Time courses of calcium and calcium-bound buffers following calcium influx in a model cell

Fixed and diffusible calcium (Ca) buffers shape the spatial and temporal distribution of free Ca following Ca entry through voltage-gated ion channels. This modeling study explores intracellular Ca levels achieved near the membrane and in deeper locations following typical Ca currents obtained with patch clamp experiments. Ca ion diffusion sets an upper limit on the maximal average Ca concentration achieved near the membrane. Fixed buffers restrict Ca elevation spatially to the outermost areas of the cell and slow Ca equilibration. Fixed buffer bound with Ca near the membrane can act as Ca source after termination of Ca influx. The relative contribution of fixed versus diffusible buffers to shaping the Ca transient is determined to a large extent by the binding rate of each buffer, with diffusible buffer dominating at equal binding rates. In the presence of fixed buffers, diffusible buffers speed Ca equilibration throughout the cell. The concentration profile of Ca-bound diffusible buffer differs from the concentration profile of free Ca, reflecting theoretical limits on the temporal resolution which can be achieved with commonly used diffusible Ca indicators. A Ca indicator which is fixed to an intracellular component might more accurately report local Ca concentrations.     

Nutrient intakes of adolescents with phenylketonuria and infants and children with maple syrup urine disease on semisynthetic diets

Adequacy of nutrient intakes of adolescents with and without phenylketonuria (PKU) and infants and children with and without maple syrup urine disease (MSUD) were assessed using 3-day diet records sorted by disease and by age of the subject. Mean intakes of all nutrients were greater than two-thirds of the Recommended Dietary Allowances (RDA) or Estimated Safe and Adequate Daily Dietary Intakes (ESADDI) for all adolescents studied, with the exception of selenium (Se) in PKU adolescents, which averaged 27.8 micrograms. For adolescents with PKU, > 50% of the RDA or ESADDI for all nutrients was provided by elemental or modified protein hydrolysate medical foods, except for vitamin A in children aged 11-15 years and Se in children 11-18 years. Mean nutrient intakes of all infants and children were greater than two-thirds of the RDA or ESADDI for all nutrients except Se in MSUD children aged 1-11 years, where intakes ranged from 6.4 to 13.2 micrograms (21-66% of the RDA). The medical foods provided for most of the RDA and ESADDI recommendations, with the exception of Se in MSUD children.     

Further characterization of the alpha-actinin-actin interface and comparison with filamin-binding sites on actin

The interaction between alpha-actinin and actin was further characterized using natural and synthetic peptides of actin together with anti-actin antibodies of known specificity. We demonstrated that two alpha-actinin binding sequences on actin are located within residues 112-125 and 360-372. Each peptide was shown to directly bind alpha-actinin and was able to dissociate the alpha-actinin-actin complex using solid phase binding assays and cosedimentation experiments. Taking into account the three-dimensional structure of actin (Kabsch, W., Mannherz, H. G., Suck, D., Pai, E. F., and Holmes, K. C. (1990) Nature 347, 37-44), we postulate that these two segments, proximal in the actin structure, are part of the same site. In addition, we compared these two segments with those recently found for filamin (Méjean, C., Lebart, M. C., Boyer, M., Roustan, C., and Benyamin, Y. (1992) Eur. J. Biochem. 209, 555-562), Egan, S., Stewart, M., Stossel, T. P., Kwiatkowski, D. J., and Hartwig, J. H. (1990) J. Cell Biol. 111, 1089-1105), and concluded that the two actin-binding proteins interact with closely spaced or overlapping but not identical sequences of actin subdomain 1.     

Effects of ketotifen on human lymphocytes in vitro and in vivo

The effects of the antiasthmatic drug ketotifen (CAS 34580-13-7) on human mononuclear leukocytes were studied in vitro and in vivo. In vitro ketotifen concentration-dependently inhibited mitogen-stimulated lymphocyte proliferation. High ketotifen concentrations also inhibited T-lymphocyte mitogen- and adenosine triphosphate stimulated increases in intracellular Ca2+ in lymphocytes and the U937 human monocyte precursor cell line, respectively; this involved inhibition of both Ca2+ influx and intracellular mobilization. In in vivo experiments, treatment of healthy volunteers with 1 mg ketotifen b.i.d. for 7 d did not alter the number or subset composition of circulating lymphocytes. Moreover, the mitogen-stimulated in vitro proliferation of lymphocytes obtained before and after ketotifen treatment in vivo was similar. It is concluded that high ketotifen concentrations can inhibit the activation of resting lymphocytes in vitro but standard ketotifen treatment does not notably affect the number of function of circulating lymphocytes in vivo.     

Brainstem auditory evoked responses in patients with tinnitus

Brainstem auditory evoked responses of 355 patients with uni- or bilateral tinnitus were recorded in order to evaluate the effect of tinnitus on the central auditory system. The amplitudes of waves I, III and V and the latencies of each wave and interpeak latencies were compared to those of a group of 129 controls with normal hearing. The study of the control group initially identified a certain number of concurrent parameters. The brainstem evoked responses of men and women evolved differently from the age of 30 years, latencies of I-III and I-V in men lengthening with age and those of women tending to shorten. The patient groups were therefore compared to a control group of the same sex ratio or of the same sex, half being between 30 and 56 years of age. The tinnitus patients were divided into three groups according to the side affected by tinnitus. Latencies and amplitudes in these groups differed significantly from those of the control group. In order to eliminate hearing loss, the most difficult concurrent factor and almost always associated with tinnitus, the results of individuals with symmetrical hearing loss were compared to those of the control group. Tinnitus was always associated with significant lengthening of 0-I and I-V latencies on the tinnitus-affected side, with a significant reduction in amplitudes of waves I and III, and sometimes of wave V, particularly in the group with left-sided tinnitus. Comparison of tinnitus patients with symmetrical and asymmetrical hearing by sex showed that tinnitus patients of all groups had lengthening of right and left 0-I latencies, apart from the women in the group with right-sided tinnitus, and significant reduction in amplitudes of waves I and III in women and of left III only in men. When hearing loss was asymmetrical and on the tinnitus-affected side, there was also lengthening of 0-I latencies on the tinnitus-affected side in both sexes and of ipsi- and contralateral I-V latencies in women. Right- and left-sided tinnitus was associated with additional differences between the three groups. Correlation coefficient study confirmed that 0-I, I- III and I-V latencies were independent of the mean degree of deafness, deafness at high frequencies and at frequencies around the tinnitus, up to a threshold of hearing loss of 40 dB, above which 0-I and 0-V lengthened in addition to tinnitus. On the other hand, whatever the frequency, tinnitus involved significant lengthening of wave I latencies and modification of the previously recorded amplitudes. Two groups of tinnitus patients could be distinguished: the first, with symmetrical hearing loss, with symmetrical normal latencies, apart from 0-I latencies and the amplitude of the wave on the tinnitus side, and the second with hearing loss predominant on the tinnitus-affected side, with different latencies on each side, 0-I being shorter on the unaffected side, I-III and I-V being lengthened on the unaffected side and 0-I being lengthened on the tinnitus-affected side. Moreover, as disturbances of brainstem evoked responses caused by tinnitus particularly affected waves I and III, the hypothesis of possible involvement of the efferent systems could be proposed.     

Comparing the power of linkage detection by the transmission disequilibrium test and the identity-by-descent test

The purpose of this study was to identify new trends in the changing indications for penetrating keratoplasty. We retrospectively reviewed the clinical and pathologic diagnoses of 1,104 corneal buttons that had been submitted to the Estelle Doheny Eye Pathology Laboratory, Los Angeles, during the 5-year period 1989-1993. The leading indications, in order of decreasing frequency, were pseudophakic corneal edema (24.8%), regrafts (21.3%), scarring with or without chronic inflammation (11.1%), keratoconus (7.1%), aphakic corneal edema (6.4%), and ulcerative conditions (5.8%). The incidence of pseudophakic corneal edema remained stable over the study period and was actually surpassed by regraft in the last year of the study. Although pseudophakic corneal edema remains the predominant indication for penetrating keratoplasty, our findings suggest that its occurrence rate has begun to level off.     

Endgroup analysis of polyethylene glycol polymers by matrix-assisted laser desorption/ionization Fourier-transform ion cyclotron resonance mass spectrometry

Fourier-transform ion cyclotron resonance mass spectrometry (FTICR-MS) by external injection of matrix-assisted laser desorbed and ionized (MALDI) polymers offers good possibilities for characterization of low molecular weight homopolymers (MW range up to 10 kDa). The molecular masses of the molecular weight distribution (MWD) components of underivatized and derivatized (dimethyl, dipropyl, dibutyl and diacetyl) polyethylene glycol (PEG) 1000 and 4000 were measured by MALDI-FTICR-MS. These measurements have been performed using a commercial FTICR spectrometer with a home-built external ion source. MALDI of the samples with a 2,5-dihydroxybenzoic acid matrix in a 1000:1 matrix-to-analyte molar ratio produces sodiated molecules in a sufficient yield to trap the ions in the ICR cell. The masses of the molecular weight distribution of PEG components were measured in broad-band mode with a mass accuracy of < 5 ppm in the mass range around 1000 u and within 40 ppm accuracy around 4000 u. From these measurements, the endgroup mass of the polymer was determined by correlation of the measured component mass with the degree of polymerization. The masses of the PEG endgroups have been determined within a deviation of 3-10 millimass units for the PEG1000 derivatives and 10-100 millimass units for the PEG4000 derivatives, thus confirming the identity of the distal parts of the model compounds.     

Fatty acid composition of total lipids,neutral lipids and phospholipids in wild and farmed matrinxã (Brycon cephalus) in the Brazilian Amazon area

BACKGROUND: The chemical composition of Amazonian fish is extremely variable, being influenced by the season and the type and amount of food. A special interest in the fish oil composition has been developed owing to the presence of essential fatty acids, since this is directly related to human health. This study aimed to investigate the fatty acid composition (FAC) of the total lipid (TL), neutral lipid (NL) and phospholipid (PL) fractions of the dorsal muscle and orbital cavity of farmed and wild matrinxã in the Amazon area captured in different seasons. RESULTS: Fatty acids (FA) were analysed by high‐resolution gas chromatography/mass spectrometry. Sixty‐five FA were detected in the TL, 66 in the NL and 55 in the PL. The main FA found in farmed and wild fish were oleic, palmitic, stearic and linoleic acids. No distinctions in the quality or quantity of these fractions between dorsal muscle and orbital cavity were found. CONCLUSION: The season had a significant influence on the TL and FAC. Fish captured during the dry season showed lower levels of lipid and a higher percentage of long‐chain polyunsaturated fatty acids. Matrinxã farmed in a semi‐intensive system showed a nutritional quality comparable to that of wild matrinxã captured in the wet season. Copyright © 2007 Society of Chemical Industry