Omics Analysis of Blood-Responsive Regulon in Bordetella pertussis Identifies a Novel Essential T3SS Substrate

Bacterial pathogens sense specific cues associated with different host niches and integrate these signals to appropriately adjust the global gene expression. Bordetella pertussis is a Gram-negative, strictly human pathogen of the respiratory tract and the etiological agent of whooping cough (pertussis). Though B. pertussis does not cause invasive infections, previous results indicated that this reemerging pathogen responds to blood exposure. Here, omics RNA-seq and LC–MS/MS techniques were applied to determine the blood-responsive regulon of B. pertussis. These analyses revealed that direct contact with blood rewired global gene expression profiles in B. pertussis as the expression of almost 20% of all genes was significantly modulated. However, upon loss of contact with blood, the majority of blood-specific effects vanished, with the exception of several genes encoding the T3SS-secreted substrates. For the first time, the T3SS regulator BtrA was identified in culture supernatants of B. pertussis. Furthermore, proteomic analysis identified BP2259 protein as a novel secreted T3SS substrate, which is required for T3SS functionality. Collectively, presented data indicate that contact with blood represents an important cue for B. pertussis cells.     

Salivary Proteome Changes in Response to Acute Psychological Stress Due to an Oral Exam Simulation in University Students: Effect of an Olfactory Stimulus

The autonomic nervous system (ANS) plays a crucial role both in acute and chronic psychological stress eliciting changes in many local and systemic physiological and biochemical processes. Salivary secretion is also regulated by ANS. In this study, we explored salivary proteome changes produced in thirty-eight University students by a test stress, which simulated an oral exam. Students underwent a relaxation phase followed by the stress test during which an electrocardiogram was recorded. To evaluate the effect of an olfactory stimulus, half of the students were exposed to a pleasant odor diffused in the room throughout the whole session. Saliva samples were collected after the relaxation phase (T0) and the stress test (T1). State anxiety was also evaluated at T0 and T1. Salivary proteins were separated by two-dimensional electrophoresis, and patterns at different times were compared. Spots differentially expressed were trypsin digested and identified by mass spectrometry. Western blot analysis was used to validate proteomic results. Anxiety scores and heart rate changes indicated that the fake exam induced anxiety. Significant changes of α-amylase, polymeric immunoglobulin receptor (PIGR), and immunoglobulin α chain (IGHA) secretion were observed after the stress test was performed in the two conditions. Moreover, the presence of pleasant odor reduced the acute social stress affecting salivary proteome changes. Therefore, saliva proteomic analysis was a useful approach to evaluate the rapid responses associated to an acute stress test also highlighting known biomarkers.     

A new family of sugar-inducible expression vectors for Escherichia coli

A set of 11 expression vectors was constructed, each of themharbouring a cloning cassette under the control of the promoter.Some of these vectors enable expression of foreign proteinsin the cytoplasm, while others include a synthetic sequencecoding for a very efficient secretion signal sequence. Otherfeatures are an fl origin of replication (in plus or minus orientation)and a promoter^up mutation that enhances the already very highlevel of expression from these vectors. With such a versatilevector family, cloning, sequencing and sitedirected mutagenesiscan be performed on the same vector, and the level of expressioncan be defined according to the specific constraints of a givenprotein.     

齿科微晶玻璃的腐蚀机理及溶胶-凝胶防腐蚀涂层研究

利用体外试验方法以及SEM／EDAX、ICP、FTIR等分析技术，对氟硅碱钙石(fluorcanasite,K2Na4Ca5Si12O30F4)齿科微晶玻璃的腐蚀机理，以及溶胶-凝胶Al2O3，、TiO2及ZrO2涂层改善微晶玻璃抗侵蚀的效果进行了比较分析．研究表明，口腔溶液对该齿科微晶玻璃的侵蚀源于溶液中的H3O 离子与微晶玻璃表面的Na 及K 发生迅速的离子交换，从而形成结构疏松的表面富SiO2层，其Si-O网络进而被H2O分子所断开，使SiO2以可溶性硅胶形式溶解于溶液中．此反应导致大量的表面结构缺陷，从而降低了材料的强度和使用寿命。浸泡25天后微晶玻璃表面最终的反应产物是一层低结晶度的颗粒状碳酸羟基磷灰石Cal0(PO4)6(2OH-，CO3^2-)．溶胶-凝胶A12O3涂层具有显著的改善齿科微晶玻璃抗唾液腐蚀的作用。     

Dental amalgam – the effect of the technology of alloy powder preparation on the corrosion behaviour and the release of mercury

Dental amalgams are based on a broad spectrum of materials differing in their chemical composition, metallurgical treatment, and in the way the initial alloys powders are prepared. In addition to their chemical composition, amalgams based on various powders differ in both their microstructure and the amount of mercury needed for preparation. All these facts may affect electrochemical processes occurring during their interaction with oral fluids, and also mercury release. While verifying the effect of the technology used for the preparation of the high‐copper ternary alloy powder on the properties of resulting amalgams, this study aimed at the mechanism of their interaction with a model saliva solution as well as mercury release was included. Measurements were done in a model saliva solution using standard electrochemical methods and exposition measurements. The interaction of individual types of amalgams with artificial saliva did not reveal any significant differences. The free corrosion potential of all these amalgams in an aerated solution settled in the range of values in which tin oxidation, resulting in a layer of insoluble corrosion producsts, turned out to be the dominant anodic process. The rate of mercury release was the lowest for amalgams based on a gas‐atomized alloy. The highest rate of mercury release, and also its dependence on time, was exhibited by lathe‐cut powder based amalgam. In addition to different volume fraction of the Ag‐Hg phase and the level of its tin alloying, this different behaviour may be explained by differences in the rate at which a layer of tin corrosion products acting as a barrier to mercury release is formed.     

Conventional and Unconventional Mechanisms by which Exocytosis Proteins Oversee β-cell Function and Protection

Type 2 diabetes (T2D) is one of the prominent causes of morbidity and mortality in the United States and beyond, reaching global pandemic proportions. One hallmark of T2D is dysfunctional glucose-stimulated insulin secretion from the pancreatic β-cell. Insulin is secreted via the recruitment of insulin secretory granules to the plasma membrane, where the soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) and SNARE regulators work together to dock the secretory granules and release insulin into the circulation. SNARE proteins and their regulators include the Syntaxins, SNAPs, Sec1/Munc18, VAMPs, and double C2-domain proteins. Recent studies using genomics, proteomics, and biochemical approaches have linked deficiencies of exocytosis proteins with the onset and progression of T2D. Promising results are also emerging wherein restoration or enhancement of certain exocytosis proteins to β-cells improves whole-body glucose homeostasis, enhances β-cell function, and surprisingly, protection of β-cell mass. Intriguingly, overexpression and knockout studies have revealed novel functions of certain exocytosis proteins, like Syntaxin 4, suggesting that exocytosis proteins can impact a variety of pathways, including inflammatory signaling and aging. In this review, we present the conventional and unconventional functions of β-cell exocytosis proteins in normal physiology and T2D and describe how these insights might improve clinical care for T2D.     

Effect of volatiles collected above fecal pellets on behavior of the rabbit,Oryctolagus cuniculus,tested in an experimental chamber

Gas chromatographic techniques were used to trap and fractionate the volatile substances emanating from the fecal pellets of wild rabbits,Oryctolagus cuniculus, which incorporate the odor of the anal gland secretion. The volatiles were bubbled through water, trapped on Chromosorb 105, transferred to Silicone SF96 traps, passed through a GC capillary column, and retrapped on SF96. The behavioral effect of trapped and fractionated volatiles was determined in four series of bioassays involving 51 adult, male, wild-type rabbits in 112, ten-minute tests. The bioassay was based on the demonstration of territorial confidence by the rabbits under the influence of their own odor. In the bioassays of the unfractionated volatiles eluted from Chromosorb 105 and SF96 traps the animals were significantly more confident in the presence of the volatiles from their own fecal pellets. One of the two fractions of the total volatiles was ineffective while the effect of the other was less than that of the unfractionated odor. The results demonstrate that the gas chromatographic techniques can be used to manipulate complex mammalian odors with precision. Combined with a discriminative bioassay, this opens up the possibility of identifying the specific combinations of volatile substances involved in the formation of olfactory signals.     

A generalist herbivore in a specialist mode Metabolic,sequestrative, and defensive consequences

Adults of a generalist herbivore, the lubber grasshopper,Romalea guttata, can be converted to functional specialists by feeding them exclusively on catnip,Nepeta cataria. No obvious adverse effects on adult development resulted from this enforced monophagy. Notwithstanding the fact thatR. guttata has had no coevolutionary relationship with this Eurasian mint, it readily sequesters compounds that are identical to or derived from the terpenoid lactones that are characteristic ofN. cataria. R. guttata appears to both biomagnify minor allelochemicals and to sequester metabolites of theNepeta terpenes in its paired defensive glands. The levels of autogenously produced phenolics are not affected by feeding onN. cataria and the defensive secretions of catnip-fed grasshoppers are more repellent to ants than those of wild-fed acridids. Metabolites of theN. cataria monoterpenes are sequestered in the defensive glands when catnip is added to the natural diet ofR. guttata. The ability of a generalist,R. guttata, to facilely bioaccumulate a potpourri of foreign allelochemicals when feeding in a specialist mode is analyzed in terms of its biochemical, physiological, and functional significance. Sequestration is examined as a response to the enteric effronteries represented by the phytochemicals that can be characteristic of the overload in a monophagous diet.