Prospective Clinical Application of Thioredoxin Reductase as a Novel Diagnostic Tumor Marker

Background: Developing a novel, efficient biomarker for detecting malignant tumors is essential for the early diagnosis of cancers. Our aim was to assess the diagnostic value of a potential plasma tumor marker, thioredoxin reductase (TR), which is expressed in many types of malignant tumor, for the non-invasive detection of cancers. Methods: The plasma activities of TR were measured in 1513 patients with common clinical diseases, 59 patients with benign tumors, and 154 patients with cancers and 586 healthy controls. The area under the ROC curve (AUC) of TR and logistic regression results of different groups were compared by sensitivity, specificity and Youden’s index. Diagnostic cut-offs and clinical reference intervals were established via ROC curve analysis. Results: The logistic regression indicated that TR activity can discriminate between cancers and benign tumors or other common diseases very well (p < 0.0001), with an area under the curve from the receiver-operator characteristics between 0.91 and 0.96. The positive critical value was 2.51 and the cancer critical value was 9.90. The diagnostic gray zone (2.51 - 9.90) may be associated with benign tumors and some common clinical diseases. Conclusions: As a novel potential marker of malignant tumors with quantitative evaluation of proliferation, TR activity detection has an excellent diagnostic potential for early-stage malignant tumors. Impact: The convenient, economical, relatively non-invasive, and reproducible detection method of TR activity makes it suitable for routine clinical practice.

Website: https://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Catalytically Important Residues in E. coli 1-Deoxy-D-Xylulose 5-Phosphate Synthase

1-deoxy-D-xylulose 5-phosphate synthase (DXS) catalyzes the initial step of the 2-C-methyl-D- erythritol 4-phosphate (MEP) pathway consisting in the condensation of (hydroxiethyl)thiamin derived from pyruvate with D-glyceraldehyde 3-phosphate (GAP) to yield 1-deoxy-D-xylulose 5-phosphate (DXP). The role of the conserved residues H49, E370, D427 and H431 of E. coli DXS was examined by site-directed mutagenesis and kinetic analysis of the purified recombinant enzyme mutants. Mutants at position H49 showed a severe reduction in their specific activities with a decrease of the kcat/KM ratio by two orders of magnitude lower than the wild-type DXS. According to available structural data residue H49 is perfectly positioned to abstract a proton from the donor substrate. Mutations in DXS E370 showed that this residue is also essential for catalytic activity. Three-dimensional structure supports its involvement in cofactor deprotonation, the first step in enzymatic thiamin catalysis. Results obtained with H431 mutant enzymes indicate that this residue plays a role contributing to transition state stabilization. Finally, mutants at position D427 also showed a severe specific activity decrease with a reduction of the kcat/KM ratio. A role in binding the substrate and selecting the stereoisomer is proposed for D427.

Website: https://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Enzyme Kinetic Equations of Irreversible and Reversible Reactions in Metabolism

This paper compares the irreversible and reversible rate equations from several uni-uni kinetic mechanisms (Michaelis-Menten, Hill and Adair equations) and bi-bi mechanisms (single- and double- displacement equations). In reversible reactions, Haldane relationship is considered to be identical for all mechanisms considered and reversible equations can be also obtained from this rela- tionship. Some reversible reactions of the metabolism are also presented, with their equilibrium constant.

Website: https://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Data Classification with Modified Density Weighted Distance Measure for Diffusion Maps

Clinical data analysis is of fundamental importance, as classifications and detailed characterizations of diseases help physicians decide suitable management for patients, individually. In our study, we adopt diffusion maps to embed the data into corresponding lower dimensional representation, which integrate the information of potentially nonlinear progressions of the diseases. To deal with nonuniformaity of the data, we also consider an alternative distance measure based on the estimated local density. Performance of this modification is assessed using artificially generated data. Another clinical dataset that comprises metabolite concentrations measured with magnetic resonance spectroscopy was also classified. The algorithm shows improved results compared with conventional Euclidean distance measure.

Website: https://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Attention Drawing of Movie Trailers Revealed by Electroencephography Using Sample Entropy

A movie trailer is a common advertising tool in the entertainment industry. Detection of a viewer’s brain responses to a movie trailer can help film producers to tailor a more appealing trailer of a movie. In this study, we acquired electroencephalographic (EEG) signals from subjects when they watched movie trailers (labeled as Movie session), and compared with their resting state session (labeled as Resting session) or when they watch nature scenes (labeled as Nature session). We used Sample Entropy (SampEn) to analyze the EEG signals between different sessions. Results showed that the complexity ratios at Fp1, Fp2 and Fz channels derived from Movie session were significantly lower than that in Resting state or when subjects watched Nature session (p < 0.001). Our results suggest that the brain status can affect the complexity of their EEG. Further, the attraction of attention of a movie trailer can be observed from the change of EEG.

Website: https://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Think globally, research locally: Paradigms and place in agroecological research

Conducting science for practical ends implicates scientists, whether they wish it or not, as agents in social-ecological systems, raising ethical, economic, environmental, and political issues. Considering these issues helps scientists to increase the relevance and sustainability of research outcomes. As we rise to the worthy call to connect basic research with food production, scientists have the opportunity to evaluate alternative food production paradigms and consider how our research funds and efforts are best employed. In this contribution, we review some of the problems produced by science conducted in service of industrial agriculture and its associated economic growth paradigm. We discuss whether the new concept of “ecological intensification” can rescue the industrial agriculture/growth paradigm and present an emerging alternative paradigm of decentralized, localized, biodiversity-promoting agriculture for a steady-state economy. This “custom fit” agriculture engages constructively with complex and highly localized ecosystems, and we draw from examples of published work to demonstrate how ecologists can contribute by using approaches that acknowledge local agricultural practices and draw on community participation

Website: https://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Wildlife Ecotoxicology of Pesticides: Can We Track Effects to the Population Level and Beyond?

During the past 50 years, the human population has more than doubled and global agricultural production has similarly risen. However, the productive arable area has increased by just 10%; thus the increased use of pesticides has been a consequence of the demands of human population growth, and its impact has reached global significance. Although we often know a pesticide′s mode of action in the target species, we still largely do not understand the full impact of unintended side effects on wildlife, particularly at higher levels of biological organization: populations, communities, and ecosystems. In these times of regional and global species declines, we are challenged with the task of causally linking knowledge about the molecular actions of pesticides to their possible interference with biological processes, in order to develop reliable predictions about the consequences of pesticide use, and misuse, in a rapidly changing world.

Website: https://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Energy Use and Greenhouse Gas Emissions from Crop Production Using the Farm Energy Analysis Tool

Using the Farm Energy Analysis Tool (FEAT), we compare energy use and greenhouse gas (GHG) emissions from the cultivation of different crops, highlight the role of sustainable management practices, and discuss the impact of soil nitrous oxide (N2O) emissions and the uncertainty associated with denitrification estimates in the northeastern United States. FEAT is a transparent, open-source model that allows users to choose parameter estimates from an evolving database. The results show that nitrogen fertilizer and N2O emissions accounted for the majority of differences between crop energy use and GHG emissions, respectively. Integrating sustainable practices such as no tillage and a legume cover crop reduced energy use and GHG emissions from corn production by 37% and 42%, respectively. Our comparisons of diverse crops and management practices illustrate important trade-offs and can inform decisions about agriculture. We also compared methods of estimating N2O emissions and suggest additional research on this potent GHG.

Website: https://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Navigating a Critical Juncture for Sustainable Weed Management

Agricultural weed management has become entrenched in a single tactic—herbicide-resistant crops—and needs greater emphasis on integrated practices that are sustainable over the long term. In response to the outbreak of glyphosate-resistant weeds, the seed and agrichemical industries are developing crops that are genetically modified to have combined resistance to glyphosate and synthetic auxin herbicides. This technology will allow these herbicides to be used over vastly expanded areas and will likely create three interrelated challenges for sustainable weed management. First, crops with stacked herbicide resistance are likely to increase the severity of resistant weeds. Second, these crops will facilitate a significant increase in herbicide use, with potential negative consequences for environmental quality. Finally, the short-term fix provided by the new traits will encourage continued neglect of public research and extension in integrated weed management. Here, we discuss the risks to sustainable agriculture from the new resistant crops and present alternatives for research and policy.

Website: https://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Water pollution by agriculture

Agriculture disrupts all freshwater systems hugely from their pristine states. The former reductionist concept of pollution was of examining individual effects of particular substances on individual taxa or sub-communities in freshwater systems, an essentially ecotoxicological concept. It is now less useful than a more holistic approach that treats the impacts on the system as a whole and includes physical impacts such as drainage and physical modification of river channels and modification of the catchment as well as nutrient, particulate and biocide pollution. The European Water Framework Directive implicitly recognizes this in requiring restoration of water bodies to ‘good ecological quality’, which is defined as only slightly different from pristine state. The implications for the management of agriculture are far more profound than is currently widely realized.

Website: https://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Regenerative, Semiclosed Systems: A Priority for Twenty-First-Century Agriculture

Current systems cause overconsumption of environmental resources, contribute to climate change, rely on increasingly expensive fossil fuel, and result in environmental (e.g., groundwater) contamination. Moreover, the agronomic–urban interface is growing, as are markets for ecologically friendly produce, the need for low-input farming systems in low-income regions, and disenchantment with the subsidization of conventional agriculture. There is reasonable biological and economic evidence to support advocacy for a shift to regenerative systems. Such a shift presents challenges—for example, although higher labor input enhances community well-being and rural social capital, it is costly. It also offers opportunities—for example, to adapt technologies to monitor and minimize wastage. Shifting to semiclosed systems would be accelerated by (a) routine life cycle analysis and costing; (b) calculation of the full costs to society of farm inputs such as pesticides; (c) food labeling and standards that draw attention to energy and other inputs; (d) government grants supporting the transition to semiclosed systems; (e) changing priorities for agronomic research; and (f) greater engagement of urban societies in agriculture through recreation and philanthropy.

Website: https://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Pico Kit for Gene Expression Array Target Prep

For use with GeneChip Whole-Transcriptome (WT) Arrays, the new WT Pico Kit is compatible with small sample isolation techniques, including flow cytometry, laser capture microdissection, and fine needle aspiration. The kit prepares targets from multiple sample types, including fresh and fresh frozen tissues, cultured cells, FFPE specimens, and whole blood samples without a globin mRNA reduction step. The use of a single kit for multiple sample types improves the ability to better compare data from different samples, especially important for translational research.

Website: https://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Rare Genetic Mutations Occur More Often in Schizophrenia Patients

“While we cannot point to specific mutations that play a causal role in schizophrenia, we show that schizophrenia patients collectively have more of these mutations than unaffected individuals,” said Loes Olde Loohuis, the study’s first author and a postdoctoral fellow at UCLA’s Center for Neurobehavioral Genetics. The center is part of the university’s Jane and Terry Semel Institute for Neuroscience and Human Behavior.

Website: https://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Mutations in Single Gene Underlie Vulnerability to Two Unrelated Types of Infections

When a genetic error weakens a child’s immunity, otherwise nonthreatening microbes can sicken and sometimes kill. In work published July 9 in Science, researchers at The Rockefeller University and their colleagues identify one surprising case in which mutations in a single gene render children vulnerable to two very different diseases: aggravating, but treatable fungal infections, as well as invasive and potentially fatal bacterial disease.

Website: https://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Digital Dispenser Now Offers Aqueous Liquid Dispensing

The  HP D300 Digital Dispenser – available from Tecan – now allows customers to titrate low volumes of biomolecules in aqueous solutions, offering greater flexibility for drug discovery workflows. Users can choose between dispensing DMSO, for small molecule studies, and aqueous solutions in combination with a surfactant, for the investigation of proteins, antibodies, enzymes and nucleic acids.

Website: https://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Combination of Acacetin with Antibiotics against Methicillin Resistant Staphylococcus aureus Isolated from Clinical Specimens

Methicillin-restitant Staphylococcus aureus (MRSA) is very dangerous bacteria and one of the most feared nosocomial germs. In this study, acacetin was evaluated against 20 clinical isolates of MRSA, either alone or in combination with antibiotics. The acacetin exhibited a good activity against isolates MRSA with MICs/MBCs ranged between 10-80/20-160 μg/mL, for ampicillin 64-1024/128-2048 μg/mL, and for oxacillin 8-32/16-64 μg/mL. The combination of acacetin plus oxacillin or ampicillin was reduced by ≥4-fold against isolates MRSA tested, evidencing a synergistic effect as defined by a FICI of ≤0.5. Furthermore, a time-kill study evaluating the growth of the tested bacteria was completely attenuated after 2-5 h of treatment with the 1/2 MIC of acacetin, regardless of whether it was administered alone or with oxacillin (1/2 MIC) or ampicillin (1/2 MIC). In conclusion, acacetin exerted synergistic effects when administered with oxacillin or ampicillin and the antibacterial activity and resistant regulation of acacetin against clinical isolates of MRSA might be useful in controlling MRSA infections.

Website: http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Biochemical Estimation of Glucose 6 Phosphate Dehydrogenase Deficiency in Saudi Adults: Different Methods and Its Rationalization

Heterozygous females go undetected in a commonly used method. The aim of the study was to identify & rationalize different biochemical methods for detections of G6PD deficiency. Methods: Cross section retrospective study was conducted on 1584 (800 males, 784 females) blood samples collected from King Abdulaziz University Hospital (KAUH) and King Fahd Armed force hospital (KFAFH) in Jeddah, Western Saudi Arabia. Blood samples were screened for G6PD activity by fluorescence spot test, semi quantitative color reduction test and spectrometric quantitative evaluation. Hemoglobin (Hb) was measured on the same sample by BC-3200 Auto hematology Analyser. G6PD activity was recorded as U/g Hb. Samples identified as deficient with cutoff ≤4.6 U/gHb. Results: The prevalence of G6PD deficiency identified by fluorescence spot test was 73(4.6%) and all were deficient male. By semi quantitative method, the prevalence rate was 51(3.2%) and again all were male deficit patients. However, when quantitative spectrometric method was used, the prevalence was found in 90(5.7%), where in 73(4.6%) deficient patients were males and 17(1.1%) were females. Conclusion: Since the fluorescence spot test did not miss any G6PD deficient male, it should be restricted to males and quantitative test should be done on females. Each ethnic group should cultivate their own cutoff value for categorization of deficient patients.


Website: http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Obtaining Bacteriocins by Chromatographic Methods

Bacteriocins can be widely used as preservatives and antibiotic alternatives in medicine. Nisin, a Streptococcus lactis-derived bacteriocin, has been in practice in food industry for a long time. A relevant product contains about 2.5% of nisin. For medical use (e.g., when injected into the blood stream), highly purified drugs are required. However, the yield of bacteriocins accounts for no more than a few percents from the total activity in the culture liquid. In this paper, we propose methods (by example of two B. subtilis strains), allowing to increase the yield up to ~80%. It is believed that other bacteriocins may be purified by these methods and with the same yield.

Website: http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

A Systems Approach for Determining Gene Expression from Experimental Observation of Compound Presence and Absence

Based on complete knowledge of the structure of the metabolic network and all the reactions taking place in the cell, elementary modes (EMs) and minimal cut sets (MCSs) can relate compounds observed in a tissue, to the genes being expressed by respectively providing the full set of non-decomposable routes of reactions and compounds that lead to the synthesis of external products, and the full set of possible target genes for blocking the synthesis of external products. So, for a particular tissue, only the EMs containing the reactions that are related to the genes being expressed in those tissues, are active for the production of the corresponding compounds. This concept is used to develop an algorithm for determining a matrix of reactions (which can be related to corresponding genes) taking place in a tissue, using experimental observations of compounds in a tissue. The program is applied to the Arabidopsis flower and identified 20 core reactions occurring in all the viable EMs. They originate from the trans-cinnamate compound and lead to the formation of kaempferol and quercetin compounds and their derivatives, as well as anthocyanin compounds. Analyses of the patterns in the matrix identify reaction sets related to certain functions such as the formation of derivatives of the two anthocyanin compounds present, as well as the reactions leading from the network’s external substrate erythrose-4P to L-Phenylalanine, cinnamyl-alc to trans-cinnamate and so on. The program can be used to successfully determine genes taking place in a tissue, and the patterns in the resulting matrix can be analysed to determine gene sets and the state of the tissue.

Website: http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Optimum Conditions for Bioethanol Production

Addition of a small amount of alpha-amylase enzyme to potato solution was found to enhance the potato starch degradation and made the fermentation process quicker. This study observed that 1750 unit alpha-amylase is enough to degrade the starch in 15% of 500 ml potato starch solution. From fermentation time study, 6-day incubation time was found to be enough to complete the fermentation process and optimum production of bioethanol form potato starch. Suitable concentration of potato in fermentation process was determined using five different potato solutions (5%, 10%, 15%, 20% and 30%). A highest production of bioethanol was found in  20% potato treatment. Therefore, 20% potato solution is recommended for high-scale production of bioethanol from potato starch.

Website: http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Prevalence and Perceptions about Migraine among Students and Patients in Khyber Pakhtunkhwa Province

To assess the migraine, its frequency, symptoms, usual triggers and patients’ perceptions, the present study was conducted. For this purpose a prospective study was carried out based on properly designed questionnaire. The data were collected from 7 educational institutes, including students of Islamia College Peshawar, University of Peshawar, Khyber Medical College, Engineering University Peshawar, Fatima Jinnah College Peshawar, Commerce College Peshawar and Agriculture University Peshawar. Part of data was also collected from patients visiting Khyber Teaching Hospital Peshawar for their headache problems. A total of 270 questionnaires were filled up. About 81 (30.00%) headache cases were recorded, fulfilling the International criteria of Headache. Out of 81 recorded cases, 31 (38.3%) patients were of migraine with aura and 50 (61.7%) were of migraine without aura condition. The prevalence of migraine in the study population was found to be 30.00%. The frequency of female migraine patients was higher i.e. 31 (34.1%) than the male sufferers 50 (27.9%). The present study clearly showed that the higher frequency (65.0%) of migraine exists in the age group of above 30 years age. The most prevalent trigger was found to be tension, which was in parallel followed by the lack of sleep or oversleeping. A family history for migraine was found in migraine patients. Usually the condition was found to limit the routine activities of patients. The present study recorded that most of the migraine patients (40.2%) did not visit doctors at hospital and preferably go for self-medication, which really needs a public awareness in order to properly cope this neurological disorder, which may pose future medical complications as well.

Website: http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Probability of Osteoporotic Vertebral Fractures Assessment Based on DXA Measurements and Finite Element Simulation

The model is intended for simulating the degenerative process in the skeleton, with the consequent lost of bone mass and hence the decrease of its mechanical resistance which enables the fracture due to different traumatisms. Clinical studies were chosen, both in non-treatment conditions and receiving drug therapy, and fitted to specific patients according their actual bone mineral density measures. The predictive model is applied in a finite element simulation of the lumbar spine. The fracture zone would be determined according loading scenario (fall, impact, accidental loads, etc.), using the mechanical properties of bone obtained from the evolutionary model corresponding to the considered time. Bone mineral density evolution in untreated patients and in those under different treatments was analyzed. Evolutionary curves of fracture probability were obtained from the evolution of mechanical damage. The evolutionary curve of the untreated group of patients presented a marked increase of the fracture probability, while the curves of patients under drug treatment showed variable decreased risks, depending on the therapy type. The finite element model allowed obtaining detailed maps of damage and fracture probability, identifying high-risk local zones at vertebral body, which are the usual localization of osteoporotic vertebral fractures. The developed model is suitable for being used in individualized cases. The model might better identify at-risk individuals in early stages of osteoporosis and might be helpful for treatment decisions.

Website: http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Application of Green Fluorescent Protein in Immunoassays

Because of such characteristics, the usage of GFP is widespread in both in vivo and in vitro applications. In addition, recent advances in biotechnology have enabled GFP to be expressed in various hosts, including bacteria, yeast, plants, animals, and even living-cells, for multiple purposes. Currently, GFP is a subject of great interest in the analytical sciences, especially in immunoassays for qualitative and quantitative analyses, when it is fused with an antibody because of the high sensitivity of GFP and antigen-binding specificity of antibodies. Recently the fluobody, which is a fusion protein of GFP with single-chain variable fragment antibody (scFv), has become a useful tool in various fields. We review here the applications of GFP as fluobodies in immunoassays.

Website: http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Effect of Copper and Iron on Acidogenic Biomass in an Anaerobic Packed Bed Reactor

Copper and iron were fed to the reactor at concentrations of 100 and 300 mg·L^-1, respectively. Both metal ions had insignificant inhibitory effect over the metabolism of the biomass, specifically, on substrate consumption and production of volatile fatty acids (VFAs). The microstructural characterization of the biofilm by Scanning Electron Microscopy showed no effect on the morphology of the microorganisms after the metals treatment. Copper and iron removal was also measured in the reactor, achieving a breakthrough time of 3 days, during which removal efficiencies were higher than 90%. It was also observed that the biomass had a greater affinity for copper. The results indicate that acidogenic biomass can be used effectively as a sorbent agent. The pollution of river-streams with heavy metals—mainly copper and iron—was one of the most compelling motivations for conducting this investigation. The San Pedro River Basin, a trans-boundary river that originates near the mining town of Cananea, Sonora, México, and flows to Arizona, USA, exemplifies this environmental issue.

Website: http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Revisiting the X-Chromosome Inactivation and Its Impact on Female Longevity

Given that the probability of mutant X-linked allele expression decreases as a result of the inactivation, it has been proposed that females have biological advantages relative to males. These advantages have grabbed the attention of the scientific community in recent years and have focused it on this topic and its clinical implications. To shed some new light on this intriguing phenomenon, this article reviews the most relevant molecular events involved in this process. These events include the role of Xist, the selection mechanism for future X-chromosome inactivation, the age-related inactivation skewing, and the relationship between inactivation and the emergence of X-linked diseases, possible treatments, and longevity.

Website: http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Basidiocarp Production of Stropharia melanosperma (Bull.) Gillet in Nutrient Agar Media

The present work studies the fruiting body production of S. melanosperma (Bull.) Gillet using pairing test in nutrient agar media. Fourteen monokaryotic strains from single fruit body were chosen for the pairing test. Ninety-one crossings were made and fruit bodies were found in 20 crossings. Fully mature basidiocarps were produced in a ring-like fashion in one or both colonies. The patterns of the fruit body production in crossing single spore isolates are similar to those common in heterothallic tetrapolar species. The conducted work shows that S. melanosperma can serve as a model organism for the study of developmental processes in the basidiomycetes.

Website: http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Spontaneous and Bleomycin-Induced γH2AX Signals in CHO9 Metaphase Chromosomes

We detected γH2AX signals in CHO9 chromosomes of controls which significantly increase after BLM-exposure. γH2AX signals were uniformly distributed in chromosomes of controls. However, the γH2AX signal distribution in BLM exposed cells was significantly different between chromosomes and among chromosome regions, with few signals near the centromeres and a tendency to increase towards the telomeres. Interestingly, both basal and BLM-induced γH2AX signal distribution were statistically equal between CHO9 homologous chromosome regions. Our results suggest that BLM exerts an effect on H2AX139 phosphorylation, prevailing towards acetylated and gene-rich distal chromosome segments. The comparable H2AX139 phosphorylation of homologous regions puts forward its dependence on chromatin structure or function and its independence of the position in the karyotype.

website: http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Improved Recovery of Erythropoietin and Darbepoetin from Equine Plasma by the Application of a Wheat Germ Agglutinin Mediated Pre-Extraction Prior to Immunoaffinity Chromatography

We describe a two-step method that uses wheat germ agglutinin immobilized on Sepharose gel followed by immunoaffinity chromatography (IAC) to extract recombinant human erythropoietin and Darbepoetin from equine plasma. Lectin affinity chromatography was shown to be an effective approach for isolating these epoetins from plasma and in combination with IAC; this method gave superior recovery when compared to the use of the latter technique alone. Moreover, due to the ease with which it can be scaled up, it is particularly well suited for pre-concentrating larger volumes of samples prior to IAC and this provides a facile way of improving the overall sensitivity with which these foreign proteins can be detected in equine plasma.

Website: http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Purification and Biochemical Characterization of a Protease Inhibitor II Family from Jalapeño Pepper

The purpose of this paper is to report the isolation and purification of a novo-member of a protease inhibitor from jalapeño pepper (Capsicum annuum L.) (PIJP). The molecular weight of PIJP inhibitor is 5.95 kDa with 56 amino acids and 6 Cys residues with high inhibitory activity to trypsin with a Ki value of 95 nM. This inhibitor according to the alignment with homologous from NCBI and Pfam databases is a member of proteinase inhibitors II. It is worthwhile to mention a major compositional difference between the proteinase inhibitor II families which have 8 Cys residues. PIJP is the first purified proteinase inhibitor, member of this family with only 6 Cys residues.

Website: http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Application of High-Throughput Sequencing: Discovery of Informative SNPs to Subtype Bacillus anthracis

Single Nucleotide Polymorphisms (SNPs) are the most common and abundant genetic variation found in the genome of any living species, from bacteria to humans. In bacterial genotyping, these evolutionarily stable point mutations represent important DNA markers that can be used to elucidate deep phylogenetic relationships among worldwide strains, but also to discriminate closely related strains. With the advent of next generation sequencing (NGS) technologies, affordable solutions are now available to get access to the complete genome sequence of an organism. Sequencing efforts of an increasing number of strains provide an unprecedented opportunity to create comprehensive species phylogenies. In this study, a comparative analysis of 161 genomes of Bacillus anthracis has being conducted to discover new informative SNPs that further resolves B. anthracis SNP-based phylogenetic tree. Nine previously unpublished SNPs that define major groups or sub-groups within the B. anthracis species were selected and developed into SNP discriminative assays. We report here a cost-effective high-resolution melting-based genotyping method for the screening of 27 canonical SNPs that includes these new diagnostic markers. The present assays are useful to rapidly assign an isolate to one sub-lineages or sub-groups and determine its phylogenetic placement on the B. anthracis substructure population.

Website: http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Aptamer-Based Extraction of Ergot Alkaloids from Ergot Contaminated Rye Feed

The ingestion of ergot contaminated cereal crops can lead to a severe poisoning known as ergotism. For food and feed safety purposes, the extraction of ergot alkaloids from ergot contaminated flour was investigated. For the specific recognition of ergot alkaloids, DNA aptamer ligands specially selected for ergot alkaloids were grafted onto silica gel in order to construct a specific solid phase extraction system. The aptamer-functionalized silica gels were used to extract ergot alkaloids from a contaminated rye feed sample. The presence of ergot alkaloids eluted from the aptamer-functionalized silica gels was analyzed using LC-QTOF-MS. By using this simple system, it was possible to specifically extract ergosine, ergokryptine and ergocornine from an ergot contaminated rye feed sample. This aptamer-based extraction tool shows the applicability of aptamers for the specific extraction of toxins or natural compounds from turbid matrices in a one-step procedure.

Website:  http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Genetic and Metabolic Variability between Two Subspecies of Chamaeleo chamaeleon (Reptilia: Chamaeleonidae) in Egypt

Total lipids and proteins of liver and muscle tissues in both species were analyzed. Three Ldh isoforms were recorded for both subspecies and the activity. Rate of flow (RF) of Ldh-1 seemed to be higher in C. chamaeleon chamaeleon than in C. chamaeleon musae. This high activity could be supported by the significant increase in the total lipids and proteins in liver and muscle tissues of this species. It may thus be reasonable to suppose that C. chamaeleon chamaeleon is more active, energetic and adaptable in its habitat than C. chamaeleon musae. The α-Est showed four fractions in both subspecies. The null variations in the activity of α-Ests in the studied tissue may indicate, to some extent, the safety of the diet applied to both subspecies of chameleons.

Website: http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Recent Advances in the Chemistry of Oripavine and Its Derivatives

It is an important intermediate in the biosynthesis of morphine alkaloids. Recently, new Papaver somniferum strains have been developed which accumulate thebaine and oripavine, but not morphine and codeine. Therefore, the chemistry of oripavine has been studied intensively to synthesize opioid pharmaceuticals such as oxymorphone, naloxone and buprenorphine.


Website: http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

Study on Cadmium Resistant-Bacteria Isolated from Hospital Wastewaters

However, isolated bacteria from hospital effluents have high resistance to antimicrobial agents and therefore the ability to uptake cadmium and other metals. So, their use is recommended for wastewater bioremediation. In this study, twelve cadmium-resistant bacteria were isolated from hospital wastewaters and identified. The cadmium-resistant isolates characterized include both Gram-negative [75%] and Gram-positive [25%]. The Minimum Inhibitory Concentration of hospital wastewaters isolates was determined in solid media and was ranged from 250 to 950 micrograms/milliliters. All the isolates showed co-resistance to zinc and mercury. They were also antibiotic resistant and tolerated up a significant concentration of NaCl. The cadmium-resistant species Klebsiella pneumoniae and Pseudomonas aeruginosa were the most resistant to cadmium. The species Klebsiella pneumonia also resists to the highest concentrations of zinc and mercury. The results of toxicity tests on Vibrio fischeri, showed that the DI50 [15 min] as low as 0.07 carried away 50% luminescence inhibition.

Website: http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/

A Novel Cellular Autoaggregative Developmentally CRP Regulated Behaviour Generates Massively Chondrule-Like Formations over Surface of Old Escherichia coli K-12 Macrocolony Biofilms

How Escherichia coli bacteria develop a particular colonial, 3-D biofilm morphological pattern is still a poorly understood process. Recently, we reported a new E. coli K-12 morphotype exhibited by old macrocolonies described as volcano-like. The formative developmental process of this morphotype has been presented as a suitable experimental model for the study of 3D patterning in macrocolony biofilms. Here, we report the optical microscopy observations and genetic analysis that have unveiled the existence of a novel autoaggregative behaviour which generates massive lumpiness over the surface of the volcano-like macrocolonies. These lumpy formations are generated by the autoaggregation and strong interaction of tightly packed bacterial cells in structures with a chondrule-like appearance which give the colony’s surface its characteristic microscopic lumpy phenotype. Furthermore, they exhibit different levels of maturation from the edge to the center of the colony. Hence, its generation appears to follow a spatiotemporal program of development during the macrocolony’s morphogenesis. Interestingly, the agar’s hardness influences the morphology exhibited by these formations, with high agar concentration (1.5%, 15 g/L) suppressing its development.

Websites: http://www.arjonline.org/biosciences/american-research-journal-of-biosciences/