Somatostatin analogue (octreotide) inhibits bile duct epithelial cell proliferation and fibrosis after extrahepatic biliary obstruction.

Extrahepatic biliary obstruction leads to bile duct epithelial cell proliferation. Somatostatin and its analogue, octreotide, have been shown to inhibit DNA synthesis and proliferation in hepatocytes. We investigated the effect of octreotide on the biliary epithelial cell proliferative responses to biliary obstruction. Male Sprague-Dawley rats underwent common bile duct ligation and subcutaneous injection of either saline or octreotide (6 micrograms/kg) twice daily for 7 days. Morphometric analysis of hepatocytes, bile duct epithelial cells, and periportal connective tissue was performed by computerized point counting. Hepatocyte volume was preserved with octreotide treatment, which also significantly decreased bile duct proliferation and periportal extracellular matrix deposition in response to biliary obstruction compared with saline treated, duct-ligated animals. These results indicate that octreotide prevents the morphological changes that accompany extrahepatic biliary obstruction.     

Aggregation‐induced changes in the chemical exchange saturation transfer (CEST) signals of proteins

Chemical exchange saturation transfer (CEST) is an MRI technique that allows mapping of biomolecules (small metabolites, proteins) with nearly the sensitivity of conventional water proton MRI. In living organisms, several tissue‐specific CEST effects have been observed and successfully applied to diagnostic imaging. In these studies, particularly the signals of proteins showed a distinct correlation with pathological changes. However, as CEST effects depend on various properties that determine and affect the chemical exchange processes, the origins of the observed signal changes remain to be understood. In this study, protein aggregation was identified as an additional process that is encoded in the CEST signals of proteins. Investigation of distinct proteins that are involved in pathological disorders, namely amyloid beta and huntingtin, revealed a significant decrease of all protein CEST signals upon controlled aggregation. This finding is of particular interest with regard to diagnostic imaging of patients with neurodegenerative diseases that involve amyloidogenesis, such as Alzheimer's or Huntington's disease. To investigate whether the observed CEST signal decrease also occurs in heterogeneous mixtures of aggregated cellular proteins, and thus prospectively in tissue, heat‐shocked yeast cell lysates were employed. Additionally, investigation of different cell compartments verified the assignment of the protein CEST signals to the soluble part of the proteome. The results of in vitro experiments demonstrate that aggregation affects the CEST signals of proteins. This observation can enable hypotheses for CEST imaging as a non‐invasive diagnostic tool for monitoring pathological alterations of the proteome in vivo.     

Diversification of Clonal Complex 5 Methicillin-Resistant Staphylococcus aureus Strains (Rhine-Hesse Clone) within Germany

Since 1995, a methicillin-resistant Staphylococcus aureus (MRSA) clone has spread in southern Germany. The strain was assigned to the Rhine-Hesse pulsed-field gel electrophoresis (PFGE) type by the staphylococcal reference center and was highly similar to epidemic clones known to belong to clonal complex 5 (CC5; USA100) based on multilocus sequence typing (MLST). Here we analyzed a defined collection of strains assigned to the Rhine-Hesse/USA100 PFGE type. Using sequence-based typing methods (MLST, spa), the isolates were divided into two distinct clusters, ST5 and its single-locus variant ST225. These two lineages are not distinguishable by PFGE or phage typing. Most of the ST5 isolates were derived from patients and volunteers from the Tübingen area in southwest Germany, whereas the ST225 isolates were mostly from other locations in Germany. The locally restricted ST5 isolates were shown to contain different SSCmec islands and exhibited different antibiotic resistance profiles. In contrast, the ST225 isolates form a highly homogenous group and are emerging all over Germany. The two lineages are clearly distinguishable by their phage content and spa type: ST5 strains from Tübingen are characterized by a Sa7int phage that carries the virulence gene sak, which codes for staphylokinase, and ST225 isolates are characterized by a Sa1int phage. In conclusion, based on sequence typing and phage content, CC5 strains can be subdivided into two distinct lineages with different epidemicities.     

Direct determination of surface tension in the lung.

We have used the spreading behavior of small drops of several fluorocarbon fluids and silicone oil on air-liquid interfaces to measure the surface tension of lungs in situ. The test fluids were calibrated in a surface balance at 37 degrees on monolayers of dipalmitoylphosphatidylcholine. At particular surface tensions characteristic of each fluid used, an increase in the tension of 1 mN/m or less caused the droplets to spread reversibly from a sphere to a lens shape. Using micropipettes we placed such droplets on the alveolar surfaces of excised rat lungs held at functional residual capacity and 37 degrees and found that the surface tension remained below 9 mN/m for at least 30 min. The surface tension-volume relationship was linear for tensions ranging from 9 to 20 mN/m.     

The regioselectivity of glutathione adduct formation with flavonoid quinone/quinone methides is pH-dependent

In the present study, the formation of glutathionyl adducts from a series of 3',4'-dihydroxy flavonoid o-quinone/p-quinone methides was investigated with special emphasis on the regioselectivity of the glutathione addition as a function of pH. The flavonoid o-quinones were generated using horseradish peroxidase, and upon purification by HPLC, the glutathionyl adducts were identified by LC/MS as well as (1)H and (13)C NMR. The major pH effect observed for the glutathione conjugation of taxifolin and luteolin quinone is on the rate of taxifolin and luteolin conversion and, as a result, on the ratio of mono- to diglutathione adduct formation. With fisetin, 3,3',4'-trihydroxyflavone, and quercetin, decreasing the pH results in a pathway in which glutathionyl adduct formation occurs in the C ring of the flavonoid, being initiated by hydration of the quinone and H(2)O adduct formation also in the C ring of the flavonoid. With increasing pH, for fisetin and 3,3',4'-trihydroxyflavone glutathione adduct formation of the quinone occurs in the B ring at C2' as the preferential site. For quercetin, the adduct formation of its quinone/quinone methide shifts from the C ring at pH 3.5, to the A ring at pH 7.0, to the B ring at pH 9.5, indicating a significant influence of the pH and deprotonation state on the chemical electrophilic behavior of quercetin quinone/quinone methide. Together the results of the present study elucidate the mechanism of the pH-dependent electrophilic behavior of B ring catechol flavonoids, which appears more straightforward than previously foreseen.     

Polyethylene glycol-surfactant for lavage lung injury in rats

Addition of ionic and nonionic water-soluble polymers to pulmonary surfactants in the presence of inactivating substances prevents surfactant inactivation in vitro and improves lung function in several models of lung injury. However, a recent report found opposite effects when surfactant plus polyethylene glycol (PEG) was used to treat lung injury caused by saline lung lavage. Therefore, we examined the reasons why the polymer effect is less evident in the saline lung lavage lung injury model. We treated rats with lavage lung injury with a commercial lung surfactant extract derived from bovine lung (Survanta) with or without addition of PEG. Groups treated with Survanta + PEG had significantly higher static post mortem lung volumes than groups treated with Survanta. However, groups treated with Survanta + PEG had more tracheal fluid and no significant benefit in arterial oxygenation compared with the group treated with Survanta, despite our use of measures to reduce pulmonary edema. Measurements after intravascular injections of (125)I-labeled albumin confirmed that addition of PEG increased extravascular lung water and that this effect is mitigated by furosemide. We conclude that surfactant + PEG mixtures are less effective in lavage injury than in other forms of lung injury because of increased extravascular lung water.