Purification of the human blood platelet thromboxane A2/prostaglandin H2 receptor protein.

The human platelet thromboxane A2/prostaglandin H2 receptor has been purified 6100-fold to apparent homogeneity by a three-step chromatographic procedure with an overall yield of 6%. A 6-fold purification of the receptor was first achieved by chromatography of 3-[(3-cholamidopropyl)dimethyl-ammonio]-1-propanesulfonate (CHAPS)-solubilized membrane proteins from human platelets on a diethylaminoethyl (DEAE)-Sepharose column. The DEAE eluate fractions containing receptor activity were then applied to a newly developed affinity column using the cyclohexyl derivative of SQ30,741 (SQ31,491) as the immobilized ligand. Elution of the receptor from the affinity column with BM13.177 yielded a further purification of 1700-fold. An additional 4-fold receptor purification from the affinity column eluate was achieved by HPLC using GPC 500 and GPC 100 columns connected in tandem. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining of the HPLC eluate containing purified receptor revealed a single, distinct band with a molecular weight of 55,000. The receptor binding activity was detected with [3H]SQ29,548 using a newly developed binding assay which involved immobilization of the receptor on polyethyleneimine-treated glass fiber filters. The binding of [3H]SQ29,548 to the purified receptor was time dependent, saturable, reversible and highly specific. Unlabeled SQ29,548, BM13.505, and U46619 (but not thromboxane B2 or 6-keto prostaglandin F1 alpha) competed for [3H]SQ29,548 binding to the purified receptor in a concentration-dependent manner. Scatchard analysis of [3H]SQ29,548 binding to the purified receptor revealed the presence of a single class of high-affinity binding sites, with a Kd of 4 nM and a Bmax of 17 nmol/mg protein.     

Expression of neuronal intermediate filament proteins ON1 and ON2 during goldfish optic nerve regeneration: effect of tectal ablation

Goldfish retinal explants were used to study optic tectum participation in the regulation of intermediate filament protein synthesis in retinal ganglion cells during optic nerve regeneration. Retinas were explanted at various times after removal of the contralateral optic tectum. The synthesis of the intermediate filament proteins ON1 and ON2 in the cultures was quantitated by labeling with [35S]methionine, followed by two-dimensional gel electrophoresis, autoradiography, and densitometry. Neuritic growth from the explants was quantitated based on fiber length and density. In retinal explants placed in culture after 23 days of optic nerve regeneration, the synthesis of ON1 and ON2 was reduced when the tectum had been ablated. In contrast, synthesis of these proteins in explants placed in culture at an earlier stage of regeneration was not affected by tectal ablation. At all time points tested, neuritic outgrowth from retinal explants was stimulated by tectal ablation. These findings indicate that the synthesis of the ON1 and ON2 intermediate filament proteins during regeneration is not directly regulated by axonal volume. Further, our findings suggest that interaction between growing axons and tectum is important for sustained expression of these proteins during the later stages of optic nerve regeneration.     

Infantile fibrosarcoma: radiological and clinical features

Two cases of infantile fibrosarcoma are described. This rare childhood malignancy of mesodermal origin usually affects the lower limbs, as it did in both of our cases. Previously, the only treatment option available involved some form of radical and often mutilating surgery. More recently, combination chemotherapy has given good results, with the effect that various imaging modalities have become important in assessing both the initial extent of disease and the response to treatment. Computed tomography has the advantage of demonstrating the amount of osseous involvement, but at the expense of a considerable dose of ionizing radiation. On the other hand, magnetic resonance imaging, with its multiplanar capacity, gives superior demonstration of breaching of tissue planes, which has important implications for planning of surgery. However, as in other soft tissue tumours, changes in signal characteristics with treatment have proved less specific than was originally anticipated.     

cAMP production via the adenylyl cyclase pathway is reduced in RCS rat RPE.

cAMP production was investigated in retinal pigment epithelium (RPE) cells isolated from normal rats and from rats with an inherited retinal dystrophy (Rdy/p+). In normal RPE cells, 5'-[N-Ethylcarboxamido]-adenosine (A2 receptors) produced a fivefold increase in the level of cyclic adenosine monophosphate (cAMP) over basal levels. However, only a onefold increase in cAMP was observed in dystrophic cells. cAMP production by prostaglandins E1 and E2 (prostaglandin receptors) in dystrophic RPE cells was only 29-38% of the level observed in normal cells. Direct stimulation of adenylyl cyclase by 10 mumol/l forskolin increased cAMP levels in normal RPE cells by 90 fold over basal, but only by sixfold in the dystrophic cells. These data suggest there may be a defect in the adenylyl cyclase signaling pathway in dystrophic RPE cells.     

Monoclonal antibody for avian thymic hormone.

Avian thymic hormone (ATH), a parvalbumin with immunomodulating properties, was used as antigen for development of a hybridoma cell line. Hybridoma screening by enzyme-linked immunosorbent assay (ELISA) led to the culture, purification, and characterization of an hybridoma, designated 4A6, which secretes IgG1 that is specific for ATH. The monoclonal antibody (MAb) was characterized by techniques involving indirect competitive and non-competitive ELISA, Western Blot analysis, and immunohistological staining. The MAb was highly specific for ATH with no cross-reactivity to other chicken tissue extracts or commercially available parvalbumins.     

Symptomatic treatment in the fragile X-associated tremor/ataxia syndrome.

There is no established treatment for the neurological features of the recently discovered fragile X-associated tremor/ataxia syndrome (FXTAS). Fifty-six patients with FXTAS completed a questionnaire to determine whether any medications had been effective for neurological symptoms. Of 11 subjects with definite FXTAS, 8 (70%) were on medications for their neurological symptoms, whereas most subjects with possible or probable FXTAS, 31 (70%) of 45 subjects, were not on medications. Although no therapy was uniformly effective for intention tremor, ataxia, Parkinsonism, memory loss, or anxiety, some subjects with intention tremor or Parkinsonism reported improvement with medications frequently used in other movement disorders. Overall, all 22 subjects on medications reported improvement in one or more symptoms. Lack of insight, recall bias, and cognitive impairment may have resulted in an underestimation of the beneficial effect of medical therapy. This study suggests that patients with FXTAS can derive improvement from medication treatment for some of their symptoms.     

The Community Links of a Sample of People with Intellectual Disabilities

Background In 1995, an unpublished study (S. Hewson & C. Waters) showed that the community links of residents in 11 local National Health Services (NHS) trust houses were meagre, despite the service's stated commitment to community presence and participation. The 1995 study was repeated for the same 11 houses in 2002 to examine whether any changes had occurred. Method A test–retest design, with repeated measures was used, involving, as closely as possible, the same participants at two time points. Community links referred to the time residents spent outside their house, and the time unpaid visitors spent inside the house in the presence of the residents. Results The community links of the people studied were no better in 2002 than in 1995. Conclusions These findings question whether current service provision can deliver policy objectives for social inclusion for people with intellectual disabilities.     

Diversity in Clinical Psychology

Clinical psychology is not as diverse as society, and the generalizability of clinical psychology to diverse groups has largely been untested. Some progress has been made in increasing the number of ethnic minorities receiving PhD degrees in clinical psychology and in increasing ethnic minority representation in clinical trials. Nevertheless, clinical psychologists and clinical psychology research are not diverse. A stages of change model may be useful in motivating clinical psychologists to diversify. A future scenario involving inertia and a second involving change are offered.     

Corrosion potential variation of aged dental amalgam restorations over time.

OBJECTIVE: The corrosion potential of a dental amalgam restoration is generally determined using a single measurement, even though environmental factors and abrasion can continuously alter the surface state and reactivity of this alloy. It was, therefore, the purpose of this study to determine the maximum variability of the corrosion potential of aged dental amalgam restorations, for 28 days. METHODS: The corrosion potentials of 148 aged dental amalgam restorations in 12 human subjects were measured at t = 0 and 4 h, and 4, 7, 14, 21 and 28 days. Measurements were made with a high impedance voltmeter and a Ag/AgCl micro-reference electrode. The subjects were instructed not to alter their usual eating and oral hygiene routines. RESULTS: Corrosion potential changes occurred throughout the 28 days. They were both positive and negative for the same restoration, and were sometimes very large. Only 4 h after the initial measurement, the absolute value of the corrosion potential changes ranged between 18 and 287 mV for 50% of the restorations. The largest maximum absolute corrosion potential change for each subject's restorations ranged between 85 and 329 mV. Statistical analysis showed that the overall mean maximum absolute corrosion potential change for the subjects' restorations was 74 mV. SIGNIFICANCE: It was shown that the corrosion potential of aged dental amalgam restorations varies substantially over time, and that a single measurement is not representative of short- or long-term electrochemical behavior. This finding has implications regarding the corrosion rate of dental amalgam restorations, particularly those that are part of a galvanic couple.