Enhancing the cyclodextrin production by synchronous utilization of isoamylase and α-CGTase

Cyclodextrins (CD) are cyclic α-1,4-glucans composed of glucose units, and they have multiple applications in food, pharmaceuticals, cosmetics, agriculture, chemicals, etc. CD are usually produced by cyclodextrin glycosyltransferase (CGTase) from starch. In the present study, a simultaneous conversion approach was developed to improve the yield of CD from starch by conjunction use of isoamylase with α-CGTase. The isoamylase of Thermobifida fusca was cloned and expressed in Escherichia coli BL21(DE3). The biochemical characterization of the enzyme showed that the optimum temperature and pH of the recombinant enzyme was 50 °C and 5.5, respectively, and it maintained 60 %, 85 % and 78 % relative activity at 30 °C, 40 °C and 60 °C, respectively. When the recombinant isoamylase and α-CGTase were used simultaneously to convert potato starch (15 %, w/v) into CD, the optimum conditions were found to be: 10 U of α-CGTase and 48 U of isoamylase per gram of substrate, with reaction temperature of 30 °C and pH 5.6. On the optimum condition, the total yield of CD reached 84.6 % (w/w) after 24 h, which was 31.2 % higher than transformation with α-CGTase alone. This is the first report of synchronous bioconversion of CD by both α-CGTase and isoamylase, and represents the highest efficiency of CD production reported so far.     

The effect of teaching the nature of science on students’ acceptance and understanding of evolution: myth or reality?

The results of studies of the nature of science (NOS) as a factor that enhances students’ understanding of evolution have been inconclusive. Therefore, the main purpose of this study was to test the role of NOS instruction in enhancing students’ learning about evolution. We used a quasi-experimental design with pre- and post-tests to investigate the impact of teaching evolution with and without NOS in two classes with 15–16-year-old students, who were randomly assigned to these two classes. To measure their understanding of NOS and their acceptance and understanding of evolution, we used three different instruments that have been shown to generate reliable and valid inferences in comparable populations. The main results of this study were that, in the class in which the teaching of evolution included NOS instruction, the students’ understanding of NOS and their acceptance of evolution significantly improved. However, irrespective of the use of NOS instruction, both classes increased their understanding of evolution. These results support the claim that NOS instruction may influence students’ acceptance of evolution but not their understanding of evolution and natural selection.     

Enhancing fluxes through the mevalonate pathway in Saccharomyces cerevisiae by engineering the HMGR and β-alanine metabolism

Mevalonate (MVA) pathway is the core for terpene and sterol biosynthesis, whose metabolic flux influences the synthesis efficiency of such compounds. Saccharomyces cerevisiae is an attractive chassis for the native active MVA pathway. Here, the truncated form of Enterococcus faecalis MvaE with only 3-Hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) activity was found to be the most effective enzyme for MVA pathway flux using squalene as the metabolic marker, resulting in 431-fold and 9-fold increases of squalene content in haploid and industrial yeast strains respectively. Furthermore, a positive correlation between MVA metabolic flux and β-alanine metabolic activity was found based on a metabolomic analysis. An industrial strain SQ3-4 with high MVA metabolic flux was constructed by combined engineering HMGR activity, NADPH regeneration, cytosolic acetyl-CoA supply and β-alanine metabolism. The strain was further evaluated as the chassis for terpenoids production. Strain SQ3-4-CPS generated from expressing β-caryophyllene synthase in SQ3-4 produced 11.86 ± 0.09 mg l⁻¹ β-caryophyllene, while strain SQ3-5 resulted from down-regulation of ERG1 in SQ3-4 produced 408.88 ± 0.09 mg l⁻¹ squalene in shake flask cultivations. Strain SQ3-5 produced 4.94 g l⁻¹ squalene in fed-batch fermentation in cane molasses medium, indicating the promising potential for cost-effective production of squalene.     

Optimization of α-amylase production by Bacillus subtilis RSKK96: using the Taguchi experimental design approach

In this study, the Taguchi experimental design was applied to optimize the conditions for α-amylase production by Bacillus subtilis RSKK96, which was purchased from Refik Saydam Hifzissihha Industry (RSHM). Four factors, namely, carbon source, nitrogen source, amino acid, and fermentation time, each at four levels, were selected, and an orthogonal array layout of L(16) (4(5)) was performed. The model equation obtained was validated experimentally at maximum casein (1%), corn meal (1%), and glutamic acid (0.01%) concentrations with incubation time to 72 h in the presence of 1% inoculum density. Point prediction of the design showed that maximum α-amylase production of 503.26 U/mg was achieved under optimal experimental conditions.     

Sorting and Processing of the Alzheimer’s Disease Amyloid Precursor Protein Mediated by the AP-4 Complex

Proteolytic processing of the transmembrane amyloid precursor protein (APP) to aggregation-prone amyloid-β (Aβ) peptide underlies the development of Alzheimer’s disease.     

Gifted and non-gifted students’ diurnal preference and the relationship between personality,sleep, and sleep quality

The current study had two main objectives. First, we examined gifted and non-gifted students’ diurnal preferences. Secondly, we examined the relationships among age, gender, personality, sleep quality, and chronotype of gifted students. Data were gathered from 276 gifted students and 1921 non-gifted students whose ages range between 7 and 17 years old, in same three cities in Turkey using the Composite Scale of Morningness (CSM) to assess diurnal preference, the Big Five Inventory (BIG-5) to assess personality and the Pittsburgh Sleep Quality Index (PSQI) to measure sleep quality. The first result indicated that gifted students were more morning-oriented compared to non-gifted students. The other main result was that the conscientiousness was the best predictor of CSM scores in gifted students. Additionally, conscientiousness, age, and global PSQI predicted CSM scores, respectively. Moreover agreeableness, conscientiousness, and emotional stability were positively related to morning orientation, while age, sleep quality, sleep latency, daytime dysfunction, and global PSQI were negatively related with chronotype in gifted students.     

Stimulation of interferons and endorphins/enkephalins by electro-aerosol inhalation? An experimental approach for testing an expanded hypothesis

The biological effects of endorphins/enkephalins and of interferons closely resemble those attributed to air ions and electro-aerosols. Air ions/electro-aerosols have been reported to affect brain functions and feelings of well-being; to have sedative and analgesic effects; to be therapeutically effective in certain viral (e.g., upper respiratory) infections; and to have tumor-attenuating effects. It is, therefore, conceivable that endorphins/enkephalins and interferons might be the mediators of these air ion/electro-aerosol effects.An experimental approach for testing this hypothesis is described. It calls for mice to be challenged with a suitable agent and to be exposed under appropriate conditions to a negatively charged aerosol of physiological saline 6 hours/day for up to 3 weeks; for the serial sacrifice of subgroups of these mice; for collecting blood and brains of the sacrificed animals; for the bioassay of the sera for interferon; and for radioimmunoassays of brains for endorphins/enkephalins. Special considerations, necessitated by the nature of the experiment, are discussed.     

Is it always possible to distinguish two- and three-state systems by evaluating the van't Hoff enthalpy?

Many small globular proteins are traditionally classified as thermodynamical two-state systems, i.e., the protein is either in the native, active state (folded) or in the denatured state (unfolded). We challenge this view and show that there may exist (protein) systems for which a van't Hoff analysis of experimental data cannot determine whether the system corresponds to two or three thermodynamical states when only temperatures in a narrow temperature region around the transition are considered. We generalize a widely employed two-state protein folding model to include a third, transition state. For this three-state system we systematically study the deviation of the calorimetric enthalpy (heat of transition) from the van't Hoff enthalpy, a measure of the two-stateness of a transition. We show that under certain conditions the heat capacity of the three-state system can be almost indistinguishable from the heat capacity for the two-state system over a broad temperature interval. The consequence may be that some three-state (or even more than three-states) systems have been misinterpreted as two-state systems when the conclusion is drawn solely upon the van't Hoff enthalpy. These findings are important not only for proteins, but also for the interpretation of thermodynamical systems in general.     

Estimating position and velocity of a submerged moving object by the clawed frog Xenopus and by fish—A cybernetic approach

The lateral-line system is a unique facility of aquatic animals to locate predator, prey, or conspecifics. We present a detailed model of how the clawed frog Xenopus, or fish, can localize submerged moving objects in three dimensions by using their lateral-line system. In so doing we develop two models of a slightly different nature. First, we exploit the characteristic properties of the velocity field, such as zeros and maxima or minima, that a moving object generates at the lateral-line organs and that are directly accessible neuronally, in the context of a simplified geometry. In addition, we show that the associated neuronal model is robust with respect to noise. Though we focus on the superficial neuromasts of Xenopus the same arguments apply mutatis mutandis to the canal lateral-line system of fish. Second, we present a full-blown three-dimensional reconstruction of the source on the basis of a maximum likelihood argument.     

High-school students in university research labs? Implementing an outreach model based on the ‘science as inquiry’ approach

In this study we designed, implemented, and evaluated an outreach programme for high-school biology students rooted in the ‘science as inquiry’ approach. Accordingly, students learn about science from experts in the field, as well as through in-class exposure to the history and philosophy of science. Our sample consisted of 11th graders (n?=?497), ages 16–17, attending advanced biology classes. Our goal was to determine whether this programme had a significant effect on students’ understanding of the ‘nature of science’ (NOS) and on their attitudes towards science. Using a controlled pre-post research design, we asked participants to complete a Likert-like questionnaire. Also, we conducted post-programme semi-structured interviews with 35 of the participants. Results show that completion of the programme significantly enhanced participants’ NOS understanding and improved their attitudes towards science. Participants expressed a deep level of NOS understanding and explicitly stated that the field visits to experts’ labs had changed their attitude towards science. We believe that our outreach programme can be adapted for teaching other sciences and for societies worldwide, as long as there is access to university laboratories and researchers willing to interact with young citizens and potential future scientists.     

Investigation of the binding interactions of Bisdemethoxycurcumin,Diacetylcurcumin and Diacetylbisdemethoxycurcumin with bovine α-lactalbumin by experimental and theoretical analysis

It was reported that bovine α-lactalbumin (BLA) as an important whey protein can be utilized as valuable vehicle for metal ions. The goal of this study was to investigate the interaction of BLA with bisdemethoxycurcumin (BDMC), Diacetylcurcumin (DAC), and diacetylbisdemethoxycurcumin (DABC) as three bioactive compounds by fluorescence quenching measurements and docking studies. It was observed that these ligands come closer to tryptophan residues and quench their emission without any change in their micro region polarity. The Stern–Volmer equation which is the best model to provide information about the interaction between small bioactive molecules and proteins was used to obtain the binding constants and the binding stoichiometry. Information about the extent of resonance energy transfer and Förster’s distance between donor and acceptor was estimated. Thermodynamic parameters confirmed that the final BDMC–BLA complex was stabilized by hydrogen bonds, whereas the final DABC–BLA and DAC–BLA complexes were stabilized by hydrophobic bonds which are in accordance with their chemical structures. Both the synchronous and docking studies verified that theTrp-26 which is the most exposed Tryptophan residue has the most contribution in the binding process. The Förster’s distances between bound ligands and tryptophans were in agreement with the measured distances by docking studies. The obtained achievements confirmed that there are considerable binding interactions between these curcuminoids and BLA.     

Plant use by the q’eqchi’ maya of belize in ethnopsychiatry and neurological pathology

Plant Use by the Q’eqchi’ Maya of Belize in Ethnopsychiatry and Neurological Pathology. Economic Botany 59(4):326–336, 2005. Neurological and mental health disorders are extremely debilitating, and sufferers in developing countries often rely on traditional practitioners for these medical needs. Few studies have focused on botanical remedies used in this context, although these offer interesting treatment alternatives. The present study investigated plant use for the treatment of neurological and mental health disorders by Q'eqchi' Maya healers of southern Belize. We found that these healers generally recognize and treat epilepsy/seizures, headache, madness, fright (susto), depression, numbness, insomnia, and stress with herbal remedies. Quantitative analyses showed that there is selection for the use of certain species and botanical families, namely for species of the Pteridophyta division and for plants from the Piperaceae family. This study denotes the importance and selectivity of plant use by Q'eqchi' healers of southern Belize for the treatment of neurological and mental conditions and points to a possible underrepresentation of these disorders in the ethnobotanical literature.     

Bacterial Pu(V) reduction in the absence and presence of Fe(III)–NTA: modeling and experimental approach

Plutonium (Pu), a key contaminant at sites associated with the manufacture of nuclear weapons and with nuclear-energy wastes, can be precipitated to “immobilized” plutonium phases in systems that promote bioreduction. Ferric iron (Fe³⁺) is often present in contaminated sites, and its bioreduction to ferrous iron (Fe²⁺) may be involved in the reduction of Pu to forms that precipitate. Alternately, Pu can be reduced directly by the bacteria. Besides Fe, contaminated sites often contain strong complexing ligands, such as nitrilotriacetic acid (NTA). We used biogeochemical modeling to interpret the experimental fate of Pu in the absence and presence of ferric iron (Fe³⁺) and NTA under anaerobic conditions. In all cases, Shewanella alga BrY (S. alga) reduced Pu(V)(PuO₂ ⁺) to Pu(III), and experimental evidence indicates that Pu(III) precipitated as PuPO_4(am). In the absence of Fe³⁺ and NTA, reduction of PuO₂ ⁺ was directly biotic, but modeling simulations support that PuO₂ ⁺ reduction in the presence of Fe³⁺ and NTA was due to an abiotic stepwise reduction of PuO₂ ⁺ to Pu⁴⁺, followed by reduction of Pu⁴⁺ to Pu³⁺, both through biogenically produced Fe²⁺. This means that PuO₂ ⁺ reduction was slowed by first having Fe³⁺ reduced to Fe²⁺. Modeling results also show that the degree of PuPO_4(am) precipitation depends on the NTA concentration. While precipitation out-competes complexation when NTA is present at the same or lower concentration than Pu, excess NTA can prevent precipitation of PuPO_4(am).     

Central-to-axial chirality transfer revealed by liquid crystals: a combined experimental and computational approach for the determination of absolute configuration of carboxylic acids with an α chirality centre

The conversion into 6,7-dihydro-5H-dibenz[c,e]azepine (DAZ) N-protected amides is a viable route for the determination of the absolute configuration of chiral 2-substituted carboxylic acids. The biphenyl moiety of DAZ, besides being a probe of chirality for the electronic circular dichroism (ECD) spectroscopy, makes these systems suitable for configuration assignment by exploiting the chirality amplification which occurs in nematic liquid crystals. To assess the reliability of the liquid crystal method in detecting the absolute stereochemistry of chiral amides bound to a biphenyl group, we measured the helical twisting power of a series of DAZ-N-protected amides and compared these data with the results obtained from ECD measurements. We will show that the liquid crystal method, corroborated by HTP predictions, is trustworthy with our biphenyl derivatives, even when ECD spectra are ambiguous for the presence of aryl moieties displaying strong UV absorptions in the same range of the biphenyl chromophore.     

Structure–activity relationships for ferriprotoporphyrin IX association and β-hematin inhibition by 4-aminoquinolines using experimental and ab initio methods

In order to probe structure–activity relationships of association with ferriprotoporphyrin IX (log K) and inhibition of β-hematin formation, a series of 4-aminoquinolines with varying substituents at the 7-position (X) have been synthesized. These have been further elaborated by introduction of two different R groups on the 4-amino nitrogen atom in the form of methyl (R = Me) and ethylamine (R = EtNH₂) side chains. Data for a previously investigated series containing an N,N-diethyl-ethylamine side chain were also compared with the findings of this study. Experimentally, log K values for the simple 4-aminoquinoline series (R = H) were found to correlate with the hydrophobicity constant (π) of the group X. The log K values for the series with R = Me and EtNH₂ were found to correlate with those of the series with R = H. The log of the 50% β-hematin inhibitory activity (log BHIA₅₀) was found to correlate with log K and either meta (σ_m) or para (σ_p) Hammett constants for the series with R = Me and EtNH₂, but not the simple series with R = H. To further improve predictability, correlations with ab initio electrostatic parameters, namely Mulliken and CHelpG charges were investigated. The best correlations were found with CHelpG charges which indicated that log K values can be predicted from the charges on atom H-8 and the group X in the quinolinium species computed in vacuum, while log BHIA₅₀ values can be predicted from the CHelpG charges on C-7, C-8 and N-1 for the neutral species in vacuum. These correlations indicate that association and inhibition of β-hematin formation are separately determined. They also suggest that electron withdrawing groups at the 7-position, but not necessarily hydrophobic groups are required for hemozoin inhibition. The upshot is that the correlations imply that considerably more hydrophilic hemozoin inhibitors are feasible.