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Messenger RNA has been isolated from rat lens tissue. The mRNA species which codes for the A2 chain of alpha-crystallin, revealed the same extremely high sedimentation value (14S) as the corresponding messenger from calf lens. However, it has been shown that in rat lens the 14-S messenger preparation directs the synthesis of an additional alpha-crystallin chain, designated as alpha-X with an approximate molecular weight of 24000. For comparative purpose crystallin synthesis has also been studied as well in cultured rat lens cells as in the rat lens cell-free system.

This study tested the effects of life stress and variables hypothesized to moderate these effects using a multivariate hierarchical regression model. College undergraduates served as subjects. The variables of life stress, dispositional locus of control beliefs, perceptions of control over life event outcomes and psychological disorder were measured at two points in time. At both measurement periods, negative life events were significantly related to psychological disorder. This relationship was found even when initial psychological disorder was statistically controlled. Dispositional locus of control beliefs and control perceptions failed to moderate the effects of life events at either measurement period. Control perceptions appeared to be more a function of event characteristics than subjects' dispositional characteristics, given that these perceptions were unrelated to locus of control beliefs and that the outcomes of negative events were consistently viewed as less controllable than those of positive events. The findings support the relationship between life stress and psychological disorder but they call into question the moderating effects of locus of control beliefs and perceptions of control over life event outcomes.

Fifty-three teen-agers with spina bifida participated in a mail survey and completed measures of recent life events, perceived family environment, and psychological distress. Low levels of perceived family conflict and control served as life stress buffers in the prediction of distress, whereas a high level of perceived independence served as a life stress exacerbator. These interaction effects differ from those obtained for a normal sample of adolescents in the lone previous study (Burt, Cohen, & Bjorck, 1988) that reported comparable analyses. The results suggest that the process by which family environments moderate stress adjustment differs for able-bodied vs. spina bifida adolescents.

Administered measures of coping, life events, and anxiety and depression to junior high and senior high school samples on two occasions, separated by a 5-month interval. Factor analyses supported the creation of coping subscales for problem solving, cognitive coping, social entertainment, physical exercise, and peer support. A one-item index of parental support was also included in the analyses. The coping subscales showed moderate temporal stability. Mothers' reports of their children's coping provided only marginal support for the validity of the adolescents' self-reports. Prospective regression analyses of the early adolescent data revealed that problem-solving coping was negatively related to depression, and that social entertainment coping was negatively related to anxiety. The prospective effects for the middle adolescents' coping were nonsignificant. The findings are discussed in terms of their implications for the measurement of adolescent coping strategies and research on the relation between these strategies and psychological functioning.

14-S mRNA from rat lens codes for two subunits of alpha-crystallin, A2 (Mr 20 000) and AIns (Mr 24 000, previously referred to as alphaX). Structural relationship between both translation products has been proved by immunoprecipitation with antisera directed against the different crystallin classes. Competition immunoprecipitation showed that the 14-S mRNA translation products are precipitated by common antibodies, specific for the A subunit of alpha-crystallin. Two-dimensional gel electrophoresis and peptide analysis provided further evidence that the 24 000-Mr polypeptide, synthesized in vitro under direction of 14-S mRNA, is identical with native alphaAIns. Although the structures of alphaA2 and alphaAIns are very similar, no precursor-product relationship exists between both 14-S-mRNA-encoded polypeptides.

Squalene synthetase activity in liver microsomes from rats sacrificed at three different times of the diurnal cycle showed no significant differences. Addition of 4% cholestyramine to the food resulted in a marked increase in activity (280% of control), independent of the time of killing. 3-Hydroxy-3-methylglutaryl coenzyme A reductase and cholesterol 7 alpha-hydroxylase activity, determined as positive controls, were also found to be elevated by cholestyramine and additionally showed a diurnal variation. On the other hand, five control enzyme activities, not directly related to cholesterol metabolism, i.e. glutamate dehydrogenase, NADPH cytochrome-c reductase, beta-hexosaminidase, catalase and acyl coenzyme A oxidase, showed neither an influence of cholestyramine feeding nor a time of sacrifice dependent variation.

The defect in urinary acidification induced by amphotericin B (AMB) was further characterized in turtle bladder. Since AMB has been shown to increase the hydrogen ion (H+) permeability of this epithelium in the absence of exogenous bicarbonate ions (HCO3-), we explored the permeability characteristics in the presence of imposed bicarbonate ion gradients, comparable to those occurring in vivo. With mucosal (M) pH lowered to the point of zero net hydrogen ion secretion, the transepithelial flow of bicarbonate ions (JHCO3) from serosa (S) to M was 0.91 +/- .06 y mole/hr in response to a 20 mM HCO3- gradient. After AMB addition to M, back diffusion of hydrogen ions from M to S (-JH) increased from zero to 0.36+/-0.05 micronmole/hr, whereas bicarbonate ion transport from S to M (JHCO3) failed to increase (0.91+/-0.06 before and 0.82+/-0.09 micronmole/hr after AMB). In contrast to M addition, S addition of AMB had no effect on either -JH or JHCO3. The defect in urinary acidification induced by AMB is characterized by a large increase in the permeability for hydrogen ions rather than that for bicarbonate ions and depends on direct exposure of the luminal cell membrane to AMB. The permeability increase is cation selective, not only for hydrogen ions but also, as shown previously, for potassium ions, and to a lesser extent, for sodium ions. The results are consistent with the formation by AMB of aqueous half pores in the luminal membrane. Although the passive permeabilities for bicarbonate and chloride ions are not affected primarily, they may increase after prolonged exposure, probably as a results of paracellular leaks that are not specific for AMB.

Rat lens alpha-crystallin contains, besides the usual alphaA and alphaB subunits, an additional minor chain. This subunit was purified by ion-exchange chromatography and its primary structure studied. It appeared to be an elongated alphaA-like chain, having an insertion of 22 residues between position 63 and 64 of an otherwise normal alphaA2 chain. Therefore this subunit was called alphaAIns, i.e. an alphaA chain with an inserted sequence. This inserted region, which contains three methionyl, five basic and no acidic residues, apparently results in an adequately functioning alphaAIns chain. The alphaAIns chain may be the product of a gene which has originated, after duplication of the alphaA gene, by insertion in one of the copies of a stretch of 66 nucleotides of unknown origin, or alternatively be the result of unusual transcription or processing of precursor mRNA (pre-mRNA), leaving an extra 66 nucleotides internally in the mRNA to be translated.

The structure of the gene coding for ribosomal protein S33, - a protein which escapes the coordinate control of ribosomal protein synthesis in rna 2 mutant cells -, was determined by sequence analysis. The gene comprises an uninterrupted coding region of 204 nucleotides encoding a protein of 8.9 kD. Like for other yeast ribosomal protein genes that have been sequenced so far, a relatively strong codon bias was observed. By S1 nuclease mapping the 5' end of the S33 mRNA was shown to be located at 11 to 15 nucleotides upstream from the initiation codon.

A colony bank of yeast dna obtained by cloning HindIII-generated fragments of total yeast nuclear DNA in Escherichia coli K-12 with the vector pBR322, was screened with a radioactive RNA probe enriched for a subset of ribosomal protein mRNAs. The selected recombinant DNA molecules were hybridized with poly(A)-containing mRNA under R-loop conditions. From the DNA-RNA hybrids the respective mRNAs were melted off and translated in vitro in a rabbit reticulocyte cell-free system. The translational products were analyzed by immunoprecipitation with antibodies raised against ribosomal proteins. The identity of the ribosomal protein gene products was further established by electrophoresis on two-dimensional gels. At least 15 recombinant DNA molecules were shown to contain ribosomal protein genes. Four of them, i.e. Y65, Y89, Y113 and Y138, have been characterized preliminarily.

We show that in sea urchin embryos, the daughter cells of the small micromeres become part of the coelomic sacs, in contrast to the long-held view that these sacs are purely of macromere origin. In addition, after prolonged mitotic quiescence, and following their incorporation into the coelomic sacs, these cells resume dividing, contrary to the previous view that they do not divide. Since coelomic sac cells give rise to much of the adult urchin, our results indicate that the small micromeres are founders of cell lineages involved in the formation of adult tissues. The setting aside of these cells in a nondividing state may be analogous to a phenomenon in Drosophila development, in which primordial imaginal and germ cells divide approximately once after the blastoderm stage and do not resume dividing until the larval stage.

The first of these two studies examined the relationship between adolescents' scores on a social desirability questionnaire and scores of negative, positive, and total life events. Only number of positive events was significantly related to social desirability. The second study compared various scoring strategies with respect to their intercorrelation and their ability to predict adolescents' maladjustment (depression, anxiety, and number of missed school days). The results demonstrated that (a) total number of events and readjustment-weighted life change scores were equally predictive of the maladjustment criteria; (b) negative events, but not positive events, however scored, were significantly related to the maladjustment criteria; (c) indices based on psychologist-judges' ratings of event desirability were not more predictive of the maladjustment criteria than were indices based on the adolescents' self-reports; and (d) uncontrollable negative events and controllable negative events were equally predictive of the maladjustment criteria.

College undergraduates classified as high (n = 25) and low (n = 25) on recent life stress participated in an experiment involving a novel laboratory stressor. Heart rate and pulse arrival time (PAT) were measured during baseline, anticipation, testing, and recovery periods of the experiment. The results did not replicate those obtained by Pardine and Napoli in that high and low life stress subjects did not show differential physiological reactions. In addition, regression analyses failed to demonstrate that physiological reactivity moderated the relationship between life stress and subsequent self-reported psychiatric or physical health symptomatology. The present findings demonstrated neither the stress-buffering effects of physiological reactivity nor a relationship between life stress and reactivity when the latter was conceptualized as an outcome.

Bicarbonate is transferred across the serosal (S) membrane of the epithelial cells of the turtle bladder in two directions. Cellular HCO3- generated behind the H+ pump moves this membrane into the serosal solution. This efflux of HCO3- is inhibited by SITS (4-isothiocyano-4'-acetamido-2,2'-disulfonic stilbene). When HCO3- is added to the serosal solution it is transported across the epithelium in exchange for absorbed Cl-. This secretory HCO3- flow traverses the serosal cell membrane in the opposite direction. In this study the effects of serosal addition of 5 x 10(-4) M SITS on HCO3- secretion and Cl- absorption were examined. The rate of H+ secretion was brought to zero by an opposing pH gradient, and 20 mM HCO3- was added to S. HCO3- secretion, measured by pH stat titration, was equivalent to the increase in M leads to S Cl- flux after HCO3- addition. Neither the S leads to M flux of HCO3- nor the M leads to S flux of Cl- were affected by SITS. In the absence of electrochemical gradients, net Cl- absorption was observed only in the presence of HCO3- in the media; under such conditions, unidirectional and net fluxes of Cl- were not altered by serosal or mucosal SITS. H+ secretion, however, measured simultaneously as the short-circuit current in ouabain-treated bladders decreased markedly after serosal SITS. The inhibition of the efflux of HCO3- in series with the H+ pump and the failure of SITS to affect HCO3- secretion and Cl- absorption suggest that the epithelium contains at least two types of transport systems for bicarbonate in the serosal membrane.

From previous studies on cloned yeast ribosomal protein genes we obtained evidence that a large number of them contain an intron [Bollen et al. (1982) Gene 18, 29-38]. In the temperature-sensitive rna2-mutant transcription of these genes leads to the accumulation of precursor RNAs at the restrictive temperature. These precursor mRNAs are several hundreds of nucleotides longer than the respective mature mRNAs. The split character of one of these ribosomal protein genes, viz. the gene coding for the major phosphorylated small-subunit protein S10, was further established by sequence analysis. The intervening sequence interrupts the coding sequence after the second codon and has a length of 352 nucleotides. Genomic Southern hybridizations with a DNA fragment carrying part of the S10-gene revealed that this gene is duplicated on the yeast genome. The molecular weight of S10 as deduced from the sequence analysis was estimated to be 31462 dal. Comparison of the N-terminal aminoacid sequence of the yeast ribosomal protein S10 with that of ribosomal protein S6 from rat liver revealed a striking homology between both proteins. Moreover, at the C-terminal end of the yeast ribosomal protein the sequence Arg-Ala-Ser-Ser-Leu-Lys is present which is very similar to the phosphorylation site of the rat liver protein S6.

Streptokinase was injected directly into the arterial tree following balloon-catheter embolectomy on 13 occasions to remove residual thrombus that could not be mechanically retrieved in 12 patients with imminent limb (ten patients) or kidney (two patients) necrosis. Effective lysis, confirmed by arteriography, pulse return, and increased ankle pressures, was achieved in 11 trials (85%). Bleeding complications, minor in three patients and severe in two patients, were ascribed to systemic lysis although other factors were contributory. One of five deaths was related to therapy. Six limbs were salvaged. The average total dose of streptokinase used, 110,000 units, was given in intermittent boluses of 25,000 to 50,000 units injected below a clamp placed to temporarily occlude distal circulation. Safe application of this technique requires intraoperative monitoring of coagulation parameters, aggressive replacement therapy, and prudent patient selection. This preliminary experience suggests that intraoperative lytic therapy (1) is an effective method for clearing thrombus not amenable to mechanical extraction and (2) may improve patency and tissue salvage.

This study tested the relation between mood (depressed [D], elated [E], or neutral [N]), induced by the Velten (1968) procedure, and college students' responses on a subjectively scored life events questionnaire and measures of perceived and received social support. A manipulation check showed that the mood manipulation was successful. There was a significant mood effect on the number of self-reported negative life events, with E subjects reporting the fewest. However, mood had no significant effect on the number of self-reported positive life events or the rated intensity of negative and positive events. Mood had a significant effect on perceived social support, with D subjects scoring the lowest. Self-report of received social support, however, was not affected by the mood manipulation. The findings challenge the widespread use of life event and perceived social support questionnaires whose independence from a mood-related response bias has not been adequately demonstrated. The findings also challenge causal interpretation of significant effects for self-reported life stress and perceived social support obtained in cross-sectional prediction studies of concurrent psychological distress.

Used a longitudinal design to test the main and stress-buffering effects of instrumentality of measures of trait anxiety, depression, and self-esteem. Ss were early (junior high school) and middle (senior high school) adolescent boys and girls. The hypothesis that instrumentality would be negatively related to psychological distress was supported by the cross-sectional and longitudinal regression analyses of every subgroup of Ss. The hypothesized stress-buffering effect of instrumentality was found in the cross-sectional and longitudinal analyses of senior high girls, specifically in the context of interpersonal negative events (relationship stress). The analyses for junior high girls revealed that instrumentality was beneficial only at a low level of relationship stress. There were no significant interaction effects in the regression analyses of boys. The findings suggest that, when experiencing relationship stress, instrumentality is helpful for middle adolescent girls but not for early adolescent girls. Therefore, future research on adolescents' life-stress adjustment should consider the roles of gender, age, personality, and type of life stress.

From a colony bank of HindIII-generated yeast DNA fragments we have isolated a number of recombinant DNAs carrying genes for ribosomal proteins (e.g., S10, S16A, S20, S24, S31, S33, L16, L25 and L34) of the yeast Saccharomyces carlsbergensis. By electron microscopic analysis of the R-loops formed between various DNA fragments and yeast mRNA, we could locate the ribosomal protein genes on the physical maps of the cloned DNA fragments. The R-loop structures observed indicate that a number of the ribosomal protein genes contain an intervening sequence.

The histone 1 (H1) fraction of HeLa S-3 cells contains two principal subtypes, H1A (Mr approximately 21 000) and H1B (Mr approximately 22 000). In G1 cells, the H1 molecules are distributed among several phosphorylation states, most H1A molecules containing 0 or 1 phosphate groups and most H1B molecules containing 0, 1, 2, or 3 phosphate groups. Both subtypes undergo a general increase in phosphorylation levels of approximately 1 P/mol during the S phase and a further increase or 3--4 P/mol during mitosis. These two increases affect most of the H1 molecules and thus reflect phosphorylations occurring widely throughout the chromatin, presumably in association with replication and mitotic chromosome condensation. During all these periods, multiple phosphorylation levels of H1 molecules persist, as does the phosphorylation differential between H1A and H1B. Thus, there appear to be phosphorylation states that only some of the H1 molecules occupy, a fact that may be related to the conformational diversity in interphase and mitotic chromatin. The existence of differences between H1A and H1B phosphorylation states throughout the cell cycle, and within a single cell type, is in accord with the hypothesis that the H1 subtypes are functionally distinct, such that subtype-specific phosphorylations contribute to the control of chromatin organization.

At least two mechanisms generate tissue differences in the histone subtype composition during development: subtype dilution and subtype replacement. Subtype dilution, which occurs when cells continue dividing after having ceased to synthesize one more histone subtypes, allows the elimination of stable subtypes. It is the major mechanism generating cell differences in histone composition in sea urchin embryogenesis. Subtype replacement has been observed in mammalian tissues, both in the intact animal and in cultured cells. It is most evident in nondividing cells but occurs to some extent in dividing cells as well. Examples of the two mechanisms are presented and their possible biological significance, as well as that of the differences they produce, is discussed.

We have used immunofluorescent staining of sea urchin embryos to study how histone H1 alpha is distributed among progeny cells formed after the cessation of its synthesis. Our results are consistent with H1 alpha being distributed to both daughter cells at mitosis, resulting in it being most concentrated in cells that stop dividing shortly after H1 alpha synthesis ends, while cells that continue to divide dilute their H1 alpha content in proportion to the number of cell divisions. This rules out our earlier suggestion that H1 alpha becomes segregated in dividing cells. In addition, our results show that most dividing cells of the 3-day embryo contain predominantly H1 beta and H1 gamma. Since these subtypes are known not to undergo phosphorylation, this finding has implications regarding the roles of H1 phosphorylation in the cell cycle.

Methods for the mass isolation of diverse organs from small animals are described. They involve novel devices: a mechanical dissecting system, a centrifugal agitator for the separation of fibrillar from globular particles, and a settling chamber for the fractionation at unit gravity of particles with sedimentation velocities above the useful range for centrifugation. The application of these methods to the isolation of polytene and nonpolytene nuclei from Drosophila melanogaster larvae is described.