Liquid chromatographic determination of ketoconazole, a potent inhibitor of CYP3A4-mediated metabolism

A high-performance liquid chromatographic assay with UV detection has been developed for the determination of ketoconazole in human plasma. Quantitative extraction was achieved by a single solvent extraction involving a mixture of acetonitrile–n-butyl chloride (1:4, v/v). Ketoconazole and the internal standard (clotrimazole) were separated on a column packed with Inertsil ODS-80A material and a mobile phase composed of water–acetonitrile–tetrahydrofuran–ammonium hydroxide–triethylamine (45:50.2:2.5:0.1:0.1, v/v). The column effluent was monitored at a wavelength of 206 nm with a detector range set at 0.5. The calibration graph was linear in the range of 20–2000 ng/ml, with a lower limit of quantitation of 20.0 ng/ml. The extraction recoveries for ketoconazole and clotrimazole in human plasma were 93±9.7% and 83±10.0%, respectively. The developed method has been successfully applied to a clinical study to examine the pharmacokinetics of ketoconazole in a cancer patient.     

Immunocytochemical localization of muscarinic acetylcholine receptors in the rat endocrine pancreas

Summary Immunocytochemical application of the antimuscarinic acetylcholine receptor antibody M35 to pancreas tissue revealed the target areas for the parasympathetic nervous system. Immunoreactivity in the endocrine pancreas was much higher than that in the exocrine part. Moreover, the endocrine cells at the periphery of the islets of Langerhans displayed the highest level of immunoreactivity. Based on these findings in the mantle of the islets, two types of islets have been distinguished: type-I islets with intensely stained mantle cells, and type-II islets with a much lower concentration of these cells. On average, type-I islets were larger (244.8 m±6.1 SEM) than type-II islets (121.5 m±3.8 SEM). M35-immunoreactivity was present on the majority of D cells, which were characterized by their immunoreactivity to somatostatin [of 446 D cells 356 (79.8%) were M35-immunopositive]. However, only a small proportion of the intensely stained mantle cells belonged to the D cell population. Therefore, it is concluded that the majority of the intensely stained mantle cells represent glucagon-secreting A and/or pancreatic polypeptide-secreting F cells. The intensity of M35-immunoreactivity at the periphery and central core of the islets paralleled the density of cholinergic innervation, suggesting a positive correlation between the intensity of cholinergic transmission and the number of muscarinic acetylcholine receptors at the target structures. The present study further revealed some striking parallels for the muscarinic acetylcholine receptor characteristics between the (endocrine) pancreas and the central nervous system.     

Feeding and temperature responses to intravenous leptin infusion are differential predictors of obesity in rats

Obesity is frequently associated with leptin resistance. The present study investigated whether leptin resistance in rats is present before obesity develops, and thus could underlie obesity induced by 16 wk exposure to a liquid, palatable, high-energy diet (HED). Before HED exposure, male Wistar rats (weighing between 330 and 360 g) received intravenous infusions of 20 microg leptin 2 h before dark (approximately 57 microg/kg rat). Relative to saline infusion, this caused a highly variable effect on food intake (ranging between -94 and +129%), with food intake suppression that appeared negatively correlated with HED-induced increases in body weight gain, caloric intake, adiposity, and plasma leptin levels. In contrast, leptin's thermogenic response was positively correlated to body weight gain linked to weights of viscera, but not to adiposity. Before HED exposure, leptin unexpectedly increased food intake in some rats (fi+, n = 8), whereas others displayed the normal reduction in food intake (fi-, n = 7). HED-exposed fi+ rats had higher plasma leptin levels, retroperitoneal fat pad weight, HED intake, and body weight gain than fi- and chow-fed rats. These parameters were also higher in HED-exposed fi-rats relative to chow rats, except for plasma leptin concentrations. It is concluded that leptin's reduced efficacy to suppress food intake could predict obesity on an HED. An unexpected orexigenic effect of leptin might potentially contribute to this as well.     

Neocortical Dialysate Monoamines of Rats After Acute, Subacute, and Chronic Liver Shunt

Abstract: Intracerebral microdialysis was applied to monitor the neocortical extracellular levels of the aromatic amino acids phenylalanine, tyrosine, and tryptophan, the neurotransmitters dopamine (DA), noradrenaline (NA), and serotonin (5-HT), and the metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and 5-hydroxyindole-3-acetic acid (5-HIAA) in rats with various forms of experimental hepatic encephalopathy (HE). The extracellular aromatic amino acid levels were clearly increased in acute, subacute, and chronic HE. No changes compared with controls in the neocortical DA release could be detected in the three experimental HE rat models investigated. The NA release showed a significant increase only in the subacute HE group. These data suggest that HE may not be associated with any major reduction of neocortical DA or NA release as previously suggested. In acute and subacute HE, decreased extracellular DOPAC but elevated 5-HIAA concentrations were seen. In chronic HE, elevations of both DOPAC and 5-HIAA were observed. Neocortical 5-HT release did not change in subacute and chronic HE, whereas it decreased in acute HE compared with control values. Significant increase in extracellular concentrations of 5-HIAA and of the 5-HIAA/5-HT ratio in the present study are in agreement with previously reported increases in 5-HT turnover in experimental HE. However, a substantially increased 5-HT turnover in experimental HE does not appear to be related to an increase in neuronal neocortical 5-HT release.     

Influence of internal loading on phosphorus concentration in shallow lakes before and after reduction of the external loading

An analysis of data from 49 shallow lakes showed, that the parameters of empirical models between phosphorus loading and concentration in the lake (e.g. Vollenweider type of relations) differ significantly for lakes without or with a reduced external loading. For lakes without a reduction of the external loading the summer phosphorus concentration is determined by the external phosphorus loading and the hydraulic loading. For these lakes the classical models suffice; deviations between calculations and measurements are partly due to errors made in the determination of the loading.In contrast, for lakes where the external loading was reduced, the measured internal loading explains most of the variation in the summer lake concentration. The external loading is of minor importance and the classical models cannot be applied. The internal loading measured before reduction of the external loading is not useful in predicting the concentration afterwards. Instead of the internal loading, the sediment composition can be used. The advantage of using sediment composition is that these variables are easier to determine and vary less in time. The most promising variable is the ratio between total P and total Fe in the sediment.Abbreviations: Q_s hydraulic loading (m y^-1) - hydraulic retention time (y) - L_ext external phosphorus loading (gP m^-2 y^-1) - L_int internal phosphorus loading (gP m^-2 y^-1) - P_lake phosphorus concentration in the lake (gP m^-3) - P_inlet phosphorus concentration in the inlet water (gP m^-3) - P_sed phosphorus content on the sediment (gP kg^-1 d.w.) - Fe_sed iron content of the sediment (gFe kg^-1 d.w.) - Y dependent variable multiple regression calculations - X1, X2 independent variables multiple regressions calculations - a, a1 constants - a2, b constants     

A gravel-covered iceberg provides an offshore breeding site for ivory gulls <Emphasis Type="Italic">Pagophila eburnea</Emphasis> off Northeast Greenland

The ivory gull Pagophila eburnea is an Arctic seabird species whose distribution is tightly coupled to the availability of sea ice. During the last decades, strong declines have been reported for breeding colonies in Canada and Greenland, which are usually located on nunataks or remote coastal islands. Here, we report the observation of a colony of ivory gulls breeding on a gravel-covered iceberg 70 km off Northeast Greenland in August 2014. It concerned approximately 60 adults, including two ringed individuals, and many chicks. This represents an unusual breeding site for the species, to be compared with a few cases of colonies on gravel-covered sea ice. Breeding on an offshore iceberg may be advantageous since it provides ultimate protection from predators. Furthermore, the proximity to the productive North East Water polynya may have been attractive to these gulls. As a consequence of this and previous observations, colony surveys should not solely focus on inland and coastal breeding habitats, but should be extended towards the ocean.     

Anoxygenic photosynthetic hydrogen production and electron transport in the cyanobacterium oscillatoria limnetica

The induction of anoxygenic photosynthesis in the cyanobacterium Oscillatoria limnetica by sulfide was shown to involve the synthesis of a sulfide oxidizing factor; this factor, partly adsorbed on the thylakoid membrane, can be recovered in the soluble phase and is active also on membranes from oxygenically grown cells. The factor is required for sulfide dependent light-induced hydrogen evolution. It accelerates electron transport from sulfide to the electron donor of photosystem I, P700, in membranes from cells in which anoxygenic photosynthesis is induced. The plastiquinone analogue DBMIB does not inhibit electron transport to P700 but accelerates it. The analogue might promote cyclic electron transport involving P700, thus preventing electrons to reach hydrogenase.Abbrevlations chl chlorophyll - DBMIB 3,5-dibromo-3-methyl-6-isopropyl-8-benzoquinone - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - EDTA ethylenediamine tetraacetate - FAD flavin adenine dinucleotide - HEPES N-2-hydroxyethyl-piperazine N-2-ethane sulfonic acid - PS photosystem Dedicated to Prof. L.N.M. Duysens on the occasion of his retirement     

Technological Analysis of the World’s Earliest Shamanic Costume: A Multi-Scalar,Experimental Study of a Red Deer Headdress from the Early Holocene Site of Star Carr,North Yorkshire,UK

Shamanic belief systems represent the first form of religious practice visible within the global archaeological record. Here we report on the earliest known evidence of shamanic costume: modified red deer crania headdresses from the Early Holocene site of Star Carr (c. 11 kya). More than 90% of the examples from prehistoric Europe come from this one site, establishing it as a place of outstanding shamanistic/cosmological significance. Our work, involving a programme of experimental replication, analysis of macroscopic traces, organic residue analysis and 3D image acquisition, metrology and visualisation, represents the first attempt to understand the manufacturing processes used to create these artefacts. The results produced were unexpected—rather than being carefully crafted objects, elements of their production can only be described as expedient.