The importance preoperative and postoperative determination sepsis biomarkers for diagnosis and prognosis intraabdominal Infections

Značaj preoperativnog i postoperativnog određivanja biomarkera sepse za dijagnozu i prognozu intraabdominalnih infekcija Sepsa je klinička manifestacija generalizovane inflamatorne reakcije domaćina na infekciju. Odgovor domaćina na mikroorganizam koji u njega prodire obuhvata brzo umnožavanje sinhronizovanih signala i odgovora koji se mogu širiti iz zahvaćenog tkiva. Imunološki mehanizmi obično uspevaju da potisnu sepsu. Kada ovi mehanizmi budu nadjačani, i to onda kada mikroorganizam krene iz lokalnog žarišta u cirkulaciju, homeostaza može zatajiti i tada nastaje stanje teške sepse. Sepsa se može razviti kao odgovor na infekciju izazvanu mikroorganizmima, kao što su Gram-pozitivne i/ili Gram-negativne bakterije koje pokreću imuno-metaboličku reakciju organizma poznatu kao "sistemski inflamatorni odgovor" tj. SIRS. Prema ACCP/SCCM, sepsa se definiše kao sistemski odgovor organizma na infekciju sa najmanje dve odlike za SIRS i potvrđenim prisustvom infektivnog agensa. Ako ovakvom stanju pridodamo i disfunkciju organa, praćenu hipoperfuzijom i hipotenzijom, govorimo o teškoj sepsi. Odgovor organizma u sepsi podrazumeva oslobađanje mnogih biomarkera, i to je činjenica koja nas navodi da se neki od ovih biomarkera mogu koristiti kao markeri infekcije ili ozbiljnosti sepse. U poslednjoj deceniji prokalcitonin (PCT) je potvrdio svoju ulogu kao marker sepse, a zajedno sa drugim rutinskim parametrima (broj leukocita (WBC) i koncentracija C-reaktivnog proteina (CRP)) omogućava brzu i pravovremenu dijagnozu infektivnih stanja. U poslednjih 4–5 godina, kao pouzdan marker sepse sve više se izdvaja i presepsin. Cilj istraživanja bio je da se ispitaju dijagnostički i prognostički značaj biomarkera sepse kod akutnih abdominalnih stanja, kao i da se ispita korelacija između parametara. Uzorci krvi su sakupljani u vreme prijema, preoperativno, i nakon hirurške intervencije. Određivani su sledeći parametri: presepsin, prokalcitonin (PCT), CRP, IL- 6, LBP i rutinski biohemijski, hematološki i parametri hemostaze. Ispitivanje je sprovedeno u Laboratoriji Centra za medicinsku biohemiju koja primenjuje standardne metode po instrukcijama proizvođača. Rezultati su statistički analizirani. ROC analiza je korišćena da se ispita sposobnost ispitivanih markera za dijagnozu sepse. Istraživanje je obuhvatilo dve studije. Prva studija obuhvatila je 98 pacijenata sa abdominalnim bolom i 90 zdravih dobrovoljaca (kao kontrolna grupa). Nakon hirurškog zahvata, obrazovane su grupe na osnovu pozitivnog (sepsa grupa, N=58) tj. negativnog (SIRS grupa, N=40) mikrobiološkog nalaza o intraabdominalnoj infekciji. Koncentracije prokalcitonina (izražene kao medijana) u serumu zdravih pojedinaca, bolesnika sa dijagnozom SIRS i bolesnika sa sepsom, na prijemu, su iznosile 0,21 ng/mL, 0,45 ng/mL i 2,32 ng/mL.The importance preoperative and postoperative determination sepsis biomarkers for diagnosis and prognosis intraabdominal infections Sepsis is a clinical manifestation of the general inflammatory reaction of the host-body to the infection. The reaction of the host-body to the microorganism that has entered into it implies the rapid multiplication of synchronized signals and responses issuing from the infected tissue. Immunological mechanisms usually manage to suppress sepsis. However, if these mechanisms are overwhelmed, which usually happens due to the invasion of the bloodstream by the microorganisms from a focus of infection, homeostasis is likely to be broken and a severe case of sepsis occurs. Sepsis may develop as a response to the infection caused by any microorganism, by various Gram-positive and Gram-negative bacteria which trigger the immunological reaction of the organism – the systemic inflammation response syndrome (SIRS). According to ACCP/SCCM, sepsis can be defined as the systemic response of the organism to the infection if the presence of at least two SIRS symptoms and an infectious agent is confirmed. Should we add organ failure, hypoperfusion and hypotension to the aforementioned symptoms, we are referring to the severe case of sepsis. The sepsis response involves the release many biomarkers, a fact that leads to the suggestion that some of these biomarkers could be used as markers of infection or sepsis severity. In the last decade Procalcitonin (PCT) has confirmed its role as marker of sepsis, and along with other routine parameters (white blood cell (WBC) count and C-reactive protein (CRP) concentration) allow rapid and timely diagnosis of infectious conditions. In the last 4–5 years, presepsin became a valuable marker in sepsis qualification. The aim of study was to identify the diagnostic and prognostic significance sepsis biomarkers in acute abdominal conditions and also to examine the correlation between parameters. Samples of blood for measurement each markers were collected at the time of admission and after medical treatment. We determined the following parameters, markers of sepsis: presepsin, prokalcitonin (PCT), CRP, IL-6, LBP and routine biochemical, hematological and hemostatic parameters. Sepsis biomarkers were determined at the central laboratory Center of Medical Biochemistry using commercial available methods following the instructions of the manufacturers. Results were statisticaly analyzed. ROC analyses were used to examine the capability of markers to diagnose sepsis. Research was based on two study. First study comprised 98 patients suffering from acute abdominal conditions, and 90 healthy volunteers as the control group..

MimoDB 2.0: a mimotope database and beyond

Mimotopes are peptides with affinities to given targets. They are readily obtained through biopanning against combinatorial peptide libraries constructed by phage display and other display technologies such as mRNA display, ribosome display, bacterial display and yeast display. Mimotopes have been used to infer the protein interaction sites and networks; they are also ideal candidates for developing new diagnostics, therapeutics and vaccines. However, such valuable peptides are not collected in the central data resources such as UniProt and NCBI GenPept due to their ‘unnatural’ short sequences. The MimoDB database is an information portal to biopanning results of random libraries. In version 2.0, it has 15 633 peptides collected from 849 papers and grouped into 1818 sets. Besides the core data on panning experiments and their results, broad background information on target, template, library and structure is included. An accompanied benchmark has also been compiled for bioinformaticians to develop and evaluate their new models, algorithms and programs. In addition, the MimoDB database provides tools for simple and advanced searches, structure visualization, BLAST and alignment view on the fly. The experimental biologists can easily use the database as a virtual control to exclude possible target-unrelated peptides. The MimoDB database is freely available at http://immunet.cn/mimodb

Looking for leakage or monitoring for public assurance?

Monitoring is a regulatory requirement for all carbon dioxide capture and geological storage (CCS) projects to verify containment of injected carbon dioxide (CO2) within a licensed geological storage complex. Carbon markets require CO2 storage to be verified. The public wants assurances CCS projects will not cause any harm to themselves, the environment or other natural resources. In the unlikely event that CO2 leaks from a storage complex, and into groundwater, to the surface, atmosphere or ocean, then monitoring methods will be required to locate, assess and quantify the leak, and to inform the community about the risks and impacts on health, safety and the environment. This paper considers strategies to improve the efficiency of monitoring the large surface area overlying onshore storage complexes. We provide a synthesis of findings from monitoring for CO2 leakage at geological storage sites both natural and engineered, and from monitoring controlled releases of CO2 at four shallow release facilities – ZERT (USA), Ginninderra (Australia), Ressacada (Brazil) and CO2 field lab (Norway)

Thermodynamic study of interactions between ZnO and ZnO binding peptides using isothermal titration calorimetry

Whilst material specific peptide binding sequences have been identified using a combination of combinato-rial methods and computational modelling tools, a deep molecular level understanding of the fundamental principles through which these interactions occur and in some instances modify the morphology of inorganic materials is far from being fully realized. Understanding the thermodynamic changes that occur during peptide-inorganic interactions and correlating these to structural modifications of the inorganic materials could be the key to achieving and mastering con-trol over material formation processes. This study is a detailed investigation applying isothermal titration calorimetry (ITC) to directly probe thermodynamic changes that occur during interaction of ZnO binding peptides (ZnO-BPs) and ZnO. The ZnO-BPs used are reported sequences G-12 (GLHVMHKVAPPR), GT-16 (GLHVMHKVAPPR-GGGC) and alanine mutants of G-12 (G-12A6, G-12A11 and G-12A12) whose interaction with ZnO during solution synthesis studies have been extensively investigated. The interactions of the ZnO-BPs with ZnO yielded biphasic isotherms comprising both an endo-thermic and an exothermic event. Qualitative differences were observed in the isothermal profiles of the different pep-tides and ZnO particles studied. Measured ΔG values were between -6 and -8.5 kcal/mol and high adsorption affinity val-ues indicated the occurrence of favourable ZnO-BP-ZnO interactions. ITC has great potential in its use to understand peptide-inorganic interactions and with continued development, the knowledge gained may be instrumental for simplifi-cation of selection processes of organic molecules for the advancement of material synthesis and design

Examining the Interactome of Huperzine A by Magnetic Biopanning

Huperzine A is a bioactive compound derived from traditional Chinese medicine plant Qian Ceng Ta (Huperzia serrata), and was found to have multiple neuroprotective effects. In addition to being a potent acetylcholinesterase inhibitor, it was thought to act through other mechanisms such as antioxidation, antiapoptosis, etc. However, the molecular targets involved with these mechanisms were not identified. In this study, we attempted to exam the interactome of Huperzine A using a cDNA phage display library and also mammalian brain tissue extracts. The drugs were chemically linked on the surface of magnetic particles and the interactive phages or proteins were collected and analyzed. Among the various cDNA expressing phages selected, one was identified to encode the mitochondria NADH dehydrogenase subunit 1. Specific bindings between the drug and the target phages and target proteins were confirmed. Another enriched phage clone was identified as mitochondria ATP synthase, which was also panned out from the proteome of mouse brain tissue lysate. These data indicated the possible involvement of mitochondrial respiratory chain matrix enzymes in Huperzine A's pharmacological effects. Such involvement had been suggested by previous studies based on enzyme activity changes. Our data supported the new mechanism. Overall we demonstrated the feasibility of using magnetic biopanning as a simple and viable method for investigating the complex molecular mechanisms of bioactive molecules

Bimodal dynamics of primary metabolism-related responses in tolerant potato-Potato virus Y interaction

BACKGROUND: Potato virus Y (PVY) is a major pathogen that causes substantial economic losses in worldwide potato production. Different potato cultivars differ in resistance to PVY, from severe susceptibility, through tolerance, to complete resistance. The aim of this study was to better define the mechanisms underlying tolerant responses of potato to infection by the particularly aggressive PVY(NTN) strain. We focused on the dynamics of the primary metabolism-related processes during PVY(NTN) infection. RESULTS: A comprehensive analysis of the dynamic changes in primary metabolism was performed, which included whole transcriptome analysis, nontargeted proteomics, and photosynthetic activity measurements in potato cv. Désirée and its transgenic counterpart depleted for accumulation of salicylic acid (NahG-Désirée). Faster multiplication of virus occurred in the NahG-Désirée, with these plants developing strong disease symptoms. We show that while the dynamics of responses at the transcriptional level are extensive and bimodal, this is only partially translated to the protein level, and to the final functional outcome. Photosynthesis-related genes are transiently induced before viral multiplication is detected and it is down-regulated later on. This is reflected as a deficiency of the photosynthetic apparatus at the onset of viral multiplication only. Interestingly, specific and constant up-regulation of some RuBisCO transcripts was detected in Désirée plants, which might be important, as these proteins have been shown to interact with viral proteins. In SA-deficient and more sensitive NahG-Désirée plants, consistent down-regulation of photosynthesis-related genes was detected. A constant reduction in the photochemical efficiency from the onset of viral multiplication was identified; in nontransgenic plants this decrease was only transient. The transient reduction in net photosynthetic rate occurred in both genotypes with the same timing, and coincided with changes in stomatal conductivity. CONCLUSIONS: Down-regulation of photosynthesis-related gene expression and decreased photosynthetic activity is in line with other studies that have reported the effects of biotic stress on photosynthesis. Here, we additionally detected induction of light-reaction components in the early stages of PVY(NTN) infection of tolerant interaction. As some of these components have already been shown to interact with viral proteins, their overproduction might contribute to the absence of symptoms in cv. Désirée. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-015-1925-2) contains supplementary material, which is available to authorized users

Simvastatin Sodium Salt and Fluvastatin Interact with Human Gap Junction Gamma-3 Protein

Finding pleiomorphic targets for drugs allows new indications or warnings for treatment to be identified. As test of concept, we applied a new chemical genomics approach to uncover additional targets for the widely prescribed lipid-lowering pro-drug simvastatin. We used mRNA extracted from internal mammary artery from patients undergoing coronary artery surgery to prepare a viral cardiovascular protein library, using T7 bacteriophage. We then studied interactions of clones of the bacteriophage, each expressing a different cardiovascular polypeptide, with surface-bound simvastatin in 96-well plates. To maximise likelihood of identifying meaningful interactions between simvastatin and vascular peptides, we used a validated photo-immobilisation method to apply a series of different chemical linkers to bind simvastatin so as to present multiple orientations of its constituent components to potential targets. Three rounds of biopanning identified consistent interaction with the clone expressing part of the gene GJC3, which maps to Homo sapiens chromosome 7, and codes for gap junction gamma-3 protein, also known as connexin 30.2/31.3 (mouse connexin Cx29). Further analysis indicated the binding site to be for the N-terminal domain putatively ‘regulating’ connexin hemichannel and gap junction pores. Using immunohistochemistry we found connexin 30.2/31.3 to be present in samples of artery similar to those used to prepare the bacteriophage library. Surface plasmon resonance revealed that a 25 amino acid synthetic peptide representing the discovered N-terminus did not interact with simvastatin lactone, but did bind to the hydrolysed HMG CoA inhibitor, simvastatin acid. This interaction was also seen for fluvastatin. The gap junction blockers carbenoxolone and flufenamic acid also interacted with the same peptide providing insight into potential site of binding. These findings raise key questions about the functional significance of GJC3 transcripts in the vasculature and other tissues, and this connexin’s role in therapeutic and adverse effects of statins in a range of disease states

Thermal studies of silver- poly(methylmethacrylate) nanocomposites

The novel nanocomposite material based on surface-modified silver nanoparticles (Np) with average diameter of 6 nm and poly(methylmethacrylate) (PMMA) matrix was synthesized. The effect of silver loading on the thermal properties of PMMA was studied. The obtained transparent nanocomposites Ag/PMMA films were characterized by IR spectroscopy and thermal techniques (TGA, DSC). The chemical composition of the PMMA matrix is not changed in the presence of the metal nanoparticles, but the thermal stability of the nanocomposites was found to be better than thermal stability of the pure polymer. Also, Ag nanoparticles have pronounced effect on a glass transition temperature (Tg) of PMMA