Spectroscopic study of the interaction of cationic dyes with mitochondria

Data are presented on the changes of absorption spectra of two cation dyes: nils blue and acridine orange while binding them with intact mitochondria of the rat liver in various functional states. It is found that when the dyes are bound with mitochondria characteristic changes proceed in the absorption spectra of dyes. These changes show that the formation of the dye molecule dimers is more intensive on the membrane than in solution. Energisation of mitochondria leads to a more intensive formation of acridine orange dimers, but the formation of nils blue dimers becomes less intensive. A suggestion is made about different nature of the interaction between these dyes and active groups of the mitochondria membrane.     

Quantitative parameters for amino acid-base interaction: implications for prediction of protein-DNA binding sites

Inspection of the amino acid-base interactions in protein-DNA complexes is essential to the understanding of specific recognition of DNA target sites by regulatory proteins. The accumulation of information on protein-DNA co-crystals challenges the derivation of quantitative parameters for amino acid-base interaction based on these data. Here we use the coordinates of 53 solved protein-DNA complexes to extract all non-homologous pairs of amino acid-base that are in close contact, including hydrogen bonds and hydrophobic interactions. By comparing the frequency distribution of the different pairs to a theoretical distribution and calculating the log odds, a quantitative measure that expresses the likelihood of interaction for each pair of amino acid-base could be extracted. A score that reflects the compatibility between a protein and its DNA target can be calculated by summing up the individual measures of the pairs of amino acid-base involved in the complex, assuming additivity in their contributions to binding. This score enables ranking of different DNA binding sites given a protein binding site and vice versa and can be used in molecular design protocols. We demonstrate its validity by comparing the predictions using this score with experimental binding results of sequence variants of zif268 zinc fingers and their DNA binding sites.     

Open-space schools: Anticipation of peer interaction and development of cooperative interdependence.

42 same-sex pairs of varied racial composition were randomly selected from 8th graders in each of 2 matched schools, with open- vs closed-space architectural styles. Open-space Ss were more likely to develop cooperative interdependence in a mixed-motive game (a decomposed prisoner's dilemma) and were more inclined to make proximal seating choices indicative of anticipated peer interaction. A Sex by Race by Trial Blocks interaction effect reflected different patterns of responding for males and females. Females of either race learned to cooperate in same-race pairs and to compete in mixed-race pairs. White males learned to cooperate and Black males to compete independent of their partner's race. External scores on Rotter's Internal–External Locus of Control Scale were not related to schools but were, as expected, higher for Black than for White students. (14 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Astrocytoma cell interaction with elastin substrates: implications for astrocytoma invasive potential

Elastin has been identified within the meninges and the microvasculature of the normal human brain. However, the role that elastin plays in either facilitating astrocytoma cell attachment to these structures or modulating astrocytoma invasion has not been previously characterized. We have recently shown that astrocytoma cell lines and specimens produce tropoelastin, and express the 67 kDa elastin binding protein (EBP). In the present report, we have established that astrocytoma cells attach to elastin as a substrate in vitro. The U87 MG astrocytoma cell line demonstrated the greatest degree of adhesion. In addition, all astrocytoma cell lines examined were capable of penetrating and migrating through an intact elastin membrane, and of degrading tritiated-elastin, a process that could be prevented by the pre-incubation of astrocytoma cells with EDTA, but not with alpha1-antitrypsin. Astrocytoma cells were also capable of penetrating 1 mm sections of human brain tissue maintained as organotypic cultures. Interestingly, the invasive potential of cultured astrocytoma cells plated on organotypic cultures of human brain was significantly increased after exposure to elastin degradation products (kappa-elastin), which interact with astrocytoma cell surface EBP. Our data show that astrocytoma cells express a functional 67 kDa EBP, enabling them to potentially recognize and attach to elastin as a substrate. These data also suggest that this elastin receptor may be involved in processes which regulate regional astrocytoma invasion.     

Spectroscopic evidence for interaction between transmembrane helices 3 and 5 in rhodopsin

Recent molecular models of rhodopsin (Rho) propose a specific interaction between transmembrane (TM) helices 3 and 5, which appears to be mediated by amino acid residues Glu122 and His211 on TM helices 3 and 5, respectively. To test this proposed interaction, four single-site histidine replacement mutants (H100N, H152N, H211N, and H211F), two single-site glutamic acid replacement mutants (E122Q and E122A), and three double-site replacement mutants (E122Q/H211F, E122Q/H211N, and E122A/H211F) of Rho were prepared. The expressed mutant pigments reconstituted into membranes were studied by FTIR difference spectroscopy addressing especially the transition to metarhodopsin I (MI). It is shown that the lipid environment influences bands typical of the MI state. Spectra of mutants with substituted Glu122 allowed assignments of the C=O stretch of protonated Glu122 in the dark state and in MI of Rho. Mutation of His211, but not of other histidine residues, affects these vibrational modes assigned to Glu122. In addition, replacements of His211 affect protein modes that are proposed to arise from a third, hydroxyl-bearing group, which also interacts with Glu122. These modes are influenced as well when Glu122 is replaced by Ala in mutant E122A but not when it is replaced by Gln in mutant E122Q. These results provide direct experimental evidence for an interaction between TM helices 3 and 5 in Rho, which is mediated by Glu122 and His211.     

Trefoil peptide protection of intestinal epithelial barrier function: cooperative interaction with mucin glycoprotein

BACKGROUND & AIMS: Goblet cells secrete a combination of trefoil peptides and mucin glycoproteins to form a continuous gel on the mucosal surface. The functional effects of these products remain uncertain. METHODS: Trefoil peptides and/or mucin glycoproteins were added to Transwell monolayers of the human colonic cancer-derived T84 cell line. Intact monolayers permitted penetration of < 4% of the inert marker [3H]mannitol at 4 hours. Exposure to the toxic lectin phytohemagglutinin (1 mg/mL), oleic acid (8 mmol/L) and taurocholic acid (12 mmol/L), or Clostridium difficile toxin A (0.7 microgram/mL) resulted in loss of barrier function with 36%, 62%, and 45% of [3H]mannitol penetration, respectively. RESULTS: Addition of recombinant human intestinal trefoil factor in physiological concentrations (1-5 micrograms/microL) resulted in attenuation of the damage to monolayer integrity by up to 52%. Protection was enhanced (up to 95%) by the copresence of human colonic mucin glycoproteins. Similar effects were observed when rat intestinal trefoil factor or human spasmolysin, another human trefoil peptide, were added alone or in the presence of human mucin glycoproteins. Conversely, mucin glycoproteins isolated from the rat colon or stomach facilitated protection when added with human spasmolysin or human intestinal trefoil factor. CONCLUSIONS: Trefoil peptides and mucin glycoproteins protect gastrointestinal mucosa from a variety of insults.     

SA-210C高压锅炉用无缝钢管的开发

介绍了新钢公司SA-210C高压锅炉用无缝钢管的开发过程,以及采用的技术标准和原则生产工艺。通过试制,获得成功。     

Initial study of 210Pb in indoor air

An exploratory program was undertaken to determine the feasibility of measuring the indoor air concentration of 210Pb. Low flow rate pumps and membrane filters were used to collect samples in an office. The air filter samples were stored for 1.6 y and 210Po was chemically separated and measured by alpha spectrometry. Unsupported 210Po was also determined in air samples collected at a later time. The average air concentrations were 150 microBq 210 Pb m-3 and 12 microBq 210Po m-3. The average indoor to outdoor ratio of 210Pb in air is estimated to be 0.25.     

Effects of cooperative and individualistic learning experiences on interethnic interaction.

51 4th graders were assigned to conditions on a stratified random basis controlling for ethnic membership, ability, and sex. Ss participated in an instructional unit for 45 min/day for 16 instructional days. Behavioral measures were taken for cross-ethnic interaction within the instructional situation and during daily free-time periods. A number of attitude measures were also administered. Results indicate that cooperative learning experiences, compared with individualistic ones, promote more cross-ethnic interaction in both instructional and free-time activities. (16 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

General hospital psychiatry: implications for occupational therapy--one case study

The hospitalization policy in general hospital psychiatric wards is for crisis intervention. The high cost in such a ward in Israel (in comparison with the cost in psychiatric hospitals) is justified since the patient is treated both physically and mentally. Some cases include patients who suffer from chronic diseases and aggravation of their physical condition causes severe mental reactions. In these cases, the short-term psychiatric admission is often in conflict with the condition of the patient and the desired treatment programme. The occupational therapist whose professional values are founded on the quality of life and his or her holistic approach to the patient, often has to deal with a serious dilemma: hospital policy on one hand and the patient's needs on the other. This dilemma is presented in its most acute form in the case study discussed in this article.     

Biological variation of cystatin C: implications for the assessment of glomerular filtration rate

To assess the inherent potential for detecting mild to moderate reductions in glomerular filtration rate, this study determined the biological variability of serum cystatin C and creatinine in 12 healthy subjects. After accounting for analytical variation, interindividual variance accounted for 93% and intraindividual variance accounted for 7% of serum creatinine biological variation. As such, to lie outside the assay reference interval, some subjects must exceed 13 SD from their usual mean value, whereas in others, a change of only 2 SD would be sufficient. For cystatin C, interindividual variation explained 25% and intraindividual variance explained 75% of biological variability. Therefore, the upper limit of the population reference interval for cystatin C is seldom more than 3-4 SD from the mean value of any healthy individual. The critical difference for sequential values significant at P < or = 0.05 was calculated as 37% for serum cystatin C and 14% for serum creatinine. We conclude that cystatin C is potentially a better marker for detecting impaired renal function than serum creatinine, but serum creatinine is probably still the better marker for detecting temporal changes of renal function in individuals with established renal disease.     

Effects of metacognitive strategy training within a cooperative group learning context on computer achievement and anxiety: An aptitude–treatment interaction study.

Two aptitude–treatment interaction studies examined the comparative effects of metacognitive strategy training in self-questioning within a cooperative group learning context and a traditional direct instruction approach, on the acquisition of computing competencies, learning anxiety, and positive cognitions. When prior competence in using computers is controlled, students' initial aptitudes interact significantly with teaching method. Cooperative groups scored significantly better on achievement tests, self-concept, and sense of control-mastery than did the direct instruction groups. Paradoxically, for the initially high-anxious learners, some aspects of computing anxiety remained high in the cooperative group relative to the direct instruction group, suggesting that anxiety may facilitate learning. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Persistence of the interaction of calmodulin with adenylyl cyclase: implications for integration of transient calcium stimuli

Ca2+/calmodulin-sensitive adenylyl cyclase plays a role in several forms of synaptic plasticity and learning. To understand how cellular signals from neuronal activity during behavioral stimuli might be integrated by adenylyl cyclase, we have characterized the response of type I adenylyl cyclase to transient Ca2+ stimuli. Stimulation by a several second Ca2+ stimulus is delayed, rising to a peak after the Ca2+ stimulus has ended. We attempted to identify the site of the persistent Ca2+ signal that enabled adenylyl cyclase stimulation to increase after free Ca2+ had declined. Free calmodulin itself displayed no persistent activation by Ca2+ and was unable to activate adenylyl cyclase if exposed to low Ca2+ solution <1 s before reaching adenylyl cyclase. In contrast, activation of the calmodulin-adenylyl cyclase complex persisted for seconds after Ca2+ stimulus. Activation decayed with a time constant of 6 or 13 s depending on assay conditions. These results suggest that the calmodulin-adenylyl cyclase complex can serve as a site of cellular memory for a Ca2+ transient that has ended even before adenylyl cyclase is fully activated.     

An excitatory scorpion toxin with a distinctive feature: an additional alpha helix at the C terminus and its implications for interaction with insect sodium channels

BACKGROUND: Scorpion neurotoxins, which bind and modulate sodium channels, have been divided into two groups, the alpha and beta toxins, according to their activities. The beta-toxin class includes the groups of excitatory and depressant toxins, which differ in their mode of action and are highly specific against insects. The three-dimensional structures of several alpha and beta toxins have been determined at high resolution, but no detailed 3D structure of an excitatory toxin has been presented so far. RESULTS: The crystal structure of an anti-insect excitatory toxin from the scorpion Buthotus judaicus, Bj-xtrIT, has been determined at 2.1 A resolution and refined to an R factor of 0.209. The first 59 residues form a closely packed module, structurally similar to the conserved alpha and beta toxins ('long toxins') affecting sodium channels. The last 17 residues form a C-terminal extension not previously seen in scorpion toxins. It comprises a short alpha helix anchored to the N-terminal module by a disulfide bridge and is followed by a highly mobile stretch of seven residues, of which only four are seen in the electron-density map. This mobile peptide covers part of a conserved hydrophobic surface that is thought to be essential for interaction with the channel in several long toxins. CONCLUSIONS: Replacement of the last seven residues by a single glycine abolishes the activity of Bj-xtrIT, strongly suggesting that these residues are intimately involved in the interaction with the channel. Taken together with the partial shielding of the conserved hydrophobic surface and the proximity of the C terminus to an adjacent surface rich in charged residues, it seems likely that the bioactive surface of Bj-xtrIT is formed by residues surrounding the C terminus. The 3D structure and a recently developed expression system for Bj-xtrIT pave the way for identifying the structural determinants involved in the bioactivity and anti-insect specificity of excitatory toxins.     

Localization of Niemann-Pick C1 protein in astrocytes: implications for neuronal degeneration in Niemann- Pick type C disease

Niemann-Pick type C disease (NP-C) is an inherited neurovisceral lipid storage disorder characterized by progressive neurodegeneration. Most cases of NP-C result from inactivating mutations of NPC1, a recently identified member of a family of genes encoding membrane-bound proteins containing putative sterol sensing domains. By using a specific antipeptide antibody to human NPC1, we have here investigated the cellular and subcellular localization and regulation of NPC1. By light and electron microscopic immunocytochemistry of monkey brain, NPC1 was expressed predominantly in perisynaptic astrocytic glial processes. At a subcellular level, NPC1 localized to vesicles with the morphological characteristics of lysosomes and to sites near the plasma membrane. Analysis of the temporal and spatial pattern of neurodegeneration in the NP-C mouse, a spontaneous mutant model of human NP-C, by amino-cupric-silver staining, showed that the terminal fields of axons and dendrites are the earliest sites of degeneration that occur well before the appearance of a neurological phenotype. Western blots of cultured human fibroblasts and monkey brain homogenates revealed NPC1 as a 165-kDa protein. NPC1 levels in cultured fibroblasts were unchanged by incubation with low density lipoproteins or oxysterols but were increased 2- to 3-fold by the drugs progesterone and U-18666A, which block cholesterol transport out of lysosomes, and by the lysosomotropic agent NH4Cl. These studies show that NPC1 in brain is predominantly a glial protein present in astrocytic processes closely associated with nerve terminals, the earliest site of degeneration in NP-C. Given the vesicular localization of NPC1 and its proposed role in mediating retroendocytic trafficking of cholesterol and other lysosomal cargo, these results suggest that disruption of NPC1-mediated vesicular trafficking in astrocytes may be linked to neuronal degeneration in NP-C.