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We describe a method for identifying genes encoding proteins with stereospecific intracellular localizations in the fission yeast Schizosaccharomyces pombe. Yeast are transformed with a gene library in which S. pombe genomic sequences are fused to the gene encoding the Aequorea victoria green fluorescent protein (GFP), and intracellular localizations are subsequently identified by rapid fluorescence screening in vivo. In a model application of these methods to the fission yeast nucleus, we have identified several novel genes whose products are found in specific nuclear regions, including chromatin, the nucleolus, and the mitotic spindle, and sequence similarities between some of these genes and previously identified genes encoding nuclear proteins have validated the approach. These methods will be useful in identifying additional components of the S. pombe nucleus, and further extensions of this approach should also be applicable to a more comprehensive identification of the elements of intracellular architecture in fission yeast.  相似文献   
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Prostate cancer impacts on the daily lives of men, particularly their physical and emotional health, relationships and social life. This paper highlights how men cope with disease and treatment and the strategies they employ to manage their diagnosis alongside daily life. Twenty‐seven men were interviewed at different stages in their disease pathway: nine men prior to radiotherapy, eight men at 6–8 months post radiotherapy and 10 men at 12–18 months post radiotherapy. A grounded theory approach was used to collect and analyse the data. Regardless of the point at which they were interviewed four areas emerged as important to the men: the pathway to diagnosis; the diagnosis; the impact of prostate cancer and its treatment on daily life; and living with prostate cancer. Prostate cancer was diagnosed using the prostate‐specific antigen (PSA) test, rectal examination and biopsy. Many men did not understand the consequences of a high PSA reading before they undertook the test. Painful investigative biopsies were viewed as the worst part of the disease experience. Radiotherapy was considered less invasive than other treatments, although preparatory regimes were associated with stress and inconvenience. Men used various strategies to deal with treatment‐induced threats to their masculinity in the long term.  相似文献   
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A Stea  C A Nurse 《Neuroscience》1992,47(3):727-736
In this study we use whole-cell recording to characterize at least two distinct populations of cultured neurons from perinatal rat petrosal or petrosal/jugular ganglia based on differential sensitivity of the transient inward Na+ current to tetrodotoxin. These ganglia supply chemoreceptor and baroreceptor afferents which mediate several cardiovascular reflexes. Approximately 50% of the neurons sampled had Na+ currents that were virtually unaffected by bath addition of tetrodotoxin (0.5-2.0 microM) but were abolished by choline substitution for external Na+. The majority of the remaining neurons had Na+ currents that were rapidly and reversibly blocked by 500 nM tetrodotoxin. A few cells had both tetrodotoxin-resistant and tetrodotoxin-sensitive Na+ currents. All neurons had similar voltage-activated Ca2+ and K+ currents. The inward Ca2+ current had no obvious fast transient or T-type component and appeared to be due mainly to the presence of long-lasting L-type Ca2+ channels. The outward currents consisted largely of a delayed rectifying K+ current (IKdr) and a Ca(2+)-activated K+ current (IKca), but no obvious fast transient K+ current (IA) was observed. Exposure to a chemosensory stimulus, hypoxia (PO2 approximately 20 Torr), had no effect on these neurons, in contrast to the pronounced decrease in K+ current it produces in cultured glomus cells, the presumed chemoreceptors and normal targets for a subset of petrosal neurons in vivo. Current-clamp recordings indicated that some neurons gave single spikes while others gave multiple spikes in response to long-depolarizing stimuli. No correlation between spiking behaviour and tetrodotoxin-sensitivity was observed. Thus, cultures enriched in petrosal neurons contain subpopulations with differential sensitivities to tetrodotoxin. Since many of these neurons innervate a single chemosensory target organ, the carotid body, it is of interest to know whether one or both subtypes can form functional synapses with glomus cells of the carotid body and mediate a chemoreceptor reflex.  相似文献   
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