illumina R (NEB, USA), according to the manufacturer’s instructions. The quality of samples were checked using fastqc and multiQC. Results were compared RA vs Controls and OA vs. Controls. Target gene prediction and functional annotation of the deregulated miRNAs were done using Mienturnet. The top deregulated miRNAs in each disease were selected for further validation using qRT-PCR. Results: The average number of sequencing reads per sample exceeded 2.2 million, of which approximately 57% were mapped to the human reference genome. The top DEMs in RA vs controls were miR-6724-5p, miR-1469, miR-194-3p (up), miR-1468-5p, miR-486-3p (down). In comparison, the top DEMs in OA vs controls were miR-1908-3p, miR-122b-3p, miR-3960 (up), miR-1468-5p, miR-15b-3p (down). The functional enrichment of the selected top deregulated miRNAs revealed the highly enriched KEGG pathways and GO terms. Six of the deregulated miRNAs (miR-15b, -128, -194, -328, -542 and -3180) had multiple target genes in the RA pathway, so they are more likely to affect the RA pathogenesis. Conclusion: Six of our studied deregulated miRNAs (miR-15b, -128, -194, -328, -542 and -3180) might be highly involved in the disease pathogenesis. Further functional studies are crucial to assess their functions and actual target genes.

Keywords: next generation sequencing, mirnas, rheumatoid arthritis, osteoarthritis

Procedia PDF Downloads 64

Authors: Isaac Quiros-Fernandez, Angel Cid-Arregui

Abstract:

Tumor-reactive T cell receptors (TCRs) are being subject of intense investigation since they offer great potential in adoptive cell therapies against cancer. However, the identification of tumor-specific TCRs has proven challenging, for instance, due to the limited expansion capacity of tumor-infiltrating T cells (TILs) and the extremely low frequencies of tumor-reactive T cells in the repertoire of patients and healthy donors. We have developed an approach for rapid identification and characterization of neoepitope-reactive TCRs from the T cell repertoire of healthy donors. CD8 T cells isolated from multiple donors are subjected to a first sorting step after staining with HLA multimers carrying the peptide of interest. The isolated cells are expanded for two weeks, after which a second sorting is performed using the same peptide-HLA multimers. The cells isolated in this way are then processed for single-cell sequencing of their TCR alpha and beta chains. Newly identified TCRs are cloned in appropriate expression vectors for functional analysis on Jurkat, NK92, and primary CD8 T cells and tumor cells expressing the appropriate antigen. We have identified TCRs specifically binding HLA-A2 presenting epitopes of tumor antigens, which are capable of inducing TCR-mediated cell activation and cytotoxicity in target cancer cell lines. This method allows the identification of tumor-reactive TCRs in about two to three weeks, starting from peripheral blood samples of readily available healthy donors.

Keywords: cancer, TCR, tumor antigens, immunotherapy

Procedia PDF Downloads 51

Authors: Wikanda Phaphan

Abstract:

Mixture generalized gamma distribution is a combination of two distributions: generalized gamma distribution and length biased generalized gamma distribution. These two distributions were presented by Suksaengrakcharoen and Bodhisuwan in 2014. The findings showed that probability density function (pdf) had fairly complexities, so it made problems in estimating parameters. The problem occurred in parameter estimation was that we were unable to calculate estimators in the form of critical expression. Thus, we will use numerical estimation to find the estimators. In this study, we presented a new method of the parameter estimation by using the expectation – maximization algorithm (EM), the conjugate gradient method, and the quasi-Newton method. The data was generated by acceptance-rejection method which is used for estimating α, β, λ and p. λ is the scale parameter, p is the weight parameter, α and β are the shape parameters. We will use Monte Carlo technique to find the estimator's performance. Determining the size of sample equals 10, 30, 100; the simulations were repeated 20 times in each case. We evaluated the effectiveness of the estimators which was introduced by considering values of the mean squared errors and the bias. The findings revealed that the EM-algorithm had proximity to the actual values determined. Also, the maximum likelihood estimators via the conjugate gradient and the quasi-Newton method are less precision than the maximum likelihood estimators via the EM-algorithm.

Keywords: conjugate gradient method, quasi-Newton method, EM-algorithm, generalized gamma distribution, length biased generalized gamma distribution, maximum likelihood method

Procedia PDF Downloads 206

Authors: Yetti Marlida, Harnentis, Yuliaty Shafan Nur

Abstract:

Background: Tempoyak is a dish derived from fermented durian fruit. Tempoyak is a food consumed as a side dish when eating rice. Besides being eaten with rice, tempoyak can also be eaten directly. But this is rarely done because many cannot stand the sour taste and aroma of the tempoyak itself. In addition, tempoyak can also be used as a seasoning. The taste of tempoyak is acidic, this occurs because of the fermentation process in durian fruit meat which is the raw material. Tempoyak is already very well known in Indonesia, especially in Padang, Bengkulu, Palembang, Lampung, and Kalimantan. Besides that, this food is also famous in Malaysia. The purpose of this research is to improvement production of γ-aminobutyric acid (GABA) by Lactobacillus plantarum isolated from indigenous fermented durian (tempoyak). Selected Lactic Acid Bacteria (LAB) previously isolated from indigenous fermented durian (tempoyak) that have ability to produce γ-aminobutyric acid (GABA). The study was started with identification of selected LAB by 16 S RNA, followed optimation of GABA production by culture condition using different initial pH, temperature, glutamate concentration, incubation time, carbon and nitrogen sources. Results: The result from indentification used polymerase chain reaction of 16S rRNA gene sequences and phylogenetic analysis was Lactobacillus plantarum (coded as Y3) with a sequenced length of 1400bp. The improvement of Gaba production was found highest at pH: 6.0; temperature: 30 °C; glutamate concentration: 0.4%; incubation time: 60 h; glucose and yeast extract as carbon and nitrogen sources. Conclusions: GABA can be produced with the optimum condition fermentation were 66.06 mM.

Keywords: lactic acid bacteria, γ-amino butyric acid, indigenous fermented durian, PCR

Procedia PDF Downloads 128

Authors: Hawon Lee, Young-Pil Kim

Abstract:

Visualizing matrix metalloproteinases (MMPs) activity is necessary for understanding cancer metastasis because they are implicated in cell migration and invasion by degrading the extracellular matrix (ECM). While much effort has been made to sense the MMP activity, but extracellularly long-term monitoring of MMP activity still remains challenging. Here, we report a collagen-bound fluorescent bioprobe for the detection of MMP-2 activity in the extracellular environment. This bioprobe consists of ECM-immobilized part (including collagen-bound protein) and MMP-sensing part (including peptide substrate linked with fluorescence resonance energy transfer (FRET) coupler between donor green fluorescent protein (GFP) and acceptor TAMRA dye), which was constructed through intein-mediated self-splicing conjugation. Upon being immobilized on the collagen-coated surface, this bioprobe enabled efficient long-lasting observation of MMP-2 activity in the cultured cells without affecting cell growth and viability. As a result, the FRET ratio (acceptor/donor) decreased as the MMP2 activity increased in cultured cancer cells. Furthermore, unlike wild-type MMP-2, mutated MMP-2 expression (Y580A in the hemopexin region) gave rise to lowering the secretion of MMP-2 in HeLa. Conclusively, our method is anticipated to find applications for tracing and visualizing enzyme activity.

Keywords: collagen, ECM, FRET, MMP

Procedia PDF Downloads 187

Authors: Pragya Paneru, Prativa Paneru

Abstract:

This paper explores the attitude of high school Nepalese students toward scientists using a famous method named as Draw-A-Scientist-Test (DAST). A total of 145 students from Grade 11 and Grade 12 took part in this research and drew images of scientists. The findings indicated gender imbalance with male dominance in the images of scientists. The result also showed some usual stereotypes relating to hair, equipment, objects, use of eyeglasses, and lab coat in the drawings of scientists. Moreover, the influence of some mainstream western male scientists was widely seen in the drawings implying the exposure of limited male scientists to the students. In contrast to this, no real-life female scientists were mentioned by the participants demonstrating limited exposure of female scientists contributing to the gendered attitude toward the scientists. However, some of the findings also challenged the previous findings and depicted scientists with local features, positive expression, and working outdoors. Moreover, participants’ awareness that scientists could be anyone with an inquisitive mind was indicated by the variations in the characters in their drawings. The drawings indicated that scientists could be someone like a mother, themselves, a fashion icon, Buddha, or a crazy-looking person. This study recommends the inclusion of participants’ interviews, and exploration of their textbooks’ depiction of scientists to uncover additional details regarding their understanding of scientists. Also, a critical discussion of the stereotypical attitudes about scientists in class could help challenge the stereotypical assumptions of scientists.

Keywords: scientists, drawings, stereotypes, gender, high school students

Procedia PDF Downloads 61

Authors: Gyati Shilakari Asthana, Abhay Asthana, Dharm Veer Kohli, Suresh Prasad Vyas

Abstract:

Purpose: The therapeutic potential of oligonucleotide (ODN) is primarily dependent upon its safe and efficient delivery to specific cells overcoming degradation and maximizing cellular uptake in vivo. The present study is focused to design low molecular weight chitosan nanoconstructs to meet the requirements of safe and effectual delivery of ODNs. LMW-chitosan is a biodegradable, water soluble, biocompatible polymer and is useful as a non-viral vector for gene delivery due to its better stability in water. Methods: LMW chitosan ODN nanoparticles (CHODN NPs) were formulated by self-assembled method using various N/P ratios (moles ratio of amine groups of CH to phosphate moieties of ODNs; 0.5:1, 1:1, 3:1, 5:1, and 7:1) of CH to ODN. The developed CHODN NPs were evaluated with respect to gel retardation assay, particle size, zeta potential and cytotoxicity and transfection efficiency. Results: Complete complexation of CH/ODN was achieved at the charge ratio of 0.5:1 or above and CHODN NPs displayed resistance against DNase I. On increasing the N/P ratio of CH/ODN, the particle size of the NPs decreased whereas zeta potential (ZV) value increased. No significant toxicity was observed at all CH concentrations. The transfection efficiency was increased on increasing N/P ratio from 1:1 to 3:1, whereas it was decreased with further increment in N/P ratio upto 7:1. Maximum transfection of CHODN NPs with both the cell lines (Raw 267.4 cells and Hela cells) was achieved at N/P ratio of 3:1. The results suggest that transfection efficiency of CHODN NPs is dependent on N/P ratio. Conclusion: Thus the present study states that LMW chitosan nanoparticulate carriers would be acceptable choice to improve transfection efficiency in vitro as well as in vivo delivery of oligonucleotide.

Keywords: LMW-chitosan, chitosan nanoparticles, biocompatibility, cytotoxicity study, transfection efficiency, oligonucleotide

Procedia PDF Downloads 831

Authors: Carmen M. Gonzalez-Henriquez, Mauricio A. Sarabia-Vallejos, Juan Rodriguez-Hernandez

Abstract:

DMAEMA, as a monomer, has been widely studied and used in several application fields due to their pH-sensitive capacity (tertiary amine protonation), being relevant in the biomedical area as a potential carrier for drugs focused on the treatment of genetic or acquired diseases (efficient gene transfection), among others. Additionally, the inhibition of bacterial growth and, therefore, their antimicrobial activity, can be used as dual-functional antifogging/antimicrobial polymer coatings. According to their interesting physicochemical characteristics and biocompatible properties, DMAEMA was used as a monomer to synthesize a smart pH-sensitive hydrogel, namely poly(HEMA-co-PEGDA575-co-DMAEMA). Thus, different mole ratios (ranging from 5:1:0 to 0:1:5, according to the mole ratio between HEMA, PEGDA, and DEAEMA, respectively) were used in this research. The surface patterns formed via a two-step polymerization (redox- and photo-polymerization) were first chemically studied via 1H-NMR and elemental analysis. Secondly, the samples were morphologically analyzed by using Field-Emission Scanning Electron Microscopy (FE-SEM) and Atomic Force Microscopy (AFM) techniques. Then, a particular relation between HEMA, PEGDA, and DEAEMA (0:1:5) was also characterized at three different pH (5.4, 7.4 and 8.3). The hydrodynamic radius and zeta potential of the micro-hydrogel particles (emulsion) were carried out as a possible control for morphology, exploring the effect that produces hydrogel micelle dimensions in the wavelength, height, and roughness of the wrinkled patterns. Finally, contact angle and cross-hatch adhesion test was carried out for the hydrogels supported on glass using TSM-silanized surfaces in order to measure their mechanical properties.

Keywords: wrinkled patterns, smart pH-sensitive hydrogels, hydrogel micelle diameter, adhesion tests

Procedia PDF Downloads 191

Authors: Ammar Said Ahmed, M. Reissmann, R. Bortfeldt, G. A. Brockmann

Abstract:

Milk protein genetic variants are of interest for characterizing domesticated mammalian species and breeds, and for studying associations with economic traits. The aim of this work was to analyze milk protein genetic variation in the Sudanese native cattle breeds, which have been gradually declining in numbers over the last years due to the breed substitution, and indiscriminate crossbreeding. The genetic variation at three milk protein genes αS1-casein (CSN1S1), αS2-casein (CSN1S2) and ƙ-casein (CSN3) was investigated in 250 animals belonging to five Bos indicus cattle breeds of Sudan (Butana, Kenana, White-nile, Erashy and Elgash). Allele specific primers were designed for five SNPs determine the CSN1S1 variants B and C, the CSN1S2 variants A and B, the CSN3 variants A, B and H. Allele, haplotype frequencies and genetic distances (D) were calculated and the phylogenetic tree was constructed. All breeds were found to be polymorphic for the studied genes. The CSN1S1*C variant was found very frequently (>0.63) in all analyzed breeds with highest frequency (0.82) in White-nile cattle. The CSN1S2*A variant (0.77) and CSN3*A variant (0.79) had highest frequency in Kenana cattle. Eleven haplotypes in casein gene cluster were inferred. Six of all haplotypes occurred in all breeds with remarkably deferent frequencies. The estimated D ranged from 0.004 to 0.049. The most distant breeds were White-nile and Kenana (D 0.0479). The results presented contribute to the genetic knowledge of indigenous cattle and can be used for proper definition and classification of the Sudanese cattle breeds as well as breeding, utilization, and potential development of conservation strategies for local breeds.

Keywords: milk protein, genetic variation, casein haplotype, Bos indicus

Procedia PDF Downloads 420

Authors: Sang-Sun Yoon, Yea-Hyun Leem, Sangmee Ahn Jo

Abstract:

The N-acetylated-alpha-linked-acidic (NAAG) peptidase inhibitor 2-phosphonomethyl pentanedioic acid (2-PMPA) has demonstrated to be neuroprotective against glutamate-mediated neuron degeneration and neurological disorders such as ischemia. Several studies have demonstrated impaired psychiatric function by altered glutamate carboxypeptidase II expression, although 2-PMPA has not yet been studied. Thus, we investigated effect of 2-PMPA on depressive-like phenotype using C57BL/6 mice. Treatment of 2-PMPA (10 mg/kg for 6 days/daily, ip injection) on C57BL/6 naïve mice showed depressive-like symptoms such as decreased social preference, but did not affect the immobility measured by tail suspension test. Reduction of phosphorylated cAMP-responsive element binding (p-CREB) known as a representative marker of depressive-like behavior was observed in layer 1 and piriform cortex subregions of the prefrontal cortex of 2-PMPA-treated mice. The immunoreactivity of metabotropic glutamate receptors 5 (mGluR5) that mediate phosphorylation of CREB was also decreased in layer 1 and piriform cortex subregions of the prefrontal cortex of 2-PMPA injected mice. Thus, our results suggest that dysregulation of the GCPII or NAAG by 2-PMPA treatment is likely to be associated with pathogenesis of depression and further studies are needed to understand whether the reduced NAAG level or enhanced glutamate level in the brain is involved in this response.

Keywords: depression, GCPII, 2-PMPA, p-CREB, mGluR5

Procedia PDF Downloads 249

Authors: Rabson Banda

Abstract:

This abstract delves into the transformative journey of Rabson Banda, a visionary leader in Zambia who has harnessed the power of photovoice, mental fitness, and entrepreneurship to combat substance use disorders in the face of limited job opportunities in Africa. Rabson's personal battle with alcohol-induced disorder serves as the foundation for his pioneering work in establishing Elevated Initiatives, a grassroots organization dedicated to empowering individuals struggling with addiction through innovative approaches. Through the lens of photovoice, Rabson Banda amplifies the voices of those affected by substance abuse, providing a platform for self-expression and healing. By intertwining mental fitness techniques and entrepreneurial skills, he equips individuals with the tools needed to break free from the cycle of addiction and chart a new path toward holistic wellness and economic independence. Rabson Banda's story embodies resilience, hope, and the unwavering belief in the transformative power of community-driven initiatives. His work not only addresses the immediate challenges of substance use disorders but also addresses the root causes of addiction by advocating for increased job opportunities and economic empowerment in Zambia and across Africa for decent job creation and Entrepreneurship: A Lived Experience of Alcohol-Induced Disorder and Recovery in Zambia.

Keywords: mental fitness, entrepreneurship, photovoice, descent jobs

Procedia PDF Downloads 23

Authors: Sayanti Kar, Amitava Ghosh, Pritam Aitch, Gupinath Bhandari

Abstract:

An extensive seasonal analysis of wastewater had been done from outfalls of river Ganga in Howrah, Hooghly, 24 PGS (N) District, West Bengal, India during 2017. The morphological parameters of Bitter gourd (Momordica charantia L.) were estimated under wastewater treatment. An approach to study the activity within the range of low molecular weight peptide 3-0.5 kDa were taken through its extraction and purification by ion exchange resin column, cation, and anion exchanger. HPLC analysis had been done for both in wastewater treated and untreated plants. The antioxidant activity by using DPPH and germination percentage in control and treated plants were also determined in relation to wastewater effect. The inhibition of growth and its parameters were maximum in pre-monsoon in comparing to post-monsoon and monsoon season. The study also helped to explore the effect of wastewater on the peptidome of Bitter gourd (Momordica charantia L.). Some of these low molecular weight peptide(s) (3-0.5 kDa) also inhibited during wastewater treatment. Expression of particular peptide(s) or absence of some peptide(s) in chromatogram indicated the adverse effects on plants which may be the indication of stressful condition. Pre monsoon waste water was found to create more impact than other two.

Keywords: bitter gourd (Momordica charantia l.), low molecular weight peptide, river ganga, waste water

Procedia PDF Downloads 109

Authors: Rofida Gamal, Mostafa Mohammed, Mariam Adel, Marwa Gamal, Marwa kamal, Ayat Saber, Maha Mamdouh, Amira Emad, Mai Ramadan

Abstract:

Lynch Syndrome is an inherited genetic condition associated with an increased risk of colorectal and other cancers. Detecting Lynch Syndrome in individuals is crucial for early intervention and preventive measures. This study proposes a computational pipeline for Lynch Syndrome detection by integrating alignment, variant calling, and annotation. The pipeline leverages popular tools such as FastQC, Trimmomatic, BWA, bcftools, and ANNOVAR to process the input FASTQ file, perform quality trimming, align reads to the reference genome, call variants, and annotate them. It is believed that the computational pipeline was applied to a dataset of Lynch Syndrome cases, and its performance was evaluated. It is believed that the quality check step ensured the integrity of the sequencing data, while the trimming process is thought to have removed low-quality bases and adaptors. In the alignment step, it is believed that the reads were accurately mapped to the reference genome, and the subsequent variant calling step is believed to have identified potential genetic variants. The annotation step is believed to have provided functional insights into the detected variants, including their effects on known Lynch Syndrome-associated genes. The results obtained from the pipeline revealed Lynch Syndrome-related positions in the genome, providing valuable information for further investigation and clinical decision-making. The pipeline's effectiveness was demonstrated through its ability to streamline the analysis workflow and identify potential genetic markers associated with Lynch Syndrome. It is believed that the computational pipeline presents a comprehensive and efficient approach to Lynch Syndrome detection, contributing to early diagnosis and intervention. The modularity and flexibility of the pipeline are believed to enable customization and adaptation to various datasets and research settings. Further optimization and validation are believed to be necessary to enhance performance and applicability across diverse populations.

Keywords: Lynch Syndrome, computational pipeline, alignment, variant calling, annotation, genetic markers

Procedia PDF Downloads 55

Authors: Migena Mana, Ahmed Khalid Syed, Abdul Malik, Nikhil Cherian

Abstract:

In recent years, advancements in deep learning enabled researchers to tackle the problem of self-driving cars. Car companies use huge datasets to train their deep learning models to make autonomous cars a reality. However, this approach has certain drawbacks in that the state space of possible actions for a car is so huge that there cannot be a dataset for every possible road scenario. To overcome this problem, the concept of reinforcement learning (RL) is being investigated in this research. Since the problem of autonomous driving can be modeled in a simulation, it lends itself naturally to the domain of reinforcement learning. The advantage of this approach is that we can model different and complex road scenarios in a simulation without having to deploy in the real world. The autonomous agent can learn to drive by finding the optimal policy. This learned model can then be easily deployed in a real-world setting. In this project, we focus on three RL algorithms: Q-learning, Deep Deterministic Policy Gradient (DDPG), and Proximal Policy Optimization (PPO). To model the environment, we have used TORCS (The Open Racing Car Simulator), which provides us with a strong foundation to test our model. The inputs to the algorithms are the sensor data provided by the simulator such as velocity, distance from side pavement, etc. The outcome of this research project is a comparative analysis of these algorithms. Based on the comparison, the PPO algorithm gives the best results. When using PPO algorithm, the reward is greater, and the acceleration, steering angle and braking are more stable compared to the other algorithms, which means that the agent learns to drive in a better and more efficient way in this case. Additionally, we have come up with a dataset taken from the training of the agent with DDPG and PPO algorithms. It contains all the steps of the agent during one full training in the form: (all input values, acceleration, steering angle, break, loss, reward). This study can serve as a base for further complex road scenarios. Furthermore, it can be enlarged in the field of computer vision, using the images to find the best policy.

Keywords: autonomous driving, DDPG (deep deterministic policy gradient), PPO (proximal policy optimization), reinforcement learning

Procedia PDF Downloads 127

Authors: Belaynesh Tsegay Beyene, Teklay Gebrecherkos, Atsebaha Gebrekidan Kahsay, Mahmud Abdulkader

Abstract:

Background: Hepatitis B and C viruses are of important health and socioeconomic problem of the globe with remarkable diseases and deaths in Sub-Saharan African countries. The burden of hepatitis is unknown in the prison settings of Tigrai. Therefore, we aimed to describe the seroprevalence and associated factors of hepatitis B and C viruses among prisoners of Tigrai, Ethiopia. Methods: A cross-sectional study was carried out from February 2020 to May 2020 at the prison facilities of Tigrai. Demographics and associated factors were collected from 315 prisoners prospectively. Five milliliter of blood was collected and tested using rapid tests kits of HBsAg (Zhejiang orient Gene Biotech Co., Ltd., China) and HCV antibodies (Volkan Kozmetik Sanayi Ve Ticaret Ltd. STI, Turkey). Positive samples were confirmed using enzyme-linked immunosorbent assay (ELISA) (Beijing Wantai Biological Pharmacy Enterprise Co. Ltd). Data were analyzed using Statistical Package for Social Sciences (SPSS) version 20 and p < 0.05 was considered statistically significant. Results: The overall seroprevalence of HBV and HCV were 25 (7.9%) and 1(0.3%), respectively. The majority of hepatitis B viral infections were identified from the age groups of 18-25 years (10.7%) and unmarried prisoners (11.8%). Prisoners greater than 100 per cell [AOR =3.95, 95% CI= (1.15, 13.6, p =0.029)] and having history of alcohol consumption [AOR =3.01, 95% CI= (1.17, 7.74, p =0.022)] were significantly associated with HBV infections. Conclusions: The seroprevalence of HBV among prisoners was nearly high or borderline (7.9%) with a very low HCV prevalence (0.3%). HBV was most prevalent among young adults, large number of prisoners per cell and those who had history of alcohol consumption. This study recommends that there should be prison-focused intervention including regular health education by emphasis on the mode of transmission and introducing HBV screening policy for prisoners especially when they enter to the prison.

Keywords: seroprevalence, HBV, HCV, prisoners, Tigrai

Procedia PDF Downloads 58

Authors: Sara Przetocka, Krzysztof Żak, Grzegorz Dubin, Tadeusz Holak

Abstract:

Toll-like receptors (TLRs) play central role in the innate immune response and inflammation by recognizing pathogen-associated molecular patterns (PAMPs). A fundamental basis of TLR signalling is dependent upon the recruitment and association of adaptor molecules that contain the structurally conserved Toll/interleukin-1 receptor (TIR) domain. MyD88 (myeloid differentiation primary response gene 88) is the universal adaptor for TLRs and cooperates with Mal (MyD88 adapter-like protein, also known as TIRAP) in TLR4 response which is predominantly used in inflammation, host defence and carcinogenesis. Up to date two possible models of MyD88, Mal and TLR4 interactions have been proposed. The aim of our studies is to confirm or abolish presented models and accomplish the full structural characterisation of TIR domains interaction. Using molecular cloning methods we obtained several construct of MyD88 and Mal TIR domain with GST or 6xHis tag. Gel filtration method as well as pull-down analysis confirmed that recombinant TIR domains from MyD88 and Mal are binding in complexes. To examine whether obtained complexes are homo- or heterodimers we carried out cross-linking reaction of TIR domains with BS3 compound combined with mass spectrometry. To investigate which amino acid residues are involved in this interaction the NMR titration experiments were performed. 15N MyD88-TIR solution was complemented with non-labelled Mal-TIR. The results undoubtedly indicate that MyD88-TIR interact with Mal-TIR. Moreover 2D spectra demonstrated that simultaneously Mal-TIR self-dimerization occurs which is necessary to create proper scaffold for Mal-TIR and MyD88-TIR interaction. Final step of this study will be crystallization of MyD88 and Mal TIR domains complex. This crystal structure and characterisation of its interface will have an impact in understanding the TLR signalling pathway and possibly will be used in development of new anti-cancer treatment.

Keywords: cancer, MyD88, TIR domains, Toll-like receptors

Procedia PDF Downloads 275

Authors: Farnaz Yusuf, Naseem A. Gaur

Abstract:

The increase in environmental concerns, rapid depletion of fossil fuel reserves and intense interest in achieving energy security has led to a global research effort towards developing renewable sources of fuels. Second generation biofuels have attracted more attention recently as the use of lignocellulosic biomass can reduce fossil fuel dependence and is environment-friendly. Xylose is the main pentose and second most abundant sugar after glucose in lignocelluloses. Saccharomyces cerevisiae does not readily uptake and use pentose sugars. For an economically feasible biofuel production, both hexose and pentose sugars must be fermented to ethanol. Therefore, it is important to develop S. cerevisiae host platforms with more efficient xylose utilization. This work aims to construct a xylose fermenting yeast strains with engineered oxido-reductative pathway for xylose metabolism. Engineered strain was further improved by adaptive evolutionary engineering approach. The engineered strain is able to grow on xylose as sole carbon source with the maximum ethanol yield of 0.39g/g xylose and productivity of 0.139g/l/h at 96 hours. The further improvement in strain development involves over expression of pentose phosphate pathway and protein engineering of xylose reductase/xylitol dehydrogenase to change their cofactor specificity in order to reduce xylitol accumulation.

Keywords: biofuel, lignocellulosic biomass, saccharomyces cerevisiae, xylose

Procedia PDF Downloads 194

Authors: Rosa Buxeda, Lorenzo Saliceti-Piazza, Rodolfo J. Romañach, Luis Ríos, Sandra L. Maldonado-Ramírez

Abstract:

Biopharmaceuticals manufacturing is one of the major economic activities worldwide. Ninety-three percent of the workforce in a biomanufacturing environment concentrates in production-related areas. As a result, strategic collaborations between industry and academia are crucial to ensure the availability of knowledgeable workforce needed in an economic region to become competitive in biomanufacturing. In the past decade, our institution has been a key strategic partner with multinational biotechnology companies in supplying science and engineering graduates in the field of industrial biotechnology. Initiatives addressing all levels of the educational pipeline, from K-12 to college to continued education for company employees have been established along a ten-year span. The Amgen BioTalents Program was designed to provide undergraduate science and engineering students with training in biomanufacturing. The areas targeted by this educational program enhance their academic development, since these topics are not part of their traditional science and engineering curricula. The educational curriculum involved the process of producing a biomolecule from the genetic engineering of cells to the production of an especially targeted polypeptide, protein expression and purification, to quality control, and validation. This paper will report and describe the implementation details and outcomes of the first sessions of the program.

Keywords: biomanufacturing curriculum, interdisciplinary learning, workforce development, industry-academia partnering

Procedia PDF Downloads 269