GPCR kinase knockout cells reveal the impact of individual GRKs on arrestin binding and GPCR regulation.

G protein-coupled receptors (GPCRs) activate G proteins and undergo a complex regulation by interaction with GPCR kinases (GRKs) and the formation of receptor-arrestin complexes. However, the impact of individual GRKs on arrestin binding is not clear. We report the creation of eleven combinatorial HEK293 knockout cell clones lacking GRK2/3/5/6, including single, double, triple and the quadruple GRK knockout. Analysis of ß-arrestin1/2 interactions for twelve GPCRs in our GRK knockout cells enables the differentiation of two main receptor subsets: GRK2/3-regulated and GRK2/3/5/6-regulated receptors. Furthermore, we identify GPCRs that interact with ß-arrestins via the overexpression of specific GRKs even in the absence of agonists. Finally, using GRK knockout cells, PKC inhibitors and ß-arrestin mutants, we present evidence for differential receptor-ß-arrestin1/2 complex configurations mediated by selective engagement of kinases. We anticipate our GRK knockout platform to facilitate the elucidation of previously unappreciated details of GRK-specific GPCR regulation and ß-arrestin complex formation.

Sphingolipid signaling in epidermal homeostasis. Current knowledge and new therapeutic approaches in dermatology.

In the present review we have attempted to give an overview of the role of sphingolipids in skin homoeostasis. Sphingolipid metabolites are emerging as potent second messengers in diverse cellular signaling pathways. In the skin little is known about sphingolipids in signaling events. In various cell populations it has been shown that different sphingolipid metabolites have opposing effects on the biological outcome of a stimulus. Therefore, the term 'sphingolipid rheostat' has been established and has also been shown to exist in skin-derived cell populations. In many cells ceramide is a mediator of proliferation inhibition and apoptosis, whereas sphingosine-1-phosphate acts more like a growth factor and reverses ceramide effects. In keratinocytes extracellular and intracellular ceramides play important roles. Extracellular ceramides are necessary for the water retention capacity and for maintaining the permeability barrier of the skin. Intracellular ceramides cause differentiation of keratinocytes. Until now less is known about the effect of other sphingolipid metabolites in the skin.

Chemosensory event-related potentials change with age.

The study examined age-related changes in the perception of olfactory and trigeminal chemical stimuli using chemosensory event-related potentials (CSERP). Three groups of healthy volunteers, each comprised of 8 men and 8 women, were tested (age ranges 15-34, 35-54, and 55-74 years). Subjects underwent extensive psychological testing focusing on impairments of memory and attention. In addition, odor identification and discrimination ability was evaluated, as well as detection threshold sensitivity for two odorants. Odor discrimination scores exhibited a significant age-related decrease. Significant age-related changes were also observed for CSERP N1P2 and P2 amplitudes, and for the N1 peak latency. The age-related decrease of CSERP amplitudes appeared to follow a different time course for responses to trigeminal and olfactory stimulants.

Olfactory function in acute rhinitis.

This study was performed to investigate the effects of the common cold on olfactory function, which was assessed using chemosensory event-related potentials (CSERP, in response to both olfactory [H2S] and trigeminal [CO2] stimuli) and psychophysical measures (intensity ratings, odor discrimination, butanol threshold); nasal volume was assessed by means of acoustic rhinometry. The investigation was performed in 36 subjects (18 women, 18 men). After onset of the rhinitis (day 0) measurements were performed on days 2, 4, 6 and 35. The cold produced a decrease of the volume of the anterior nasal cavity accompanied by an increase of mucus secretion, an increase of olfactory thresholds, a decrease of intensity ratings and a decrease of N1 CSERP amplitudes to olfactory and trigeminal stimuli. When mucus secretion of the contralateral nasal cavity was controlled with oxymetazoline, N1 amplitudes to olfactory stimuli were still affected by the cold as indicated by the significant increase of amplitudes as subjects recovered; this phenomenenon was not found for responses to trigeminal stimuli. This indicates that the common cold has a small effect on olfactory function which may be independent of nasal congestion.

Chemosensory event-related potentials in response to trigeminal and olfactory stimulation in idiopathic Parkinson's disease.

Decrease of olfactory function in patients with Parkinson's disease (PD) has been reported by several authors. The current study investigated olfaction in PD patients using olfactory event-related potentials (OERPs) as an electrophysiologic correlate of olfactory function in combination with psychophysical testing. A specific focus was the influence of antiparkinsonian drugs. We investigated PD patients treated with antiparkinsonian drugs (n = 13) and PD patients who received no pharmacologic treatment (n = 18). They were compared to age- and sex-matched control subjects (n = 38). To obtain OERPs, stimulants were chosen to stimulate specifically the olfactory nerve (2.1 ppm vanillin, 0.8 ppm H2S). In addition, chemosomatosensory event-related potentials were recorded after trigeminal stimulation with 52% v/v CO2. Moreover, the subjects' ability to identify and to discriminate odorants was tested by means of a "squeeze bottle" technique. The study yielded the following major results: (1) Odor identification was impaired in PD patients. It was not influenced by treatment with antiparkinsonian drugs. (2) The OERP latencies were prolonged in both PD patients taking and not taking antiparkinsonian drugs; however, this effect was more pronounced in PD patients taking antiparkinsonian drugs. (3) The intranasal chemosensory trigeminal system seemingly was neither affected by the neuronal degeneration seen in PD nor by treatment with antiparkinsonian drugs.

"Sniffin' sticks": screening of olfactory performance.

"Sniffin' Sticks" is a new test of nasal chemosensory performance based on pen-like odour-dispensing devices. This portable test is suited for repetitive, inexpensive screening of odour identification. The test includes a forced odour-identification task for seven odours performed by means of a list of four items (multiple-choice). In 146 subjects the basic screening test was compared to a down-scaled version of the UPSIT (CC-SIT). Sniffin' Sticks exhibited a relatively higher coefficient of correlation with the subjects' age; they also demonstrated the women's superior olfactory sensitivity more pronounced when compared to men. In addition, the coefficient of correlation between age and olfactory performance was slightly higher when the sticks were used. Preliminary investigations in nine patients with impaired olfactory function (i.e., anosmic or hyposmic patients) revealed significantly lower scores in patients compared to healthy controls matched for age and sex (p < 0.001). It is concluded that Sniffin' Sticks may be useful in the routine clinical assessment of olfactory performance where both time and costs matter.

Heavy ion results from different locations on LDEF.

Heavy ions (Z = 8-26) with energies far below the geomagnetic cutoff energy were measured in three different plastic nuclear track detector experiments on the 28.5 degrees inclination orbit of the NASA satellite Long Duration Exposure Facility (LDEF) at 460 km mean altitude. The results of the three experiments M0002 (Kiel University, Germany), A0015 (Deutsche Gesellschaft fur Luft- und Raumfahrt (= DLR) Koln, Germany) and HIIS (Naval Research Laboratory (= NRL) Washington, DC, U.S.A.), which were exposed at different satellite locations, agree with each other. At E < or = 70 MeV/nuc all particle groups showed a steeply falling energy spectrum and an anisotropic arrival direction distribution. The results were consistent with magnetically trapped particles registered in the South Atlantic. The detected particles originated from the anomalous cosmic rays (O, Ne, Ar) and from an unknown source (Mg, Si, Fe). At E > or = 70 MeV/nuc measured particles of the Fe-group showed a flattening energy spectrum.

Loss of olfactory function leads to a decrease of trigeminal sensitivity.

Healthy controls were compared to patients with decreased olfactory sensitivity (n = 32) to investigate interactions between the olfactory and trigeminal systems. Amplitudes of chemo-somatosensory event-related potentials in response to suprathreshold trigeminal stimuli (CO2) were found to be smaller in patients (P < 0.05) indicating a decrease of trigeminally mediated sensations.

On the origin of trapped heavy ions at L = 1.4-1.6.

Aboard the NASA satellite Long Duration Exposure Facility (LDEF) heavy ions of nuclear charge Z = 8-26 were detected with energies between 15 and 50 MeV/nuc which are far below the cutoff energy required of fully stripped ions to reach the LDEF orbit. The arrival directions and the falling energy spectra of these particles are consistent with a trapped component incident in the South Atlantic Anomaly at L = 1.4-1.6. The trapped oxygen, neon and argon ions probably originate from the anomalous cosmic rays, whereas the origin of the other particles like magnesium, silicon and iron is not yet solved but may be associated with the October 89 solar energetic particle events.