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Background: Intima-media thickness (IMT) is a useful marker in early detection of atherosclerotic lesions in carotid arteries in Type-2 Diabetes Mellitus (T2DM) patients. The study was conducted to determine the association of IMT with various physical and biochemical parameters in the Indian population. Objective: To compare IMT of common and internal carotid arteries in T2DM patients and healthy individuals and study the correlation of IMT with physical and biochemical parameters. Methods: Case group with T2DM (n=27) were matched with Control group consisting of healthy individuals (n=27) of similar age and sex were included. Physical parameters and routine biochemical data were collected and compared. Ultrasonographical imaging of the common and internal carotid arteries of both sides were performed presence of plaque was observed. Data was analyzed by software IBM® SPSS Statistics V20.0. For all analyses, P

The current study was based on Computed Tomography (CT) findings and results. We found 63% of patients with acute interstitial edematous pancreatitis, and we found 37% of patients with acute necrotic pancreatitis in 46 cases. Based on our findings, we observed bulky pancreas among 26 cases (56.52%), 17 cases (36.95%) of pancreatic necrosis, 9 cases of pancreatic atrophy, 11 cases of pseudocyst, and 9 cases of peri-pancreatic fluid. When we evaluated the Modified CT Severity index among the study cases, we observed that 19.56% cases have mild index, the majority (56.52% cases) have moderate index while 23.91% have severe index. We also found that majority of the cases were clinically cured and discharged, i.e., 47.82%. 19.56% of the total study subjects reported recurrence of episodes of pancreatitis, while 30.43% of the total patients turned into chronic pancreatitis. This study presents a cross-sectional hospital-based analytical study carried out among 46 cases of AP referred to under the radiology department for further diagnostic evaluation, in a tertiary healthcare facility in Maharashtra. The present study Data Source included all the AP patients referred to the radiological department in a tertiary health research center. They met the standards for inclusiveness, which assented to the analysis. Type of study is Hospital analytical based study. The study duration is 18 months. Data collection was done through semi-structured, pre-designed, and pre-validated proformas in patients meeting inclusion requirements that included disease history, clinical outcomes, investigative records, and descriptions of surgical interventions.

Cell-penetrating peptides (CPPs) have distinct properties to translocate across cell envelope. The key property of CPPs to translocation with attached molecules has been utilized as vehicles for the delivery of several potential drug candidates that illustrate the significant effect in in-vitro experiment but fail in in-vivo experiment due to selectively permeable nature of cell envelop. Penetratin, a well-known CPP identified from the third α-helix of Antennapedia homeodomain of Drosophila, has been widely used and studied for the delivery of bioactive molecules to treat cancers, stroke, and infections caused by pathogenic organisms. Few studies have demonstrated that penetratin directly possesses antimicrobial activities against bacterial and fungal pathogens; however, the mechanism is unknown. In this study, we have utilized the power of high-throughput Saccharomyces cerevisiae proteome microarrays to screen all the potential protein targets of penetratin. Saccharomyces cerevisiae proteome microarrays assays of penetratin followed by statistical analysis depicted 123 Saccharomyces cerevisiae proteins as the protein targets of penetratin out of ~5800 Saccharomyces cerevisiae proteins. To understand the target patterns of penetratin, enrichment analyses were conducted using 123 protein targets. In biological process: ribonucleoprotein complex biogenesis, nucleic acid metabolic process, actin filament-based process, transcription, DNA-templated, and negative regulation of gene expression are a few significantly enriched terms. Cytoplasm, nucleus, and cell-organelles are enriched terms for cellular component. Protein-protein interactions network depicted ribonucleoprotein complex biogenesis, cortical cytoskeleton, and histone binding, which represent the major enriched terms for the 123 protein targets of penetratin. We also compared the protein targets of penetratin and intracellular protein targets of antifungal AMPs (Lfcin B, Histatin-5, and Sub-5). The comparison results showed few unique proteins between penetratin and AMPs. Nucleic acid metabolic process and cellular component disassembly were the common enrichment terms for penetratin and three AMPs. Penetratin shows unique enrichment items that are related to DNA biological process. Moreover, motif enrichment analysis depicted different enriched motifs in the protein targets of penetratin, LfcinB, Histatin-5, and Sub-5.

With their wide repertoire of mechanisms, antimicrobial peptides (AMPs) are promising alternatives to fight against varied pathogenic microorganisms (bacteria, fungi, viruses, parasites, etc.). AMPs, novel components of the innate immune defense system, are secreted by all organisms. The aquatic environment represents a huge population and an enormous source of varied AMPs. Polyphemusin-I, a marine AMP isolated from hemocytes of an American horseshoe crab, possesses high antimicrobial activities. Studies on polyphemusin-I have verified the intracellular mechanisms of action, however, its intracellular targets are not yet explored. In this study, we employed Escherichia coli proteome microarrays to systematically screen the entire intracellular protein targets of polyphemusin-I. A total of 97 protein targets of polyphemusin-I were statistically analyzed from the quadruplicate Escherichia coli proteome microarrays assays. Among these identified protein targets, 56 proteins had cellular location inside the cell (i.e., cytoplasm), one in the plasma membrane, one in the periplasm and the rest 39 proteins had no specified cellular location. The bioinformatics analysis of these identified protein targets of polyphemusin-I in gene ontology (GO) enrichment category of molecular function revealed significant enrichment in nucleic acid related GO terms i.e., “RNA binding”, “nucleotide binding”, “nuclease activities”, “uracil DNA N-glycosylase activities” and others. Moreover, enrichment in GO category of biological process also depicted enrichment in nucleic acid related GO terms, such as “nucleic acid phosphodiester bond hydrolysis”, “deoxyribonucleotide metabolism”, and others. In accordance to GO enrichment analysis, protein families (PFAM) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways enrichment analysis also showed significant enrichment in nucleic acid terms. These enrichment results suggest that polyphemusin-I targets nucleic acid-associated proteins. Furthermore, to provide a comprehensive study, we compared the identified protein targets of polyphemusin-I with previously identified protein targets of four AMPs (P-Der, Lfcin B, PR-39, and Bac 7) using Escherichia coli proteome microarrays. The comparison study of five AMPs (polyhemusin-I, P-Der, Lfcin B, PR-39, and Bac 7) showed only nine common protein targets in all the five AMPs, whereas a total of 39 and 43 common protein targets were identified among the two marine AMPs (polyphemusin-I and P-Der) and three terrestrial AMPs (Lfcin B, PR-39 and Bac7), respectively. To further reveal the target pattern of marine and terrestrial AMPs, the enrichment results obtained from common protein targets of marine AMPs with terrestrial AMPs were compared. The comparison result indicated that AMPs have unique mechanism of action among marine or terrestrial AMPs. Hence, in this study, we have not only identified the intracellular protein targets of polyphemusin-I, but also revealed the protein target differences between marine AMPs and terrestrial AMPs.

We used yeast proteome microarrays (∼5800 purified proteins) to conduct a high-throughput and systematic screening of PI5P-interacting proteins with PI5P-tagged fluorescent liposomal nanovesicles. Lissamine rhodamine B-dipalmitoyl phosphatidylethanol was incorporated into the liposome bilayer to provide the nanovesicles with fluorescence without any encapsulants, which not only made the liposome fabrication much easier without the need for purification but also improved the chip-probing quality. A special chip assay was washed very gently without the traditional spin-dry step. Forty-five PI5P-interacting proteins were identified in triplicate with this special chip assay. Subsequently, we used flow cytometry to validate these interactions, and a total of 41 PI5P-interacting proteins were confirmed. Enrichment analysis revealed that these proteins have significant functions associated with ribosome biogenesis, rRNA processing, ribosome binding, GTP binding, and hydrolase activity. Their component enrichment is located in the nucleolus. The InterPro domain analysis indicated that PI5P-interacting proteins are enriched in the P-loop containing nucleoside triphosphate hydrolases domain (P-loop). Additionally, using the MEME program, we identified a consensus motif (IVGPAGTGKSTLF) that contains the Walker A sequence, a well-known nucleotide-binding motif. Furthermore, using a quartz crystal microbalance, both the consensus motif and Walker A motif showed strong affinities to PI5P-containing liposomes but not to PI5P-deprived liposomes or PI-containing liposomes. Additionally, the glycine (G6) and lysine (K7) residues of the Walker A motif (-GPAGTG6K7S-) were found to be critical to the PI5P-binding ability. This study not only identified an additional set of PI5P-interacting proteins but also revealed the strong PI5P-binding affinity (Kd = 1.81 × 10-7 M) of the Walker A motif beyond the motif's nucleotide-binding characteristic.

Antimicrobial peptides (AMPs) are intensively studied in terms of alternative drugs. Sub5 is a synthetic 12-mer AMP with substitutions of five amino acids of bactenecin 2A (Bac2A), a linear-ized bactenecin variant of bovine. Sub5 is highly effective against fungi with an ability to trans-locate cell membrane, but its targets are unknown. Systematic analysis of Sub5 targets will facil-itate our understanding on its mechanism of action. In this study, we used high-throughput Saccharomyces cerevisiae proteome microarrays to explore the potential protein targets of Sub5. The screening results showed 128 potential protein targets of Sub5. Bioinformatics analysis of protein targets of Sub5 revealed significant gene ontology (GO) enrichment in actin related pro-cess of “actin filament-based process”, “actin filament organization”, “actin cortical patch or-ganization”, regulation of “actin filament bundle assembly”. Moreover, the other enriched cat-egories in GO enrichment mostly contained actin associate proteins. In total, 11 actin-associated proteins were identified in the protein targets of Sub5. Protein family (PFAM) enrichment anal-ysis shows protein domain enriched in actin binding, i.e., “Cytoskeletal-regulatory complex EF hand (helix E-loop-helix F motif)”. Being consistent with GO analysis, Search Tool for the Re-trieval of Interacting Genes/Proteins (STRING) analysis of the protein targets of Sub5 showed ac-tin network with involvement of 15 protein targets. Along with actin-network, STRING analysis showed protein–protein interaction network in ribonucleoprotein, transcription and translation, chromosome, histone, and ubiquitin related, DNA repair, and chaperone. Multiple Expression motifs for Motif Elicitation (MEME) suite provided a consensus binding motif of [ED][ED]EEE[ED][ED][ED][ED][ED], in total of 75 protein targets of Sub5. This motif was present in 9 out of 15 actin-related proteins identified among protein targets of Sub5.

This is the twelfth chapter of the guideline "Calculated initial parenteral treatment of bacterial infections in adults - update 2018" in the 2Dies ist das zwölfte Kapitel der von der Paul-Ehrlich-Gesellschaft für Chemotherapie e.V. (PEG) herausgegebenen S2k Leitlinie „Kalkulierte parenterale Initialtherapie bakterieller Erkrankungen bei Erwachsenen – Update 2018“ in der 2. aktualisierten Fassung.Die bakterielle Endokarditis ist gekennzeichnet durch eine gleichbleibende Inzidenz, aber eine Veränderung in der betroffenen Patientenpopulation durch den Einsatz von Klappenprothesen, Fremdmaterialien wie Schrittmachern und den zunehmenden Einsatz invasiver medizinischer Maßnahmen. Damit verbunden ist eine Veränderung des Erregerspektrums hin zu Staphylokokken auch bei der Nativklappen-Endokarditis. Dieses Kapitel gibt Empfehlungen für das interdsiziplinäre Management der bakteriellen Endokarditis von der Diagnostik über die Prävention zur Therapie mit dem Fokus auf der Antibiotika-Therapie und evaluiert die aktuellen Empfehlungen der ESC aus deutscher Sicht.

Antimicrobial peptides (AMPs) have potential antifungal activities, however, their intracellular protein targets are poorly reported. Proteome microarray is an effective tool with high-throughput and rapid platform that systematically identifies the protein targets. In this study, we have used yeast proteome microarrays for systematical identification of the yeast protein targets of Lactoferricin B (Lfcin B) and Histatin-5. A total of 140 and 137 protein targets were identified from the triplicate yeast proteome microarray assays for Lfcin B and Histatin-5, respectively. The Gene Ontology (GO) enrichment analysis showed that Lfcin B targeted more enrichment categories than Histatin-5 did in all GO biological processes, molecular functions, and cellular components. This might be one of the reasons that Lfcin B has a lower minimum inhibitory concentration (MIC) than Histatin-5. Moreover, pairwise essential proteins that have lethal effects on yeast were analyzed through synthetic lethality. A total of 11 synthetic lethal pairs were identified within the protein targets of Lfcin B. However, only three synthetic lethal pairs were identified within the protein targets of Histatin-5. The higher number of synthetic lethal pairs identified within the protein targets of Lfcin B might also be the reason for Lfcin B to have lower MIC than Histatin-5. Furthermore, two synthetic lethal pairs were identified between the unique protein targets of Lfcin B and Histatin-5. Both the identified synthetic lethal pairs proteins are part of the Spt-Ada-Gcn5 acetyltransferase (SAGA) protein complex that regulates gene expression via histone modification. Identification of synthetic lethal pairs between Lfcin B and Histatin-5 and their involvement in the same protein complex indicated synergistic combination between Lfcin B and Histatin-5. This hypothesis was experimentally confirmed by growth inhibition assay.

Antimicrobial peptides (AMPs) act either through membrane lysis or by attacking intracellular targets. Intracellular targeting AMPs are a resource for antimicrobial agent development. Several AMPs have been identified as intracellular targeting peptides; however, the intracellular targets of many of these peptides remain unknown. In the present study, we used an Escherichia coli proteome microarray to systematically identify the protein targets of three intracellular targeting AMPs: bactenecin 7 (Bac7), a hybrid of pleurocidin and dermaseptin (P-Der), and proline-arginine-rich peptide (PR-39). In addition, we also included the data of lactoferricin B (LfcinB) from our previous study for a more comprehensive analysis. We analyzed the unique protein hits of each AMP in the Kyoto Encyclopedia of Genes and Genomes. The results indicated that Bac7 targets purine metabolism and histidine kinase, LfcinB attacks the transcription-related activities and several cellular carbohydrate biosynthetic processes, P-Der affects several catabolic processes of small molecules, and PR-39 preferentially recognizes proteins involved in RNA- and folate-metabolism-related cellular processes. Moreover, both Bac7 and LfcinB target purine metabolism, whereas LfcinB and PR-39 target lipopolysaccharide biosynthesis. This suggested that LfcinB and Bac7 as well as LfcinB and PR-39 have a synergistic effect on antimicrobial activity, which was validated through antimicrobial assays. Furthermore, common hits of all four AMPs indicated that all of them target arginine decarboxylase, which is a crucial enzyme for Escherichia coli survival in extremely acidic environments. Thus, these AMPs may display greater inhibition to bacterial growth in extremely acidic environments. We have also confirmed this finding in bacterial growth inhibition assays. In conclusion, this comprehensive identification and systematic analysis of intracellular targeting AMPs reveals crucial insights into the intracellular mechanisms of the action of AMPs.

Antimicrobial peptides have been considered well-deserving candidates to fight the battle against microorganisms due to their broad-spectrum antimicrobial activities. Several studies have suggested that membrane disruption is the basic mechanism of AMPs that leads to killing or inhibiting microorganisms. Also, AMPs have been reported to interact with macromolecules inside the microbial cells such as nucleic acids (DNA/RNA), protein synthesis, essential enzymes, membrane septum formation and cell wall synthesis. Proteins are associated with many intracellular mechanisms of cells, thus protein targets may be specifically involved in mechanisms of action of AMPs. AMPs like pyrrhocoricin, drosocin, apidecin and Bac 7 are documented to have protein targets, DnaK and GroEL. Moreover, the intracellular targeting AMPs are reported to influence more than one protein targets inside the cell, suggesting for the multiple modes of actions. This complex mechanism of intracellular targeting AMPs makes them more difficult for the development of resistance. Herein, we have summarized the current status of AMPs in terms of their mode of actions, entry to cytoplasm and inhibition of macromolecules. To reveal the mechanism of action, we have focused on AMPs with intracellular protein targets. We have also included the use of high-throughput proteome microarray to determine the unidentified AMP protein targets in this review.

Cordyceps militaris hot water extract (CMHW) containing cordycepin (cordycepin-CMHW) was used to study the anti-cancer effects in human A549 non-small cell lung carcinoma cells. Our results showed cordycepin-CMHW can inhibit cell proliferations in A549 cells by activating A3 adenosine receptor (A3AR) via the inactivation of Akt pathways. Cordycepin-CMHW can also induce apoptosis in the A549 cells by enhancing DNA fragmentation and chromatin condensation. We further observed that cordycepin-CMHW up-regulated caspase-9 and increased cleavage of caspase-3 and poly ADP ribose polymerase (PARP) in A549 cells. The results suggested cordycepin-CMHW is a highly selective treatment to de-regulation of the cell proliferation and apoptosis in non-small cell lung carcinoma via signaling pathways generated by A3AR activation.

Crude glycerol, an inevitable byproduct during biodiesel production, is emerging as a potential feedstock for fermentation, due to its availability and a reasonable price. Biological utilization of abundant crude glycerol to several value added products is contemporary research area with beneficial features. Solving the problem of proper disposal and raising economic viability of biodiesel industries. Several researches have been directed toward the production of numerous products by using Escherichia coli, an ideal organism for heterologous expression of various foreign proteins. In this fashion, recombinant E. coli strains were constructed for the simultaneous production of poly 3-hydroxybutyrate (P3HB) and bioethanol from crude glycerol. The incorporation of aldehyde reductase (Alrd) and aldehyde dehydrogenase (AldH) in recombinant strain showed 2-fold increment in crude glycerol utilization under aerobic condition. Moreover, these two enzymes introduced an alternative pathway leading toward the potential production of bioethanol which was more than redox-balancing steps. Acetate was accumulated as an intermediate product. Subsequently, acetate was utilized as substrate in the second pathway, which directly converted acetyl-CoA to P3HB. This strategy demonstrated a potential production manner of bioethanol as an extracellular product and P3HB as water insoluble inclusion bodies inside E. coli. The maximum production of bioethanol and P3HB in the recombinant strain was 0.8 g L(-1) (17.4 mmol L(-1)) and 30.2% (w/w dry cell weight), respectively, which were higher than the parental strain.

Biotin is an essential vitamin which plays an important role for maintaining normal physiological function. A rapid, sensitive and simple method is necessary to monitor the biotin level. Here, we reported a replacement assay for the detection of biotin using a replaceable liposomal aptamer. Replacement assay is a competitive assay where a sample analyte replaces the labeled competitor of analyte out of its biorecognition element on a surface. It is user friendly and time-saving because of washing free. We used aptamer as a competitor, not a biorecognition element as tradition. To label aptamers, we used cholesterol-conjugated aptamers to tag signal-amplifying-liposomes. Without the need of conjugation procedure, aptamers can be easily incorporated into the surface of dye-encapsulating liposomes. Two aptamers as competitors of biotin, ST-21 and ST-21M with different affinities to streptavidin, were studied in parallel for the detection of biotin using replacement assays. ST-21 and ST-21M aptamers reached to limits of detection of 1.32 pg/80 μl and 0.47 pg/80 μl, respectively. The dynamic ranges of our assays using ST-21 and ST-21M aptamers were seven and four orders of magnitude, respectively. This assay can be completed in 20 minutes without washing steps. These results were overall better than previous reported assays.

Autotrophic ammonia-oxidizing bacteria are a critical factor of the microbial community in industrial wastewater treatment systems. We evaluated the diversity and community composition of β-proteobacterial ammonia-oxidizing bacteria in two full-scale treatment reactors - a sand filter and a biological aerated filter - receive an identical wastewater. Polymerase chain reaction of the 16S rRNA gene fragments of ammonia-oxidizing bacteria-selective primers was merged with denaturing gradient gel electrophoresis to allow the comparative analysis of the dominant ammonia-oxidizing bacteria populations. The phylogenetic affinities of the dominant ammonia-oxidizing bacteria were verified by cloning and sequencing of polymerase chain reaction-amplified 16S rRNA gene fragments. Denaturing gradient gel electrophoresis profiles were evaluated using a probability-based similarity index. An exploitation of a probabilistic index of similarity permitted us to consider the differences and similarities observed in ammonia-oxidizing bacteria community structure in different samples were statistically significant or could be accounted for random matching of bands in denaturing gradient gel electrophoresis profiles that would propose random colonization of the reactors at different ammonia-oxidizing bacteria. All Possibly-like sequences recognized, grouped within the Nitrosomonas genus. A greater diversity of ammonia-oxidizing bacteria were detected in trickling filters than the BAF on all samples analyzed were initiate to be dominated by ammonia oxidizing bacteria most closely linked to Nitrosococcus mobilis. Numerical investigation of the denaturing gradient gel electrophoresis profiles indicated that the ammonia-oxidizing bacteria community in depth profiles from the filter beds was selected in a non-random manner.

Administration of access control is a major issue in large-scale computer systems. Many such computer systems proposed over recent years aim at reducing the effort required to govern access. Role-based access control (RBAC) systems are a huge benefit to this point. They reduce the tasks of an administrator or authorities when users take on different roles in an organization and need to be assigned different access rights or privileges based on these roles. RBAC is a very expressive and flexible access control mechanism that makes it possible to have security policies based on the principle of least privilege, static and dynamic separation of duties, conflicts between roles and permissions, and many more. This research proposes the use of negative permissions and remote authorization for improving the scalability of an RBAC implementation.

Rad je kritički pregled uloge dezinfekcije površina u širenju bolničkih infekcija i osvrt na nove preporuke CDC za rutinsku dezinfekciju svih površina u bolnici.