Elizabeth S. Maxwell

Genomic Survey of Bipolar Illness in the NIMH Genetics Initiative Pedigrees: A Preliminary Report

NIMH Genetics Initiative Bipolar Group: John I. Nurnberger, Jr.* (Chair), J. Raymond DePaulo,Elliot S. Gershon, Theodore Reich, Mary C. Blehar, and collaborators from Indiana University(Howard J. Edenberg, Tatiana Foroud, Marvin Miller, Elizabeth Bowman, Aimee Mayeda, N. LeelaRau, Carrie Smiley, and P. Michael Conneally), Johns Hopkins University (Francis Mc-Mahon, Deborah Meyers, Sylvia Simpson, Melvin McInnis, and O. Colin Stine), NIMH IntramuralResearch Program (Sevilla Detera-Wadleigh, Lynn Goldin, Juliet Guroff, Elizabeth Max-well, Diane Kazuba, Pablo V. Gejman, Judith Badner, and Alan Sanders), and WashingtonUniversity of St. Louis (John Rice, Laura Bierut, and Alison Goate).Four sites collaborated with the NIMH todevelop a resource for the genetic study ofbipolar (BP) illness. Common methods of as-certainment and assessment were devel-oped in 1989. A series of families with a bi-polar I (BPI) proband and at least one BPIor schizoaffective, bipolar type (SA/BP)first-degree relative has been studied. Wenow report initial data from a genomic sur-vey with an average intermarker interval of10 cM on 540 subjects from 97 families. Thisis the largest commonly ascertained and as-sessed linkage sample for bipolar illness re-ported to date; it includes 232 subjects withBPI, 32 SA/BP, 72 bipolar II (BPII), and 88unipolar, recurrent (UPR). Nonparametricmethods of analysis were employed, with allsites using affected sib pair analysis. Thestrongest findings thus far appear to be onchromosomes 1, 6, 7, 10, 16, and 22. Supporthas also been found for some previously re-ported linkages, including 21q and possiblyXq26. All these areas (as well as others) willbe followed up with additional markers andfurther analyses. No locus tested thus farmeets stringent criteria for an initial find-ing of significant linkage. Am. J. Med. Genet.74:227–237, 1997.

Family-based association of FKBP5 in bipolar disorder.

The FKBP5 gene product forms part of a complex with the glucocorticoid receptor and can modulate cortisol-binding affinity. Variations in the gene have been associated with increased recurrence of depression and with rapid response to antidepressant treatment. We sought to determine whether common FKBP5 variants confer risk for bipolar disorder. We genotyped seven tag single-nucleotide polymorphisms (SNPs) in FKBP5, plus two SNPs previously associated with illness, in 317 families with 554 bipolar offspring, derived primarily from two studies. Single marker and haplotypic analyses were carried out with FBAT and EATDT employing the standard bipolar phenotype. Association analyses were also conducted using 11 disease-related variables as covariates. Under an additive genetic model, rs4713902 showed significant overtransmission of the major allele (P=0.0001), which was consistent across the two sample sets (P=0.004 and 0.006). rs7757037 showed evidence of association that was strongest under the dominant model (P=0.001). This result was consistent across the two datasets (P=0.017 and 0.019). The dominant model yielded modest evidence for association (P<0.05) for three additional markers. Covariate-based analyses suggested that genetic variation within FKBP5 may influence attempted suicide and number of depressive episodes in bipolar subjects. Our results are consistent with the well-established relationship between the hypothalamic–pituitary–adrenal (HPA) axis, which mediates the stress response through regulation of cortisol, and mood disorders. Ongoing whole-genome association studies in bipolar disorder and major depression should further clarify the role of FKBP5 and other HPA genes in these illnesses.

Nucleoside monophosphate kinases: I. Transphosphorylation between adenosine triphosphate and nucleoside monophosphates

Abstract 1. 1. The nucleoside monophosphate kinase catalyzing the ATP-UMP reaction has been purified 40–45-fold from an extract of calf liver acetone powder. The ATP-CMP kinase followed the fractionation closely but the ATP-AMP kinase was partially removed while the ATP-GMP kinase and the kinases for reactions between nucleoside triphosphate and AMP were completely removed. 2. 2. Some of the properties of this ATP-nucleoside monophosphate kinase preparation are presented.

Yeast uridine diphosphogalactose-4-epimerase, correlation between activity and fluorescence

Abstract Yeast grown on galactose contains a strikingly fluorescent protein in the fractions which contain UDP-galactose-4-epimerase. UDP-galactose-4-epimerase from yeast, unlike the corresponding enzyme from liver, does not require addition of diphosphopyridine nucleotide for activity. Further purification for the yeast enzyme gives a close correlation between enzyme activity and fluorescence. Addition of p -chloromercuribenzoate brings about a disappearance of fluorescence as well as activity. The latter can be restored by addition of diphosphopyridine nucleotide. A discussion of possible mechanisms of the enzymic interconversion poses a number of new problems.

Nucleoside monophosphate kinases: II. Transphosphorylation between adenosine monophosphate and nucleoside triphosphates

Abstract 1. 1. A nucleoside monophosphate kinase has been fractionated from aqueous extracts of calf liver acetone powder. It catalyzes transphosphorylations between adenosine 5′-phosphate and any nucleoside triphosphate to yield adenosine 5′-diphosphate and another nucleoside diphosphate. The reactions are reversible. 2. 2. The enzyme fraction has been freed of phosphatase activity and contains no nucleoside diphosphate kinase. It has also been separated from another nucleoside monophosphate kinase which is specific for ATP but reacts with any of several nucleoside monophosphates.

[20] Enzymes of the Leloir pathway

Publisher Summary This chapter focuses on the enzymes of the leloir pathway, and describes the quantitative determination of enzymatic activity in broken cell preparations. Specific enzymatic assays designed especially for use with crude bacterial extracts are described. Galactokinase catalyzes the reaction ATP + Gal→ ADP +α -Gal-l-P. The Gal-1-P produced in a preincubation mixture is quantitatively determined. The Gal-l-P formed is determined in the proteinfree filtrate by an analytical incubation mixture. The absolute molar amount of TPNH formed is a direct measure of Gal-l-P and is thus a measure of galacto-kinase activity. The rate of G-1-P formation in Gal-l-P uridyl transferase is analyzed by adding the two indicator enzymes and coenzymes in excess, phosphoglueomutase (plus glucose-l,6-diphosphate) and G-6-P dehydrogenase plus TPN. In UDPGal-4-epimerase, UDPG formed is determined in a protein-free filtrate by means of a DPN-dependent specific dehydrogenase (UDPG dehydrogenase). The chapter also describes the purification of galactose-l-phosphate uridyl transferase, and purification of UDPgalactose-4-epimerase.

[143A] Determination of UDPG and UTP by means of UDPG dehydrogenase

Publisher Summary UDPG can be oxidized to UDP-glucuronic acid (UDPGA) in the presence of DPN and the enzyme UDPG dehydrogenase. Two moles of DPN is reduced per mole of UDPG oxidized. 1 The enzyme is highly specific is stable, and can be purified to a rather high degree. Enzymatic determination of UTP is based on the principle that UTP, in the presence of yeast UDPG pyrophosphorylase (PP-uridyl transferase), can react with α-glucose-1-phosphate to give UDPG. If DPN and UDPG dehydrogenase are also added to the digest, the transformation of UTP to UDPGlycosyl compound (UDP-glucuronic acid, UDPGA) is carried to completion. Moreover the reaction can be followed spectrophotometrieally at 340 mμ, owing to the accompanying DPN reduction. If glucose-1-phosphate is in excess, the amount of UDPG formed will be governed by the amount of UTP present. ATP is practically inactive in this reaction.

Helix formation between polyribonucleotides and purine nucleosides: III. The effects of purine nucleosides in cell-free amino acid-incorporating systems

The preceding paper in this series demonstrated that 2-aminoadenosine and adenosine form regular ordered structures with poly U or poly UC. The extent of interaction is dependent upon temperature and upon the concentration of the reactants. In the present paper we report that complementary ribosides influence the capacity of poly U or poly UC to direct amino acids into protein in cell-free systems from rat liver and Escherichia coli. Polyphenylalanine synthesis directed by poly U is inhibited in both systems. The incorporation of amino acids directed by poly UC is inhibited by 2-aminoadenosine in the system from rat liver but is stimulated in the system from E. coli. Both the inhibition and the enhancement of the capacity of the polymer to serve as messenger BNA depend upon temperature and upon the concentration of riboside, suggesting that both effects are due to the formation of ordered structure. 2-Aminoadenosine brings about a small but temperature-dependent inhibition of the incorporation of amino acids directed by natural endogenous messenger KNA in rat liver polysomes. 2-Aminoadenosine decreases the rate of degradation of poly U in the complete incorporating system from liver. The nucleoside also interferes with the binding of poly U to ribosomes. These opposing influences offer possible explanations for the observed effects of complementary nucleosides on polyribonucleotide-directed incorporation of amino acid into protein.

Some Steroid Hormone-like Effects of Menthol

Menthol or menthone, or both, like progesterone, have been shown to have the following biological activities: (i) an inhibitory action on liver and kidney aldehyde dehydrogenase activity which, under certain circumstances, is reflected in an increased rate of oxidation of D-galactose, and (ii) a stimulatory effect on the oxidation of D-galactose by two prepubertal congenitally galactosemic subjects.

A genomic survey of bipolar illness in the NIMH genetics initiative pedigrees: a preliminary report

Four sites collaborated with the NIMH to develop a resource for the genetic study of bipolar (BP) illness. Common methods of ascertainment and assessment were developed in 1989. A series of families with a bipolar I (BPI) proband and at least one BPI or schizoaffective, bipolar type (SA/BP) first-degree relative has been studied. We now report initial data from a genomic survey with an average intermarker interval of 10 cM on 540 subjects from 97 families. This is the largest commonly ascertained and assessed linkage sample for bipolar illness reported to date; it includes 232 subjects with BPI, 32 SA/BP, 72 bipolar II (BPII), and 88 unipolar, recurrent (UPR). Nonparametric methods of analysis were employed, with all sites using affected sib pair analysis. The strongest findings thus far appear to be on chromosomes 1, 6, 7, 10, 16, and 22. Support has also been found for some previously reported linkages, including 21q and possibly Xq26. All these areas (as well as others) will be followed up with additional markers and further analyses. No locus tested thus far meets stringent criteria for an initial finding of significant linkage.