Separation of Dihydro-Isocoumarins and Dihydro-Stilbenoids from Hydrangea macrophylla ssp. serrata by Use of Counter-Current Chromatography.

Previously, different Hydrangea macrophylla ssp. serrata cultivars were investigated by untargeted LC-MS analysis. From this, a list of tentatively identified and unknown compounds that differ significantly between these cultivars was obtained. Due to the lack of reference compounds, especially for dihydro-isocoumarins, we aimed to isolate and structurally characterise these compounds from the cultivar 'Yae-no-amacha' using NMR and LC-MS methods. For purification and isolation, counter-current chromatography was used in combination with reversed-phase preparative HPLC as an orthogonal and enhanced purification workflow. Thirteen dihydro-isocoumarins in combination with other metabolites could be isolated and structurally identified. Particularly interesting was the clarification of dihydrostilbenoid glycosides, which were described for the first time in H. macrophylla ssp. serrata. These results will help us in further studies on the biological interpretation of our data.

Discovery of Novel Bacterial Chalcone Isomerases by a Sequence-Structure-Function-Evolution Strategy for Enzymatic Synthesis of (S)-Flavanones.

Chalcone isomerase (CHI) is a key enzyme in the biosynthesis of flavonoids in plants. The first bacterial CHI (CHIera ) was identified from Eubacterium ramulus, but its distribution, evolutionary source, substrate scope, and stereoselectivity are still unclear. Here, we describe the identification of 66 novel bacterial CHIs from Genbank using a novel Sequence-Structure-Function-Evolution (SSFE) strategy. These novel bacterial CHIs show diversity in substrate specificity towards various hydroxylated and methoxylated chalcones. The mutagenesis of CHIera according to the substrate binding models of these novel bacterial CHIs resulted in several variants with greatly improved activity towards these chalcones. Furthermore, the preparative scale conversion catalyzed by bacterial CHIs has been performed for five chalcones and revealed (S)-selectivity with up to 96 % ee, which provides an alternative biocatalytic route for the synthesis of (S)-flavanones in high yields.

Personal Finance Primer for the Future Orthopaedic Surgeon: A Starting Point.

The level of financial literacy varies among orthopaedic trainees. Personal finance ideally should be taught before accepting student loans; however, when this has not happened, it is imperative that trainees start taking their personal finances seriously. Many trainees are faced with large amounts of student debt and struggle with how to manage a large salary increase in their first job. This can lead to poor financial decisions including insufficient savings. The authors provide a comprehensive viewpoint on personal finance for the orthopaedic trainee. In this article, we provide future orthopaedic surgeons with a framework for personal financial management as a starting point to understanding the financial concepts of budgeting, investment, debt management, mortgage, disability insurance, and life insurance.

First measurements of the radiation dose on the lunar surface.

Human exploration of the Moon is associated with substantial risks to astronauts from space radiation. On the surface of the Moon, this consists of the chronic exposure to galactic cosmic rays and sporadic solar particle events. The interaction of this radiation field with the lunar soil leads to a third component that consists of neutral particles, i.e., neutrons and gamma radiation. The Lunar Lander Neutrons and Dosimetry experiment aboard China's Chang'E 4 lander has made the first ever measurements of the radiation exposure to both charged and neutral particles on the lunar surface. We measured an average total absorbed dose rate in silicon of 13.2 ± 1 µGy/hour and a neutral particle dose rate of 3.1 ± 0.5 µGy/hour.

Investigations into the Natural Occurrence of 1-Phenylethyl Acetate (Styrallyl Acetate).

So far, the occurrence of the flavor constituent 1-phenylethyl acetate in a natural source has not been unambiguously confirmed. The present work provides the detailed identification of 1-phenylethyl acetate from clove (Syzygium aromaticum (L.) Merr. & L.M. Perry) buds. In addition, headspace solid-phase microextraction-gas chromatography/mass spectrometry (GC/MS) analysis revealed further occurrence of 1-phenylethyl acetate in cocoa pulp and grape hyacinth flowers. A total of 15.2 g of essential oil was recovered from 7.2 kg of clove buds by simultaneous distillation-extraction followed by vacuum distillation. The distillate obtained was fractionated by silica column chromatography, whereby a significant enrichment of 1-phenylethyl acetate was achieved. The fraction containing the target analyte was further purified by preparative high-performance liquid chromatography, resulting in a final purity of ∼93.0%, yielding a total of 1 to 2 mg of 1-phenylethyl acetate. Identification of the isolated compound was achieved by GC/MS, infrared spectroscopy, enantioselective GC, isotope ratio MS, and nuclear magnetic resonance spectroscopy. Enantioselective GC/MS analysis revealed an enantiomeric excess of 60% (1S)-(-)-1-phenylethyl acetate in the isolate. The δ13CV-PDB value of -32.5 ± 0.5‰ was in accordance with that of C3-plants and other constituents found in genuine clove extracts.

The charged particle radiation environment on Mars measured by MSL/RAD from November 15, 2015 to January 15, 2016.

The Radiation Assessment Detector (RAD) on board the Mars Science Laboratory (MSL) Curiosity rover has been measuring the radiation environment in Gale crater on Mars since August, 2012. These first in-situ measurements provide an important data set for assessing the radiation-associated health risks for future manned missions to Mars. Mainly, the radiation field on the Martian surface stems from Galactic Cosmic Rays (GCRs) and secondary particles created by the GCRs' interactions with the Martian atmosphere and soil. RAD is capable of measuring differential particle fluxes for lower-energy ions and isotopes of hydrogen and helium (up to hundreds of MeV/nuc). Additionally, RAD also measures integral particle fluxes for higher energies of these ions. Besides providing insight on the current Martian radiation environment, these fluxes also present an essential input for particle transport codes that are used to model the radiation to be encountered during future manned missions to Mars. Comparing simulation results with actual ground-truth measurements helps to validate these transport codes and identify potential areas of improvements in the underlying physics of these codes. At the First Mars Radiation Modeling Workshop (June 2016 in Boulder, CO), different groups of modelers were asked to calculate the Martian surface radiation environment for the time of November 15, 2015 to January 15, 2016. These model results can then be compared with in-situ measurements of MSL/RAD conducted during the same time frame. In this publication, we focus on presenting the charged particle fluxes measured by RAD between November 15, 2015 and January 15, 2016, providing the necessary data set for the comparison to model outputs from the modeling workshop. We also compare the fluxes to initial GCR intensities, as well as to RAD measurements from an earlier time period (August 2012 to January 2013). Furthermore, we describe how changes and updates in RAD on board processing and the on ground analysis tools effect and improve the flux calculations. An in-depth comparison of modeling results from the workshop and RAD fluxes of this publication is presented elsewhere in this issue (Matthiä et al., 2017).

Smart, programmable and responsive injectable hydrogels for controlled release of cargo osteoporosis drugs.

Easy-to-prepare drug delivery systems, based on smart, silica gels have been synthesized, characterized, and studied as hosts in the controlled release of bisphosphonates. They exhibit variable release rates and final % release, depending on the nature of bisphosphonate (side-chain length, hydro-philicity/-phobicity, water-solubility), cations present, pH and temperature. These gels are robust, injectable, re-loadable and re-usable.

Iron incorporation in biosilica of the marine diatom Stephanopyxis turris: dispersed or clustered?

Iron incorporation into diatom biosilica was investigated for the species Stephanopyxis turris. It is known that several "foreign" elements (e.g., germanium, titanium, aluminum, zinc, iron) can be incorporated into the siliceous cell walls of diatoms in addition to silicon dioxide (SiO2). In order to examine the amount and form of iron incorporation, the iron content in the growth medium was varied during cultivation. Fe:Si ratios of isolated cell walls were measured by ICP-OES. SEM studies were performed to examine of a possible influence of excess iron during diatom growth upon cell wall formation. The chemical state of biosilica-attached iron was characterized by a combination of infrared, 29Si MAS NMR, and EPR spectroscopy. For comparison, synthetic silicagels of variable iron content were studied. Our investigations show that iron incorporation in biosilica is limited. More than 95% of biosilica-attached iron is found in the form of iron clusters/nanoparticles. In contrast, iron is preferentially dispersedly incorporated within the silica framework in synthetic silicagels leading to Si-O-Fe bond formation.

Charged particle spectra measured during the transit to Mars with the Mars Science Laboratory Radiation Assessment Detector (MSL/RAD).

The Mars Science Laboratory (MSL) started its 253-day cruise to Mars on November 26, 2011. During cruise the Radiation Assessment Detector (RAD), situated on board the Curiosity rover, conducted measurements of the energetic-particle radiation environment inside the spacecraft. This environment consists mainly of galactic cosmic rays (GCRs), as well as secondary particles created by interactions of these GCRs with the spacecraft. The RAD measurements can serve as a proxy for the radiation environment a human crew would encounter during a transit to Mars, for a given part of the solar cycle, assuming that a crewed vehicle would have comparable shielding. The measurements of radiological quantities made by RAD are important in themselves, and, the same data set allow for detailed analysis of GCR-induced particle spectra inside the spacecraft. This provides important inputs for the evaluation of current transport models used to model the free-space (and spacecraft) radiation environment for different spacecraft shielding and different times in the solar cycle. Changes in these conditions can lead to significantly different radiation fields and, thus, potential health risks, emphasizing the need for validated transport codes. Here, we present the first measurements of charged particle fluxes inside a spacecraft during the transit from Earth to Mars. Using data obtained during the last two month of the cruise to Mars (June 11-July 14, 2012), we have derived detailed energy spectra for low-Z particles stopping in the instrument's detectors, as well as integral fluxes for penetrating particles with higher energies. Furthermore, we analyze the temporal changes in measured proton fluxes during quiet solar periods (i.e., when no solar energetic particle events occurred) over the duration of the transit (December 9, 2011-July 14, 2012) and correlate them with changing heliospheric conditions.

Is altered behavior linked to cellular energy regulation in a freshwater mussel (Elliptio complanata) exposed to triclosan?

Environmental stress may alter the bioenergetic balance of organisms by resulting in greater energy investment into detoxification processes, which diverts energy from other biological functions. Here, we examine responses to triclosan (TCS) exposure in a freshwater mussel across multiple biological levels: behavioral (e.g., burrowing and movement activity), organismal (e.g., metabolic rate and heart rate), and subcellular (e.g., gene expression and protein abundance/activity). At the subcellular level, we employed both energetic (i.e., AMP-activated protein kinase (AMPK)) and traditional (i.e., heat shock protein (HSP70), superoxide dismutase (SOD), glutathione-S-transferase (GST)) biomarkers. We found a significant reduction in burrowing and movement behaviors, a 1.8-fold increase in total-AMPK protein abundance, and a 2.8-fold increase in AMPK activity after 21d. GST activity increased after 4d, but not after 21d. Our findings suggest that TCS exposure results in an energetic tradeoff between detoxification at the cellular level and whole-animal activity.

The effects of precipitation, river discharge, land use and coastal circulation on water quality in coastal Maine.

Faecal pollution in stormwater, wastewater and direct run-off can carry zoonotic pathogens to streams, rivers and the ocean, reduce water quality, and affect both recreational and commercial fishing areas of the coastal ocean. Typically, the closure of beaches and commercial fishing areas is governed by the testing for the presence of faecal bacteria, which requires an 18-24 h period for sample incubation. As water quality can change during this testing period, the need for accurate and timely predictions of coastal water quality has become acute. In this study, we: (i) examine the relationship between water quality, precipitation and river discharge at several locations within the Gulf of Maine, and (ii) use multiple linear regression models based on readily obtainable hydrometeorological measurements to predict water quality events at five coastal locations. Analysis of a 12 year dataset revealed that high river discharge and/or precipitation events can lead to reduced water quality; however, the use of only these two parameters to predict water quality can result in a number of errors. Analysis of a higher frequency, 2 year study using multiple linear regression models revealed that precipitation, salinity, river discharge, winds, seasonality and coastal circulation correlate with variations in water quality. Although there has been extensive development of regression models for freshwater, this is one of the first attempts to create a mechanistic model to predict water quality in coastal marine waters. Model performance is similar to that of efforts in other regions, which have incorporated models into water resource managers' decisions, indicating that the use of a mechanistic model in coastal Maine is feasible.

(1)H-(13)C-(29)Si triple resonance and REDOR solid-state NMR-A tool to study interactions between biosilica and organic molecules in diatom cell walls.

Triple resonance solid-state NMR experiments using the spin combination (1)H-(13)C-(29)Si are still rarely found in the literature. This is due to the low natural abundance of the two heteronuclei. Such experiments are, however, increasingly important to study hybrid materials such as biosilica and others. A suitable model substance, ideally labeled with both (13)C and (29)Si, is thus very useful to optimize the experiments before applying them to studies of more complex samples such as biosilica. Tetraphenoxysilane could be synthesized in an easy, two-step synthesis including double isotope labelling. Using tetraphenoxysilane, we established a (1)H-(13)C-(29)Si double CP-based HETCOR experiment and applied it to diatom biosilica from the diatom species Thalassiosira pseudonana. Furthermore, we carried out (1)H-(13)C{(29)Si} CP-REDOR experiments in order to estimate the distance between the organic matrix and the biosilica. Our experiments on diatom biosilica strongly indicate a close contact between polyamine-containing parts of the organic matrix and the silica. This corroborates the assumption that the organic matrix is essential for the control of the cell wall formation.

AMP-activated protein kinase is a biomarker of energetic status in freshwater mussels exposed to municipal effluents.

Although biomarkers are frequently used to assess sublethal effects of contaminants, a lack of mechanistic linkages to higher-level effects limits the predictive power of biomarkers. Bioenergetics has been proposed as a framework for linking cellular effects to whole-animal effects. We investigated sublethal effects of exposure to wastewater treatment facility effluent in freshwater mussels in situ, thereby capturing ecologically relevant exposure conditions. Our study focused on the energetic biomarker AMP-activated protein kinase (AMPK), while also considering more traditional biomarkers like heat shock proteins (HSP70), and antioxidant enzymes (i.e., superoxide dismutase (SOD), glutathione-S-transferase (GST)). We examined biomarkers at mRNA and protein levels. Effluent exposure caused a reduction in total-AMPK protein abundance (p=0.05) and AMPK mRNA expression (p=0.02). Conversely, AMPK activity increased at downstream sites by 2.2-fold (p=0.05), indicating increased cellular energy consumption. HSP70 protein abundance was lower at downstream sites (p<0.05), while SOD and GST activity levels significantly increased. By using various biomarkers, we demonstrate that exposure to municipal effluent creates an energetically taxing situation. This is the first study to use AMPK to evaluate the effects of contamination in situ, and our results suggest that energetic biomarkers, like AMPK, complement traditional biomarkers and may help establish functional links between cellular and whole-animal effects.

Changes in body composition in patients with chronic obstructive pulmonary disease: do they influence patient-related outcomes?

AIMS: The follow-up of the ECLIPSE study, a prospective longitudinal study to identify and define parameters that predict disease progression over 3 years in chronic obstructive pulmonary disease (COPD), allows the examination of the effect of body composition changes on COPD-related outcomes. METHODS: Body composition and health status were established in 2,115 COPD patients, 327 smoking and 239 nonsmoking controls at baseline and 3 years, while mortality was recorded in year 2 and 3 in the COPD patients. Associations between fat free mass index (FFMI) and fat mass index (FMI) changes to deterioration in health status and mortality were determined. RESULTS: Change in FFMI and FMI over 3 years was small and comparable between the groups. Underweight and obese patients had the worst health status. Worsening health status was associated with FFMI decrease in underweight patients and FMI increase in overweight/obese patients. While overweight patients had the lowest mortality, FFMI or FMI decrease was associated with a higher mortality. CONCLUSION: Changes in body composition over 3 years were small and comparable in COPD patients and control subjects. Nevertheless, muscle mass decline in underweight and fat mass increase in overweight/obese patients is associated with worsening health status. Overweight is associated with decreased mortality, but muscle mass and fat mass decline are detrimental for mortality.

CHEK2 I157T and endometrial cancer.

Endometrial cancer (EC) is the most common malignancy of the female reproductive system in the industrialized world. Similar to other common diseases, gene variations are believed to be able to alter an individual's predisposition to developing the disease. The CHEK2 gene encodes a tumor suppressor that takes part in various cell processes, including cell cycle regulation, DNA repair, and apoptosis. The polymorphic variant Ile157Thr in exon 3 of the gene has been demonstrated to enhance the risk of several types of cancer and at the same time to reduce the risk for developing other cancer types. To study the significance of CHEK2 I157T for EC, we have genotyped 268 patients and 449 female controls. We found carriers of I157T more often among controls than we did among patients (2.45% vs. 1.75%), but the difference was not statistically significant. Case-only analysis revealed that the variant is overrepresented in patients diagnosed at 75 or more years of age (9.09%, p = 0.05) and in those with deep myometrial invasion (3.85%, p = 0.06). The highest frequency was observed in patients with both the aforementioned characteristics (20%, p = 0.01). Tumors of I157T carriers showed endometrioid, clear cell, and mucinous morphology, which suggested that the variant may not be restricted to a certain histotype of the disease and could even be overrepresented in rare ones. This study is the first to explore the association between germline CHEK2 I157T and EC. It suggests the need for further large-scale evaluation of the role this variant plays in endometrial carcinogenesis.

[Case report. Restrictive cardiomyopathy with chylothorax in a cat: the pathogenesis]. / Fallbericht: Restriktive Kardiomyopathie mit Chylothorax bei einer Katze: Zur Pathogenese.

A 8.5 year old castrated female domestic short-haired cat was presented because of progressive dyspnea, inappetence and weight loss. Special examinations revealed a chylothorax. In addition a restrictive cardiomyopathy was suspected. The cat was euthanized. The histopathologic examination of the heart confirmed the diagnosis of restrictive cardiomyopathy. The examination of the ductus thoracicus showed a intramural fibrosis with additional edema and a interstitial partly perivascular inflammation. The coherence between feline cardiopathy and chylothorax was discussed speculatively in previous reports. In this report we show a possible aetiopathology.

[The effect of hypothyroidism on cardiac function in dogs]. / Einfluss der Hypothyreose auf die kardiale Funktion beim Hund.

The thyroid hormones have direct and indirect effects on the heart. So it is possible that depression of left ventricular function is associated with hypothyroidism. This publication describes cardiac findings (auscultation, electrocardiography, echocardiography) in ten hypothyroid dogs. Low heart rates, reduced R-amplitudes and bradycardic arrhythmias (first and second-degree AV block) were found on the electrocardiogram before treatment. On the echocardiograms most of the dogs showed reduced contractillity and reduced left ventricular wall thickness. Seven dogs were reexamined after levothyroxine supplementation. Effects of treatment were increased heart rates and R-amplitudes as well as disappearance of the bradycardic arrhythmias in electrocardiographic examination. The echocardiographic examination showed increased contractility and increased left ventricular wall thickness.

Response of dogs to short-term infusions of carbohydrate- or lipid-based parenteral nutrition.

Parenteral nutrition (PN) is used to support intensive care patients. The risk for adverse metabolic effects depends on the composition of infused solutions and the duration of application. The present study in dogs compares metabolic and endocrine effects of two infusion solutions, with either triglycerides or glucose being the major energy sources, administered in a comparatively short infusion period (10 h/day). PN was administered for 9 days to two groups of five adult dogs to meet energy maintenance requirements. In group PN-LIP 61% of the total energy was derived from lipids and 22% from carbohydrates, compared with 21 and 62% in group PN-GLUC. Among routine haematology and clinical chemistry the plasma levels of glucose, triglycerides, insulin, insulin-like growth factor-I (IGF-I), glucagon, 3,5,3'-triiodothyronine and thyroxin were measured in non-infused dogs and at 2, 4, 6, and 8 h after the start of infusion at days 2 and 8 of the study. Infusions protocols did not cause gross metabolic aberrations. During the actual infusions glucose, triglyceride and insulin concentrations were elevated, each depending on the infusion solution. Concentrations of IGF-I, glucagon, 3,5,3'-triiodothyronine, thyroxin and cortisol did not change significantly. In conclusion short infusion periods of 10 h per day were tolerated by healthy dogs without adverse signs, which could improve practicability of PN also in clinical cases.

Embolized mesothelial cells within mediastinal lymph nodes of three dogs with idiopathic haemorrhagic pericardial effusion.

Embolized mesothelial-like cells were detected within pericardial lymphatics and mediastinal lymph nodes of three golden retrievers with idiopathic haemorrhagic pericardial effusion. Morphological, cytochemical, and immunohistochemical investigations indicated that the embolized cells most likely originated from the pericardial mesothelium. None of the dogs showed evidence of an underlying neoplastic disorder. Such findings have not been reported previously in animals, but so-called "benign mesothelial cell inclusions" have been reported in mediastinal lymph nodes of human patients with pleuritis and pericarditis but no history of neoplasia. The present findings in dogs indicate the need to distinguish between lymphatic emboli arising from reactive mesothelial cells and metastases arising from a mesothelioma.