Formation of proteasome-PA700 complexes directly correlates with activation of peptidase activity.

The proteolytic activity of the eukaryotic 20S proteasome is stimulated by a multisubunit activator, PA700, which forms both 1:1 and 2:1 complexes with the proteasome. Formation of the complexes is enhanced by an additional protein assembly called modulator, which also stimulates the enzymatic activity of the proteasome only in the presence of PA700. Here we show that the binding of PA700 to the proteasome is cooperative, as is the activation of the proteasome's intrinsic peptidase activity.

Psychometric properties of the grade 4 reading level Multiple Affect Adjective Check List-Revised with offenders.

The reliability and validity of the Grade 4 reading level Multiple Affect Adjective Check List was assessed with offenders in four settings within the criminal justice system. With the exception of the Sensation Seeking Scale, the MAACL-R4 scales showed good internal consistency and test-retest reliability and adequate convergent and discriminant validity, but not for the Depression scale for 53 female arrestees and the Hostility scale with 51 male arrestees. Correlations with self-ratings of health and stress and with the Family Environment scales were in expected directions.

Validation of a self-report measure of somatic health.

The primary purpose was to assess the validity of a new self-report inventory of general somatic health status. In addition, we sought to examine the influence of social desirability and negative affectivity on health reports and to learn whether asking respondents to report concrete, memorable aspects of illness episodes such as seeing a doctor or staying home in bed for all or most of a day would enhance validity. The somatic component of the Cornell Medical Index was used as a criterion measure of health status because that Index has been shown to reflect medical records with great accuracy.

Amide modes of the alpha-helix: Raman spectroscopy of filamentous virus fd containing peptide 13C and 2H labels in coat protein subunits.

The filamentous virus fd consists of a single-stranded DNA genome sheathed by 2700 copies of a 50-residue alpha-helical subunit (protein pVIII) and serves as a model assembly of alpha-helices. To advance vibrational assignments for the alpha-helix, we have investigated Raman spectra of fd virions containing 13C and 2H (deuterium) labels at various main-chain sites of the pVIII subunits. 13C was introduced at specific peptide carbonyls, while deuterium was introduced at selected alpha-carbon (Calpha) and amide nitrogen positions.

Construction of validity scales for the Multiple Affect Adjective Check List-Revised.

Three scales, the Fake Bad Scale, the Fake Good Scale, and the Fake Bad-Fake Good scales were developed and evaluated with respect to their capacity to detect response manipulation on the Multiple Affect Adjective Check List-Revised. Cutting scores for each scale were cross-validated in two samples consisting of three groups: (1) college students simulating either "fake good" or "fake bad," (2) college students under standard instructions, and (3) psychiatric patients. Cutting scores on the three scales were compared with cutting scores established for the MAACL-R Dysphoria and Positive Affect plus Sensation Seeking.

Structural basis of DNA recognition and mechanism of quadruplex formation by the beta subunit of the Oxytricha telomere binding protein.

Interactions of the beta subunit of the Oxytricha nova telomere binding protein with the telomeric DNA sequences, d(T4G4)2 and dT6(T4G4)2, have been investigated in vitro using Raman and fluorescence spectroscopies. Raman difference spectra show that the beta subunit binds to both d(T4G4)2 and dT6(T4G4)2 but promotes the formation of a parallel-stranded quadruplex only in dT6(T4G4)2, thus demonstrating the importance of the telomeric 5' tail for in vitro recognition and guanine quadruplex formation. While d(T4G4)2 is not a suitable substrate for quadruplex promotion by the beta subunit, the Raman spectra reveal other structural rearrangements of this DNA strand upon beta subunit binding, including changes in guanine glycosyl torsion angles from syn to anti and disruption of carbonyl hydrogen-bonding interactions.

Risk adjustment of nursing home quality indicators.

The purpose of this study was to develop a method for risk adjusting nursing home quality indicators (QI's). The QI's measure incidence and prevalence of resident-level care processes and outcomes which are indicative of care quality. Risk adjustment was carried out by stratifying residents into risk groups (high and low), calculating QI rates within groups, and then drawing comparisons across facilities.

Dopamine D2 receptor stimulation alters G-protein expression in rat pituitary intermediate lobe melanotropes.

Stimulation of dopamine D2 receptors inhibits melanotrope pro-opiomelanocortin (POMC) biosynthesis and alpha-melanocyte-stimulating hormone (MSH) secretion. These effects are mediated by G-protein alpha i- and alpha o-subunits and are reversed by stimulating receptors linked to activation of G alpha s protein. Melanotrope activity is increased by haloperidol, a D2 receptor antagonist, and decreased by bromocriptine, a D2 receptor agonist.

Writing Amish culture into genes: biological reductionism in a study of manic depression.

Critical realism is used to explore the problem of reductionism in a classic (the Amish Study) and widely-cited study of manic depression. Along with related ideas drawn from the works of R.C.

Spfkh1 encodes a transcription factor implicated in gut formation during sea urchin development.

A member of the forkhead class of transcription factors from sea urchins (Spfkh1) that is expressed specifically in the endoderm of developing embryos has been identified. Spfkh1 was expressed transiently in the embryo, with peak levels of messenger ribonucleic acid (mRNA) accumulating at the time endoderm invaginated into the interior of the embryo. Expression was limited to the invaginating endoderm in the early gastrula, then became further restricted to the base of the invaginating gut at the mid-gastrula stage.

Role of heme-hemopexin in human T-lymphocyte proliferation.

Heme-hemopexin supports and stimulates proliferation of human acute T-lymphoblastic (MOLT-3) cells, suggesting the participation of heme in cell growth and division. MOLT-3 cells express approximately 58,000 hemopexin receptors per cell (apparent Kd 20 nM), of which about 20% are on the cell surface. Binding is dose- and temperature-dependent, and growth in serum-free IMDM medium is stimulated by 100-1000 nM heme-hemopexin, consistent with the high affinity of the receptor for hemopexin, and maximal growth is seen in response to 500 nM complex.

Synthesis and beta-adrenergic activities of R-fluoronaphthyloxypropanolamine.

Purpose: Many biogenic amines where an aromatic proton is substituted with fluorine have exhibited pharmacological properties that are dependent on the position of fluorine on the aromatic ring. For example, 6-fluoroepinephrine is selective for alpha-adrenergic receptors whereas the 2-fluoroisomer is selective for beta-receptors. Aryloxypropanolamines are beta-receptor agonists or antagonists, depending on the aryl group and its substituents.

GABAA receptor subunit expression changes in the rat cerebellum and cerebral cortex during aging.

Significant aging-related decreased expression of various GABAAR subunit mRNAs (alpha 1, gamma 2, beta 2, beta 3 and sigma) was found in both cerebellum and cerebral cortex using quantitative dot blot and in situ hybridization techniques. Contrary to the other subunits, the alpha 6 mRNA expression was significantly increased in the aged cerebellum. Parallel age-related changes in protein expression for gamma 2 and beta 2/3 (decrease) and alpha 6 (increase) were revealed in cerebellum by quantitative immunocytochemistry.

The effect of pravastatin on hepatic 3-hydroxy-3-methylglutaryl CoA reductase obtained from poloxamer 407-induced hyperlipidemic rats.

A single 300-mg intraperitoneal injection of poloxamer 407 (P-407) to rats produces a marked hypercholesterolemia for a minimum of 96 hours and increases the activity of hepatic 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase compared with the enzyme activity found in microsomal homogenates of control animals. We attempted to determine whether inhibition of microsomal HMG-CoA reductase by pravastatin sodium would yield similar values for the maximum reaction in velocity (Vmax) and the HMG-CoA reductase-pravastatin dissociation constant (Ki) when the enzyme was in the activated state compared with the control state. Knowledge of the respective values for Vmax and Ki would allow us to determine whether P-407-induced hypercholesterolemia in the rat was refractory to pravastatin treatment.

A developmental perspective on family risk factors in somatization.

It is widely believed that the family plays an etiologically significant role in somatization. Although research indicates many high symptom reporters come from dysfunctional homes, very little is known about the more specific aspects of daily family functioning that may heighten the risk of physical symptomatology at different developmental levels. Accordingly, we report findings on the relationship between ten dimensions of family-of-origin environment and physical health complaints in 933 children, adolescents, and young adults in junior high, high, and undergraduate school (grades 6-16).

Rural mental health and aging: implication for case management.

Older adults are underrepresented as recipients of mental health services, despite evidence that preventive interventions are effective and complement physical health interventions. Barriers to using these services include the stigma associated with mental health problems, lack of trained professionals and general barriers of availability and accessibility in rural service delivery. Case managers can play an important role in breaking down some of these barriers and assisting older adults to access t-ese services.

Growth factor control of cultured rat uterine stromal cell proliferation is progesterone dependent.

Uterine stromal cells undergo mitosis and differentiate into the decidua just prior to the expected time of implantation in humans and rodents. We have utilized a culture system that will be suitable for study of the molecular mechanisms regulating stromal cell proliferation. Stromal cells were isolated from the uteri of ovariectomized rats and were cultured in chemically defined medium.

Vibrational analysis of nucleic acids. IV. Normal modes of the DNA phosphodiester structure modeled by diethyl phosphate.

Raman and ir spectra are reported for diethyl phosphate [(CH3CH2O)2PO2-] and diethyl phosphate isotopomers incorporating carbon-13 at methylene group sites [(CH313CH2O)2PO2-] and deuterium substituents on methyl and methylene carbons [(CH3CD2O)2PO2-, (CD3CH2O)2PO2-, (CD3CD2O)2PO2-]. The vibrational spectra are analyzed to develop a consistent set of assignments for the C-C-O-P(O2-)-O-C-C network, which serves as a model for the nucleic acid phosphodiester backbone. The present study resolves previously conflicting vibrational assignments for the phosphodiester skeleton and provides a firm empirical basis for interpreting conformationally sensitive modes of DNA and RNA.

Surface spectroscopy of apatitic materials: limitations and concerns.

The use of x-ray photoelectron spectroscopy for the study of biological materials has provided mixed results. The experimental difficulties associated with the application of this technique to the study of biological materials include the effects of ultra-high vacuum, x-ray exposure, surface charging, and, for insulating materials, the inherent lack of an intrinsic energy reference. Since most biological materials are insulators, the lack of an intrinsic energy reference complicates one's ability to draw direct comparisons between different studies.

Vibrational analysis of nucleic acids. V. Force field and conformation-dependent modes of the phosphodiester backbone modeled by diethyl phosphate.

A generalized valence force field is derived for the diethyl phosphate anion [(CH3CH2O)2PO2-] and its deuterium [(CH3CD2O)2PO2-, (CD3CH2O)2PO2- and (CD3CD2O)2PO2-] and carbon-13 [(CH3 13CH2O)2PO2-] derivatives in the stable trans-gauche-gauche-trans conformation. Normal coordinate analysis of the trans-gauche-gauche-trans conformer, which serves as a structural analog of the nucleic acid phosphodiester group, is based on comprehensive infrared and Raman spectroscopic data and vibrational assignments obtained for the diethyl phosphate anion. The generalized valence force field is in good agreement with the scaled ab initio force field of diethyl phosphate and represents significant improvement over earlier modeling of the phosphodiester moiety with dimethyl phosphate.

Aging-related subunit expression changes of the GABAA receptor in the rat hippocampus.

Aging-related changes in the subunit expression of some hippocampal GABAA receptors have been found. Quantitative in situ hybridization has revealed that alpha 1, subunit messenger RNA expression was significantly increased in the hippocampus (34%) of old rats. The largest increases were observed in the dentate gyrus (76%) and in the CA1 field (30%).

Heme binding by a bacterial repressor protein, the gene product of the ferric uptake regulation (fur) gene of Escherichia coli.

The fur gene product, Fur, of Escherichia coli is a repressor when it binds Fe(II). Since heme and iron metabolism are closely linked and Fur is rich in histidine, a ligand for heme, the binding of heme to Fur was investigated. The oxidized Fur-heme complex is stable and low spin with a Soret maximum at 404 nm and no 620-nm band.

Raman spectroscopy of the Ff gene V protein and complexes with poly(dA): nonspecific DNA recognition and binding.

Raman spectra of crystals and solutions of the single-stranded DNA binding protein of bacteriophage Ff (gene V protein, gVp) and of solution complexes of gVp with single-stranded poly-(deoxyadenylic acid) [poly(dA)] reveal the following: (i) The gVp secondary and tertiary structures are similar in solution and in the crystal and are dominated by beta-sheet domains, in agreement with NMR and X-ray findings. (ii) Subunit conformation and side chain environments of gVp are virtually unchanged over a wide range of salt concentration (0 < [NaCl] < 100 mM); however, the solution conformation of poly(dA) exhibits sensitivity to added salt. The perturbed Raman markers indicate subtle changes in helix backbone geometry with accompanying small differences in base stacking as the concentration of NaCl is changed.

Raman signature of the four-stranded intercalated cytosine motif in crystal and solution structures of DNA deoxycytidylates d(CCCT) and d(C8).

The Raman spectral signature of the four-stranded cytosine structure formed by intercalation of two hemiprotonated and parallel-stranded oligodeoxycytidylate duplexes (so-called i motif) has been obtained from the crystal structure of d(CCCT) [Kang, C.H., Berger, I.

Structural transitions in the scaffolding and coat proteins of P22 virus during assembly and disassembly.

An in vitro system for investigating the assembly of the Salmonella phage P22 has been exploited to elucidate the structural basis of recognition between scaffolding protein (gp8) and coat protein (gp5) subunits of the viral procapsid. Raman spectroscopy and circular dichroism have been employed to examine structural thermostabilities of both gp8 and gp5 in native procapsids, and to characterize structural changes accompanying scaffolding exit, procapsid expansion, and shell disassembly. It is found that the secondary structure of the isolated gp8 subunit is rich in alpha-helix (approximately 40%), is highly thermolabile, and is characterized by noncooperative unfolding (Tm approximately 49 degrees C).