3D Cultivation Techniques for Primary Human Hepatocytes

One of the main challenges in drug development is the prediction of in vivo toxicity based on in vitro data. The standard cultivation system for primary human hepatocytes is based on monolayer cultures, even if it is known that these conditions result in a loss of hepatocyte morphology and of liver-specific functions, such as drug-metabolizing enzymes and transporters. As it has been demonstrated that hepatocytes embedded between two sheets of collagen maintain their function, various hydrogels and scaffolds for the 3D cultivation of hepatocytes have been developed. To further improve or maintain hepatic functions, 3D cultivation has been combined with perfusion. In this manuscript, we discuss the benefits and drawbacks of different 3D microfluidic devices. For most systems that are currently available, the main issues are the requirement of large cell numbers, the low throughput, and expensive equipment, which render these devices unattractive for research and the drug-developing industry. A higher acceptance of these devices could be achieved by their simplification and their compatibility with high-throughput, as both aspects are of major importance for a user-friendly devic

Identification of the Transcriptional Regulator NcrB in the Nickel Resistance Determinant of Leptospirillum ferriphilum UBK03

The nickel resistance determinant ncrABCY was identified in Leptospirillum ferriphilum UBK03. Within this operon, ncrA and ncrC encode two membrane proteins that form an efflux system, and ncrB encodes NcrB, which belongs to an uncharacterized family (DUF156) of proteins. How this determinant is regulated remains unknown. Our data indicate that expression of the nickel resistance determinant is induced by nickel. The promoter of ncrA, designated pncrA, was cloned into the promoter probe vector pPR9TT, and co-transformed with either a wild-type or mutant nickel resistance determinant. The results revealed that ncrB encoded a transcriptional regulator that could regulate the expression of ncrA, ncrB, and ncrC. A GC-rich inverted repeat sequence was identified in the promoter pncrA. Electrophoretic mobility shift assays (EMSAs) and footprinting assays showed that purified NcrB could specifically bind to the inverted repeat sequence of pncrA in vitro; this was confirmed by bacterial one-hybrid analysis. Moreover, this binding was inhibited in the presence of nickel ions. Thus, we classified NcrB as a transcriptional regulator that recognizes the inverted repeat sequence binding motif to regulate the expression of the key nickel resistance gene, ncrA

Isomeric excitation energy for 99^{99}Inm^{m} from mass spectrometry reveals constant trend next to doubly magic 100^{100}Sn

The excitation energy of the 1/2$^-$ isomer in $^{99}$In at ${N=50}$ is measured to be 671(37) keV and the mass uncertainty of the 9/2$^+$ ground state is significantly reduced using the ISOLTRAP mass spectrometer at ISOLDE/CERN. The measurements exploit a major improvement in the resolution of the multi-reflection time-of-flight mass spectrometer. The results reveal an intriguing constancy of the $1/2^-$ isomer excitation energies in neutron-deficient indium that persists down to the $N = 50$ shell closure, even when all neutrons are removed from the valence shell. This trend is used to test large-scale shell model, \textit{ab initio}, and density functional theory calculations. The models have difficulties describing both the isomer excitation energies and ground-state electromagnetic moments along the indium chain.Comment: 13 pages, 4 figure

Cesium, iodine and tritium in NW Pacific waters - a comparison of the Fukushima impact with global fallout

Radionuclide impact of the Fukushima Dai-ichi nuclear power plant accident on the distribution of radionuclides in seawater of the NW Pacific Ocean is compared with global fallout from atmospheric tests of nuclear weapons. Surface and water column samples collected during the <i>Ka'imikai-o-Kanaloa</i> (<i>KOK</i>) international expedition carried out in June 2011 were analyzed for ¹³⁴Cs, ¹³⁷Cs, ¹²⁹I and ³H. The ¹³⁷Cs, ¹²⁹I and ³H levels in surface seawater offshore Fukushima varied between 0.002–3.5 Bq L⁻¹, 0.01–0.8 μBq L⁻¹, and 0.05–0.15 Bq L⁻¹, respectively. At the sampling site about 40 km from the coast, where all three radionuclides were analyzed, the Fukushima impact on the levels of these three radionuclides represents an increase above the global fallout background by factors of about 1000, 50 and 3, respectively. The water column data indicate that the transport of Fukushima-derived radionuclides downward to the depth of 300 m has already occurred. The observed ¹³⁷Cs levels in surface waters and in the water column are compared with predictions obtained from the ocean general circulation model, which indicates that the Kuroshio Current acts as a southern boundary for the transport of the radionuclides, which have been transported from the Fukushima coast eastward in the NW Pacific Ocean. The ¹³⁷Cs inventory in the water column is estimated to be about 2.2 PBq, what can be regarded as a lower limit of the direct liquid discharges into the sea as the seawater sampling was carried out only in the area from 34 to 37° N, and from 142 to 147° E. About 4.6 GBq of ¹²⁹I was deposited in the NW Pacific Ocean, and 2.4–7 GBq of ¹²⁹I was directly discharged as liquid wastes into the sea offshore Fukushima. The total amount of ³H released and deposited over the NW Pacific Ocean was estimated to be 0.1–0.5 PBq. These estimations depend, however, on the evaluation of the total ¹³⁷Cs activities released as liquid wastes directly into the sea, which should improve when more data are available. Due to a suitable residence time in the ocean, Fukushima-derived radionuclides will provide useful tracers for isotope oceanography studies on the transport of water masses during the next decades in the NW Pacific Ocean

Ototoksične tvari na radnom mjestu: kratak uvid u stanje

Ototoxic chemicals can impair the sense of hearing and balance. Lately, efforts have been intensifi ed to compile evidence-based lists of workplace agents with ototoxic properties. This article gives a rough overview of the latest relevant publications, which confirm that toluene, styrene, and lead should receive particular attention as ototoxic substances at the workplace. Moreover, there is sufficient evidence that occupational exposure to trichloroethylene, mercury, carbon monoxide, and carbon disulfide can affect the ear. Based on the existing information, industrial hygienists should make sure that occupational health professionals and the workforce are made aware of the risks posed by ototoxic substances; support their replacement or new technical measures to reduce exposure; make these substances a part of regular screening, develop tools that can early diagnose chemically induced hearing impairment, and investigate further into the ototoxic properties of these substances. Further research should focus on quantifying the combined effects of ototoxic substances and noise.Ototoksične kemikalije mogu narušiti osjetilo sluha i ravnotežu. Nedavno su uloženi dodatni napori u izradu znanstveno utemeljenih popisa tvari koje su prisutne na radnom mjestu, a koje imaju ototoksična svojstva. Ovaj rad daje kratak uvid u najnovije publikacije objavljene na ovu temu. Usporedba navedenih publikacija potvrđuje da bi toluen, stiren i olovo trebalo razmatrati kao izrazito bitne ototoksične tvari koje postoje na radnom mjestu. Nadalje, postoje dovoljni dokazi koji potvrđuju da ototoksične tvari poput trikloretilena, žive, ugljikova monoksida i disulfida u radnom okruženju mogu oštetiti sluh. Temeljem postojećih informacija stručnjaci u području higijene rada trebali bi upozoravati stručnjake u području medicine rada i same radnike na rizike koje ototoksične tvari predstavljaju; poticati ih na zamjenu takvih tvari ili uvođenje novih mjera za smanjenje izlaganja; uključiti ototoksične tvari u redoviti program praćenja i osmisliti mjere za rano otkrivanje oštećenja sluha zbog izloženosti kemijskim tvarima; dodatno istražiti ototoksična svojstva ovih tvari. Buduća istraživanja trebala bi se usredotočiti na izračun ukupnih učinaka ototoksičnih tvari i buke

Proton Pump Inhibitors Inhibit Metformin Uptake by Organic Cation Transporters (OCTs)

Metformin, an oral insulin-sensitizing drug, is actively transported into cells by organic cation transporters (OCT) 1, 2, and 3 (encoded by SLC22A1, SLC22A2, or SLC22A3), which are tissue specifically expressed at significant levels in various organs such as liver, muscle, and kidney. Because metformin does not undergo hepatic metabolism, drug-drug interaction by inhibition of OCT transporters may be important. So far, comprehensive data on the interaction of proton pump inhibitors (PPIs) with OCTs are missing although PPIs are frequently used in metformin-treated patients. Using in silico modeling and computational analyses, we derived pharmacophore models indicating that PPIs (i.e. omeprazole, pantoprazole, lansoprazole, rabeprazole, and tenatoprazole) are potent OCT inhibitors. We then established stably transfected cell lines expressing the human uptake transporters OCT1, OCT2, or OCT3 and tested whether these PPIs inhibit OCT-mediated metformin uptake in vitro. All tested PPIs significantly inhibited metformin uptake by OCT1, OCT2, and OCT3 in a concentration-dependent manner. Half-maximal inhibitory concentration values (IC50) were in the low micromolar range (3–36 µM) and thereby in the range of IC50 values of other potent OCT drug inhibitors. Finally, we tested whether the PPIs are also transported by OCTs, but did not identify PPIs as OCT substrates. In conclusion, PPIs are potent inhibitors of the OCT-mediated metformin transport in vitro. Further studies are needed to elucidate the clinical relevance of this drug-drug interaction with potential consequences on metformin disposition and/or efficacy

Search for CP violation in D+→K−K+π+D^{+} \to K^{-}K^{+}\pi^{+} decays

A model-independent search for direct CP violation in the Cabibbo suppressed decay $D^+ \to K^- K^+\pi^+$ in a sample of approximately 370,000 decays is carried out. The data were collected by the LHCb experiment in 2010 and correspond to an integrated luminosity of 35 pb$^{-1}$. The normalized Dalitz plot distributions for $D^+$ and $D^-$ are compared using four different binning schemes that are sensitive to different manifestations of CP violation. No evidence for CP asymmetry is found.Comment: 13 pages, 8 figures, submitted to Phys. Rev.

Effects of automated alerts on unnecessarily repeated serology tests in a cardiovascular surgery department: a time series analysis

<p>Abstract</p> <p>Background</p> <p>Laboratory testing is frequently unnecessary, particularly repetitive testing. Among the interventions proposed to reduce unnecessary testing, Computerized Decision Support Systems (CDSS) have been shown to be effective, but their impact depends on their technical characteristics. The objective of the study was to evaluate the impact of a Serology-CDSS providing point of care reminders of previous existing serology results, embedded in a Computerized Physician Order Entry at a university teaching hospital in Paris, France.</p> <p>Methods</p> <p>A CDSS was implemented in the Cardiovascular Surgery department of the hospital in order to decrease inappropriate repetitions of viral serology tests (HBV).</p> <p>A time series analysis was performed to assess the impact of the alert on physicians' practices. The study took place between January 2004 and December 2007. The primary outcome was the proportion of unnecessarily repeated HBs antigen tests over the periods of the study. A test was considered unnecessary when it was ordered within 90 days after a previous test for the same patient. A secondary outcome was the proportion of potentially unnecessary HBs antigen test orders cancelled after an alert display.</p> <p>Results</p> <p>In the pre-intervention period, 3,480 viral serology tests were ordered, of which 538 (15.5%) were unnecessarily repeated. During the intervention period, of the 2,095 HBs antigen tests performed, 330 unnecessary repetitions (15.8%) were observed. Before the intervention, the mean proportion of unnecessarily repeated HBs antigen tests increased by 0.4% per month (absolute increase, 95% CI 0.2% to 0.6%, <it>p </it>< 0.001). After the intervention, a significant trend change occurred, with a monthly difference estimated at -0.4% (95% CI -0.7% to -0.1%, <it>p </it>= 0.02) resulting in a stable proportion of unnecessarily repeated HBs antigen tests. A total of 380 unnecessary tests were ordered among 500 alerts displayed (compliance rate 24%).</p> <p>Conclusions</p> <p>The proportion of unnecessarily repeated tests immediately dropped after CDSS implementation and remained stable, contrasting with the significant continuous increase observed before. The compliance rate confirmed the effect of the alerts. It is necessary to continue experimentation with dedicated systems in order to improve understanding of the diversity of CDSS and their impact on clinical practice.</p