Small Molecular Mass Research Articles

Modelling the supernova-driven ISM in different environments

We use hydrodynamical simulations in a $(256\;{\rm pc})^3$ periodic box to model the impact of supernova (SN) explosions on the multi-phase interstellar medium (ISM) for initial densities $n = 0.5-30$ cm$^{-3}$ and SN rates $1-720$ Myr$^{-1}$. We include radiative cooling, diffuse heating, and the formation of molecular gas using a chemical network. The SNe explode either at random positions, at density peaks, or both. We further present a model combining thermal energy for resolved and momentum input for unresolved SNe. Random driving at high SN rates results in hot gas ($T\gtrsim 10^6$ K) filling $> 90$% of the volume. This gas reaches high pressures ($10^4 < P/k_\mathrm{B} < 10^7$ K cm$^{-3}$) due to the combination of SN explosions in the hot, low density medium and confinement in the periodic box. These pressures move the gas from a two-phase equilibrium to the single-phase, cold branch of the cooling curve. The molecular hydrogen dominates the mass ($>50$%), residing in small, dense clumps. Such a model might resemble the dense ISM in high-redshift galaxies. Peak driving results in huge radiative losses, producing a filamentary ISM with virtually no hot gas, and a small molecular hydrogen mass fraction ($\ll 1$%). Varying the ratio of peak to random SNe yields ISM properties in between the two extremes, with a sharp transition for equal contributions. The velocity dispersion in HI remains $\lesssim 10$ km s$^{-1}$ in all cases. For peak driving the velocity dispersion in H$_\alpha$ can be as high as $70$ km s$^{-1}$ due to the contribution from young, embedded SN remnants.

Synergistic effect of anionic and nonionic monomers on the synthesis of high solid content waterborne polyurethane

High solid content waterborne polyurethane dispersions (PUDs) were synthesized via to the combination of ionic and nonionic monomers, in which the effect of the molecular mass of poly (ethylene glycol) (PEG) and its connection relationships of those hydrophilic groups in polyurethane with dimethylol propionic acid (DMPA) (through changed the synthetic schemes of PUDs) were researched. It is found that the molecular mass of PEG has obvious effect on combining with DMPA to synthesize PUDs. The incorporation of small molecular mass PEG200 (Mn = 200) had little effect on improving the dispersion and stability properties of anionic PUDs. But as the molecular mass of PEG increasing, they (PEG400, PEG600, PEG1000 and PEG2000) have obvious effect on improving the dispersion and stability properties, and can decrease the required dosage of DMPA to stabilize PUDs, moreover, cam improve the solid content of the latex. However, the effect of PEG2000 is less prominent than that of PEG400, PEG600 and PEG1000. The connection relationships of PEG with DMPA also have a certain effect on the property of the latex. When the small molecular mass one (PEG400) directly connects with DMPA, the synergistic effort is enhanced and the required dosage of DMPA to stabilize latex decreases to 2.0% (using DMPA alone needs 2.4%). As a result, the solid content of PUDs increased from 42% (using DMPA alone) to 52%, and the water resistance of PUDs is also improved. The thermal, dynamic mechanical and tensile-strength properties of derived films of PUDs were also analyzed, and the results show the incorporating PEG into polyurethane decrease the glass transition temperature of soft segments (Tgs), as well decreases the crystallinity and the extent of phase separation, which both cause the decrease of heat resistance. But through the combination, the synthetic formulas of PUDs with better tensile strength and elongation ratio than using DMPA alone were obtained. Besides, it is found that the molecular mass of PEG and the connection relationships of PEG with DMPA also had some influence on the properties of the derived films.

An alkaloid of Norway spruce (Picea abies) possesses powerful antibacterial and antifungal potential

Norway spruce, Picea abies, along with other species of Picea contains a number of small molecular mass alkaloids [1]. To the best of our knowledge there exists no earlier publications on the antimicrobial effects of spruce alkaloids. On the other hand, there are a number of publications on the antibacterial effects of essential oils and oil rich fractions of spruce [2]. Spruce needle oil and decoctions of young spruce are traditionally used for treatment of cold, bronchitis, symptoms of fever and stomach ailments, among others. The present study was carried out to evaluate the antimicrobial effects of alkaloid EpI found in P. abies. Focus was put on bacterial strains causing food poisonings as well as other common human pathogenic bacteria and two Candida species. The antimicrobial effects were evaluated using microplate methods. The lowest minimum inhibitory concentration of 5.37 µg/ml was recorded against Pseudomonas aeruginosa, Enterococcus faecalis, Candida glabrata and Candida albicans. For comparison, the conventionally used tetracycline gave a MIC of 15.6 µg/ml against P. aeruginosa and amphotericin B a MIC of 1.95 µg/ml against C. albicans. Very promising MIC value of 10.75 µg/ml was recorded against Staphylococcus aureus and Bacillus cereus, both common food poisoning bacteria. In addition promising MIC of 43 µg/ml was recorded against Salmonella enterica. Further studies will be performed on testing this antimicrobially promising alkaloid EpI for cytotoxicity against fibroblasts and other normal cell lines.

Microfluidic Western blotting of low-molecular-mass proteins.

We describe a microfluidic Western blot assay (μWestern) using a Tris tricine discontinuous buffer system suitable for analyses of a wide molecular mass range (6.5–116 kDa). The Tris tricine μWestern is completed in an enclosed, straight glass microfluidic channel housing a photopatterned polyacrylamide gel that incorporates a photoactive benzophenone methacrylamide monomer. Upon brief ultraviolet (UV) light exposure, the hydrogel toggles from molecular sieving for size-based separation to a covalent immobilization scaffold for in situ antibody probing. Electrophoresis controls all assay stages, affording purely electronic operation with no pumps or valves needed for fluid control. Electrophoretic introduction of antibody into and along the molecular sieving gel requires that the probe must traverse through (i) a discontinuous gel interface central to the transient isotachophoresis used to achieve high-performance separations and (ii) the full axial length of the separation gel. In-channel antibody probing of small molecular mass species is especially challenging, since the gel must effectively sieve small proteins while permitting effective probing with large-molecular-mass antibodies. To create a well-controlled gel interface, we introduce a fabrication method that relies on a hydrostatic pressure mismatch between the buffer and polymer precursor solution to eliminate the interfacial pore-size control issues that arise when a polymerizing polymer abuts a nonpolymerizing polymer solution. Combined with a new swept antibody probe plug delivery scheme, the Tris tricine μWestern blot enables 40% higher separation resolution as compared to a Tris glycine system, destacking of proteins down to 6.5 kDa, and a 100-fold better signal-to-noise ratio (SNR) for small pore gels, expanding the range of applicable biological targets.

Poly(N-isopropylacrylamide-co-tris-nitrilotriacetic acid acrylamide) for a combined study of molecular recognition and spatial constraints in protein binding and interactions.

Many biological processes require precise regulation and synergy of proteins, and consequently involve molecular recognition and spatial constraints between biomolecules. Here, a library of poly(N-isopropylacrylamide-co-tris-nitrilotriacetic acid acrylamide) (PNTs) has been synthesized and complexed with Cu(2+) in order to serve as models for investigation of the combined effects of molecular recognition and spatial constraints in biomolecular interactions. The average distance between Cu(2+)-trisNTA binding sites in PNTs polymers was varied from 4.3 to 31.5 nm by adjusting their trisNTA contents. His tag (His6), His-tagged enhanced yellow fluorescent protein (His6-eYFP), and His6-tagged collagenase G (His6-ColG), with sizes ranging from 1 to 11 nm, were used as models to assess whether the binding ability is influenced by a cooperative topology based on molecular recognition interactions with Cu(2+)-trisNTA binding sites, and spatial constraints created by decreasing average distance between trisNTAs. His-tagged molecules bound to all PNTs polymers due to their molecular recognition interaction involving histidines and Cu(2+)-trisNTA pockets, but with a binding ability that was highly modulated by the average distance between the trisNTA binding sites. Small molecular mass molecules (His6) exhibit a high binding ability to all PNTs polymers, whereas his-tagged proteins bind to PNTs efficiently only when the average distance between trisNTA binding sites is larger than the protein dimensions.

Isoforms of wild type proteins often appear as low molecular weight bands on SDS-PAGE.

Immunoblotting, after polyacrylamide gel electrophoresis with sodium dodecyl sulfate (SDS-PAGE), is a technique commonly used to detect specific proteins. SDS-PAGE often results in the visualization of protein band(s) in addition to the one expected based on the theoretical molecular mass (TMM) of the protein of interest. To determine the likelihood of additional band(s) being nonspecific, we used liquid chromatography - mass spectrometry to identify proteins that were extracted from bands with the apparent molecular mass (MM) of 40 and 26 kD, originating from protein extracts derived from non-malignant HEK293 and cancerous MDA-MB231 (MB231) cells separated using SDS-PAGE. In total, approximately 57% and 21% of the MS/MS spectra were annotated as peptides in the two cell samples, respectively. Moreover, approximately 24% and 36.2% of the identified proteins from HEK293 and MB231 cells matched their TMMs. Of the identified proteins, 8% from HEK293 and 26% from MB231 had apparent MMs that were larger than predicted, and 67% from HEK293 and 37% from MB231 exhibited smaller MM values than predicted. These revelations suggest that interpretation of the positive bands of immunoblots should be conducted with caution. This study also shows that protein identification performed by mass spectrometry on bands excised from SDS-PAGE gels could make valuable contributions to the identification of cancer biomarkers, and to cancer-therapy studies.

Bacillus subtilis Fermentation for Enhancement of Feed Nutritive Value of Soybean Meal

Abstract In order to increase the nutritional quality of soybeanmeal (SBM) as an animal feed, Bacillus subtilis TP6, a previouslyisolated strain from an Indonesian traditional fermented soybeanfood, Tempeh, was used as a starter organism for solid-statefermentation. In the pre-treated SBM with water content of 60%(v/w), B. subtilis TP6 was grown to a maximum viable cellnumber of 3.5×10 9 CFU/g. Compared to control, crude protein inBacillus fermented SBM was increased by 16%, while raffinose,stachyose, and trypsin inhibitors were reduced by 31, 37, and90%, respectively. The Sodium dodecyl sulfate-polyacrylamidegel electrophoresis analysis showed that proteins in the fermentedSBM were remarkably hydrolyzed into smaller molecular masses,resulting in a decrease in large sized proteins. Our data suggestedthat B. subtilis fermentation could increase the nutritive value ofSBM through reduction of anti-nutritive factors and improvementof protein quality by hydrolysis of soy protein. In addition, B.subtilis TP6 produced a functional ingredient, poly-γ-glutamicacid which has various health benefits.Keywords animal feed · Bacillus subtilis · soybean meal · trypsin inhibitorIntroductionSoybean (Glycine max) is an oilseed crop belonging to the familyLeguminosae and one of the world’s largest and most efficientsources of plant protein. Soybean meal (SBM), which is abyproduct remaining after extraction of oil from soybean, hasbecome the most important source of protein for poultry and otherlivestock, due to its high concentration of protein and excellentprofile of essential amino acids in comparison to other plant proteinsources (Park, 1987; Adeola, 1996; Emmert and Baker, 1997;Boonyaratpalin et al., 1998). SBM contains around 40−50%protein and is a rich source of the amino acids including lysine,tryptophan, threonine, isoleucine, and valine that are deficient incorn, grain sorghum and other cereal grains used for animal feed(Emmert and Baker, 1997; Kim et al., 2012).Nutritive value of SBM is limited by several anti-nutritionalfactors such as protease inhibitor, oligosaccharide, and phytic acidthat interfere with digestion, absorption, and utilization of nutrientsin the diet (Liener, 1981; Grant, 1989). SBM possesses high contentof protease inhibitors that adversely affect protein digestibility andamino acid availability. Especially, the presence of trypsin inhibitorsin animal feed has been implicated in growth inhibition andpancreatic hypertrophy (Liener, 1981; Grant, 1989; Gumbmann etal., 1989; Friedman et al., 1991). Oligosaccharides such as raffinoseand stachyose are also poorly utilized by some species of animals,and fermentation of these oligosaccharides in the lower gut causesthe production of gastrointestinal gases (Krause et al., 1994). Mostphosphorus present in SBM is in the form of phytic acid, whichis generally indigestible by monogastric animals such as poultryand swine (Grant, 1989). The level of anti-nutritional factors canbe reduced by heating during the processing of soybean (Ratnerand Crawford, 1955; Friedman et al., 1991). However, an excessof heat may increase Maillard reactions between amino acids and

Advantages and Disadvantages of Zinc-Ethanol-Formaldehyde as a Fixative for Immunocytochemical Studies and Confocal Laser Microscopy

The paper presents an analysis of authors' results obtained using fixation of various tissues in zink-ethanol-formaldehyde (ZEF). It was found that fixation in ZEF, in comparison with other methods of fixation, allowed to achieve higher sensitivity of immunocytochemical reaction for a large number of antigens studied and, in many cases, to avoid heat unmasking of antigens. It also provided high resolution of the images obtained using the fluorescent and confocal laser microscopy. However, the studies of antigens with small molecular mass revealed the antigen diffusion from the site of original localization. The data obtained suggest that fixation of a material in ZEF fixative is promising for both immunocytochemical studies, including those using the fluorescent and confocal laser microscopy, and general histological practice.

Discovery of a cell wall porin in the mycolic‐acid‐containing actinomycete Dietzia marisDSM 43672

The cell wall of the Gram-positive mycolic-acid-containing actinomycete Dietziamaris DSM 43672 was found to contain a pore-forming protein, as observed from reconstitution experiments with artificial lipid bilayer experiments in the presence of cell wall extracts. The cell wall porin was purified to homogeneity using different biochemical methods and had an apparent molecular mass of about 120 kDa on tricine-containing SDS/PAGE. The 120 kDa protein dissociated into subunits with a molecular mass of about 35 kDa when it was heated to 100°C in 8m urea. The 120 kDa protein, here named PorADm , formed ion-permeable channels in lipid bilayer membranes with a high single-channel conductance of about 5.8 nS in 1m KCl. Asymmetric addition of PorADm to lipid bilayer membranes resulted in an asymmetric voltage dependence. Zero-current membrane potential measurements with different salt solutions suggested that the porin of D.maris is cation-selective because of negative charges localized at the channel mouth. Analysis of the single-channel conductance using non-electrolytes with known hydrodynamic radii indicated that the diameter of the cell wall channel is about 2nm. The channel characteristics of the cell wall porin of D.maris are compared with those of other members of the mycolata. They share some common features because they are composed of small molecular mass subunits and form large and water-filled channels. The porin was subjected to protein analysis by mass spectrometry but its sequence had no significant homology to any known porin sequences.

Preliminary study of a novel R718 refrigeration cycle with single stage centrifugal compressor and two-phase ejector

The paper describes the investigations of a novel R718 refrigeration cycle with single stage centrifugal compressor and two-phase ejector as a second stage compression device. The limitations of the R718 centrifugal stage pressure ratio, connected with small molecular mass of the water and high isentropic exponent, result on high impeller peripheral speed, high Mach number and high discharge temperature. Reynolds number is low and dimensions of the compressor stage are large. The complex thermal and flow phenomena inside two-phase ejector flow field are investigated and performance characteristics are estimated. Although efficiency of the concept of compression with hydraulic pump and two-phase ejectors is lower, the integration of two-phase ejectors in the R718 centrifugal unit results in low cost high capacity refrigeration system with high energy efficiency, competitive with traditional refrigeration systems by environmental, technical and economical reasons.

High resolution mass spectrometry based techniques at the crossroads of metabolic pathways

The metabolome is the set of small molecular mass compounds found in biological media, and metabolomics, which refers to as the analysis of metabolome in a given biological condition, deals with the large scale detection and quantification of metabolites in biological media. It is a data driven and multidisciplinary approach combining analytical chemistry for data acquisition, and biostatistics, informatics and biochemistry for mining and interpretation of these data. Since the middle of the 2000s, high resolution mass spectrometry is widely used in metabolomics, mainly because the detection and identification of metabolites are improved compared to low resolution instruments. As the field of HRMS is quickly and permanently evolving, the aim of this work is to review its use in different aspects of metabolomics, including data acquisition, metabolite annotation, identification and quantification. At last, we would like to show that, thanks to their versatility, HRMS instruments are the most appropriate to achieve optimal metabolome coverage, at the border of other omics fields such as lipidomics and glycomics.

Abstract 3436: Integrated profiling of p53 wild-type cell lines identifies differentially responsive populations and a gene expression signature that predicts sensitivity to SAR405838, a potent and selective disruptor of the p53-MDM2 interaction.

Abstract The tumor suppressor protein p53 functions at the center of a complex biological network that translates diverse stress signals into cell cycle arrest or apoptosis. MDM2 inhibits p53 activity by binding to the transactivation domain of p53 and by acting as a p53-specific E3 ubiquitin ligase. The disruption of the interaction between p53 and MDM2 with small molecular mass molecules has demonstrated encouraging single agent anti-tumor activity in preclinical in vitro and in vivo settings. While p53 wild-type status is necessary for response to antagonists of the p53-MDM2 protein-protein interaction (PPI), there is diversity in response to such therapeutic agents across p53 wild-type tumor models. As such, the ability to predict p53 wild-type tumor responsiveness to antagonists of the p53-MDM2 PPI is an unmet need that could significantly impact clinical development of modulators of the p53-MDM2 PPI. Here, we explored the effect of SAR405838, a selective p53-MDM2 antagonist, on a panel of 65 p53 wild-type cell lines derived from different tumors of origin. Over 80% of these cell lines showed some degree of sensitivity to SAR405838 when used as a single agent. Three broad response classes were observed: non-responders, responders involving apoptosis, and responders not involving apoptosis. A small number of tissue types were consistently enriched among responders (melanoma and leukemia) and non-responders (cervical and uterine tumors). By comparing pre-treatment gene expression profiling data with the single agent sensitivity to SAR405838, we identified a signature of 39 genes that clearly predicts non-response vs response. Within these 39 genes, CDKN2A was upregulated in non-responders, and MDM2 was upregulated in responders. Gene set enrichment analysis and GO category enrichments were consistent with predicted sensitivity in melanoma and leukemia, and indicate that a gene expression signature of loss of p53 pathway function predicts lack of single agent activity of SAR405838. 3-fold cross-validation of this classifier showed an accuracy of 94%, and principle component analyses was used to down-select the signature to the smallest number of genes carrying the maximum information and derive the optimal cut-off for making prospective response predictions. These data indicate that the development of p53-MDM2 PPI modulators could be targeted to a subset of p53 wild-type cancer patients and eliminate more predicted non-responders than those harbouring somatic mutations in p53. Citation Format: James Watters, Steve Rowley, Laurent Debussche. Integrated profiling of p53 wild-type cell lines identifies differentially responsive populations and a gene expression signature that predicts sensitivity to SAR405838, a potent and selective disruptor of the p53-MDM2 interaction. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3436. doi:10.1158/1538-7445.AM2013-3436

Proteolytic elimination of N-myristoyl modifications by the Shigella virulence factor IpaJ

Protein N-myristoylation is a 14-carbon fatty-acid modification that is conserved across eukaryotic species and occurs on nearly 1% of the cellular proteome. The ability of the myristoyl group to facilitate dynamic protein-protein and protein-membrane interactions (known as the myristoyl switch) makes it an essential feature of many signal transduction systems. Thus pathogenic strategies that facilitate protein demyristoylation would markedly alter the signalling landscape of infected host cells. Here we describe an irreversible mechanism of protein demyristoylation catalysed by invasion plasmid antigen J (IpaJ), a previously uncharacterized Shigella flexneri type III effector protein with cysteine protease activity. A yeast genetic screen for IpaJ substrates identified ADP-ribosylation factor (ARF)1p and ARF2p, small molecular mass GTPases that regulate cargo transport through the Golgi apparatus. Mass spectrometry showed that IpaJ cleaved the peptide bond between N-myristoylated glycine-2 and asparagine-3 of human ARF1, thereby providing a new mechanism for host secretory inhibition by a bacterial pathogen. We further demonstrate that IpaJ cleaves an array of N-myristoylated proteins involved in cellular growth, signal transduction, autophagasome maturation and organelle function. Taken together, these findings show a previously unrecognized pathogenic mechanism for the site-specific elimination of N-myristoyl protein modification.

Expression of a functional recombinant human basic fibroblast growth factor from transgenic rice seeds.

Basic fibroblast growth factor (FGF-2) is an important member of the FGF gene family. It is widely used in clinical applications for scald and wound healing in order to stimulate cell proliferation. Further it is applied for inhibiting stem cell differentiation in cultures. Due to a shortage of plasma and low expression levels of recombinant rbFGF in conventional gene expression systems, we explored the production of recombinant rbFGF in rice grains (Oryza sativa bFGF, OsrbFGF). An expression level of up to 185.66 mg/kg in brown rice was obtained. A simple purification protocol was established with final recovery of 4.49% and resulting in a yield of OsrbFGF reaching up to 8.33 mg/kg OsrbFGF. The functional assay of OsrbFGF indicated that the stimulating cell proliferation activity on NIH/3T3 was the same as with commercialized rbFGF. Wound healing in vivo of OsrbFGF is equivalent to commercialized rbFGF. Our results indicate that rice endosperm is capable of expressing small molecular mass proteins, such as bFGF. This again demonstrates that rice endosperm is a promising system to express various biopharmaceutical proteins.

Thermal degradation behavior of cellulose-based material for gating systems in iron casting production

Summary — This study presents a fragment of research aimed at determining the thermal stability of a cellulose-based composite material used for building gating systems for large iron castings. The behavior of the material itself at elevated temperatures is of major significance for the mould filling process. In this research, thermal analysis methods (DTG, TG, DSC) were used to explain the course of the thermal degradation of the analysed material by establishing thermal effects of transformations occurring during its heating as well as its structural and mass changes. The research was conducted at the temperature range of 40—1000 °C. The degradation process was found to start at the temperature of approx. 300 °C, and the total mass loss at the range of 300—750 °C amounted to 48.8 %. The mass loss observed is accompanied by very strong exothermal effects. The remaining part of the sample mass which had not decomposed up to the temperature of 1000 °C (approx. 51.2 %) probably represents the carbon fibres and ceramic clay present in the material, because these decompose at a temperature higher than the range of the performed thermal analysis and higher than the temperature of the liquid metal fed into the gating system (about 1300— 1400 °C). In addition, volatile products of degradation were analysed using IR spectrum and the thermogravimetry (TG) method coupled online with mass spectrometry (MS). In the temperature range 300—750 °C signals for small molecular masses were noticed, which indicates that the degradation process and fragmentation of polymer chains in the cellulose occur as well as formation of small particle compounds, primarily H 2 O and CO 2 . No signals given off by aromatic compounds

Tailoring kappa/iota-hybrid carrageenan from Mastocarpus stellatus with desired gel quality through pre-extraction alkali treatment

Mastocarpus stellatus seaweeds were subjected to different alkali pre-treatments to investigate the effect of alkali type, concentration and treatment duration on the chemical structure and the gelling properties of extracted kappa/iota-hybrid carrageenan (KI). Increasing the concentration in KOH and the pre-treatment time gives KI with lower amounts of nu-carrageenan units, which have a direct impact on the improved gelling properties. However, excessive KOH concentration and pre-treatment duration give KI with smaller molecular mass and depressed gel properties. NaOH is more efficient in converting nu-carrageenan to iota-carrageenan, but no correlation between the chemical structure and gel properties was found. An interaction between the alkali concentration and the pre-treatment duration complicates the rationalization of NaOH efficiency. A set of KI with 60–75 mol% kappa-carrageenan and tailored gelling properties for a wide range of applications is delivered.

Activation of the small G-protein Rac by human rhinovirus attenuates the TLR3/IFN-α axis while promoting CCL2 release in human monocyte-lineage cells

Although rhinoviral infections, a major cause of asthma exacerbations, occur predominantly in upper airway bronchial epithelial cells, monocytic-lineage cells are implicated in establishing the inflammatory microenvironment observed during the disease. Human rhinovirus (HRV) is unique in that nearly genetically identical viruses bind either the ICAM-1 or low-density lipoprotein receptor (LDL-R). Within minutes of binding, HRV is capable of eliciting a signaling response in both epithelial cells and monocyte-derived macrophages. It is unclear whether this signaling response is important to the subsequent release of inflammatory mediators, particularly in cells not capable of supporting viral replication. We show here that the small molecular mass G-protein Rac is activated following exposure of macrophages to HRV serotypes known to be ICAM-1- and LDL-R-tropic. We demonstrate that inhibiting Rac resulted in the upregulation of TLR3 in macrophages exposed to major- and minor-group HRV, and resulted in increased release of IFN-α. Furthermore, inhibiting Rac in HRV-exposed macrophages attenuated activation of the stress kinase p38 and release of the pro-inflammatory cytokine CCL2, but inhibiting Rac did not affect release of the pro-inflammatory cytokine CCL5. These findings suggest that Rac is an important regulator in establishing the inflammatory microenvironment that is initiated in the human airway upon exposure to rhinovirus.

Aluminum Oxide Nanoparticles as Carriers and Adjuvants for Eliciting Antibodies from Non-immunogenic Haptens

Functionalized nanomaterials have applications yet to be discovered, especially in the biological field and, in particular, the in vivo production of antibodies. In this paper, we show that aluminum oxide nanoparticles that are covalently coupled to haptens (small molecular mass compounds) activate the immune system, eliciting antibodies (polyclonal and monoclonal) specific for the herbicide atrazine, the antibiotic sulfasalazine, and the vitamin biotin in mice and rabbits. The particles play the role of carrier and adjuvant, with the immune response being dependent on size and crystallinity. The affinity constants of the antibodies are similar to those reached with traditional immunization strategies based on the use of carrier proteins. This approach has not been previously described, being of scientific and practical interest, as it can lead to immunogens safer than the conventional ones, potentially applicable to human vaccination. In addition, the useful advantages of this technique include the stability of the metallic particles, the synthesis of immunogens in organic media, the versatility of particle derivatization, the ease of purification, the full chemical characterization of immunogens, the lack of a requirement for an adjuvant, the reduction of the cross reactivity, and the low cost of materials. Also, the analytical performances of the immunoassays developed using these antibodies are comparable to those obtained by the standard protocols. Analytical applications of the developed ELISAs were fully demonstrated. Characterization (IgG nature, affinity constants, etc.) of the immunoreagents were also fully assessed.

Alterations to the Bull Sperm Surface Proteins That Bind Sperm to Oviductal Epithelium1

Three Binder of SPerm proteins (BSP1, BSP3, BSP5) are secreted by bovine seminal vesicles into seminal plasma and adsorbed onto sperm. When sperm inseminated into the female reach the oviduct, the BSP proteins bind them to its epithelial lining, forming a sperm storage reservoir. Previously, we reported that binding of capacitated sperm to oviductal epithelium in vitro is lower than that of uncapacitated sperm and we proposed that reduced binding was due to loss of BSP proteins during capacitation. Because of differences in amino acid sequences, we predicted that each BSP would respond differently to capacitating conditions. To test whether all three BSP proteins were lost from sperm during capacitation and whether the kinetics of loss differed among the three BSP proteins, ejaculated bull sperm were incubated under various capacitating conditions, and then the amounts of BSP proteins remaining on the sperm were assayed by Western blotting. Capacitation was assayed by analysis of protein tyrosine phosphorylation. While loss of BSP1 was not detected, most of the BSP5 was lost from sperm during incubation in TALP medium, even without addition of the capacitation enhancers heparin and dbcAMP-IBMX. Surprisingly, a smaller molecular mass was detected by anti-BSP3 antibodies in extracts of incubated sperm. Its identity was confirmed as BSP3 by mass spectrometry, indicating that BSP3 undergoes modification on the sperm surface. These changes in the composition of BSP proteins on sperm could play a role in releasing sperm from the storage reservoir by modifying sperm interactions with the oviductal epithelium.

What will metabolomics studies mean to endocrinology?

Personalized medicine is a key topic for the development of future health-care provisions, even in endocrinology. The objective of personalized medicine is to use state-of-the-art diagnostics and novel therapeutic interventions that take into account the specific requirements and characteristics of the individual patient in order to optimize the effectiveness of treatments, avoid unwanted adverse reactions, and significantly reduce health-care costs. Identification of molecular pathways by integrated analysis of genetics and further molecular information by available OMICs technologies in combination with clinical phenotypes and biomarkers is the hallmark of this development. The majority of our current medical practice is based on extrapolated research in physiology, knowledge from randomized controlled trials or epidemiological studies of large cohorts, and expert experience. Most of those studies do not take into account the individual’s genetic, proteomic, and metabolic characteristics. It is time to bridge this gap between knowledge accumulated from basic science, discovering molecular pathways and mechanism by new OMICs technologies, and clinical research with an implementation of personalized medicine at the patient’s bedside ‐ this can be achieved by the approach of metabolomics. Metabolomics is a technique that provides a comprehensive assessment of small molecular mass organic compounds that are substrates or products of metabolic pathways as an individual fingerprint of metabolism. Recent advances in this field allow the acquisition of high-throughput metabolic profiles in different biofluids like plasma or urine in animal models or humans. The most common analytical platforms are mass spectrometry and nuclear magnetic resonance spectroscopy, which allow the detection of individual metabolic fingerprint (metabolic profiling) as well as untargeted quantitative or semiquantitative analysis for measurement of dynamic metabolites (metabolomics). The detected spectra contain a few hundred to thousands of signals related to both genetic and environmental contributions. Therefore, these techniques are the most promising tools for the identification of new biomarkers. Basically all OMICs techniques, such as genome-wide association studies (GWAS), transcriptomics, proteomics as well as metabolomics are able to identify the up to now unknown pathways or underlying mechanisms for many