Biological analysis of the deletion mutants of Staphylococcal enterotoxin C2

To investigate the functional domains involved in the biological activity of staphylococcal enterotoxin (SEC2), a series of SEC2 mutants were constructed. Deletion of the last 77 amino acids at the C-terminus of SEC2 did not affect its native superantigen and fever activities, and further removal of the C-terminal residues reduced SEC2 activities significantly. On the other hand, the mutants lacking 18 or more N-terminal residues severely impaired superantigen activity. These data indicated that the functional regions for the biological activities of SEC2 were confined to N-terminal domain, further implied that the proper three-dimensional structure of SEC2 is not needed for its biological activities. Our results deliver valuable information that it is possible to design new SEC2 immunotherapeutic agents which have the superantigen activity and low molecular weight for permeability.     

Biological activity of GLP-1-analogues with N-terminal modifications

Glucagon-like peptide-1 (GLP-1) stimulates insulin secretion and improves glycemic control in type 2 diabetes. In serum the peptide is degraded by dipeptidyl peptidase IV (DPP IV). The resulting short biological half-time limits the therapeutic use of GLP-1. Therefore, various GLP-1 analogues with alterations in cleavage positions were synthesized. GLP-1-receptor binding was investigated in RINm5F cells. Biological activity of the GLP-1 analogues was investigated in vitro by measuring cAMP production in RINm5F cells. GLP-1 analogues with modifications in position 2 were not cleaved by DPP IV and showed receptor affinity and in vitro biological activity comparable to native GLP-1. Analogues with alterations in positions 2 and 8, 2 and 9 or 8 and 9 showed a significant decrease in receptor affinity and biological activity. In vivo biological activity was tested in pigs. GLP-1 analogues were administered subcutaneously followed by an intravenous bolus injection of glucose. Plasma glucose and insulin were monitored over 4 h. Compared to native GLP-1, analogues with an altered position 2 showed similar or increased potency and biological half-time. Other GLP-1 analogues were less active. Despite the lack of degradation of these GLP-1 analogues by DPP IV in vitro, their biological action is as short as that of GLP-1, except for desamino-GLP-1, indicating that other degradation enzymes are important in vivo. Alterations of GLP-1 in positions 8 or 9 result in a loss of biological activity without extending biological half-time.     

Rate-Temperature Curves as an Unambiguous Indicator of Biological Activity in Soil

Experiments are described in which we used a mass spectrometer to monitor O₂ uptake of enclosed soil samples as a function of temperature. We found that an Arrhenius plot of the rate of O₂ uptake showed pronounced local maxima attributable to biological activity, whereas similar plots of rates obtained with abiotic soils yielded straight lines. This procedure thus provides a basis for distinguishing biological from chemical activity for reactions, such as O₂ uptake, that can occur via either biological or chemical pathways.     

Partial agonist/antagonist mouse interleukin-2 proteins indicate that a third component of the receptor complex functions in signal transduction.

Some mouse interleukin-2 (mIL-2) proteins with substitutions at residue Gln141 are unable to trigger a maximal biological response. The Asp141 protein induces the lowest maximal response. The Asp141 protein can weakly antagonize the biological activity of mIL-2 and strongly antagonizes the biological activity of active mIL-2 mutant proteins that have defects in interactions with the high affinity receptor. Residue 141 mutant proteins bind with reduced affinity to T cells expressing the high affinity IL-2 receptor, yet bind normally to transfected fibroblasts expressing only the alpha and beta chains of the receptor. These results suggest that a third receptor component is important for both binding and signal transduction.     

Carboxylic acid isosteres improve the activity of ring-fused 2-pyridones that inhibit pilus biogenesis in E. coli

Ring-fused 2-pyridones, termed pilicides, are small synthetic compounds that inhibit pilus assembly in uropathogenic Escherichia coli. Their biological activity is clearly dependent upon a carboxylic acid functionality. Here, we present the synthesis and biological evaluation of carboxylic acid isosteres, including, for example, tetrazoles, acyl sulfonamides, and hydroxamic acids of two lead 2-pyridones. Two independent biological evaluations show that acyl sulfonamides and tetrazoles significantly improve pilicide activity against uropathogenic E. coli.     

There May be Strict Empirical Laws in Biology, after All

This paper consists of four parts. Part 1 is an introduction. Part 2 evaluates arguments for the claim that there are no strict empirical laws in biology. I argue that there are two types of arguments for this claim and they are as follows: (1) Biological properties are multiply realized and they require complex processes. For this reason, it is almost impossible to formulate strict empirical laws in biology. (2) Generalizations in biology hold contingently but laws go beyond describing contingencies, so there cannot be strict laws in biology. I argue that both types of arguments fail. Part 3 considers some examples of biological laws in recent biological research and argues that they exemplify strict laws in biology. Part 4 considers the objection that the examples in part 3 may be strict laws but they are not distinctively biological laws. I argue that given a plausible account of what distinctively biological means, such laws are distinctively biological.     

Quasi-cubic magnetite/silica core-shell nanoparticles as enhanced MRI contrast agents for cancer imaging

Development of magnetic resonance imaging (MRI) contrast agents that can be readily applied for imaging of biological tissues under clinical settings is a challenging task. This is predominantly due to the expectation of an ideal MR agent being able to be synthesized in large quantities, possessing longer shelf life, reasonable biocompatibility, tolerance against its aggregation in biological fluids, and high relaxivity, resulting in better contrast during biological imaging. Although a repertoire of reports address various aforementioned issues, the previously reported results are far from optimal, which necessitates further efforts in this area. In this study, we demonstrate facile large-scale synthesis of sub-100 nm quasi-cubic magnetite and magnetite/silica core-shell (Mag@SiO2) nanoparticles and their applicability as a biocompatible T2 contrast agent for MRI of biological tissues. Our study suggests that silica-coated magnetite nanoparticles reported in this study can potentially act as improved MR contrast agents by addressing a number of aforementioned issues, including longer shelf life and stability in biological fluids. Additionally, our in vitro and in vivo studies clearly demonstrate the importance of silica coating towards improved applicability of T2 contrast agents for cancer imaging.     

Suggestions for unifying the terminology in biological control

This paper gives suggestions forunifying the terminology in biological controlacross different research disciplines, such asbiological control of arthropods, weeds andplant pathogens. It is suggested that use ofthe term `biological control' is restricted tothe use of living organisms. Four strategiesof biological control are outlined anddefined: (1) Classical biological control, (2) Inoculation biological control, (3) Inundationbiological control, and (4) Conservationbiological control. It is proposed to usethese four terms as defined, and avoid usageof the term `augmentation'. Terms for specificprocesses and modes of action (for example,`parasitoid' and `competitor') can be definedby usage within the different biologicalcontrol disciplines. Microbial control usuallyindicates biological control of invertebratesusing microbes and, as such, is a subdivisionof biological control. Use of additionalauxiliary terms such as biopesticide isdiscussed.     

2-pyrones possessing antimicrobial and cytotoxic activities

The 2-pyrone sub-unit is found in a number of natural products possessing broad spectrum biological activity. Such compounds are validated as being capable of binding to specific protein domains and able to exert a remarkable range of biological effects. In an effort to identify synthetic 2-pyrones with interesting biological effects, herein we report the synthesis and biological evaluation of 4-substituted-6-methyl-2-pyrones. Synthetic routes to 4-alkyl/alkenyl/aryl/alkynyl-6-methyl-2-pyrones have been developed utilising Sonogashira, Suzuki and Negishi cross-coupling starting from readily available 4-bromo-6-methyl-2-pyrone. Specific conditions for each organometallic protocol were required for successful cross-coupling. In particular, a triethylamine/acetonitrile--base/solvent mixture was crucial to Sonogashira alkynylation of 4-bromo-6-methyl-2-pyrone, whereas thallium carbonate was a mandatory base for the Suzuki cross-coupling of trialkylboranes. The 2-pyrones demonstrate potent inhibitory activity against Bacillus subtilis, Escherichia coli, Staphylococcus aureus, Schizosaccharomyces pombe and Botrytis cinerea. The growth inhibitory activities of selected 2-pyrones were determined in A2780 human ovarian carcinoma and K562 human chronic myelogenous leukaemia cell lines using an in vitro cell culture system (MTT assay). These studies demonstrate that 4-phenylethynyl-, 4-tetrahydropyranylpropargyl ether- and 4-ethynyl-6-methyl-2-pyrones have excellent potential as a new class of anticancer agents.     

The regional effects of CO2 and landscape change using a coupled plant and meteorological model

A meteorological model, the Regional Atmospheric Modelling System (RAMS), and a plant model, the General Energy and Mass Transfer Model (GEMTM), are coupled in this study. The integrated modelling system was used to investigate regional weather conditions in the central grasslands of the USA for three experimental scenarios: ? land cover is changed from current to potential vegetation; ? radiative forcing is changed from 1 × CO₂ to 2 × CO₂; and ? biological CO₂ partial pressures are doubled. Results indicate that the biological effect of enriched CO₂, and of land‐use change exhibit dominant effects on regional meteorological and biological fields, which were observed for daily to seasonal time scales and grid to regional spatial scales. Simulated radiation impacts of 2 × CO₂ were minimal, with interactive effects between the three experimental scenarios as large as the radiational impact alone. Model results highlight the importance of including 2 × CO₂ biological effects when simulating possible future changes in regional weather.     

Coupled biological and photo-Fenton pretreatment system for the removal of di-(2-ethylhexyl) phthalate (DEHP) from water

The photo-Fenton coupled with a biological system for the removal of di-(2-ethylhexyl) phthalate (DEHP) in wastewater was analyzed. The toxicity of DEHP-containing wastewater was found to be reduced after pretreatment by the photo-Fenton reaction. The effect of different factors, such as DEHP, Fe³⁺ and H₂O₂ concentrations and the reaction time, on degradation efficiency was investigated. The optimal time to stop the pretreatment process and introduce the effluent to the biological system was 60 min. The results show that effluent of DEHP-containing wastewater pretreated by the photo-Fenton method is biodegradable and that mineralization can be completed when the wastewater is subsequently treated in a biological system. The coupled Fenton and biological treatment system for the degradation of DEHP-containing wastewater can be successfully performed in a semi-continuous mode. These results indicate that the coupled photo-biological system is an effective and potential method for the treatment of DEHP-containing wastewater.     

Emphasis of biological research for space radiation]

The paper summarized issues, current status and the recent topics in biological research of space radiation. Researches to estimate a risk associated with space radiation exposure during a long-term manned space flight, such as in the International Space Station, is emphasized because of the large uncertainty of biological effects and a complexity of the radiation environment in space. The Issues addressed are; 1) biological effects and end points in low dose radiation, 2) biological effects under low dose rate and long-term radiation exposure, 3) modification of biological responses to radiation under space environments, 4) various aspects of biological end points vs. cellular and molecular mechanisms, 5) estimation of human risk associated with radiation exposure in space flight, 6) regulations for radiation exposure limits for space workers. The paper also summarized and introduced recent progress in space related radiation researches with various biological systems.     

Amplitude and frequency dissociation spectra of ion-protein complexes rotating in magnetic fields

A mechanism is presented that predicts new biological effects of static and sinusoidal weak magnetic fields. The model is based on an earlier proposed interference mechanism of quantum states of ions within protein cavities. The quantum dynamics of an ion is studied for the case of ion-protein complexes that rotate in magnetic fields. Both the individual molecular rotation and rotation together with a biological sample are taken into account. A formula is derived for the magnetic field-dependent part of the dissociation probability of an ion-protein in these conditions. The formula explains the unusual amplitude dependence of the known biological effect in PC-12 cells exposed to AC-DC magnetic field. The dependence had the functional motif J(2)(1)(2H(AC)/H(DC)), where J(1) is the first order Bessel function of the first kind. A good fit was obtained assuming individual rotation of the Li-protein complex in MF. The macroscopic rotation of a biological system, even at low speed 1.5-2 Hz, is predicted to reduce the biological effects of a "magnetic vacuum" and to shift the spectral peaks in the field and frequency dependencies of some magnetobiological effects.     

Synthesis of Mono-PEGylated Growth Hormone Releasing Peptide-2 and Investigation of its Biological Activity

The purpose of this study was to investigate an efficient synthetic route to the mono-PEGylated growth hormone releasing peptide-2 (GHRP-2) and its biological activity in vivo. The commercially available key PEGylating reagent, mPEG-NHS ester, was successfully utilized to the synthesis of mono-PEGylated GHRP-2, during which the PEGylation profiles of GHRP-2 were monitored by high-performance liquid chromatography (HPLC). The product was purified by cation exchange chromatography, and its biological activity was conducted in rats. The desired mono-PEGylated GHRP-2 as the major product was readily obtained in anhydrous aprotic solvent, such as dimethyl formamide (DMF) and dimethylsulfoxide (DMSO), when the molar ratio of mPEG-NHS ester to GHRP-2 was fixed to be 0.8:1. The products were characterized by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry. The evaluation of the biological activity for the products showed that the mono-PEGylated GHRP-2 gave a more stable activity than GHRP-2, suggesting that PEGylation led to the increase in the half-life of GHRP-2 in plasma without greatly impairing the biological activity. PEGylation of the GHRP-2 is a good choice for the development of the GHRP-2 applications.KEY WORDS: growth hormone, growth hormone releasing peptide-2, mono-PEGylated GHRP-2, mPEG-NHS, PEGylation     

Interesting and useful biochemical properties of lanthanides

The biological ubiquity and importance of calcium ions are matched by our lack of an overall theory of their biological functions. Lanthanide ions possess unique chemical properties which make them valuable probes of the interaction of Ca²⁺ with biological systems. Several new or improved techniques also exploit these properties which may form the basis for novel solutions to medical problems.     

Deletion of kringle domains or the N-terminal hairpin structure in hepatocyte growth factor results in marked decreases in related biological activities.

To determine the essential domain for biological activity in the hepatocyte growth factor (HGF) molecule, we prepared various mutated recombinant HGFs using site-directed mutagenesis, and examined the effects on DNA synthesis in hepatocytes, scattering of MDCK cells and the antiproliferative activity on HepG2 hepatoma cells. Native HGF and mutant HGFs, in which Gln534 and/or Tyr673 were respectively substituted for His and Ser to coincide with the catalytic triad amino acids in plasmin, markedly stimulated DNA synthesis of hepatocytes and scattering of MDCK cells but inhibited DNA synthesis of HepG2 cells. The mutant HGF deleted with the third or fourth kringle domain resulted in marked decrease of all three biological activities, while deletion of the N-terminal hairpin structure or the first or second kringle domain almost completely inactivated biological activities. We propose that the N-terminal hairpin structure and the first and second kringle domains are essential for biological activities of HGF and possibly for binding to its receptor.     

Automatic synchronization and distribution of biological databases and software over low-bandwidth networks among developing countries

Bioinformatics involves the collection, organization and analysis of large amounts of biological data, using networks of computers and databases. Developing countries in the Asia-Pacific region are just moving into this new field of information-based biotechnology. However, the computational infrastructure and network bandwidths available in these countries are still at a basic level compared to that in developed countries. In this study, we assessed the utility of a BitTorrent-based Peer-to-Peer (btP2P) file distribution model for automatic synchronization and distribution of large amounts of biological data among developing countries. The initial country-level nodes in the Asia-Pacific region comprised Thailand, Korea and Singapore. The results showed a significant improvement in download performance using btP2P--three times faster overall download performance than conventional File Transfer Protocol (FTP). This study demonstrated the reliability of btP2P in the dissemination of continuously growing multi-gigabyte biological databases across the three Asia-Pacific countries. The download performance for btP2P can be further improved by including more nodes from other countries into the network. This suggests that the btP2P technology is appropriate for automatic synchronization and distribution of biological databases and software over low-bandwidth networks among developing countries in the Asia-Pacific region. AVAILABILITY: http://everest.bic.nus.edu.sg/p2p/     

Variance-based sensitivity analysis of biological uncertainties in carbon ion therapy

PurposeBiological models to estimate the relative biological effectiveness (RBE) or the equivalent dose in 2 Gy fractions (EQD2) are needed for treatment planning and plan evaluation in carbon ion therapy. We present a model-independent, Monte Carlo based sensitivity analysis (SA) approach to quantify the impact of different uncertainties on the biological models.Methods and materialsThe Monte Carlo based SA is used for the evaluation of variations in biological parameters. The key property of this SA is the high number of simulation runs, each with randomized input parameters, allowing for a statistical variance-based ranking of the input variations. The potential of this SA is shown in a simplified one-dimensional treatment plan optimization. Physical properties of carbon ion beams (e.g. fragmentation) are simulated using the Monte Carlo code FLUKA. To estimate biological effects of ion beams compared to X-rays, we use the Local Effect Model (LEM) in the framework of the linear-quadratic (LQ) model. Currently, only uncertainties in the output of the biological models are taken into account.Results/conclusionsThe presented SA is suitable for evaluation of the impact of variations in biological parameters. Major advantages are the possibility to access and display the sensitivity of the evaluated quantity on several parameter variations at the same time. Main challenges for later use in three-dimensional treatment plan evaluation are computational time and memory usage. The presented SA can be performed with any analytical or numerical function and hence be applied to any biological model used in carbon ion therapy.     

Specialization and targeting of B-type cyclins.

The B-type cyclins of S. cerevisiae are diversified with respect to time of expression during the cell cycle as well as biological function. We replaced the early-expressed CLB5 coding sequence with the late-expressed CLB2 coding sequence, at the CLB5 locus. CLB5::CLB2 exhibited almost no rescue of clb5-specific replication defects, although it could rescue clb1 clb2 lethality, and in synchronized cells Clb2p-associated kinase activity from CLB5::CLB2 rose early in the cell cycle, similar to that of Clb5p. Mutagenesis of a potential substrate-targeting domain of CLB5 reduced biological activity without reducing Clb5p-associated kinase activity. Thus, Clb5p may have targeting domains required for CLB5-specific biological activity.     

Organelle-targeted delivery of biological macromolecules using the protein transduction domain: potential applications for Peptide aptamer delivery into the nucleus

Extensive effort is currently being expended on the innovative design and engineering of new molecular carrier systems for the organelle-targeted delivery of biological cargoes (e.g., peptide aptamers or biological proteins) as tools in cell biology and for developing novel therapeutic approaches. Although cell-permeable Tat peptides are useful carriers for delivering biological molecules into the cell, much internalized Tat-fused cargo is trapped within macropinosomes and thus not delivered into organelles. Here, we devised a novel intracellular targeting technique to deliver Tat-fused cargo into the nucleus using an endosome-disruptive peptide (hemagglutinin-2 subunit) and a nuclear localization signal peptide. We show for the first time that Tat-conjugated peptide aptamers can be selectively delivered to the nucleus by using combined hemagglutinin-2 subunit and nuclear localization signal peptides. This nuclear targeting technique resulted in marked enhancement of the cytostatic activity of a Tat-fused p53-derived peptide aptamer against human MDM2 (mouse double minute 2) that inhibits p53-MDM2 binding. Thus, our technique provides a unique methodology for the development of novel therapeutic approaches based on intracellular targeting.