Bioethics in the medical subspecialty literature

The high affinity growth hormone-binding protein (GHBP) circulates in human blood and represents the extracellular domain of the growth hormone (GH) receptor. It is well known that repetitive bouts of endurance type exercise result in increased integrated GH secretion. As the effects of chronic exercise on plasma GHBP levels have never been studied systematically, we investigated the effect of 2 weeks of intense endurance training on plasma GHBP as well as on plasma insulin-like growth factor (IGF)-I levels in 10 healthy, young, non-obese men. IGF-I was measured as an indicator of the effects of GH release. We also studied 10 control subjects matched for sex, age and activity, who were instructed not to change their customary activities. GHBP was determined by FPLC size exclusion chromatography and subsequent Scatchard plot analysis of the binding data; IGF-I levels were measured by RIA. The results showed that plasma IGF-I and GHBP levels were increased in the subjects who followed the training program. IGF-I and GHBP changed from 252 +/- 56 ng/ml and 912 +/- 59 pmol/l before training, to 344 +/- 61 ng/ml (p < 0.01) and 1020 +/- 48 pmol/l (p < 0.01), respectively. Another effect of the training was that the aerobic capacity of these subjects was better utilized and endurance was improved. In contrast, plasma IGF-I, GHBP, utilization of aerobic capacity and endurance did not change significantly in the control subjects. We conclude that two weeks of strenuous endurance training lead to increased plasma IGF-I and high affinity GHBP levels.     

The gastrin-releasing peptide receptor is differentially coupled to adenylate cyclase and phospholipase C in different tissues

A magnetization-prepared sequence, T2-Prep-IR, exploits T1, T2, and chemical shift differences to suppress background tissues relative to arterial blood. The resulting flow-independent angiograms depict vessels with any orientation and flow velocity. No extrinsic contrast agent is required. Muscle is the dominant source of background signal in normal volunteers. However, long-T2 deep venous blood and nonvascular fluids such as edema also contribute background signal in some patients. Three sets of imaging parameters are described to address patient-specific contrast requirements. A rapid, spiral-based, three-dimensional readout is utilized to generate high-resolution angiograms of the lower extremities. Comparisons with x-ray angiography and two-dimensional time-of-flight angiography indicate that this flow-independent technique has unique capabilities to accurately depict stenoses and to visualize slow flow and in-plane vessels.     

Heavy metal content of the hair of children in industrial cities

Nuchal screening has the following advantages: Non invasive. First trimester procedure. Results are obtained quickly. 80-90% detection rate for Down's Syndrome.     

Proteolytic cleavage and activation of PAK2 during UV irradiation-induced apoptosis in A431 cells

Exposure of mammalian cells to ultraviolet (UV) light elicits a cellular response and can also lead to apoptotic cell death. In this report, we show that a 36-kDa myelin basic protein (MBP) kinase detected by an in-gel kinase assay can be dramatically activated during the early stages of UV irradiation-triggered apoptosis of A431 cells. Immunoblot analysis revealed that this 36-kDa MBP kinase could be recognized by an antibody against the C-terminal regions of a family of p21Cdc42/Rac-activated kinases (PAKs). By using this antibody and a PAK2-specific antibody against the N-terminal region of PAK2 as studying tools, we further demonstrated that UV irradiation caused cleavage of PAK2 to generate a 36-kDa C-terminal catalytic fragment and a 30-kDa N-terminal fragment in A431 cells. The appearance of the 36-kDa C-terminal catalytic fragment of PAK2 matched exactly with the activation of the 36-kDa MBP kinase in A431 cells upon UV irradiation. In addition, UV irradiation also led to activation of CPP32/caspase-3, but not ICH-1L/caspase-2 and ICE/caspase-1, in A431 cells and the kinetics of activation of CPP32/caspase-3 appeared to correlate well with that of DNA fragmentation and of cleavage/activation of PAK2, respectively. Moreover, blockage of activation of CPP32/caspase-3 by pretreating the cells with two specific tetrapeptidic inhibitors for caspases (Ac-DEVD-cho and Ac-YVAD-cmk) could significantly attenuate the extent of cleavage/activation of PAK2 induced by UV irradiation. Collectively, the results demonstrate that cleavage and activation of PAK2 can be induced during the early stages of UV irradiation-triggered apoptosis and indicate the involvement of CPP32/caspase-3 in this process.     

Calcium modulation of activation and desensitization of nicotinic receptors from mouse brain

The effects of extracellular calcium on functional properties of nicotinic receptors from mouse thalamus were investigated. Previous studies have reported that calcium modulates the function of several neuronal nicotinic receptors. A 86Rb+ ion efflux assay was developed to measure nicotinic receptor function from brain tissue, and data indicate that alpha4beta2 receptors may mediate this response. Using the 86Rb+ efflux assay, calcium effects on receptor activation, desensitization induced by high, activating and low, subactivating concentrations of agonist, and recovery from desensitization were examined. Effects of calcium on the kinetics of ligand binding were also investigated. Calcium modulated receptor activation by increasing the maximal response to nicotine in a concentration-dependent manner, without affecting the EC50 of nicotine. Barium, but not magnesium, mimicked the effects of calcium on receptor activation. The increase in receptor activation could not be explained by changes in the ratio of activatable to desensitized receptors as assessed by the kinetics of ligand binding. Desensitization following activation was unaffected by calcium. Calcium, barium, and magnesium, however, increased the potency of nicotine for desensitization induced by exposure to low, subactivating concentrations of nicotine. Recovery from desensitization was not modulated by calcium. These data suggest that calcium modulates various functional aspects of nicotinic receptors from mouse brain and may do so via different mechanisms.     

Corrosion behaviour of an indigenous Ag-Sn-Cu cast dental alloy in artificial saliva

Tyrosine kinases are involved in various intracellular signalling cascades of different cells: Genistein has been shown to inhibit tyrosine kinase in INS-1 cells, an insulin-secreting cell line (Verspohl et al., 1995). It is, however, not established how specific and selective the tyrosine kinase inhibitors and their controls are. The tyrosine kinase inhibitors genistein and tyrphostin 25 increased insulin release, but not their negative controls with isoflavonoid structure (daidzein and genistin). In addition to this short-term effect a long-term effect was investigated. Genistein (100 microM) time-dependently increased insulin mRNA levels in INS-1 cells. On the other hand the tyrosine kinase inhibitors tyrphostin 25 and lavendustin A (both at 100 microM), which are structurally different from genistein, failed to increase the insulin mRNA whereas daidzein and genistin, normally used as negative controls, increased insulin mRNA as potently as genistein did. However, an examination of the incubation medium revealed that genistin was degraded to genistein by about 50% probably by nonspecific glucosidases first seen after 2 hours of incubation; genistin, therefore, does not appear to be a proper control though often used in this way. In conclusion, the suitability of the compounds used in recent studies is doubtful since other effects than the inhibition of tyrosine kinases are possible. Whereas the involvement of tyrosine kinase in a short-term effect (insulin release) is obvious and clearly substantiated by using the established pharmacological tools (negative controls), the involvement of tyrosine kinases in long-term effects is not that clear; only compounds with isoflavonoid structure are effective independent whether they normally are thought to be inhibitors or negative controls. One has to be cautious in using the above-mentioned compounds in an uncritical way.     

Polymerase chain reaction amplification of Trichomonas vaginalis DNA from Papanicolaou-stained smears

We describe 3 cases of Hodgkin's disease (HD) of unusual suppurative type, which were diagnosed on fine-needle aspirates. The smears were dominated by neutrophils, macrophages, and cellular debris. Only a few large, atypical cells of Hodgkin and Reed-Sternberg type were observed. The differential diagnoses of such smears include infectious mononucleosis, tuberculosis, metastatic lymph node involvement, non-Hodgkin's large-cell anaplastic Ki-1-positive lymphomas, T-cell-rich B-cell lymphomas, and peripheral T-cell lymphomas of mixed type. Immunocytochemistry identified the large atypical cells as CD 30 (BerH2)-positive and negative for CD 45 (LCA) in cytospin material from 2 patients, which allowed a conclusive diagnosis of HD.     

Specific detection of the genus Serpulina, S. hyodysenteriae and S. pilosicoliin porcine intestines by fluorescent rRNA in situ hybridization

PURPOSE: Our purpose was to evaluate the utility of spectral imaging for multicolor, multichromosome enumeration in human interphase cell nuclei. METHODS: Chromosome-specific probes labeled with different fluorochromes or nonfluorescent haptens were obtained commercially or prepared in-house. Metaphase spreads, interphase lymphocytes, or blastomeres cells were hybridized with either 7 or 11 distinctly different probes. Following 46 hr of hybridization, slides were washed and detected using either a filter-based quantitative image processing system (QUIPS) developed in-house or a commercial spectral imaging system. RESULTS: The filter-based fluorescence microscope system is preferred for simultaneous detection of up to seven chromosome targets because of its high sensitivity and speed. However, this approach may not be applicable to interphase cells when 11 or more targets need to be discriminated. Interferometer-based spectral imaging with a spectral resolution of approximately 10 nm allows labeling of chromosome-specific DNA probes with fluorochromes having greatly overlapping emission spectra. This leads to increases in the number of fluorochromes or fluorochrome combinations available to score unambiguously chromosomes in interphase nuclei. CONCLUSIONS: Spectral imaging provides a significant improvement over conventional filter-based microscope systems for enumeration of multiple chromosomes in interphase nuclei, although further technical development is necessary in its application to embryonic blastomeres. When applied to preconception/preimplantation genetic diagnosis, presently available probes for spectral imaging are expected to detect abnormalities responsible for 70-80% of spontaneous abortions caused by chromosomal trisomies.     

The alcohol deprivation effect in the alcohol-preferring P rat under free-drinking and operant access conditions

The alcohol-deprivation effect (ADE) was examined under 4-hr operant and 24-hr free-choice alcohol access in the alcohol-preferring (P) rat after deprivation intervals from 2 to 4 weeks. Results indicated that adult male P rats responding for 6 weeks on a concurrent FR-5/FR-1 schedule of reinforcement for alcohol and water, respectively, and then deprived of alcohol for 2 weeks, demonstrated a 40% increase in alcohol responding during the first 60 min of alcohol reinstatement. The alcohol deprivation effect was temporary, however, as responding did not differ from baseline levels on the second day of reinstatement. In a second experiment, weanling male and female P rats received 7 weeks of continuous access to alcohol, beginning at 21 days of age, and were then deprived of alcohol for 4 weeks. On the first day of alcohol reinstatement, P rats exhibited a 40% to 45% increase from baseline alcohol drinking levels, with alcohol intake returning to baseline levels by the 3rd day of reinstatement. Although alcohol intake was higher in females than in males when adjustment was made for body weight, there were no gender differences in the magnitude of the alcohol deprivation effect. Taken together, these results indicate that the ADE is a long-lasting phenomenon that occurs under both operant and continuous access conditions in the P rat, and thus these rats may be useful models for the study of factors involved in relapse of alcohol drinking.