Search results for: correlated genes

Authors: Ozlem Oral, Emre Taskin, Aysel Yuce, Serap Dokmeci, Devrim Gozuacik

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Autophagy is an evolutionary conserved lysosome-dependent catabolic pathway, responsible for the degradation of long-lived proteins, abnormal aggregates and damaged organelles which cannot be degraded by the ubiquitin-proteasome system. Lysosomes degrade the substrates through the activity of lysosomal hydrolases and lysosomal membrane-bound proteins. Mutations in the coding region of these proteins cause malfunctional lysosomes, which contributes to the pathogenesis of lysosomal storage diseases. Gaucher disease is a lysosomal storage disease resulting from the mutation of a lysosomal membrane-associated glycoprotein called glucocerebrosidase and its cofactor saposin C. The disease leads to intracellular accumulation of glucosylceramide and other glycolipids. Because of the essential role of lysosomes in autophagic degradation, Gaucher disease may directly be linked to this pathway. In this study, we investigated the expression of autophagy and/or lysosome-related genes and proteins in fibroblast cells isolated from patients with different mutations. We carried out confocal microscopy analysis and examined autophagic flux by utilizing the differential pH sensitivities of RFP and GFP in mRFP-GFP-LC3 probe. We also evaluated lysosomal pH by active lysosome staining and lysosomal enzyme activity. Beside lysosomes, we also performed proteasomal activity and cell death analysis in patient samples. Our data showed significant attenuation in the expression of key autophagy-related genes and accumulation of their proteins in mutant cells. We found decreased the ability of autophagosomes to fuse with lysosomes, associated with elevated lysosomal pH and reduced lysosomal enzyme activity. Proteasomal degradation and cell death analysis showed reduced proteolytic activity of the proteasome, which consequently leads to increased susceptibility to cell death. Our data indicate that the major degradation pathways are affected by multifunctional lysosomes in mutant patient cells and may underlie in the mechanism of clinical severity of Gaucher patients. (This project is supported by TUBITAK-3501-National Young Researchers Career Development Program, Project No: 112T130).

Keywords: autophagy, Gaucher's disease, glucocerebrosidase, mutant fibroblasts

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Authors: Leilani Forby, Farid Pazhoohi, Alan Kingstone

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There is abundant research on the nature and content of mind wandering (MW) in neurotypical (NT) adults, however, there is little to no research in these areas on autistic individuals. The objective of the current study was to uncover any differences between low and high autistic trait individuals in their MW. In particular, we examined their attitudes toward, and the themes and temporal dimensions (past, present, future) of, their MW episodes. For our online study, we recruited 518 students (394 women and 124 men), between the ages of 18 and 51 years (M = 20.93, SD = 3.40) from the undergraduate Human Subject Pool at the University of British Columbia. Participants completed the Short Imaginal Processes Inventory (SIPI), which includes the three subscales Positive-Constructive Daydreaming (SIPI-PC), Guilt and Fear of Failure Daydreaming (SIPI-GFF), and Poor Attentional Control (SIPI-PAC). Participants also completed the Past (IPI-past) and Present (IPI-present) subscales of the Imaginal Processes Inventory (IPI), the Deliberate (MW-D) and Spontaneous (MW-S) Mind Wandering scales, the Short Form Perceived Stress Scale (PSS-4), and the 10-item Autism Quotient (AQ-10). Results showed that overall, participant AQ-10 scores were significantly correlated with MW-S, SIPI-GFF, and PSS-4 scores, such that as the number of autistic traits endorsed by participants increased, so did their reports of spontaneous mind wandering, guilt and fear of failure themed day dreaming, and stress levels. This same pattern held for female participants, however, AQ-10 scores were positively correlated with only PSS-4 scores for males. These results suggest that compared to males with autistic traits, MW in females with autistic traits is more similar to individuals with low autistic traits in terms of content and intentionality. Results are discussed in terms of clinical implications, their limitations, and suggested directions for future research.

Keywords: autism, deliberate, mind wandering, spontaneous, perceived stress

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Authors: Ludmila A. Gavriliuc, Jerry McLarty, Heather E. Kleiner, J. Michael Mathis

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Abstract -- Background: The citrus coumarine Auraptene (Aur) is an effective chemopreventive agent, as manifested in many models of diseases and cancer. Nuclear factor erythroid 2-related factor (Nrf2) is an important regulator of genes induced by oxidative stress, such as glutathione S-transferases, heme oxygenase-1, and peroxiredoxin 1, by activating the antioxidant response element (ARE). Genetic and biochemical evidence has demonstrated that glutathione (GSH) and glutathione-dependent enzymes, glutathione reductase (GR), glutathione peroxidases (GPs), glutathione S-transferases (GSTs) are responsible for the control of intracellular reduction-oxidation status and participate in cellular adaptation to oxidative stress. The effect of Aur on the activity of GR, GPs (Se-GP and Se-iGP), and content of GSH in the liver, kidney, and spleen is insufficiently explored. Aim: Our goal was the examination of the Aur influence on the redox-system of GSH in Nrf2 wild type and Nrf2 knockout mice via activation of Nrf2 and ARE. Methods: Twenty female mice, 10 Nrf2 wild-type (WT) and 10 Nrf2 (-/-) knockout (KO), were bred and genotyped for our study. The activity of GR, Se-GP, Se-iGP, GST, G6PD, CytP450 reductase, catalase (Cat), and content of GSH were analyzed in the liver, kidney, and spleen using Spectrophotometry methods. The results of the specific activity of enzymes and the amount of GSH were analyzed with ANOVA and Spearman statistical methods. Results: Aur (200 mg/kg) treatment induced hepatic GST, GR, Se-GP activity and inhibited their activity in the spleen of mice, most likely via activation of the ARE through Nrf2. Activation in kidney Se-GP and G6PD by Aur is also controlled, apparently through Nrf2. Results of the non-parametric Spearman correlation analysis indicated the strong positive correlation between GR and G6PD only in the liver in WT control mice (r=+0.972; p < 0.005) and in the kidney KO control mice (r=+0.958; p < 0.005). The observed low content of GSH in the liver of KO mice indicated an increase in its participation in the neutralization of toxic substances with the absence of induction of GSH-dependent enzymes, such as GST, GR, Se-GP, and Se-iGP. Activation of CytP450 in kidney and spleen and Cat in the liver in KO mice probably revealed another regulatory mechanism for these enzymes. Conclusion: Thereby, obtained results testify that Aur can modulate the activity of genes and antioxidant enzymatic redox-system of GSH, responsible for the control of intracellular reduction-oxidation status.

Keywords: auraptene, glutathione, GST, Nrf2

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Authors: Zahra Fotovatnia, Jeffery A. Jones, Alexandra Gottardo

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Speech communication often occurs in environments where noise conceals part of a message. Listeners should compensate for the lack of auditory information by picking up distinct acoustic cues and using semantic and sentential context to recreate the speaker’s intended message. This situation seems to be more challenging in a nonnative than native language. On the other hand, early bilinguals are expected to show an advantage over the late bilingual and monolingual speakers of a language due to their better executive functioning components. In this study, English monolingual speakers were compared with early and late nonnative speakers of English to understand speech in noise processing (SIN) and the underlying neurobiological features of this phenomenon. Auditory mismatch negativities (MMNs) were recorded using a double-oddball paradigm in response to a minimal pair that differed in their middle vowel (beat/bit) at Wilfrid Laurier University in Ontario, Canada. The results did not show any significant structural and electroneural differences across groups. However, vocabulary knowledge correlated positively with performance on tests that measured SIN processing in participants who learned English after age 6. Moreover, their performance on the test negatively correlated with the integral area amplitudes in the left superior temporal gyrus (STG). In addition, the STG was engaged before the inferior frontal gyrus (IFG) in noise-free and low-noise test conditions in all groups. We infer that the pre-attentive processing of words engages temporal lobes earlier than the fronto-central areas and that vocabulary knowledge helps the nonnative perception of degraded speech.

Keywords: degraded speech perception, event-related brain potentials, mismatch negativities, brain regions

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Authors: Satwik Majumder, Dongyun Jung, Jennifer Ronholm, Saji George

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Bovine mastitis is the most common infectious disease in dairy cattle, with major economic implications for the dairy industry worldwide. Continuous monitoring for the emergence of antimicrobial resistance (AMR) among bacterial isolates from dairy farms is vital not only for animal husbandry but also for public health. In this study, the prevalence of AMR in 113 Escherichia coli isolates from cases of bovine clinical mastitis in Canada was investigated. Kirby-Bauer disk diffusion test with 18 antibiotics and microdilution method with three heavy metals (copper, zinc, and silver) was performed to determine the antibiotic and heavy-metal susceptibility. Resistant strains were assessed for efflux and ß-lactamase activities besides assessing biofilm formation and hemolysis. Whole-genome sequences for each of the isolates were examined to detect the presence of genes corresponding to the observed AMR and virulence factors. Phenotypic analysis revealed that 32 isolates were resistant to one or more antibiotics, and 107 showed resistance against at least one heavy metal. Quinolones and silver were the most efficient against the tested isolates. Among the AMR isolates, AcrAB-TolC efflux activity and ß-lactamase enzyme activities were detected in 13 and 14 isolates, respectively. All isolates produced biofilm but with different capacities, and 33 isolates showed α-hemolysin activity. A positive correlation (Pearson r = +0.89) between efflux pump activity and quantity of biofilm was observed. Genes associated with aggregation, adhesion, cyclic di-GMP, quorum sensing were detected in the AMR isolates, corroborating phenotype observations. This investigation showed the prevalence of AMR in E. coli isolates from bovine clinical mastitis. The results also suggest the inadequacy of antimicrobials with a single mode of action to curtail AMR bacteria with multiple mechanisms of resistance and virulence factors. Therefore, it calls for combinatorial therapy for the effective management of AMR infections in dairy farms and combats its potential transmission to the food supply chain through milk and dairy products.

Keywords: antimicrobial resistance, E. coli, bovine mastitis, antibiotics, heavy-metals, efflux pump, ß-lactamase enzyme, biofilm, whole-genome sequencing

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Authors: Hussain M. Al-foudari, Weizhong Chen, James M. Bishop

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Correlations between oceanographic variables and monthly catch rates of total shrimp and those of each of the major species (Penaeus semisulcatus, Metapenaeus affinis and Parapenaeopsis stylifera) showed significant differences for particular conditions. Catches of P. semisulcatus were basically positively correlated with temperature, i.e., the higher the temperature, the higher the catch rate, while those of M. affinis and P. stylifera were negatively correlated with temperature, i.e., high catch rates occurred in the low temperature waters. Thus, during the months January and April, P. semisulcatus preferred waters with high temperature, usually the offshore and southern areas, while M. affinis and P. stylifera preferred waters with low temperature, usually inshore and northern areas. The relationships between the catch rate of P. semisulcatus and salinity were not so clear. Results indicated that although salinity was one of the factors affecting the distribution of P. semisulcatus, it was not the principal factor, and impacts from other variables, such as temperature, might overshadow the correlation between the catch rates of P. semisulcatus and salinity. The relationship between shrimp catch rates and dissolved oxygen (DO) also showed mixed results. The catch rates of M. affinis increased with a decrease of surface DO in November 2013, but decreased with lower bottom DO in December. These results indicated that DO might be a factor affecting distributions of the shrimp; however; the true correlation between catch rate and DO might be easily overshadowed by other environmental variables. Catch rates of P. semisulcatus did not show any relationship with depth. P. semisulcatus is a migratory species and widely distributed in Kuwait's waters.During the shrimp season from July through December, P. semisulcatus occurs in almost all areas in Kuwait's waters irrespective of water depth. The catch rates of M. affinis and P. stylifera, however, showed clear relationships with depth. Both species had significantly higher catch rates in shallower waters, indicative of their restricted distribution.

Keywords: Kuwait, Penaeus semisulcatus, Metapenaeus affinis, Parapenaeopsis stylifera, Arabian gulf

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Authors: Wael Ali Mohammed Hadi

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Pseudomonas fluorescens B29B, which has asparaginase enzymatic activity, was isolated from the surface coastal seawater of Trivandrum, India. We report the complete Pseudomonas fluorescens B29B genome sequenced, identified, and annotated from a marine source. We find the genome at most minuscule a 7,331,508 bp single circular chromosome with a GC content of 62.19% and 6883 protein-coding genes. Three hundred forty subsystems were identified, including two predicted asparaginases from the genome analysis of P. fluorescens B29B for further investigation. This genome data will help further industrial biotechnology applications of proteins in general and asparaginase as a target.

Keywords: pseudomonas, marine, asparaginases, Kerala, whole-genome

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Authors: Anthony Khawaja, Jacques Prioux, Ghassan Maalouf, Rawad El Hage

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The regular practice of physical activities characterized by significant mechanical stresses stimulates bone formation and improves bone mineral density (BMD) in the most solicited sites. The purpose of this study was to explore the relationships between maximum power and bone variables in a group of young adult men. Identification of new determinants of BMD, bone mineral content (BMC) and hip geometric indices in young adult men, would allow screening and early management of future cases of osteopenia and osteoporosis. Fifty-three young adult men (18 – 35yr) voluntarily participated in this study. Weight and height were measured, and body mass index was calculated. Body composition, BMC and BMD were determined for each individual by Dual-energy X-ray absorptiometry (DXA; GE Healthcare, Madison, WI) at whole body (WB), lumbar spine (L1-L4), total hip (TH), and femoral neck (FN). FN cross-sectional area (CSA), strength index (SI), buckling ratio (BR), FN section modulus (Z), cross-sectional moment of inertia (CSMI) and L1-L4 TBS were also evaluated by DXA. The vertical jump was evaluated using a field test (sargent test). Two main parameters were retained: vertical jump performance (cm) and power (w). The subjects performed three jumps with 2 minutes of recovery between jumps. The highest vertical jump was selected. Maximum power (P max, in watts) was calculated. Maximum power was positively correlated to WB BMD (r = 0.41; p < 0.01), WB BMC (r = 0.65; p < 0.001), L1-L4 BMC (r = 0.54; p < 0.001), FN BMC (r = 0.35; p < 0.01), TH BMC (r = 0.50; p < 0.001), CSMI (r = 0.50; p < 0.001), CSA (r = 0.33; p < 0.05). Vertical jump was positively correlated to WB BMC (r = 0.31; p < 0.05), L1-L4 BMC (r = 0.40; p < 0.01), CSMI (r = 0.29; p < 0.05). The current study suggests that maximum power is a positive determinant of BMD, BMC and hip geometric indices in young adult men. In addition, it shows also that maximum power is a stronger positive determinant of bone variables than vertical jump in this population. Implementing strategies to increase maximum power in young adult men may be useful for preventing osteoporotic fractures later in life.

Keywords: bone variables, maximum power, osteopenia, osteoporosis, vertical jump, young adult men

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Authors: Rayanne Branco Dos Santos Lima, Lorena Pinheiro Barbosa, Kamila Ferreira Lima, Victor Manuel Tegoma Ruiz, Monyka Brito Lima Dos Santos, Maria Wendiane Gueiros Gaspar, Luzia Camila Coelho Ferreira, Leandro Cardozo Dos Santos Brito, Deyse Maria Alves Rocha

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Introduction: The lack of regulation of the baby's sleep-wake pattern in the first years of life affects the health of thousands of women. Maternal sleep deprivation can trigger or aggravate psychosomatic problems such as depression, anxiety and stress that can directly influence maternal safety, with consequences for the baby's and mother's sleep. Such conditions can affect the family's quality of life and child development. Objective: To correlate maternal security with maternal state anxiety scores and the mother's and baby's total sleep time. Method: Cross-sectional study carried out with 96 mothers of babies aged 10 to 24 months, accompanied by nursing professionals linked to a Federal University in Northeast Brazil. Study variables were maternal security, maternal state anxiety scores, infant latency and sleep time, and total nocturnal sleep time of mother and infant. Maternal safety was calculated using a four-point Likert scale (1=not at all safe, 2=somewhat safe, 3=very safe, 4=completely safe). Maternal anxiety was measured by State-Trait Anxiety Inventory, state-anxiety subscale whose scores vary from 20 to 80 points, and the higher the score, the higher the anxiety levels. Scores below 33 are considered mild; from 33 to 49, moderate and above 49, high. As for the total nocturnal sleep time, values between 7-9 hours of sleep were considered adequate for mothers, and values between 9-12 hours for the baby, according to the guidelines of the National Sleep Foundation. For the sleep latency time, a time equal to or less than 20 min was considered adequate. It is noteworthy that the latency time and the time of night sleep of the mother and the baby were obtained by the mother's subjective report. To correlate the data, Spearman's correlation was used in the statistical package R version 3.6.3. Results: 96 women and babies participated, aged 22 to 38 years (mean 30.8) and 10 to 24 months (mean 14.7), respectively. The average of maternal security was 2.89 (unsafe); Mean maternal state anxiety scores were 43.75 (moderate anxiety). The babies' average sleep latency time was 39.6 min (>20 min). The mean sleep times of the mother and baby were, respectively, 6h and 42min and 8h and 19min, both less than the recommended nocturnal sleep time. Maternal security was positively correlated with maternal state anxiety scores (rh=266, p=0.009) and negatively correlated with infant sleep latency (rh= -0.30. P=0.003). Baby sleep time was positively correlated with maternal sleep time. (rh 0.46, p<0.001). Conclusion: The more secure the mothers considered themselves, the higher the anxiety scores and the shorter the baby's sleep latency. Also, the longer the baby sleeps, the longer the mother sleeps. Thus, interventions are needed to promote the quality and efficiency of sleep for both mother and baby.

Keywords: sleep, anxiety, infant, mother-child relations

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Authors: Mihai Gheorghe

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Freelancing in IT has seen an increased popularity during the last years mainly because of the fast Internet adoption in the countries with emerging economies, correlated with the continuous seek for reduced development costs as well with the rise of online platforms which address planning, coordination, and various development tasks. This paper conducts an overview of the most relevant Freelance Marketplaces available and studies the market structure, distribution of the workforce and trends in IT freelancing.

Keywords: freelancing in IT, freelance marketplaces, freelance market structure, globalization, online staffing, trends in freelancing

Procedia PDF Downloads 207

Authors: Shazia Anwer Bukhari, Madiha Javeed Ghani, Muhammad Ibrahim Rajoka

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Objective: Biochemical, environmental, physical and genetic factors have a strong effect on the development of coronary disease (CAD). Plasma homocysteine (Hcy) level and DNA damage play a pivotal role in its development and progression. The aim of this study was to investigate the predictive strength of an oxidative stress, clinical biomarkers and total antioxidant status (TAS) in CAD patients to find the correlation of homocysteine, TOS and oxidative DNA damage with other clinical parameters. Methods: Sixty confirmed patients with CAD and 60 healthy individuals as control were included in this study. Different clinical and laboratory parameters were studied in blood samples obtained from patients and control subjects using commercially available biochemical kits and statistical software Results: As compared to healthy individuals, CAD patients had significantly higher concentrations of indices of oxidative stress: homocysteine (P=0.0001), total oxidative stress (TOS) (P=0.0001), serum cholesterol (P=0.04), low density lipoprotein cholesterol (LDL) (P=0.01), high density lipoprotein-cholesterol (HDL) (P=0.0001), and malondialdehyde (MDA) (P=0.001) than those of healthy individuals. Plasma homocysteine level and oxidative DNA damage were positively correlated with cholesterol, triglycerides, systolic blood pressure, urea, total protein and albumin (P values= 0.05). Both Hcy and oxidative DNA damage were negatively correlated with TAS and proteins. Conclusion: Coronary artery disease patients had a significant increase in homocysteine level and DNA damage due to increased oxidative stress. In conclusion, our study shows a significantly increase in lipid peroxidation, TOS, homocysteine and DNA damage in the erythrocytes of patients with CAD. A significant decrease level of HDL-C and TAS was observed only in CAD patients. Therefore these biomarkers may be useful diagnosis of patients with CAD and play an important role in the pathogenesis of CAD.

Keywords: antioxidants, coronary artery disease, DNA damage, homocysteine, oxidative stress, malondialdehyde, 8-Hydroxy-2’deoxyguanosine

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Authors: E. Al Daoud

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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.

Keywords: genome, species, blastp, conserved genes, Cronobacter

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Authors: Tai Chen, Caihuan Tian, Xiuxia Ren, Jingqi Xue, Xiuxin Zhang

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The herbaceous peony has become increasingly popular worldwide in recent years, especially as a cut flower with great economic value. However, peony has a very short vase life, only 3-5 d usually, which seriously affects its commodity value. In this study, we used the cut peony (Paeonia lactiflora cv. Sarah) as a material and found that melatonin treatment significantly improved its postharvest performance. In the control group, its vase life was 4.8 d, accompanied by petal dropping at last; melatonin treatment (40 μM) increased this time to 6.9 d without petal dropping at the end. Further study showed that melatonin treatment significantly increased the activity of antioxidant enzymes as well as reduced sugar content in petals, whereas the starch content in petals decreased. These results indicated that melatonin treatment may delay the oxidation reaction caused by aging, which also provides extra energy for maintaining flowering. Through full-length transcriptome sequencing, a total of 2819 differentially expressed genes (DEGs) between control and melatonin treatment groups were identified. KEGG enrichment analysis showed that these DEGs were mainly involved in three pathways, including melatonin synthesis, starch and sucrose conversion, and plant disease resistance. After the RT-qPCR verification, we identified three DEGs, named PlBAM3, PlWRKY22 and PlTIP1, and they should play major roles in melatonin-improved postharvest performance. One possible reason is that PlBAM3 caused maltose production (by starch degradation), maintained the proline biosynthesis, and then alleviated oxidative stress. Another reason is that both PlBAM3 and PlWRKY22 are key drought resistance regulators, which have the ability to alleviate osmotic stress and improve water absorption, which may also help to improve the postharvest quality of cut peony. In addition, PlTIP1 is involved in the sugar signal pathway, indicating sugar may also as a signal substance during this process. Our work may give new ideas for developing new ways to prolong the vase life of cut peony and improve its commodity value eventually.

Keywords: cut peony, melatonin, vase life, oxidation reaction, energy supply, differentially expressed genes

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Authors: B. Makoudi, R. Ghanimi, A. Bargaz, M. Mouradi, M. Farissi, A. Kabbaj, J. J. Drevon, C. Ghoulam

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Legume species are able to establish a nitrogen fixing symbiosis with soil rhizobia that allows them, when it operates normally, to ensure their necessary nitrogen nutrition. This biological process needs high phosphorus (P) supply and consequently it is limited under low phosphorus availability. To overcome this constraint, legume-rhizobia symbiosis develops many mechanisms to increase P availability in the rhizosphere and also the efficiency of P fertilizers. The objectives of our research works are to understand the physiological and biochemical mechanisms implemented by legume-rhizobia symbiosis to increase its P use efficiency (PUE) in order to select legume genotypes-rhizobia strains combination more performing for BNF under P deficiency. Our studies were carried out on two grain legume species, common bean (Phaseolus vulgaris) and faba bean (Vicia faba) tested in farmers’ fields and in experimental station fewer than two soil phosphorus levels. Under field conditions, the P deficiency caused a significant decrease of Plant and nodule biomasses in all of the tested varieties with a difference between them. This P limitation increased the contents of available P in the rhizospheric soils that was positively correlated with the increase of phosphatases activities in the nodules and the rhizospheric soil. Some legume genotypes showed a significant increase of their P use efficiency under P deficiency. The P solubilization test showed that some rhizobia strains isolated from Haouz region presented an important capacity to grow on solid and liquid media with tricalcium phosphate as the only P source and their P solubilizing activity was confirmed by the assay of the released P in the liquid medium. Also, this P solubilizing activity was correlated with medium acidification and the excretion of acid phosphatases and phytases in the medium. Thus, we concluded that medium acidification and excretion of phosphatases in the rhizosphere are the prominent reactions for legume-rhizobia symbiosis to improve its P nutrition.

Keywords: legume, phosphorus deficiency, rhizobia, rhizospheric soil

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Authors: Brenda Russell

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Police officers spend a great deal of time responding to domestic violence calls. Recent research has found that men and women in heterosexual and same-sex relationships are equally likely to initiate intimate partner violence IPV) and likewise susceptible to victimization, yet police training tends to focus primarily on male perpetration and female victimization. Criminal justice studies have found that male perpetrators of IPV are blamed more than female perpetrators who commit the same offense. While previous research has examined officer’s response in IPV cases with male and female heterosexual offenders, research has yet to investigate police response in same-sex relationships. This study examined officers’ decisions to arrest, perceptions of blame, perceived danger to others, disrespect, and beliefs in prosecution, guilt and sentencing. Officers in the U.S. (N = 248) were recruited using word of mouth and access to police association websites where a link to an online study was made available. Officers were provided with one of 4 experimentally manipulated scenarios depicting a male or female perpetrator (heterosexual or same-sex) in a clear domestic assault situation. Officer age, experience with IPV and IPV training were examined as possible covariates. Training in IPV was not correlated to any dependent variable of interest. Age was correlated with perpetrator arrest and blame (.14 and .16, respectively) and years of experience was correlated to arrest, offering informal advice, and mediating the incident (.14 to -.17). A 2(perpetrator gender) X 2 (victim gender) factorial design was conducted. Results revealed that officers were more likely to provide informal advice and mediate in gay male relationships, and were less likely to arrest perpetrators in same-sex relationships. When officer age and years of experience with domestic violence were statistically controlled, effects for perpetrator arrest and providing informal advice were no longer significant. Officers perceived heterosexual male perpetrators as more dangerous, blameworthy, disrespectful, and believed they would receive significantly longer sentences than all other conditions. When officer age and experience were included as covariates in the analyses perpetrator blame was no longer statistically significant. Age, experience and training in IPV were not related to perceptions of victims. Police perceived victims as more truthful and believable when the perpetrator was a male. Police also believed victims of female perpetrators were more responsible for their own victimization. Victims were more likely to be perceived as a danger to their family when the perpetrator was female. Female perpetrators in same-sex relationships and heterosexual males were considered to experience more mental illness than heterosexual female or gay male perpetrators. These results replicate previous research suggesting male perpetrators are more blameworthy and responsible for their own victimization, yet expands upon previous research by identifying potential biases in police response to IPV in same-sex relationships. This study brings to the forefront the importance of evidence-based officer training in IPV and provides insight into the need for a gender inclusive approach as well as addressing the necessity of the practical applications for police.

Keywords: domestic violence, heterosexual, intimate partner violence, officer response, police officer, same-sex

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Authors: Amr Saeb, Khalid Al-Rubeaan, Mohamed Abouelhoda, Manojkumar Selvaraju, Hamsa Tayeb

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Background: P. mirabilis is a common uropathogenic bacterium that can cause major complications in patients with long-standing indwelling catheters or patients with urinary tract anomalies. In addition, P. mirabilis is a common cause of chronic osteomyelitis in diabetic foot ulcer (DFU) patients. Methodology: P. mirabilis SCDR1 was isolated from a diabetic ulcer patient. We examined P. mirabilis SCDR1 levels of resistance against nano-silver colloids, the commercial nano-silver and silver containing bandages and commonly used antibiotics. We utilized next generation sequencing techniques (NGS), bioinformatics, phylogenetic analysis and pathogenomics in the identification and characterization of the infectious pathogen. Results: P. mirabilis SCDR1 is a multi-drug resistant isolate that also showed high levels of resistance against nano-silver colloids, nano-silver chitosan composite and the commercially available nano-silver and silver bandages. The P. mirabilis-SCDR1 genome size is 3,815,621 bp with G+C content of 38.44%. P. mirabilis-SCDR1 genome contains a total of 3,533 genes, 3,414 coding DNA sequence genes, 11, 10, 18 rRNAs (5S, 16S, and 23S), and 76 tRNAs. Our isolate contains all the required pathogenicity and virulence factors to establish a successful infection. P. mirabilis SCDR1 isolate is a potential virulent pathogen that despite its original isolation site, wound, it can establish kidney infection and its associated complications. P. mirabilis SCDR1 contains several mechanisms for antibiotics and metals resistance including, biofilm formation, swarming mobility, efflux systems, and enzymatic detoxification. Conclusion: P. mirabilis SCDR1 is the spontaneous nano-silver resistant bacterial strain. P. mirabilis SCDR1 strain contains all reported pathogenic and virulence factors characteristic for the species. In addition, it possesses several mechanisms that may lead to the observed nano-silver resistance.

Keywords: Proteus mirabilis, multi-drug resistance, silver nanoparticles, resistance, next generation sequencing techniques, genome analysis, bioinformatics, phylogeny, pathogenomics, diabetic foot ulcer, xenobiotics, multidrug resistance efflux, biofilm formation, swarming mobility, resistome, glutathione S-transferase, copper/silver efflux system, altruism

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Authors: Lina E. Homman, Alexis C. Edwards, Seung Bin Cho, Danielle M. Dick, Kenneth S. Kendler

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Research supports an association between alcohol problems and internalising symptoms, but the understanding of how the two phenotypes relate to each other is poor. It has been hypothesized that the relationship between the phenotypes is causal; however investigations in regards to direction are inconsistent. Clarity of the relationship between the two phenotypes may be provided by investigating the phenotypes developmental inter-relationships longitudinally. The objective of the study was to investigate a) changes in alcohol problems and internalising symptoms in college students across time and b) the direction of effect of growth between alcohol problems and internalising symptoms from late adolescent to emerging adulthood c) possible gender differences. The present study adds to the knowledge of comorbidity of alcohol problems and internalising symptoms by examining a longitudinal sample of college students and by examining the simultaneous development of the symptoms. A sample of college students is of particular interest as symptoms of both phenotypes often have their onset around this age. A longitudinal sample of college students from a large, urban, public university in the United States was used. Data was collected over a time period of 2 years at 3 time points. Latent growth models were applied to examine growth trajectories. Parallel process growth models were used to assess whether initial level and rate of change of one symptom affected the initial level and rate of change of the second symptom. Possible effects of gender and ethnicity were investigated. Alcohol problems significantly increased over time, whereas internalizing symptoms remained relatively stable. The two phenotypes were significantly correlated in each wave, correlations were stronger among males. Initial level of alcohol problems was significantly positively correlated with initial level of internalising symptoms. Rate of change of alcohol problems positively predicted rate of change of internalising symptoms for females but not for males. Rate of change of internalising symptoms did not predict rate of change of alcohol problems for either gender. Participants of Black and Asian ethnicities indicated significantly lower levels of alcohol problems and a lower increase of internalising symptoms across time, compared to White participants. Participants of Black ethnicity also reported significantly lower levels of internalising symptoms compared to White participants. The present findings provide additional support for a positive relationship between alcohol problems and internalising symptoms in youth. Our findings indicated that both internalising symptoms and alcohol problems increased throughout the sample and that the phenotypes were correlated. The findings mainly implied a bi-directional relationship between the phenotypes in terms of significant associations between initial levels as well as rate of change. No direction of causality was indicated in males but significant results were found in females where alcohol problems acted as the main driver for the comorbidity of alcohol problems and internalising symptoms; alcohol may have more detrimental effects in females than in males. Importantly, our study examined a population-based longitudinal sample of college students, revealing that the observed relationships are not limited to individuals with clinically diagnosed mental health or substance use problems.

Keywords: alcohol, comorbidity, internalising symptoms, longitudinal modelling

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Authors: Cyril Kaplan

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26 participants played 4 EEG neurofeedback (NF) games encouraged to find their strategies to control the specific NF parameter. Mixed method analysis of performance in the games and post-session interviews led to the identification of states of consciousness that correlated with success in the game. We found that increase in left frontal beta activity was facilitated by evoking interest in observed surroundings, by wondering what is happening behind the window or what lies in a drawer in front.

Keywords: EEG neurofeedback, states of consciousness, frontal beta activity, mixed methods

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Authors: Essam Al Daoud

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This study solves a phylogeny problem by using modified wild dog pack optimization. The least squares error is considered as a cost function that needs to be minimized. Therefore, in each iteration, new distance matrices based on the constructed trees are calculated and used to select the alpha dog. To test the suggested algorithm, ten homologous genes are selected and collected from National Center for Biotechnology Information (NCBI) databanks (i.e., 16S, 18S, 28S, Cox 1, ITS1, ITS2, ETS, ATPB, Hsp90, and STN). The data are divided into three categories: 50 taxa, 100 taxa and 500 taxa. The empirical results show that the proposed algorithm is more reliable and accurate than other implemented methods.

Keywords: least square, neighbor joining, phylogenetic tree, wild dog pack

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Authors: Liat Kulik

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The main goal of the present study was to examine gender differences in the variables that explain the experience of role conflict and well-being among Jewish working fathers and mothers in the Israel. The experience of work-family conflict arises from simultaneous pressures from the work and family domains that are mutually incompatible. In light of the expansion of women's role set following the addition of paid employment outside of the home, most of the studies dealing with the impact of multiple roles on well-being have been conducted among women. However, changes in gender roles in recent years have also affected men's role set, as reflected in the terms ‘new men’ and ‘new fathers’. Based on structural equation modeling, the study examined gender differences in variables that explain the experience of two types of role conflict – family interferes with work (FIW) and work interferes with family (WIF), as well as with the sense of well-being (positive and negative affect) among 611 employed Jewish mothers and fathers in Israel. The findings revealed that for women, both FIW and WIF conflict correlated negatively with well-being, whereas for men, a negative correlation with well-being was found only in the case of FIW conflict. For both men and women, egalitarian gender role ideology correlated with the dimension of positive effect, but the correlation was stronger for men. The findings highlight the contribution of egalitarian gender role ideology to alleviating the experience of role conflict and improving the emotional well-being of both men and women. Contrary to expectations, social support contributed more to mitigating negative effect among men than women. On the whole, the findings highlight the changes that men have experienced in the work-family system. In sum, the research findings shed new light on the masculine image in terms of the experience of FIW conflict. In contrast to the prevailing assumption that FIW role conflict is predominant among women, the findings of this study indicate that today, this type of role conflict is experienced equally by men and women whereas WIF conflict is predominant among men. Furthermore, contrary to expectations, levels of perceived social support were found to be similar for men and women, and men benefited from it even more than women did.

Keywords: FIW conflict, WIF conflict, social support, egalitarian gender role ideology, overload

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Authors: Khiem M. Nguyen, Ming C. Yang

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Chloroplast pigments are extremely important during photosynthesis since they play essential roles in light absorption and energy transfer. Therefore, understanding the efficiency of chlorophyll (Chl) biosynthesis could facilitate enhancement in photo-assimilates accumulation, and ultimately, in crop yield. The Chl-deficient mutants have been used extensively to study the Chl biosynthetic pathways and the biogenesis of the photosynthetic apparatus. Rice (Oryza sativa L.) is one of the most leading food crops, serving as staple food for many parts of the world. To author’s best knowledge, Chl b–lacking rice has been found; however the molecular mechanism of Chl biosynthesis still remains unclear compared to wild-type rice. In this study, the ultrastructure analysis, photosynthetic properties, and transcriptome profile of wild-type rice (Norin No.8, N8) and its Chl b-lacking mutant (Chlorina 1, C1) were examined. The finding concluded that total Chl content and Chl b content in the C1 leaves were strongly reduced compared to N8 leaves, suggesting that reduction in the total Chl content contributes to leaf color variation at the physiological level. Plastid ultrastructure of C1 possessed abnormal thylakoid membranes with loss of starch granule, large number of vesicles, and numerous plastoglobuli. The C1 rice also exhibited thinner stacked grana, which was caused by a reduction in the number of thylakoid membranes per granum. Thus, the different Chl a/b ratio of C1 may reflect the abnormal plastid development and function. Transcriptional analysis identified 23 differentially expressed genes (DEGs) and 671 transcription factors (TFs) that were involved in Chl metabolism, chloroplast development, cell division, and photosynthesis. The transcriptome profile and DEGs revealed that the gene encoding PsbR (PSII core protein) was down-regulated, therefore suggesting that the lower in light-harvesting complex proteins are responsible for the lower photosynthetic capacity in C1. In addition, expression level of cell division protein (FtsZ) genes were significantly reduced in C1, causing chloroplast division defect. A total of 19 DEGs were identified based on KEGG pathway assignment involving Chl biosynthesis pathway. Among these DEGs, the GluTR gene was down-regulated, whereas the UROD, CPOX, and MgCH genes were up-regulated. Observation through qPCR suggested that later stages of Chl biosynthesis were enhanced in C1, whereas the early stages were inhibited. Plastid structure analysis together with transcriptomic analysis suggested that the Chl a/b ratio was amplified both by the reduction in Chl contents accumulation, owning to abnormal chloroplast development, and by the enhanced conversion of Chl b to Chl a. Moreover, the results indicated the same Chl-cycle pattern in the wild-type and C1 rice, indicating another Chl b degradation pathway. Furthermore, the results demonstrated that normal grana stacking, along with the absence of Chl b and greatly reduced levels of Chl a in C1, provide evidence to support the conclusion that other factors along with LHCII proteins are involved in grana stacking. The findings of this study provide insight into the molecular mechanisms that underlie different Chl a/b ratios in rice.

Keywords: Chl-deficient mutant, grana stacked, photosynthesis, RNA-Seq, transcriptomic analysis

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Authors: Briony Gray, Glenn Simpson, Hajira Dambha-Miller, Andrew Farmer

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Multimorbidity may be strongly affected by a variety of conditions, factors, and variables requiring higher demands on health and social care services, infrastructure, and expenses. Holding one or more conditions increases one’s risk for development of future conditions; with patients over 65 years old at highest risk. Psychosocial factors such as anxiety and depression are rising exponentially globally, which has been amplified by the COVID19 pandemic. These are highly correlated and predict poorer outcomes when held in coexistence and increase the likelihood of comorbid physical health conditions. While possible future reform of social and healthcare systems may help to alleviate some of these mounting pressures, there remains an urgent need to better understand the potential role health behaviours and psychosocial conditions - such as anxiety and depression – may have on aging populations. Using the UK healthcare scene as a lens for analysis, this study uses big data collected in the UK Longitudinal Study of Aging (ELSA) to examine the role of anxiety and depression in ageing populations (65yrs+). Using logistic regression modelling, results identify the 10 most significant variables correlated with both anxiety and depression from data categorised into the areas of health behaviour, psychosocial, socioeconomic, and life satisfaction (each demonstrated through literature review to be of significance). These are compared with wider global research findings with the aim of better understanding the areas in which social and healthcare reform can support multimorbidity interventions, making suggestions for improved patient-centred care. Scope of future research is outlined, which includes analysis of 59 total multimorbidity variables from the ELSA dataset, going beyond anxiety and depression.

Keywords: multimorbidity, health behaviours, patient centred care, psychosocial factors

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Authors: Judy M. Obliosca, Olivia Vest, Sandra Poulos, Kelsi Smith, Tammy Ferguson, Abigail Powers Lott, Alicia K. Smith, Yang Xu, Christopher K. Tison

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Post-Traumatic Stress Disorder (PTSD) is a mental health problem that people may develop after experiencing traumatic events such as combat, natural disasters, and major emotional challenges. Tragically, the number of military personnel with PTSD correlates directly with the number of veterans who attempt suicide, with the highest rate in the Army. Research has shown epigenetic risks in those who are prone to several psychiatric dysfunctions, particularly PTSD. Once initiated in response to trauma, epigenetic alterations in particular, the DNA methylation in the form of 5-methylcytosine (5mC) alters chromatin structure and represses gene expression. Current methods to detect DNA methylation, such as bisulfite-based genomic sequencing techniques, are laborious and have massive analysis workflow while still having high error rates. A faster and simpler detection method of high sensitivity and precision would be useful in a clinical setting to confirm potential PTSD etiologies, prevent other psychiatric disorders, and improve military health. A nano-enhanced Surface Plasmon Resonance imaging (SPRi)-based assay that simultaneously detects site-specific 5mC base (termed as PTSD base) in methylated genes related to PTSD is being developed. The arrays on a sensing chip were first constructed for parallel detection of PTSD bases using synthetic and genomic DNA (gDNA) samples. For the gDNA sample extracted from the whole blood of a PTSD patient, the sample was first digested using specific restriction enzymes, and fragments were denatured to obtain single-stranded methylated target genes (ssDNA). The resulting mixture of ssDNA was then injected into the assay platform, where targets were captured by specific DNA aptamer probes previously immobilized on the surface of a sensing chip. The PTSD bases in targets were detected by anti-5-methylcytosine antibody (anti-5mC), and the resulting signals were then enhanced by the universal nanoenhancer. Preliminary results showed successful detection of a PTSD base in a gDNA sample. Brighter spot images and higher delta values (control-subtracted reflectivity signal) relative to those of the control were observed. We also implemented the in-house surface activation system for detection and developed SPRi disposable chips. Multiplexed PTSD base detection of target methylated genes in blood DNA from PTSD patients of severity conditions (asymptomatic and severe) was conducted. This diagnostic capability being developed is a platform technology, and upon successful implementation for PTSD, it could be reconfigured for the study of a wide variety of neurological disorders such as traumatic brain injury, Alzheimer’s disease, schizophrenia, and Huntington's disease and can be extended to the analyses of other sample matrices such as urine and saliva.

Keywords: epigenetic assay, DNA methylation, PTSD, whole blood, multiplexing

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Authors: Alexander A. Stakheev, Larisa V. Samokhvalova, Sergey K. Zavriev

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Fusarium fungi are among the most devastating plant pathogens distributed all over the world. Significant reduction of grain yield and quality caused by Fusarium leads to multi-billion dollar annual losses to the world agricultural production. These organisms can also cause infections in immunocompromised persons and produce the wide range of mycotoxins, such as trichothecenes, fumonisins, and zearalenone, which are hazardous to human and animal health. Identification of Fusarium fungi based on the morphology of spores and spore-forming structures, colony color and appearance on specific culture media is often very complicated due to the high similarity of these features for closely related species. Modern Fusarium taxonomy increasingly uses data of crossing experiments (biological species concept) and genetic polymorphism analysis (phylogenetic species concept). A number of novel Fusarium sibling species has been established using DNA barcoding techniques. Species recognition is best made with the combined phylogeny of intron-rich protein coding genes and ribosomal DNA sequences. However, the internal transcribed spacer of (ITS), which is considered to be universal DNA barcode for Fungi, is not suitable for genus Fusarium, because of its insufficient variability between closely related species and the presence of non-orthologous copies in the genome. Nowadays, the translation elongation factor 1 alpha (TEF1α) gene is the “gold standard” of Fusarium taxonomy, but the search for novel informative markers is still needed. In this study, we used two novel DNA markers, frataxin (FXN) and heat shock protein 90 (HSP90) to discover phylogenetic relationships between Fusarium species. Multilocus phylogenetic analysis based on partial sequences of TEF1α, FXN, HSP90, as well as intergenic spacer of ribosomal DNA (IGS), beta-tubulin (β-TUB) and phosphate permease (PHO) genes has been conducted for 120 isolates of 19 Fusarium species from different climatic zones of Russia and neighboring countries using maximum likelihood (ML) and maximum parsimony (MP) algorithms. Our analyses revealed that FXN and HSP90 genes could be considered as informative phylogenetic markers, suitable for evolutionary and taxonomic studies of Fusarium genus. It has been shown that PHO gene possesses more variable (22 %) and parsimony informative (19 %) characters than other markers, including TEF1α (12 % and 9 %, correspondingly) when used for elucidating phylogenetic relationships between F. avenaceum and its closest relatives – F. tricinctum, F. acuminatum, F. torulosum. Application of novel DNA barcodes confirmed the fact that F. arthrosporioides do not represent a separate species but only a subspecies of F. avenaceum. Phylogeny based on partial PHO and FXN sequences revealed the presence of separate cluster of four F. avenaceum strains which were closer to F. torulosum than to major F. avenaceum clade. The strain F-846 from Moldova, morphologically identified as F. poae, formed a separate lineage in all the constructed dendrograms, and could potentially be considered as a separate species, but more information is needed to confirm this conclusion. Variable sites in PHO sequences were used for the first-time development of specific qPCR-based diagnostic assays for F. acuminatum and F. torulosum. This work was supported by Russian Foundation for Basic Research (grant № 15-29-02527).

Keywords: DNA barcode, fusarium, identification, phylogenetics, taxonomy

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Authors: Omaima A. Sharaf, Wafaa M. Abd El-Rahim, Hassan Moawad, Michael J. Sadowsky

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Textile industry is one of the important industries in the worldwide. It is known that the eco-friendly industrial and agricultural activities are significant for socio-economic stability of all countries. The absence of appropriate industrial waste water treatments is essential barrier for sustainable development in food and agricultural sectors especially in developing country like Egypt. Thus, the development of enzymatic bioremediation technology for textile dye removal will enhance the collaboration between scientists who develop the technology and industry where this technology will be implemented towards the safe disposal of the textile dye wastes. Highly efficient microorganisms are of most importance in developing and using highly effective biological treatment processes. Bacterial degradation of azo dyes is generally initiated by an enzymatic step that involves cleavage of azo linkages, usually with the aid of an azoreductase as electron donor. Thus, expanding the spectrum of microorganisms with high enzymatic activities as azoreductases and discovering novel azo-dye degrading enzymes, with enhanced stability and superior catalytic properties, are necessary for many environmental and industrial applications. Consequently, the use of molecular tools has become increasingly integrated into the understanding of enzyme properties and characterization. Researchers have utilized a gene cloning and expression methods as a tool to produce recombinant protein for decolorizing dyes more efficiently. Thus, presumptive evidence for the presence of genes encoding azoreductases in the genomes of selected local, and most potent azo-degrading strains were obtained by using specific oligonucleotides primers. These potent strains have been isolated from textile industrial wastewater in Egypt and identified using 16S rRNA sequence analysis as 'Lysinibacillus macrolidesB8, Brevibacillus parabrevisB11, Bacillus coagulansB7, and B. cereusB5'. PCR products of two full length genes designated as (AZO1;621bp and AZO2;534bp) were detected. BLASTx results indicated that AZO1 gene was corresponding to predicted azoreductase from of Bacillus sp. ABP14, complete genome, multispecies azoreductase [Bacillus], It was submitted to the gene bank by an accession no., BankIt2085371 AZO1 MG923210 (621bp; 207 amino acids). AZO1 was generated from the DNA of our identified strains Lysinibacillus macrolidesB8. On the other hand, AZO2 gene was corresponding to a predicted azoreductase from Bacillus cereus strain S2-8. Gene bank accession no. was BankIt2085839 AZO2 MG932081 (534bp;178 amino acids) and it was amplified from our Bacillus coagulansB7. Both genes were successfully cloned into pCR2.1TOPO (Invitrogen) and in pET28b+ vectors, then they transformed into E. coli DH5α and BL21(DE3) cells for heterologous expression studies. Our recombinant azoreductases (AZO1&AZO2) exhibited potential enzyme activity and efficiently decolorized an azo dye (Direct violet). They exhibited pH stability between 6 and 8 with optimum temperature up to 60°C and 37 °C after induction by 1mM and 1.5mM IPTG, for both AZO1 &AZO2, respectively. These results suggested that further optimization and purification of these recombinant proteins by using different heterologous expression systems will give great potential for the sustainable utilization of these recombinant enzymes in several industrial applications especially in wastewater treatments.

Keywords: azoreductases, decolorization, enzyme activity, gene cloning and expression

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Authors: Olga V. Chervyakova, Elmira T. Tailakova, Vitaliy M. Strochkov, Kulyaisan T. Sultankulova, Nurlan T. Sandybayev, Lev G. Nemchinov, Rosemarie W. Hammond

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Sheep pox is a highly contagious infection that OIE regards to be one of the most dangerous animal diseases. It causes enormous economic losses because of death and slaughter of infected animals, lower productivity, cost of veterinary and sanitary as well as quarantine measures. To control spread of sheep pox infection the attenuated vaccines are widely used in the Republic of Kazakhstan and other Former Soviet Union countries. In spite of high efficiency of live vaccines, the possible presence of the residual virulence, potential genetic instability restricts their use in disease-free areas that leads to necessity to exploit new approaches in vaccine development involving recombinant DNA technology. Vaccines on the basis of recombinant proteins are the newest generation of prophylactic preparations. The main advantage of these vaccines is their low reactogenicity and this fact makes them widely used in medical and veterinary practice for vaccination of humans and farm animals. The objective of the study is to produce recombinant immunogenic proteins for development of the high-performance means for sheep pox prophylaxis. The SPV proteins were chosen for their homology with the known immunogenic vaccinia virus proteins. Assay of nucleotide and amino acid sequences of the target SPV protein genes. It has been shown that four proteins SPPV060 (ortholog L1), SPPV074 (ortholog H3), SPPV122 (ortholog A33) and SPPV141 (ortholog B5) possess transmembrane domains at N- or C-terminus while in amino acid sequences of SPPV095 (ortholog А 4) and SPPV117 (ortholog А 27) proteins these domains were absent. On the basis of these findings the primers were constructed. Target genes were amplified and subsequently cloned into the expression vector рЕТ26b(+) or рЕТ28b(+). Six constructions (pSPPV060ΔТМ, pSPPV074ΔТМ, pSPPV095, pSPPV117, pSPPV122ΔТМ and pSPPV141ΔТМ) were obtained for expression of the SPV genes under control of T7 promoter in Escherichia coli. To purify and detect recombinant proteins the amino acid sequences were modified by adding six histidine molecules at C-terminus. Induction of gene expression by IPTG was resulted in production of the proteins with molecular weights corresponding to the estimated values for SPPV060, SPPV074, SPPV095, SPPV117, SPPV122 and SPPV141, i.e. 22, 30, 20, 19, 17 and 22 kDa respectively. Optimal protocol of expression for each gene that ensures high yield of the recombinant protein was identified. Assay of cellular lysates by western blotting confirmed expression of the target proteins. Recombinant proteins bind specifically with antibodies to polyhistidine. Moreover all produced proteins are specifically recognized by the serum from experimentally SPV-infected sheep. The recombinant proteins SPPV060, SPPV074, SPPV117, SPPV122 and SPPV141 were also shown to induce formation of antibodies with virus-neutralizing activity. The results of the research will help to develop a new-generation high-performance means for specific sheep pox prophylaxis that is one of key moments in animal health protection. The research was conducted under the International project ISTC # K-1704 “Development of methods to construct recombinant prophylactic means for sheep pox with use of transgenic plants” and under the Grant Project RK MES G.2015/0115RK01983 "Recombinant vaccine for sheep pox prophylaxis".

Keywords: prophylactic preparation, recombinant protein, sheep pox virus, subunit vaccine

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Authors: J. W. Kim, Y. R. Kwon, Y. J. Ho, H. M. Jeon, G. M. Eom

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The aim of this study was to investigate the association of anthropometry with static balance in the elderly and their possible gender difference. Forty six subjects (23 men and 23 women) participated in this study. COP (Center of Pressure) was measured on a force-platform during quiet feet-together standing. As outcome measures, mean distance were derived from the COP. Weight was significantly correlated with postural variable only in the elderly men. This result suggests that the gender should be considered when normalizing postural variables.

Keywords: body characteristics, postural balance, elderly, gender difference

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Authors: Younies Saeed Hassan Mahmoud, Mai Mabrouk, Elsayed Sallam

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Analysing DNA microarray data sets is a great challenge, which faces the bioinformaticians due to the complication of using statistical and machine learning techniques. The challenge will be doubled if the microarray data sets contain missing data, which happens regularly because these techniques cannot deal with missing data. One of the most important data analysis process on the microarray data set is feature selection. This process finds the most important genes that affect certain disease. In this paper, we introduce a technique for imputing the missing data in microarray data sets while performing feature selection.

Keywords: DNA microarray, feature selection, missing data, bioinformatics

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Authors: Gileade P. Freitas, Helena B. Lopes, Alann T. P. Souza, Paula G. F. P. Oliveira, Adriana L. G. Almeida, Paulo G. Coelho, Marcio M. Beloti, Adalberto L. Rosa

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Bone tissue presents great capacity to regenerate when injured by trauma, infectious processes, or neoplasia. However, the extent of injury may exceed the inherent tissue regeneration capability demanding some kind of additional intervention. In this scenario, cell therapy has emerged as a promising alternative to treat challenging bone defects. This study aimed at evaluating the effect of local injection of bone marrow-derived mesenchymal stem cells (BM-MSCs) and adipose tissue-derived mesenchymal stem cells (AT-MSCs) on bone regeneration of rat calvaria defects. BM-MSCs and AT-MSCs were isolated and characterized by expression of surface markers; cell viability was evaluated after injection through a 21G needle. Defects of 5 mm in diameter were created in calvaria and after two weeks a single injection of BM-MSCs, AT-MSCs or vehicle-PBS without cells (Control) was carried out. Cells were tracked by bioluminescence and at 4 weeks post-injection bone formation was evaluated by micro-computed tomography (μCT) and histology, nanoindentation, and through gene expression of bone remodeling markers. The data were evaluated by one-way analysis of variance (p≤0.05). BM-MSCs and AT-MSCs presented characteristics of mesenchymal stem cells, kept viability after passing through a 21G needle and remained in the defects until day 14. In general, injection of both BM-MSCs and AT-MSCs resulted in higher bone formation compared to Control. Additionally, this bone tissue displayed elastic modulus and hardness similar to the pristine calvaria bone. The expression of all evaluated genes involved in bone formation was upregulated in bone tissue formed by BM-MSCs compared to AT-MSCs while genes involved in bone resorption were upregulated in AT-MSCs-formed bone. We show that cell therapy based on the local injection of BM-MSCs or AT-MSCs is effective in delivering viable cells that displayed local engraftment and induced a significant improvement in bone healing. Despite differences in the molecular cues observed between BM-MSCs and AT-MSCs, both cells were capable of forming bone tissue at comparable amounts and properties. These findings may drive cell therapy approaches toward the complete bone regeneration of challenging sites.

Keywords: cell therapy, mesenchymal stem cells, bone repair, cell culture

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Authors: Marina Passero, Tianhua Zhai, Zuyi (Jacky) Huang

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Alzheimer’s disease has become a major public health issue, as indicated by the increasing populations of Americans living with Alzheimer’s disease. After decades of extensive research in Alzheimer’s disease, only seven drugs have been approved by Food and Drug Administration (FDA) to treat Alzheimer’s disease. Five of these drugs were designed to treat the dementia symptoms, and only two drugs (i.e., Aducanumab and Lecanemab) target the progression of Alzheimer’s disease, especially the accumulation of amyloid-b plaques. However, controversial comments were raised for the accelerated approvals of either Aducanumab or Lecanemab, especially with concerns on safety and side effects of these two drugs. There is still an urgent need for further drug discovery to target the biological processes involved in the progression of Alzheimer’s disease. Excessive cholesterol has been found to accumulate in the brain of those with Alzheimer’s disease. Cholesterol can be synthesized in both the blood and the brain, but the majority of biosynthesis in the adult brain takes place in astrocytes and is then transported to the neurons via ApoE. The blood brain barrier separates cholesterol metabolism in the brain from the rest of the body. Various proteins contribute to the metabolism of cholesterol in the brain, which offer potential targets for Alzheimer’s treatment. In the astrocytes, SREBP cleavage-activating protein (SCAP) binds to Sterol Regulatory Element-binding Protein 2 (SREBP2) in order to transport the complex from the endoplasmic reticulum to the Golgi apparatus. Cholesterol is secreted out of the astrocytes by ATP-Binding Cassette A1 (ABCA1) transporter. Lipoprotein receptors such as triggering receptor expressed on myeloid cells 2 (TREM2) internalize cholesterol into the microglia, while lipoprotein receptors such as Low-density lipoprotein receptor-related protein 1 (LRP1) internalize cholesterol into the neuron. Cytochrome P450 Family 46 Subfamily A Member 1 (CYP46A1) converts excess cholesterol to 24S-hydroxycholesterol (24S-OHC). Cholesterol has been approved for its direct effect on the production of amyloid-beta and tau proteins. The addition of cholesterol to the brain promotes the activity of beta-site amyloid precursor protein cleaving enzyme 1 (BACE1), secretase, and amyloid precursor protein (APP), which all aid in amyloid-beta production. The reduction of cholesterol esters in the brain have been found to reduce phosphorylated tau levels in mice. In this work, a computational pipeline was developed to identify the protein targets involved in cholesterol regulation in brain and further to identify chemical compounds as the inhibitors of a selected protein target. Since extensive evidence shows the strong correlation between brain cholesterol regulation and Alzheimer’s disease, a detailed literature review on genes or pathways related to the brain cholesterol synthesis and regulation was first conducted in this work. An interaction network was then built for those genes so that the top gene targets were identified. The involvement of these genes in Alzheimer’s disease progression was discussed, which was followed by the investigation of existing clinical trials for those targets. A ligand-protein docking program was finally developed to screen 1.5 million chemical compounds for the selected protein target. A machine learning program was developed to evaluate and predict the binding interaction between chemical compounds and the protein target. The results from this work pave the way for further drug discovery to regulate brain cholesterol to combat Alzheimer’s disease.

Keywords: Alzheimer’s disease, drug discovery, ligand-protein docking, gene-network analysis, cholesterol regulation

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