Abstract: Graves’ Disease (GD), an autoimmune health condition caused by the over reactiveness of the thyroid, affects about 1 in 200 people worldwide. GD is not caused by one specific single nucleotide polymorphism (SNP) or gene mutation, but rather determined by multiple factors, each differing from each other. Malfunction of the genes in Human Leukocyte Antigen (HLA) family tend to play a major role in autoimmune diseases, but other genes, such as LOC101929163, have functions that still remain ambiguous. Currently, little studies were done to study GD, resulting in inconclusive results. This study serves not only to introduce background knowledge about GD, but also to organize and pinpoint the major SNPs and genes that are potentially related to the occurrence of GD in humans. Collected from multiple sources from genome-wide association studies (GWAS) Central, the potential SNPs related to the causes of GD are included in this study. This study has located the genes that are related to those SNPs and closely examines a selected sample. Using the data from this study, scientists will then be able to focus on the most expressed genes in GD patients and develop a treatment for GD.
Abstract: Analysis of the human microbiome using metagenomic
sequencing data has demonstrated high ability in discriminating
various human diseases. Raw metagenomic sequencing data require
multiple complex and computationally heavy bioinformatics steps
prior to data analysis. Such data contain millions of short sequences
read from the fragmented DNA sequences and stored as fastq files.
Conventional processing pipelines consist in multiple steps including
quality control, filtering, alignment of sequences against genomic
catalogs (genes, species, taxonomic levels, functional pathways,
etc.). These pipelines are complex to use, time consuming and
rely on a large number of parameters that often provide variability
and impact the estimation of the microbiome elements. Training
Deep Neural Networks directly from raw sequencing data is a
promising approach to bypass some of the challenges associated with
mainstream bioinformatics pipelines. Most of these methods use the
concept of word and sentence embeddings that create a meaningful
and numerical representation of DNA sequences, while extracting
features and reducing the dimensionality of the data. In this paper
we present an end-to-end approach that classifies patients into disease
groups directly from raw metagenomic reads: metagenome2vec. This
approach is composed of four steps (i) generating a vocabulary of
k-mers and learning their numerical embeddings; (ii) learning DNA
sequence (read) embeddings; (iii) identifying the genome from which
the sequence is most likely to come and (iv) training a multiple
instance learning classifier which predicts the phenotype based on
the vector representation of the raw data. An attention mechanism
is applied in the network so that the model can be interpreted,
assigning a weight to the influence of the prediction for each genome.
Using two public real-life data-sets as well a simulated one, we
demonstrated that this original approach reaches high performance,
comparable with the state-of-the-art methods applied directly on
processed data though mainstream bioinformatics workflows. These
results are encouraging for this proof of concept work. We believe
that with further dedication, the DNN models have the potential to
surpass mainstream bioinformatics workflows in disease classification
tasks.
Abstract: Hyaluronic acid (HA) consists of linear heteropolysaccharides repeat of D-glucuronic acid and N-acetyl-D-glucosamine. HA has various useful properties to maintain skin elasticity and moisture, reduce inflammation, and lubricate the movement of various body parts without causing immunogenic allergy. HA can be found in several animal tissues as well as in the capsule component of some bacteria including Pasteurella multocida. This study aimed to modify a genome-scale metabolic model of Escherichia coli using computational simulation and flux analysis methods to predict HA productivity under different carbon sources and nitrogen supplement by the addition of two enzymes (GLMU2 and HYAD) from P. multocida to improve the HA production under the specified amount of carbon sources and nitrogen supplements. Result revealed that threonine and aspartate supplement raised the HA production by 12.186%. Our analyses proposed the genome-scale metabolic model is useful for improving the HA production and narrows the number of conditions to be tested further.
Abstract: Learning from very big datasets is a significant problem for most present data mining and machine learning algorithms. MicroRNA (miRNA) is one of the important big genomic and non-coding datasets presenting the genome sequences. In this paper, a hybrid method for the classification of the miRNA data is proposed. Due to the variety of cancers and high number of genes, analyzing the miRNA dataset has been a challenging problem for researchers. The number of features corresponding to the number of samples is high and the data suffer from being imbalanced. The feature selection method has been used to select features having more ability to distinguish classes and eliminating obscures features. Afterward, a Convolutional Neural Network (CNN) classifier for classification of cancer types is utilized, which employs a Genetic Algorithm to highlight optimized hyper-parameters of CNN. In order to make the process of classification by CNN faster, Graphics Processing Unit (GPU) is recommended for calculating the mathematic equation in a parallel way. The proposed method is tested on a real-world dataset with 8,129 patients, 29 different types of tumors, and 1,046 miRNA biomarkers, taken from The Cancer Genome Atlas (TCGA) database.
Abstract: Additive Manufacturing is utilized in medical automation to optimize and integrate materials in accordance to energy source type leading to treatment gaps in industrial designs for extreme biomechanical forces in relation with vibration, fluid transfer, and multi-physics performance. Elastic/piezoelectric materials are strongly ordered inter-metallics for characterization of distinct features that can provide excellent compositional strength, ductility, and uniformity for superelastic shape memory alloy on medical devices. Several theories can be derived to analyze and interpret complex problems on the application of functionally graded materials used in medical machinery for genome architecture. Numerical principles on fluid and thermodynamics such as Reynolds number, Darcy rule, Friction Factor and Heat Rate are integrated with fundamental equation of numerical vibrations using Helmholtz equation. Simulation by Large Eddy approach and genetic modeling can be done using Physical and Chemical Vapor Deposition following various theories on Carrera’s Unified Formulations by comparing with various Classical Plate Theories, Equivalent Single Layer Theories, Layer-Wise Theories, Zig-Zag Theories, and Mixed Refined Variational Theories. The subject is approached towards the application of ethical and legal problems in order to resolve issues on consent and return of results.
Abstract: Rv3873 is a relatively large size protein (371 amino acids in length) and its gene is located in the immunodominant genomic region of difference (RD)1 that is present in the genome of Mycobacterium tuberculosis but deleted from the genomes of all the vaccine strains of Bacillus Calmette Guerin (BCG) and most other mycobacteria. However, when tested for cellular immune responses using peripheral blood mononuclear cells from tuberculosis patients and BCG-vaccinated healthy subjects, this protein was found to be a major stimulator of cell mediated immune responses in both groups of subjects. In order to further identify the sequence of immunodominant epitopes and explore their Human Leukocyte Antigen (HLA)-restriction for epitope recognition, 24 peptides (25-mers overlapping with the neighboring peptides by 10 residues) covering the sequence of Rv3873 were synthesized chemically using fluorenylmethyloxycarbonyl chemistry and tested in cell mediated immune responses. The results of these experiments helped in the identification of an immunodominant peptide P9 that was recognized by people expressing varying HLA-DR types. Furthermore, it was also predicted to be a promiscuous binder with multiple epitopes for binding to HLA-DR, HLA-DP and HLA-DQ alleles of HLA-class II molecules that present antigens to T helper cells, and to HLA-class I molecules that present antigens to T cytotoxic cells. In addition, the evaluation of peptide P9 using an immunogenicity predictor server yielded a high score (0.94), which indicated a greater probability of this peptide to elicit a protective cellular immune response. In conclusion, P9, a peptide with multiple epitopes and ability to bind several HLA class I and class II molecules for presentation to cells of the cellular immune response, may be useful as a peptide-based vaccine against tuberculosis.
Abstract: The comparisons of mycobacterial genomes have identified several Mycobacterium tuberculosis-specific genomic regions that are absent in other mycobacteria and are known as regions of differences. Due to M. tuberculosis-specificity, the peptides encoded by these regions could be useful in the specific diagnosis of tuberculosis. To explore this possibility, overlapping synthetic peptides corresponding to 39 proteins predicted to be encoded by genes present in regions of differences were tested for antibody-reactivity with sera from tuberculosis patients and healthy subjects. The results identified four immunodominant peptides corresponding to four different proteins, with three of the peptides showing significantly stronger antibody reactivity and rate of positivity with sera from tuberculosis patients than healthy subjects. The fourth peptide was recognized equally well by the sera of tuberculosis patients as well as healthy subjects. Predication of antibody epitopes by bioinformatics analyses using ABCpred server predicted multiple linear epitopes in each peptide. Furthermore, peptide sequence analysis for sequence identity using BLAST suggested M. tuberculosis-specificity for the three peptides that had preferential reactivity with sera from tuberculosis patients, but the peptide with equal reactivity with sera of TB patients and healthy subjects showed significant identity with sequences present in nob-tuberculous mycobacteria. The three identified M. tuberculosis-specific immunodominant peptides may be useful in the serological diagnosis of tuberculosis.
Abstract: The purpose of computing the similarity and the diversity in the species is to trace the process of evolution and to find the relationship between the species and discover the unique, the special, the common and the universal proteins. The proteins of the whole genome of 40 species are compared with the cronobacter genome which is used as reference genome. More than 3 billion pairwise alignments are performed using blastp. Several findings are introduced in this study, for example, we found 172 proteins in cronobacter genome which have insignificant hits in other species, 116 significant proteins in the all tested species with very high score value and 129 common proteins in the plants but have insignificant hits in mammals, birds, fishes, and insects.
Abstract: The role and relative importance of intrinsic and extrinsic factors in the development of complex diseases such as cancer still remains a controversial issue. Determining the amount of variation explained by these factors needs experimental data and statistical models. These models are nevertheless based on the occurrence and accumulation of random mutational events during stem cell division, thus rendering cancer development a stochastic outcome. We demonstrate that not only individual genome sequencing is uninformative in determining cancer risk, but also assigning a unique genome sequence to any given individual (healthy or affected) is not meaningful. Current whole-genome sequencing approaches are therefore unlikely to realize the promise of personalized medicine. In conclusion, since genome sequence differs from cell to cell and changes over time, it seems that determining the risk factor of complex diseases based on genome sequence is somewhat unrealistic, and therefore, the resulting data are likely to be inherently uninformative.
Abstract: The median problem is significantly applied to derive
the most reasonable rearrangement phylogenetic tree for many
species. More specifically, the problem is concerned with finding
a permutation that minimizes the sum of distances between itself
and a set of three signed permutations. Genomes with equal number
of genes but different order can be represented as permutations.
In this paper, an algorithm, namely BeamGA median, is proposed
that combines a heuristic search approach (local beam) as an
initialization step to generate a number of solutions, and then a
Genetic Algorithm (GA) is applied in order to refine the solutions,
aiming to achieve a better median with the smallest possible reversal
distance from the three original permutations. In this approach,
any genome rearrangement distance can be applied. In this paper,
we use the reversal distance. To the best of our knowledge, the
proposed approach was not applied before for solving the median
problem. Our approach considers true biological evolution scenario
by applying the concept of common intervals during the GA
optimization process. This allows us to imitate a true biological
behavior and enhance genetic approach time convergence. We were
able to handle permutations with a large number of genes, within
an acceptable time performance and with same or better accuracy as
compared to existing algorithms.
Abstract: T7 phage could be used as a perfect vector for peptides expression and haptens presentation. T7-3GnRH recombinant phage was constructed by inserting three copies of Gonadotrophin Releasing Hormone (GnRH) gene into the multiple cloning site of T7 Select 415-1b phage genome. The positive T7-3GnRH phage was selected by using polymerase chain reaction amplification, and the p10B-3GnRH fusion protein was verified by SDS-PAGE and Western-blotting assay. T7-3GnRH vaccine was made and immunized with 1010 pfu in 0.2 ml per dose in mice. Blood samples were collected at an interval in weeks, and anti-GnRH antibody and testosterone concentrations were detected by ELISA and radioimmunoassay, respectively. The results show that T7-3GnRH phage particles confer a high immunogenicity to the GnRH-derived epitope. Moreover, the T7-3GnRH vaccine induced higher level of anti-GnRH antibody than ImproVac®. However, the testosterone concentrations in both immunized groups were at a similar level, and the testis developments were significantly inhibited compared to controls. These findings demonstrated that the anti-GnRH antibody could neutralize the endogenous GnRH to down regulate testosterone level and limit testis development, highlighting the potential value of T7-3GnRH in the immunocastration vaccine research.
Abstract: A challenge for laboratories is to provide bacterial identification and antibiotic sensitivity results within a short time. Hence, advancement in the required technology is desirable to improve timing, accuracy and quality. Even with the current advances in methods used for both phenotypic and genotypic identification of bacteria the need is there to develop method(s) that enhance the outcome of bacteriology laboratories in accuracy and time. The hypothesis introduced here is based on the assumption that the chromosome of any bacteria contains unique sequences that can be used for its identification and typing. The outcome of a pilot study designed to test this hypothesis is reported in this manuscript. Methods: The complete chromosome sequences of several bacterial species were downloaded to use as search targets for unique sequences. Visual basic and SQL server (2014) were used to generate a complete set of 18-base long primers, a process started with reverse translation of randomly chosen 6 amino acids to limit the number of the generated primers. In addition, the software used to scan the downloaded chromosomes using the generated primers for similarities was designed, and the resulting hits were classified according to the number of similar chromosomal sequences, i.e., unique or otherwise. Results: All primers that had identical/similar sequences in the selected genome sequence(s) were classified according to the number of hits in the chromosomes search. Those that were identical to a single site on a single bacterial chromosome were referred to as unique. On the other hand, most generated primers sequences were identical to multiple sites on a single or multiple chromosomes. Following scanning, the generated primers were classified based on ability to differentiate between medically important bacterial and the initial results looks promising. Conclusion: A simple strategy that started by generating primers was introduced; the primers were used to screen bacterial genomes for match. Primer(s) that were uniquely identical to specific DNA sequence on a specific bacterial chromosome were selected. The identified unique sequence can be used in different molecular diagnostic techniques, possibly to identify bacteria. In addition, a single primer that can identify multiple sites in a single chromosome can be exploited for region or genome identification. Although genomes sequences draft of isolates of organism DNA enable high throughput primer design using alignment strategy, and this enhances diagnostic performance in comparison to traditional molecular assays. In this method the generated primers can be used to identify an organism before the draft sequence is completed. In addition, the generated primers can be used to build a bank for easy access of the primers that can be used to identify bacteria.
Abstract: Zymomonas mobilis is known as an example of the
uncoupled growth phenomenon. This microorganism also has a
unique metabolism that degrades glucose by the Entner–Doudoroff
(ED) pathway. In this paper, a genome-scale metabolic model
including 434 genes, 757 reactions and 691 metabolites was
reconstructed to simulate uncoupled growth and study its effect on
flux distribution in the central metabolism. The model properly
predicted that ATPase was activated in experimental growth yields of
Z. mobilis. Flux distribution obtained from model indicates that the
major carbon flux passed through ED pathway that resulted in the
production of ethanol. Small amounts of carbon source were entered
into pentose phosphate pathway and TCA cycle to produce biomass
precursors. Predicted flux distribution was in good agreement with
experimental data. The model results also indicated that Z. mobilis
metabolism is able to produce biomass with maximum growth yield
of 123.7 g (mol glucose)-1 if ATP synthase is coupled with growth
and produces 82 mmol ATP gDCW-1h-1. Coupling the growth and
energy reduced ethanol secretion and changed the flux distribution to
produce biomass precursors.
Abstract: Prediction of perturbations after genetic manipulation
(especially gene knockout) is one of the important challenges in
systems biology. In this paper, a new algorithm is introduced that
integrates microarray data into the metabolic model. The algorithm
was used to study the change in the cell phenotype after knockout of
Gss gene in Escherichia coli BW25113. Algorithm implementation
indicated that gene deletion resulted in more activation of the
metabolic network. Growth yield was more and less regulating gene
were identified for mutant in comparison with the wild-type strain.
Abstract: A DNA microarray technology is a collection of microscopic DNA spots attached to a solid surface. Scientists use DNA microarrays to measure the expression levels of large numbers of genes simultaneously or to genotype multiple regions of a genome. Elucidating the patterns hidden in gene expression data offers a tremendous opportunity for an enhanced understanding of functional genomics. However, the large number of genes and the complexity of biological networks greatly increase the challenges of comprehending and interpreting the resulting mass of data, which often consists of millions of measurements. It is handled by clustering which reveals the natural structures and identifying the interesting patterns in the underlying data. In this paper, gene based clustering in gene expression data is proposed using Cuckoo Search with Differential Evolution (CS-DE). The experiment results are analyzed with gene expression benchmark datasets. The results show that CS-DE outperforms CS in benchmark datasets. To find the validation of the clustering results, this work is tested with one internal and one external cluster validation indexes.
Abstract: DNA Barcode provides good sources of needed
information to classify living species. The classification problem has
to be supported with reliable methods and algorithms. To analyze
species regions or entire genomes, it becomes necessary to use the
similarity sequence methods. A large set of sequences can be
simultaneously compared using Multiple Sequence Alignment which
is known to be NP-complete. However, all the used methods are still
computationally very expensive and require significant computational
infrastructure. Our goal is to build predictive models that are highly
accurate and interpretable. In fact, our method permits to avoid the
complex problem of form and structure in different classes of
organisms. The empirical data and their classification performances
are compared with other methods. Evenly, in this study, we present
our system which is consisted of three phases. The first one, is called
transformation, is composed of three sub steps; Electron-Ion
Interaction Pseudopotential (EIIP) for the codification of DNA
Barcodes, Fourier Transform and Power Spectrum Signal Processing.
Moreover, the second phase step is an approximation; it is
empowered by the use of Multi Library Wavelet Neural Networks
(MLWNN). Finally, the third one, is called the classification of DNA
Barcodes, is realized by applying the algorithm of hierarchical
classification.
Abstract: Availability of different genetic tests after completion
of Human Genome Project increases the physicians’ responsibility to
keep themselves update on the potential implementation of these
genetic tests in their daily practice. However, due to numbers of
barriers, still many of physicians are not either aware of these tests or
are not willing to offer or refer their patients for genetic tests. This
study was conducted an anonymous, cross-sectional, mailed-based
survey to develop a primary data of Malaysian physicians’ level of
knowledge and perception of gene profiling. Questionnaire had 29
questions. Total scores on selected questions were used to assess the
level of knowledge. The highest possible score was 11. Descriptive
statistics, one way ANOVA and chi-squared test was used for
statistical analysis. Sixty three completed questionnaires were
returned by 27 general practitioners (GPs) and 36 medical specialists.
Responders’ age ranges from 24 to 55 years old (mean 30.2 ± 6.4).
About 40% of the participants rated themselves as having poor level
of knowledge in genetics in general whilst 60% believed that they
have fair level of knowledge; however, almost half (46%) of the
respondents felt that they were not knowledgeable about available
genetic tests. A majority (94%) of the responders were not aware of
any lab or company which is offering gene profiling services in
Malaysia. Only 4% of participants were aware of using gene profiling
for detection of dosage of some drugs. Respondents perceived greater
utility of gene profiling for breast cancer (38%) compared to the
colorectal familial cancer (3%). The score of knowledge ranged from
2 to 8 (mean 4.38 ± 1.67). Non- significant differences between score
of knowledge of GPs and specialists were observed, with score of
4.19 and 4.58 respectively. There was no significant association
between any demographic factors and level of knowledge. However,
those who graduated between years 2001 to 2005 had higher level of
knowledge. Overall, 83% of participants showed relatively high level
of perception on value of gene profiling to detect patient’s risk of
disease. However, low perception was observed for both statements
of using gene profiling for general population in order to alter their
lifestyle (25%) as well as having the full sequence of a patient
genome for the purpose of determining a patient’s best match for
treatment (18%). The lack of clinical guidelines, limited provider
knowledge and awareness, lack of time and resources to educate
patients, lack of evidence-based clinical information and cost of tests
were the most barriers of ordering gene profiling mentioned by
physicians. In conclusion Malaysian physicians who participate in
this study had mediocre level of knowledge and awareness in gene
profiling. The low exposure to the genetic questions and problems
might be a key predictor of lack of awareness and knowledge on
available genetic tests. Educational and training workshop might be useful in helping Malaysian physicians incorporate genetic profiling
into practice for eligible patients.
Abstract: Agriculture is the backbone of economy of Pakistan
and cotton is the major agricultural export and supreme source of raw
fiber for our textile industry. To combat severe problems of insect
and weed, combination of three genes namely Cry1Ac, Cry2A and
EPSPS genes was transferred in locally cultivated cotton variety
MNH-786 with the use of Agrobacterium mediated genetic
transformation. The present study focused on the molecular screening
of transgenic cotton plants at T3 generation in order to confirm
integration and expression of all three genes (Cry1Ac, Cry2A and
EPSP synthase) into the cotton genome. Initially, glyphosate spray
assay was used for screening of transgenic cotton plants containing
EPSP synthase gene at T3 generation. Transgenic cotton plants which
were healthy and showed no damage on leaves were selected after 07
days of spray. For molecular analysis of transgenic cotton plants in
the laboratory, the genomic DNA of these transgenic cotton plants
were isolated and subjected to amplification of the three genes. Thus,
seventeen out of twenty (Cry1Ac gene), ten out of twenty (Cry2A
gene) and all twenty (EPSP synthase gene) were produced positive
amplification. On the base of PCR amplification, ten transgenic plant
samples were subjected to protein expression analysis through
ELISA. The results showed that eight out of ten plants were actively
expressing the three transgenes. Real-time PCR was also done to
quantify the mRNA expression levels of Cry1Ac and EPSP synthase
gene. Finally, eight plants were confirmed for the presence and active
expression of all three genes at T3 generation.
Abstract: The MyD88 is an evolutionarily conserved host-expressed adaptor protein that is essential for proper TLR/ IL1R immune-response signaling. A previously identified complete cDNA (1626 bp) of OfMyD88 comprised an ORF of 867 bp encoding a protein of 288 amino acids (32.9 kDa). The gDNA (3761 bp) of OfMyD88 revealed a quinquepartite genome organization composed of 5 exons (with the sizes of 310, 132, 178, 92 and 155 bp) separated by 4 introns. All the introns displayed splice signals consistent with the consensus GT/AG rule. A bipartite domain structure with two domains namely death domain (24-103) coded by 1st exon, and TIR domain (151-288) coded by last 3 exons were identified through in silico analysis. Moreover, homology modeling of these two domains revealed a similar quaternary folding nature between human and rock bream homologs. A comprehensive comparison of vertebrate MyD88 genes showed that they possess a 5-exonic structure.In this structure, the last three exons were strongly conserved, and this suggests that a rigid structure has been maintained during vertebrate evolution.A cluster of TATA box-like sequences were found 0.25 kb upstream of cDNA starting position. In addition, putative 5'-flanking region of OfMyD88 was predicted to have TFBS implicated with TLR signaling, including copies of NFkB1, APRF/ STAT3, Sp1, IRF1 and 2 and Stat1/2. Using qPCR technique, a ubiquitous mRNA expression was detected in liver and blood. Furthermore, a significantly up-regulated transcriptional expression of OfMyD88 was detected in head kidney (12-24 h; >2-fold), spleen (6 h; 1.5-fold), liver (3 h; 1.9-fold) and intestine (24 h; ~2-fold) post-Fla challenge. These data suggest a crucial role for MyD88 in antibacterial immunity of teleosts.
Abstract: Genome rearrangement is an important area in computational biology and bioinformatics. The basic problem in genome rearrangements is to compute the edit distance, i.e., the minimum number of operations needed to transform one genome into another. Unfortunately, unsigned genome rearrangement problem is NP-hard. In this study an improved ant colony optimization algorithm to approximate the edit distance is proposed. The main idea is to convert the unsigned permutation to signed permutation and evaluate the ants by using Kaplan algorithm. Two new operations are added to the standard ant colony algorithm: Replacing the worst ants by re-sampling the ants from a new probability distribution and applying the crossover operations on the best ants. The proposed algorithm is tested and compared with the improved breakpoint reversal sort algorithm by using three datasets. The results indicate that the proposed algorithm achieves better accuracy ratio than the previous methods.