Prion Protein in Milk

BACKGROUND: Prions are known to cause transmissible spongiform encephalopathies (TSE) after accumulation in the central nervous system. There is increasing evidence that prions are also present in body fluids and that prion infection by blood transmission is possible. The low concentration of the proteinaceous agent in body fluids and its long incubation time complicate epidemiologic analysis and estimation of spreading and thus the risk of human infection. This situation is particularly unsatisfactory for food and pharmaceutical industries, given the lack of sensitive tools for monitoring the infectious agent. METHODOLOGY/PRINCIPAL FINDINGS: We have developed an adsorption matrix, Alicon PrioTrap®, which binds with high affinity and specificity to prion proteins. Thus we were able to identify prion protein (PrP(C))–the precursor of prions (PrP(Sc))–in milk from humans, cows, sheep, and goats. The absolute amount of PrP(C) differs between the species (from µg/l range in sheep to ng/l range in human milk). PrP(C) is also found in homogenised and pasteurised off-the-shelf milk, and even ultrahigh temperature treatment only partially diminishes endogenous PrP(C) concentration. CONCLUSIONS/SIGNIFICANCE: In view of a recent study showing evidence of prion replication occurring in the mammary gland of scrapie infected sheep suffering from mastitis, the appearance of PrP(C) in milk implies the possibility that milk of TSE-infected animals serves as source for PrP(Sc)

Accurate Prediction of Secreted Substrates and Identification of a Conserved Putative Secretion Signal for Type III Secretion Systems

The type III secretion system is an essential component for virulence in many Gram-negative bacteria. Though components of the secretion system apparatus are conserved, its substrates—effector proteins—are not. We have used a novel computational approach to confidently identify new secreted effectors by integrating protein sequence-based features, including evolutionary measures such as the pattern of homologs in a range of other organisms, G+C content, amino acid composition, and the N-terminal 30 residues of the protein sequence. The method was trained on known effectors from the plant pathogen Pseudomonas syringae and validated on a set of effectors from the animal pathogen Salmonella enterica serovar Typhimurium (S. Typhimurium) after eliminating effectors with detectable sequence similarity. We show that this approach can predict known secreted effectors with high specificity and sensitivity. Furthermore, by considering a large set of effectors from multiple organisms, we computationally identify a common putative secretion signal in the N-terminal 20 residues of secreted effectors. This signal can be used to discriminate 46 out of 68 total known effectors from both organisms, suggesting that it is a real, shared signal applicable to many type III secreted effectors. We use the method to make novel predictions of secreted effectors in S. Typhimurium, some of which have been experimentally validated. We also apply the method to predict secreted effectors in the genetically intractable human pathogen Chlamydia trachomatis, identifying the majority of known secreted proteins in addition to providing a number of novel predictions. This approach provides a new way to identify secreted effectors in a broad range of pathogenic bacteria for further experimental characterization and provides insight into the nature of the type III secretion signal

Impact of Different Etching Strategies on Margin Integrity of Conservative Composite Restorations in Demineralized Enamel

Good margin integrity with a tight seal of the adhesive interface is considered one of the key factors for the clinical success of composite restorations. This study investigated the effect of enamel etching with phosphoric acid on the margin integrity of self-etch bonded composite restorations in demineralized enamel. Crowns of bovine incisors were assigned into 14 groups (n = 10 per group) of which ten groups (groups 1–5 and 8–12) were demineralized (21 days, acid buffer, pH 4.95) to create artificial carious lesions. Standardized Class V cavities were prepared in all specimens. Demineralized groups were either etched with phosphoric acid for 10, 30, 60, or 120 s (groups 2–5 and 9–12), or no etching was performed (groups 1 and 8). The non-demineralized (sound) groups were etched for 10 s (groups 7 and 14) or remained non-etched (groups 6 and 13). Resin composite restorations were then placed using either a one-step (iBond Self Etch, groups 1–7) or two-step self-etch adhesive (Clearfil SE Bond, groups 8–14). Margin integrity of the restorations was assessed after thermocycling (5000×, 5–55 °C) using scanning electron microscopy, and the percentage of continuous margins (%CM) was statistically analyzed (α = 0.05). Phosphoric acid etching significantly increased %CM in both demineralized and sound enamel. For iBond Self Etch, a significant increase in %CM in demineralized enamel was observed with increased etching times. All etched groups treated with Clearfil SE Bond and those etched for 60 or 120 s and treated with iBond Self Etch showed similar %CM in demineralized enamel as in etched sound enamel, and significantly higher %CM than in non-etched sound enamel. In conclusion, enamel etching with phosphoric acid improves margin integrity of composite restorations in demineralized enamel when bonded with the examined adhesives

Methämoglobinämie unter Dapson

Die 17-jährige Patientin (60 kg Körpergewicht) wurde aufgrund einer Blauverfärbung der Lippen, der Finger und der Zehen sowie Kopfschmerzen, Schwindel und starker Müdigkeit im Spital vorstellig. Bei Eintritt betrug der Blutdruck 126/64 mm Hg bei einem Puls von 143/min. Dyspnoe, Thoraxschmerzen und Palpitationen waren bei der Patientin nicht vorhanden. Am Abend zuvor hatte sie einmalig 6 Tabletten à 50 mg von Dapson-Fatol® (entspricht 300 mg Dapson) zur Behandlung einer Akne vulgaris und 1 Kapsel Isotretinoin-­Mepha Solucaps® (Isotretinoin 20 mg) eingenommen. In der arteriellen Blutgasanalyse wurde initial ein Methämoglobinwert von 27,9% festgestellt. Unter Gabe von 15 Liter Sauerstoff betrug die Sättigung 88%. Es erfolgte die kumulative Gabe von 420 mg Methylenblau (entspricht 7 mg/kg Körpergewicht) über 3 Tage, worunter sich die Methämoglobinämie langsam besserte. Bei stets stabilem Hämoglobin und normwertigen Hämolyse­parametern konnte eine Hämolyse ausgeschlossen werden. Fünf Tage nach der Dapson-Einnahme erschien die Patientin erstmals nicht mehr zyanotisch. Die Patientin konnte nach einer Woche wieder entlassen werden. ­Informationen bzgl. einer angeborenen Störung wie Mangel an Glukose-6-Phosphat-Dehydrogenase oder Methämoglobin-Reduktase liegen nicht vor

Impact of Different Etching Strategies on Margin Integrity of Conservative Composite Restorations in Demineralized Enamel

Good margin integrity with a tight seal of the adhesive interface is considered one of the key factors for the clinical success of composite restorations. This study investigated the effect of enamel etching with phosphoric acid on the margin integrity of self-etch bonded composite restorations in demineralized enamel. Crowns of bovine incisors were assigned into 14 groups (n = 10 per group) of which ten groups (groups 1-5 and 8-12) were demineralized (21 days, acid buffer, pH 4.95) to create artificial carious lesions. Standardized Class V cavities were prepared in all specimens. Demineralized groups were either etched with phosphoric acid for 10, 30, 60, or 120 s (groups 2-5 and 9-12), or no etching was performed (groups 1 and 8). The non-demineralized (sound) groups were etched for 10 s (groups 7 and 14) or remained non-etched (groups 6 and 13). Resin composite restorations were then placed using either a one-step (iBond Self Etch, groups 1-7) or two-step self-etch adhesive (Clearfil SE Bond, groups 8-14). Margin integrity of the restorations was assessed after thermocycling (5000×, 5-55 °C) using scanning electron microscopy, and the percentage of continuous margins (%CM) was statistically analyzed (α = 0.05). Phosphoric acid etching significantly increased %CM in both demineralized and sound enamel. For iBond Self Etch, a significant increase in %CM in demineralized enamel was observed with increased etching times. All etched groups treated with Clearfil SE Bond and those etched for 60 or 120 s and treated with iBond Self Etch showed similar %CM in demineralized enamel as in etched sound enamel, and significantly higher %CM than in non-etched sound enamel. In conclusion, enamel etching with phosphoric acid improves margin integrity of composite restorations in demineralized enamel when bonded with the examined adhesives

Phytochemical Investigation of <i>Cordia africana</i> Lam. Stem Bark: Molecular Simulation Approach

Background: The current work planned to evaluate Cordia africana Lam. stem bark, a traditionally used herb in curing of different ailments in Africa such as gastritis and wound infections, based on phytochemical and antibacterial studies of two pathogenic microorganisms: methicillin-resistant Staphylococcus aureus (MRSA) and Helicobacter pylori. Methods: High performance liquid chromatography (HPLC) profiling was used for qualitative and quantitative investigation of the ethanol extract. The minimum inhibitory concentration (MIC) of the ethanolic extract and isolated compounds was estimated using the broth microdilution method and evidenced by molecular dynamics simulations. Results: Four compounds were isolated and identified for the first time: α-amyrin, β-sitosterol, rosmarinic acid (RA) and methyl rosmarinate (MR). HPLC analysis illustrated that MR was the dominant phenolic acid. MR showed the best bacterial inhibitory activity against MRSA and H. pylori with MIC 7.81 ± 1.7 μg/mL and 31.25 ± 0.6, respectively, when compared to clarithromycin and vancomycin, respectively. Conclusion: The antibacterial activity of the stem bark of Cordia africana Lam. was evidenced against MRSA and H. pylori. Computational modeling of the studied enzyme-ligands systems reveals that RA and MR can potentially inhibit both MRSA peptidoglycan transpeptidases and H. pylori urease, thereby creating a pathway via the use of a double target approach in antibacterial treatment

Profiling Metabolites and Biological Activities of Sugarcane (Saccharum officinarum Linn.) Juice and Its Product Molasses via a Multiplex Metabolomics Approach

Sugarcane (Saccharum officinarum L.) is an important perennial grass in the Poaceae family cultivated worldwide due to its economical and medicinal value. In this study, a combined approach using mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy was employed for the large-scale metabolite profiling of sugarcane juice and its by-product molasses. The polyphenols were analysed via UPLC-UV-ESI-MS, whereas the primary metabolites such as sugars and organic and amino acids were profiled using NMR spectroscopy and gas chromatography/mass spectrometry (GC/MS). UPLC/MS was more effective than NMR spectroscopy or GC/MS for determining differences among the metabolite compositions of the products. Under the optimized conditions, UPLC/MS led to the identification of 42 metabolites, including nine flavonoids, nine fatty acids, and two sterols. C/O Flavone glycosides were the main subclass detected, with tricin-7-O-deoxyhexosyl glucuronide being detected in sugarcane and molasses for the first time. Based on GC/MS analysis, disaccharides were the predominant species in the sugarcane juice and molasses, with sucrose accounting for 66% and 59%, respectively, by mass of all identified metabolites. The phenolic profiles of sugarcane and molasses were further investigated in relation to their in vitro antioxidant activities using free radical scavenging assays such as 2,2-Diphenyl-1-picrylhydrazyl free radical-scavenging ability (DPPH), Trolox equivalent antioxidant capacity (TEAC) and ferric reducing antioxidant power (FRAP). In view of its higher total phenolic content (TPC) (196 +/- 2.1 mg GAE/100 g extract) compared to that of sugarcane juice (93 +/- 2.9 mg GAE/100 g extract), molasses exhibited a substantially higher antioxidant effect. Interestingly, both extracts were also found to inhibit alpha-glucosidase and alpha-amylase enzymes, suggesting a possible antihyperglycaemic effect. These findings suggest molasses may be a new source of natural antioxidants for functional foods