Charles A. Marotta Publication List (Medline designation where available) *****AMERICAN JOURNAL OF ORTHOPSYCHIATRY***** (REFERENCE 1 OF 100) 89226043 Marotta CA Molecular biology in psychiatric research: Alzheimer's disease as a paradigm. In: Am J Orthopsychiatry (1989 Apr) 59(2):294-302 ISSN: 0002-9432 As illustration of the integration of molecular biological studies into psychiatric research, use of molecular techniques in the study of Alzheimer's disease is described, and the resultant observation of otherwise undetectable pathological changes is noted. Using postmortem Alzheimer brain messenger RNA, recombinant DNA studies led to the ability to clone and characterize the expressed genetic transcript for amyloid. Application of this methodology is discussed in terms of establishing cellular and animal models for a neuropsychiatric disease. Registry Numbers: 63231-63-0(RNA) Registry Numbers: 9007-49-2(DNA) Institutional address: Department of Psychiatry and Neuroscience Program Harvard Medical School. *****ANNALS OF THE NEW YORK ACADEMY OF SCIENCES***** (REFERENCE 2 OF 100) 97470278 Friedland RP, Kalaria R, Berridge M, Miraldi F, Hedera P, Reno J, Lyle L, Marotta CA Neuroimaging of vessel amyloid in Alzheimer's disease. In: Ann N Y Acad Sci (1997 Sep 26) 826:242-7 ISSN: 0077-8923 Despite extensive recent advances in understanding Alzheimer's disease (AD) we are unable to noninvasively establish a definite diagnosis during life and cannot monitor the cerebral deposition of amyloid beta protein (A beta) in living patients. We evaluated the use of 10H3, a monoclonal antibody Fab targeting A beta protein 1-28 labeled with Tc-99m. Six subjects with probable AD were studied using

single-photon emission computed tomography (SPECT) at times from 0-24 hours following injection. Curves of radioactivity in blood demonstrate a half-life of the injected Fab of 2-3 hours. Images show uptake around the head in the scalp or bone marrow in all subjects. There is no evidence of cerebral uptake of the antibody. Scalp biopsies in all six patients demonstrate diffuse staining with 10H3 of the scalp, a pattern indistinguishable from that found in controls. Evidence of amyloid deposition in the scalp in AD is not seen with other anti-A beta antibodies, suggesting that 10H3 is cross- reacting with another protein. Further studies with anti-A beta antibodies will require longer-lived radionuclides to detect cerebral uptake at later times after injection to allow for complete clearance from the blood. Alternately, imaging using labeled A beta itself may provide a means for noninvasive targeting of cerebral amyloid. Institutional address: Department of Neurology Case Western Reserve University School of Medicine Cleveland Ohio 44106 USA. rpf2@po.cwru.edu (REFERENCE 3 OF 100) 75053110 Forget BG, Marotta CA, Weissman SM, Verma IM, McCaffrey RP, Baltimore D Nucleotide sequences of human globin messenger RNA. In: Ann N Y Acad Sci (1974 Nov 29) 241(0):290-309 ISSN: 0077-8923 [No Abstract Available] (REFERENCE 4 OF 100) 75010408 Forget BG, Baltimore D, Benz EJ, Housman D, Lebowitz P, Marotta CA, McCaffrey RP, Skoultchi A, Swerdlow PS, Verma IM, Weissman SM Globin messenger RNA in the thalassemia syndromes. In: Ann N Y Acad Sci (1974) 232(0):76-87 ISSN: 0077-8923 *****ANNALS OF NEUROLOGY***** (REFERENCE 5 OF 100) 82066653 Selkoe DJ, Brown BA, Salazar FJ, Marotta CA Myelin basic protein in Alzheimer disease neuronal fractions and mammalian neurofilament preparations.

In: Ann Neurol (1981 Nov) 10(5):429-36 ISSN: 0364-5134 We previously reported a marked increase of a 20,000 molecular weight (MW) protein, P20, in some neuronal fractions and whole cortical homogenates isolated from affected cortex in Alzheimer disease; P20 comigrated electrophoretically with an unidentified, major 20,000 MW protein present in human neurofilament (NF) fractions. We now report that the 20,000 MW protein is a major constituent of rodent as well as human NF fractions and that it comigrates by one- and two- dimensional gel electrophoresis with purified myelin basic protein (MBP). Peptide mapping and staining with amido black confirmed the identity of the 20,000 MW protein of mammalian NF fractions as MBP. One- and two-dimensional gel electrophoresis of neuronal perikaryal fractions from human cortex indicated that the increased P20 protein in Alzheimer neuronal fractions comigrates with human MBP. Deliberate contamination of cortical samples with adjacent subcortical white matter (i.e., myelin) prior to neuronal separation did not result in an increase of P20 in the neuronal fraction. On the basis of these and additional experiments, we conclude that the increase of a 20,000 MW protein in neuronal fractions and whole homogenates from affected cortex in Alzheimer disease represents MBP of intracortical origin. *****ARCHIVES OF GENERAL PSYCHIATRY***** (REFERENCE 6 OF 100) 88048795 Benes FM, Majocha R, Bird ED, Marotta CA Increased vertical axon numbers in cingulate cortex of schizophrenics. In: Arch Gen Psychiatry (1987 Nov) 44(11):1017-21 ISSN: 0003-990X Data generated from an earlier study have suggested a model in which greater numbers of long, vertical, associative axons may occur in the anterior cingulate cortex of schizophrenic patients relative to control subjects. This hypothesis has now been tested using neuron- specific antibodies raised against the 200-kilodalton neurofilament subunit, a component of neuronal cytoskeleton, to immunostain axons of human postmortem cingulate cortex. A manual method for counting axons in the region of layer II and sublamina IIIA has been designed and applied blindly to parallel control and schizophrenic immunoprocessed specimens. The results show that there are 25% more vertical axons in the schizophrenic than in the control specimens. Preferentially higher numbers of both long vertical axons (62%) and axons associated with blood vessels (52%) have also been noted in the schizophrenic specimens. By contrast, the number of large-caliber horizontal axons was the same in the two groups; therefore, the greater number of vertical axons in schizophrenic specimens does not appear to represent a nonspecific effect. When these data are corrected for the effects of several confounding variables using

analysis of covariance, the overall pattern of the results persists. These findings suggest the possibility that there might be an increase of associative inputs into the anterior cingulate cortex of schizophrenic patients, although it is not clear at present whether the differences noted, if replicative, may be primarily or perhaps only secondarily related to the disorder. Institutional address: Department of Psychiatry Harvard Medical School Boston MA. *****BIOCHEMISTRY***** (REFERENCE 7 OF 100) 73221045 Marotta CA, Levy CC, Weissman SM, Varricchio F Preferred sites of digestion of a ribonuclease from Enterobacter sp. in the sequence analysis of Bacillus stearothermophilus 5S ribonucleic acid. In: Biochemistry (1973 Jul 17) 12(15):2901-4 ISSN: 0006-2960 [No Abstract Available] *****BRAIN RESEARCH***** (REFERENCE 8 OF 100) 96346590 Nagy JI, Li W, Hertzberg EL, Marotta CA Elevated connexin43 immunoreactivity at sites of amyloid plaques in Alzheimer's disease. In: Brain Res (1996 Apr 22) 717(1-2):173-8 ISSN: 0006-8993 The distribution of the astrocytic gap junctional protein, connexin43 (Cx43) was compared immunohistochemically with that of amyloid plaques in Alzheimer's Disease (AD) brain. By light microscopy, cortical areas containing numerous beta/A4 amyloid plaques exhibited increased immunostaining density for Cx43 and some plaques corresponded exactly to sites of intensified Cx43 immunoreactivity. By electron microscopy, Cx43 was localized to astrocytic gap junctions in AD brain. Increased Cx43 expression in AD may represent an attempt to maintain tissue homeostasis by augmented intercellular communication via gap junction formation between astrocytic processes that invest senile plaques, or alternatively, an aberrant induction of astrocytic Cx43 expression which may further compromise homeostasis and exacerbate pathological conditions in the

microenvironment of amyloid plaques. Institutional address: Department of Physiology University of Manitoba Winnipeg Canada. Nagyj@bldghsc.lan1.umanitoba.ca (REFERENCE 9 OF 100) 94036093 Maestre GE, Tate BA, Majocha RE, Marotta CA Membrane surface ruffling in cells that over-express Alzheimer amyloid beta/A4 C-terminal peptide. In: Brain Res (1993 Sep 3) 621(1):145-9 ISSN: 0006-8993 Deposition of beta/A4 amyloid in brain is a defining characteristic of Alzheimer disease (AD); however, the extent to which amyloid deposits may interfere with normal cellular processes is incompletely understood. We examined this issue by means of PC12 cells. After transfection with DNA coding for 97 amino acids of the beta/A4 C- terminal region of the amyloid precursor protein, beta/A4 antigen was visible at the cell membrane. We report that normal unstimulated PC12 cells exhibit ruffling activity at the cell surface when plated on a plastic substrate. Relative to control cells, however, those that over-expressed the beta/A4 C-terminal peptide had significantly higher levels of ruffling activity, suggesting a structural and/or functional membrane modification. Similar cellular alterations, if present, in Alzheimer brain cells, may indicate disturbances in membrane-associated functions, including intercellular communication. Institutional address: Department of Psychiatry Harvard Medical School Boston MA 02115. (REFERENCE 10 OF 100) 93153638 Maestre GE, Tate B, Majocha RE, Marotta CA Cell surface extensions associated with overexpression of Alzheimer beta/A4 amyloid. In: Brain Res (1992 Dec 18) 599(1):64-72 ISSN: 0006-8993 Deposition of beta/A4 amyloid in Alzheimer disease (AD) brain parenchyma and vasculature occurs by mechanisms that are currently undefined. Similarly the potential consequences of amyloid accumulation for disrupting cellular integrity have not been addressed in detail. To investigate the possible significance of amyloid deposits for cellular viability, PC12 cells were permanently

transfected with DNA coding for the beta/A4-C terminal region of the amyloid precursor protein. The DNA represented 97 amino acids of the amyloid precursor protein of which 40 amino acids were derived from the beta/A4 region. Transfected clonal cell lines and controls were examined at both the light and electron microscopic levels for morphological abnormalities. beta/A4 amyloid accumulated in the cell membrane where the peptide was located at cellular processes resembling blebs and microvilli. These specialized structures at the cell surface were over-abundant in transfected cells that overexpressed the beta/A4 peptide but not in controls. Membranous processes may be involved in the delivery of the beta/A4 peptide to the external surface of the cell of origin and release into the extracellular space. Similar surface features of cells in the AD brain, should they occur, may indicate a role for membrane-associated processes in the pathophysiology of the disorder. Registry Numbers: 9007-49-2(DNA) Institutional address: Department of Psychiatry and Neuroscience Program Harvard Medical School Boston MA 02115. (REFERENCE 11 OF 100) 84025604 Nixon RA, Brown BA, Marotta CA Limited proteolytic modification of a neurofilament protein involves a proteinase activated by endogenous levels of calcium. In: Brain Res (1983 Sep 26) 275(2):384-8 ISSN: 0006-8993 Posttranslational modification of a structural protein by limited proteolysis is demonstrated for the first time in the nervous system. The 145,000 dalton subunit of neurofilaments in mouse retinal ganglion cell (RGC) axons is selectively converted in vitro to the major 143,000 and 140,000 dalton neurofilament subunits by a neutral proteinase that is activated by endogenous levels of calcium and is distinguishable from other known brain proteinases. The close similarities between this in vitro process and the previously observed modification of the 145,000 dalton neurofilament protein during axoplasmic transport in vivo suggest that the same enzymatic mechanism is involved. These findings imply that limited proteolysis is an active process along central axons in vivo and that this enzyme may play a specific role in the function of the neuronal cytoskeleton. Registry Numbers: EC 3.4(Peptide Hydrolases) Registry Numbers: 7440-70-2(Calcium) (REFERENCE 12 OF 100) 81185506

Strocchi P, Marotta CA, Bonventre J, Gilbert JM The subunit composition of cerebellar tubulin: evidence for multiple beta tubulin messenger RNAs. In: Brain Res (1981 Apr 27) 211(1):206-10 ISSN: 0006-8993 Cytoplasmic proteins were isolated from adult rat forebrain and cerebellum and analyzed by two-dimensional gel electrophoresis under conditions which the major subunits of tubulin were separated. Forebrain cytoplasmic tubulin consisted of two groups of alpha subunits (alpha 1 and alpha 2) and a minimum of two beta subunits (beta 1 and beta 2). However, the rat cerebellar cytoplasmic proteins contained greatly decreased amounts of the beta 1 tubulin subunit relative to the analysis of forebrain proteins. Messenger RNA (mRNA) was purified from cerebellum and forebrain and translated in wheat germ homogenate. Analysis of the translation products of cerebellar mRNA indicated only a trace amount of the beta 1 subunit, whereas the expected amount of beta 1 was found among the translation products of forebrain mRNA. This data is consistent with the conclusion that the beta 1 and beta 2 subunits of tubulin are synthesized from different mRNAs. A decrease in beta 1 mRNA relative to other tubulin mRNAs may be one of the factors responsible for the low steady state amounts of beta 1 tubulin in the cerebellum. (REFERENCE 13 OF 100) 79210472 Marotta CA, Strocchi P, Gilbert JM Sununit structure of synaptosomal tubulin. In: Brain Res (1979 May 5) 167(1):93-106 ISSN: 0006-8993 The subunit structure of rat brain synaptosomal tubulin was examined by high resolution two-dimensional gel fractionation. Whole brain cytoplasmic tubulin consists of two groups of alpha subunits (alpha1 and alpha2), and a minimum of two beta subunits (beta1 and beta2). Both alpha subunits consist of a major relatively acidic form and minor relatively basic forms. In contrast, tubulin purified from synaptoplasm contains an additional subunit, alpha3, which has the same isoelectric point but slightly faster electrophoretic mobility than alpha1 and alpha2. All synaptosomal alpha subunits are the relatively acidic forms and the minor basic forms are absent. The synaptosomal beta subunits have electrophoretic properties similar to the corresponding cytoplasmic forms. The alpha3 synaptosomal tubulin subunit has affinity for colchicine, has a tryptic peptide map similar to whole brain cytoplasmic alpha tubulin, and can be purified by a standard tubulin purification method. *****BRAIN RESEARCH. MOLECULAR BRAIN RESEARCH***** (REFERENCE 14 OF 100) 89237765

Nixon RA, Lewis SE, Dahl D, Marotta CA, Drager UC Early posttranslational modifications of the three neurofilament subunits in mouse retinal ganglion cells: neuronal sites and time course in relation to subunit polymerization and axonal transport. In: Brain Res Mol Brain Res (1989 Mar) 5(2):93-108 ISSN: 0169-328X We have characterized stages in the posttranslational processing of the three neurofilament subunits, High (NF-H), Middle (NF-M), and Low (NF-L), in retinal ganglion cells in vivo during the interval between synthesis in cell bodies within the retina and appearance of these polypeptides in axons at the level of the optic nerve (optic axons). Neurofilament proteins pulse-labeled by injecting mice intravitreally with [35S]methionine or [32P]orthophosphate, were isolated from Triton-soluble and Triton-insoluble fractions of the retina or optic axons by immunoprecipitation or immunoaffinity chromatography. Within 2 h after [35S]methionine injection, the retina contained neurofilament-immunoreactive radiolabeled proteins with apparent molecular weights of 160, 139, and 70 kDa, which co-migrated with subunits of axonal neurofilaments that were dephosphorylated in vitro with alkaline phosphatase. The two larger polypeptides were not labeled with [32P]orthophosphate, indicating that they were relatively unmodified forms of NF-H and NF-M. About 75% of the subunits were Triton-insoluble by 2 h after isotope injection, and this percentage increased to 98% by 6 h. Labeled neurofilament polypeptides appeared in optic axons as early as 2 h after injection. These subunits exhibited apparent molecular weights of 160, 139, and 70 kDa and were Triton-insoluble. The time of appearance of fully modified polypeptide forms differed for each subunit (2 h for NF-L, 6- 18 h for NF-M, 18-24 h for NF-H) and was preceded by the transient appearance of intermediate forms. The modified radiolabeled subunits in optic axons 3 days after synthesis were heavily labeled with [32P]orthophosphate and exhibited the same apparent molecular weights as subunits of axonal neurofilaments (70 kDa, 145 and 140 kDa, and 195-210 kDa, respectively). Whole mounts of retina immunostained with monoclonal antibodies against NF-H in different states of phosphorylation demonstrated a transition from non-phosphorylated neurofilaments to predominantly phosphorylated ones within a region of the axon between 200 and 1000 microns downstream from the cell body. These experiments demonstrate that the addition of most phosphate groups to NF-M and NF-H takes place within a proximal region of the axon. The rapid appearance of modified forms of NF-L after synthesis may imply that processing of this subunit occurs at least partly in the cell body. The presence of a substantial pool of Triton-insoluble, unmodified subunits early after synthesis indicates that the heaviest incorporation of phosphate occurs after neurofilament proteins are polymerized.(ABSTRACT TRUNCATED AT 400 WORDS) Institutional address: Mailman Research Center McLean Hospital Belmont

MA 02178. *****CANADIAN JOURNAL OF BIOCHEMISTRY AND CELL BIOLOGY***** (REFERENCE 15 OF 100) 86001956 Majocha RE, Marotta CA, Benes FM Immunostaining of neurofilament protein in human postmortem cortex: a sensitive and specific approach to the pattern analysis of human cortical cytoarchitecture. In: Can J Biochem Cell Biol (1985 Jun) 63(6):577-84 ISSN: 0714-7511 Antibodies raised against the 200 000 neurofilament protein (NFP) of rat brain have been successfully applied to the staining of human postmortem cortex using a modified immunoperoxidase procedure. Human anterior cingulate cortex which has been in formaldehyde fixative for as long as 4 years shows extensive and reliable staining of axons and, to a lesser degree, apical dendrites of cortical neurons. The immunostaining of 200 000 NFP in human cortex has revealed cytoarchitectural details not generally visible with other more conventional neuroanatomical stains, particularly when counterstaining with cresyl violet is employed. Potential applications of this approach to the pattern analysis of human cortical cytoarchitecture in both health and disease are discussed. *****CELLULAR AND MOLECULAR NEUROBIOLOGY***** (REFERENCE 16 OF 100) 95346982 Majocha RE, Agrawal S, Tang JY, Humke EW, Marotta CA Modulation of the PC12 cell response to nerve growth factor by antisense oligonucleotide to amyloid precursor protein. In: Cell Mol Neurobiol (1994 Oct) 14(5):425-37 ISSN: 0272-4340 1. The amyloid precursor protein (APP) is widely distributed among eukaryotic cells, however, its precise role in cellular functioning is not fully clarified. APP is glycoprotein membrane constituent and it may facilitate membrane associated functions. 2. The aim of the present study was to examine the possibility that APP may play a role in mediating cellular trophic responses. The methods made use of an antisense oligonucleotide that was prepared to the 5' terminus of APP and shown specifically to reduce the level of APP isoforms. 3. In sequential mixing experiments it was observed that the APP antisense oligonucleotide did not significantly modify the trophic response of PC12 cells pretreated with nerve growth factor (NGF). However, pretreatment of cells with the antisense oligonucleotide diminished NGF-induced increases in cellular size and neurite length. 4. These

observations suggest that APP may play a role in modulating the trophic response. The combined use of APP antisense oligonucleotides and neurotrophic agents may find clinical utility in the treatment of Alzheimer-type dementia since it is known that NGF normally causes increases in APP levels. Institutional address: Department of Psychiatry and Human Behavior Brown University Providence Rhode Island 02912 USA. *****GERIATRICS***** (REFERENCE 17 OF 100) 94291961 Marotta C Alzheimer's disease: how research is changing primary care management [interview by Marc E. Weksler] In: Geriatrics (1994 Jul) 49(7):47-51 ISSN: 0016-867X [No Abstract Available] Registry Numbers: 321-64-2(Tacrine) Institutional address: Cornell University Medical College New York. *****JOURNAL OF THE AMERICAN GERIATRICS SOCIETY***** (REFERENCE 18 OF 100) 88228849 Sajdel-Sulkowska EM, Chou WG, Salim M, Rehman S, Majocha RE, Fulwiler CE, Zain SB, Marotta CA Genetic expression of amyloid and glial-specific protein in the Alzheimer brain. In: J Am Geriatr Soc (1988 Jun) 36(6):558-64 ISSN: 0002-8614 [No Abstract Available] Institutional address: Department of Psychiatry Harvard Medical School Boston Massachusetts.

*****JOURNAL OF BACTERIOLOGY***** (REFERENCE 19 OF 100) 76142407 Varricchio F, Marotta CA Thermal denaturation of mesophilic and thermophilic 5S ribonucleic acids. In: J Bacteriol (1976 Mar) 125(3):850-4 ISSN: 0021-9193 The Tm of Bacillus stearothermophilus 5S ribonucleic acid (RNA) is 1.5 +/- 0.5 C higher than that of 5S RNAs from B. subtilis and Escherichia coli. Melting in 50% methanol and in formaldehyde indicate that both base stacking and helical regions are involved in the slightly increased thermal stability of B. stearothermophilus 5S RNA. It is probable that the 5S RNA makes only a minor contribution to the thermostability of B. stearothermophilus 50S ribosomal subunits. *****JOURNAL OF BIOLOGICAL CHEMISTRY***** (REFERENCE 20 OF 100) 80137532 Forget BG, Marotta CA, Cohen-Solal M, Wilson JT, Weissman SM Partial nucleotide sequence of human alpha globin mRNA derived from analysis of 32P-labeled cRNA and 125I-labeled mRNA; correlation with a globin cDNA sequence. In: J Biol Chem (1980 Apr 10) 255(7):2814-5 ISSN: 0021-9258 [No Abstract Available] (REFERENCE 21 OF 100) 77207124 Marotta CA, Wilson JT, Forget BG, Weissman SM Human beta-globin messenger RNA. III. Nucleotide sequences derived from complementary DNA. In: J Biol Chem (1977 Jul 25) 252(14):5040-53 ISSN: 0021-9258 Sequences of human beta-globin mRNA were determined by analysis of complementary DNA. beta-mRNA was transcribed into double-stranded cDNA by RNA-dependent DNA polymerase. cDNA was cut by restriction endonucleases and the fragments were terminally labeled by means of

polynucleotide kinase and [gamma-32P]ATP. After purification, fragments were degraded by snake venom phosphodiesterase. Alternatively single-stranded [32P]cDNA was prepared by transcription in the presence of [alpha-32P]dCTP and actinomycin D; the product was digested by endonuclease IV and degraded by snake venom phosphodiesterase. cDNA tracts obtained by both labeling methods enabled us to construct a sequence for the translated and 3'-terminal untranslated regions of human beta-mRNA. (REFERENCE 22 OF 100) 77207123 Cohen-Solal M, Forget BG, Prensky W, Marotta CA, Weissman SM Human beta-globin messenger RNA. II. nucleotide sequences derived from 125I-labeled globin messenger RNA. In: J Biol Chem (1977 Jul 25) 252(14):5032-9 ISSN: 0021-9258 [No Abstract Available] Registry Numbers: 9004-22-2(Globins) Registry Numbers: EC 3.1.-(Ribonucleases) (REFERENCE 23 OF 100) 77207122 Marotta CA, Forget BG, Cohne-Solal M, Wilson JT, Weissman SM Human beta-globin messenger RNA. I. Nucleotide sequences derived from complementary RNA. In: J Biol Chem (1977 Jul 25) 252(14):5019-31 ISSN: 0021-9258 Sequence analysis studies were carried out on human beta-globin mRNA (beta-mRNA) prepared from alpha-thalassemic, sickle cell, and Hb A reticulocytes. Highly purified beta-mRNA served as substrate for the preparation of cDNA by RNA-dependent DNA polymerase. The cDNA was transcribed by Escherichia coli RNA polymerase and the resulting cRNA was analyzed. Over 300 nucleotides were assigned to the beta-mRNA coding region and 37 nucleotides were assigned to the 3'-terminal noncoding region. The normal termination codon is UAA which is separated by 28 nucleotides from an out of phase UAA triplet. The origin of each of the abnormally long beta-globin variants Tak and Cranston is consistent with reduplication of dinucleotides prior to the normal termination codon, and both globin variants can terminate at the out of phase UAA. Registry Numbers: 9004-22-2(Globins) (REFERENCE 24 OF 100) 76190162 Marotta CA, Varricchio F, Smith I, Weissman SM

The primary structure of Bacillus subtilis and Bacillus stearothermophilus 5 S ribonucleic acids. In: J Biol Chem (1976 May 25) 251(10):3122-7 ISSN: 0021-9258 The nucleotide of Bacillus stearothermophilus 5 S RNA is pC-C-U-A-G-U- G-A-C-A-A-U-A-G-C-G-(G-A-G-A-G-G-)-A-A-A-C-A-C-C-C-G-U-U-C-C-C-A-U-C- C-C-G-A-A-C-A-C-G-G-A-A-G-U-U-A-A-G-C-U-C-U-C-C-A-G-C-G-C-C-G-A-U-G-G- U-A-G-U-U-G-G-G-G-C-C-A-G-C-G-C-C-C-C-U-G-C-A-A-G-A-G-U-A-G-G-U-C-G-U- U-G-C-U-A-G-G-COH; the nucelotide sequence of Bacillus subtilis 5 S RNA is pU-U-U-G-G-U-G-G-C-G-A-U-A-G-C-G-A-A-G-A-G-G-U-C-A-C-A-C-C-C-G- U-U-C-C-C-A-U-A-C-C-G-A-A-C-A-C-G-G-A-A-G-U-U-A-A-G-C-U-C-U-U-C-A-G-C- G-C-C-G-A-U-G-G-U-A-G-U-C-G-G-G-G-G-U-U-U-C-C-C-C-C-U-G-U-G-A-G-A-G-U- A-G-G-A-C-G-C-C-G-C-C-A-A-G-COH. Comparison of the sequence of B. stearothermophilus 5 S RNA to the sequence of B. subtilis 5 S RNA and to that of Bacillus megaterium 5 S RNA (Pribula, C. D., Fox, G. E., Woese, C. R., Sogin, M., and Pace, N. (1974) FEBS Lett. 44, 322-323) indicates that the 5 S RNA isolated from the thermophile contains unique nucleotide sequences not found in 5 S RNAs isolated from the two mesophilic species of genus Bacillus. (REFERENCE 25 OF 100) 77609129 Marotta CA, Varricchio F, Smith I, Weissman SM THE PRIMARY STRUCTURE OF BACILLUS SUBTILIS AND BACILLUS STEAROTHERMOPHILUS 5 S RIBONUCLEIC ACIDS In: J Biol Chem (1976) 251(10):3122-3127 ISSN: 0021-9258 The nucleotide sequence of Bacillus stearothermophilus 5 S RNA is pC- C-U-A-G-U-G-A-C-A-A-U-A-G-C-G- (G-A-G-A-G-G)-A-A-A-C-A-C-C-C-G-U-U-C- C-C-A-U-C-C-C-G-A-A-C-A-C-G-G-A-A-G-U-U-A-A-G-C-U-C-U-C-C-A-G-C-G-C-C- G-A-U-G-G-U-A-G-U-U-G-G-G-G-C-c-A-G-C-G-C-C-C-C-U-G-C-A-A-G-A-G-U-A-G- G-U-C-G-U-U-G-C-U-A-G-G-COH; the nucleotide sequence of Bacillus subtilis 5 S RNA is pU-U-U-G-G-U-G-G-C-G-A- U-A-G-C-G-A-A-G-A-G-G-U-C- A-C-A-C-C-C-G-U-U-C-C-C-A-U-A-C-C--G-A-A-C-A-C-G-G-A-A-G-U-U-A-A-G-C- U-C-U-U-C-A-G-C-G-C-C-G-A-U-G-G-U-A-G-U-C-G-G-G-G-G-U-U-U-C-C-C-C-C-U- G-U-G-A-G-A-G-U-A-G-G-A-C-G-C-C-G-C-C-A-A-G-COH. Comparison of the sequence of B stearothermophilus 5 S RNA to the sequence of B subtilis 5 S RNA and to that of Bacillus megaterium 5 S RNA (Pribula, CD, Fox, GE, Woese, CR, Sogin, M, and Pace, N (1974) FEBS Lett 44, 322-323) indicates that the 5 S RNA isolated from the thermophile contains unique nucleotide sequences not found in 5 S RNAs isolated

from the two mesophilic species of genus Bacillus. (Author Abstract) Institutional address: Research 4 Massachusetts General Hospital Fruit Street Boston MA 02114. *****JOURNAL OF CELL BIOLOGY***** (REFERENCE 26 OF 100) 83007478 Nixon RA, Brown BA, Marotta CA Posttranslational modification of a neurofilament protein during axoplasmic transport: implications for regional specialization of CNS axons. In: J Cell Biol (1982 Jul) 94(1):150-8 ISSN: 0021-9525 [No Abstract Available] Registry Numbers: 7440-70-2(Calcium) (REFERENCE 27 OF 100) 83007479 Brown BA, Nixon RA, Marotta CA Posttranslational processing of alpha-tubulin during axoplasmic transport in CNS axons. In: J Cell Biol (1982 Jul) 94(1):159-64 ISSN: 0021-9525 Tubulin proteins in mouse retinal ganglion cell (RGC) neurons were analyzed to determine whether they undergo posttranslational processing during axoplasmic transport. Alpha- and beta-tubulin comprised heterogeneous proteins in the primary optic pathway (optic nerve and optic tract) when examined by two-dimensional (2D) PAGE. In addition, however, alpha-tubulin exhibited regional heterogeneity when consecutive 1.1-mm segments of the optic pathway were analyzed separately. In proximal segments, alpha-tubulin consisted of two predominant proteins separable by isoelectric point and several less abundant species. In more distal segments, these predominant proteins decreased progressively and the alpha-tubulin region of the gel was represented by less abundant multiple forms only; beta-tubulin region of the gel was represented by less abundant multiple forms only; beta- tubulin was the same in all segments. After intravitreal injection of [3H]proline to mice, radiolabeled alpha- and beta-tubulin heteroproteins were conveyed together at a rate of 0.1-0.2 mm/d in the slowest phase of axoplasmic transport. At 45 d postinjection, the

distribution of radiolabeled heterogeneous forms a alpha- and beta- tubulin in consecutive segments of optic pathway resembled the distribution of unlabeled proteins by 2D PAGE, indicating that regional heterogeneity of tubulin arises during axonal transport. Peptide mapping studies demonstrated that the progressive alteration of alpha-tubulin revealed by PAGE analysis cannot be explained by contamination of the alpha-tubulin region by other proteins on gels. The results are consistent with the posttranslational processing of alpha-tubulin during axoplasmic transport. These observations, along with the accompanying report (J. Cell Biol., 1982, 94:150-158), provide additional evidence that CNS axons may be regionally specialized. *****J CROSS CULT GERONTOL***** (REFERENCE 28 OF 100) 22982697 Salguero RH, Kohn R, Salguero LF, Marotta CA Caregivers of persons with Alzheimer's Disease: cultural differences in perceived caregiver burden in Guatemala and Rhode Island. In: J Cross Cult Gerontol (1998) 13(3):229-40 ISSN: 0169-3816 The intent of this study is to illustrate cultural differences in the amount of perceived burden for primary caregivers of persons with Alzheimer's Disease. Caregivers in Guatemala and Rhode Island were given a questionnaire exploring: caregiver well-being, available supports, traditional ideology, and perceived burden. The data indicate that Guatemalans have less institutional and more informal supports available, as compared with USA caregivers. Guatemalan caregivers brought patients to a doctor sooner after the appearance of their first symptoms (0.9+/-1.0 years versus 1.6+/-1.8 years) and had poorer perceived health than USA subjects, suggesting a higher level of caregiver burden. Cultural response bias however may account for the difference in perceived health. Institutional address: Brown University Providence RI 02906 USA. *****JOURNAL OF GERIATRIC PSYCHIATRY AND NEUROLOGY***** (REFERENCE 29 OF 100) 91128587 Tate-Ostroff B, Majocha RE, Walcott EC, Ventosa-Michelman M, Marotta CA Colocalization of amino terminal and A4 (beta-amyloid) antigens in Alzheimer plaques: evidence for coordinated processing of the amyloid precursor protein.

In: J Geriatr Psychiatry Neurol (1990 Jul-Sep) 3(3):139-45 ISSN: 0891-9887 The mechanism by which the A4 (beta-amyloid) domain of the Alzheimer amyloid precursor protein (APP) is deposited in plaques is unknown, and limited information is available concerning the extent to which other APP sites are associated with plaques. To address these issues, we prepared antiserum to a peptide adjacent to the N-terminus of the APP (referred to as N1) and examined its distribution in brain relative to A4 by double-immunostaining techniques. Anti-N1 localized to both neurons and glia in control and Alzheimer patients. In the Alzheimer brain, anti-N1 detected plaques. Quantitation revealed that 85% of thioflavin-positive plaques, and 91% of A4-positive plaques were also N1 positive. Double-staining methods directly demonstrated colocalization of distant APP sites. The data suggest that suggest that proposed mechanisms for amyloid deposition during plaque formation must take into account the extracytoplasmic domain, in addition to the A4 region, rather than be confined exclusively to the A4 site. Institutional address: Department of Psychiatry Harvard Medical School Boston MA. (REFERENCE 30 OF 100) 89302531 Majocha RE, Marotta CA Molecular and genetic investigations on Alzheimer brain amyloid. In: J Geriatr Psychiatry Neurol (1988 Apr-Jun) 1(2):65-70 ISSN: 0891-9887 Amyloid-containing plaques are a characteristic feature of the Alzheimer's disease brain and have been the object of study for decades. Only recently, however, have molecular and genetic techniques been applied to examination of amyloid in order to understand the factors that contribute to the accumulation of plaques in dementia. Current investigations have focused on the structure and properties of the amyloid protein, its corresponding messenger RNA, its cellular site of production, and its chromosomal site of origin. These data are discussed in the present review. Institutional address: Department of Psychiatry Harvard Medical School Boston MA. (REFERENCE 31 OF 100) 89302523

Benes FM, Majocha RE, Marotta CA A modular arrangement of neuronal processes in human cortex: disruption with aging and in Alzheimer's disease. In: J Geriatr Psychiatry Neurol (1988 Jan) 1(1):3-10 ISSN: 0891-9887 Studies were undertaken to assess whether or not neuron-specific immunostaining of the human brain can reveal unique cytoarchitectural features that may be affected by healthy aging and Alzheimer's disease (AD). Human prefrontal cortex (PFC) and anterior cingulate cortex (ACC) were stained with an antibody raised against the neurofilament protein 200,000 molecular weight subunit (NFP-200) using an avidin-biotin immunolocalization procedure. Immunostaining of cortex was neuron-specific and highlighted axons in particular. This staining has revealed an orderly arrangement of horizontal and vertical axon bundles which form latticelike compartments or modules throughout most of the matrix of the two cortical areas studied. The ACC shows this pattern to be intact in individuals through the ninth decade, while the PFC there was blurring of the modularity beyond the fifth decade. Irrespective of the age-related blurring of the latticelike arrangement of axons in PFC, neurologically normal elderly individuals nevertheless showed a highly organized appearance to the cortical matrix. By contrast, patients with AD showed a marked disruption of the modular arrangement of fibers in PFC, but not ACC. Differences between PFC fibers in controls and AD patients were also demonstrated with an axon-specific monoclonal antibody that reacted with phosphorylated epitopes of the NFP-200. The chaotic appearance of fiber staining in PFC seen in patients with AD was noted to be present in one subject who died in an early stage of the disease. The possible significance of this previously unknown aspect of cortical cytoarchitecture for normal cognitive functioning in humans is discussed. Institutional address: Department of Psychiatry Harvard Medical School Boston MA. *****JOURNAL OF MOLECULAR NEUROSCIENCE***** (REFERENCE 32 OF 100) 92329298 Marotta CA, Majocha RE, Tate B Molecular and cellular biology of Alzheimer amyloid. In: J Mol Neurosci (1992) 3(3):111-25 ISSN: 0895-8696 Alzheimer's Disease (AD), a disorder of unknown etiology, is the most common form of adult-onset dementia and is characterized by severe intellectual deterioration. The definitive diagnosis of AD is made by

postmortem examination of the brain, which reveals large quantities of neurofibrillary tangles (NFT) and senile plaques within the parenchyma. The NFT are composed of paired helical filaments associated with several cytoskeletal proteins. The primary protein component of senile plaques is beta/A4 amyloid, a 42-43 amino acid peptide derived from a much larger molecule, the amyloid precursor protein (APP). Vascular beta/A4 amyloidosis is also prevalent in the disease. The mechanism by which beta/A4 amyloid accumulates in the AD brain is unknown. Recent research has demonstrated that the precursor molecule, APP, is a transmembrane protein with a large extracytoplasmic domain, a membrane spanning region that includes the portion that gives rise to beta/A4 amyloid, and a short intracytoplasmic domain. The precursor has multiple forms among which are those that differ by a variable length insert within the extracytoplasmic domain. The insert has sequence homology to the family of Kunitz protease inhibitor proteins. Cellular and animal models have been developed to study the nature of APP processing and the biological and behavioral consequences of beta/A4 amyloidosis. The results of such studies indicate that the normal processing of APP involves enzymatic cleavage of the molecule within the beta/A4 amyloid region, thus preventing the accumulation of beta/A4 in the normal brain. The factors leading to abnormal processing of APP, and consequent beta/A4 amyloid accumulation within the AD brain, have yet to be identified. In cell culture, the biological effects associated with beta/A4 amyloid include neurotrophic and neurotoxic activities, while the peptide has also been shown to have dramatic behavioral effects in animal models. Institutional address: Department of Psychiatry Harvard Medical School Boston MA. *****JOURNAL OF NEUROCHEMISTRY***** (REFERENCE 33 OF 100) 89341809 Majocha RE, Jungalwala FB, Rodenrys A, Marotta CA Monoclonal antibody to embryonic CNS antigen A2B5 provides evidence for the involvement of membrane components at sites of Alzheimer degeneration and detects sulfatides as well as gangliosides. In: J Neurochem (1989 Sep) 53(3):953-61 ISSN: 0022-3042 Immunohistological and biochemical studies were initiated to determine whether or not neural membrane components were associated with degenerative changes characteristic of Alzheimer's disease (AD). Monoclonal antibody A2B5, developed against embryonic chick retinal cells and previously shown to react with neural surface gangliosides, was applied to formalin-fixed sections of control and AD brain tissue. Frontal cortex and hippocampus of AD cases exhibited high levels of A2B5 immunoreactivity within those neurons undergoing

neurofibrillary degeneration. Neuritic processes associated with senile plaques were also highly reactive with the A2B5 antibody. The amount of gangliosides and their pattern after HPTLC were the same in control and AD cases. However, the unexpected observation was made that the A2B5 antibody reacted with human brain sulfatides in addition to the expected reactivity with minor gangliosides. The average level of sulfatides in AD brain was significantly higher than in normal controls. The data support the involvement of one or more membrane components with neurodegeneration in the Alzheimer brain. Registry Numbers: 2390-54-7(thioflavin T) Institutional address: Neuroscience Program Harvard Medical School Boston Massachusetts. (REFERENCE 34 OF 100) 84241800 Nixon RA, Marotta CA Degradation of neurofilament proteins by purified human brain cathepsin D. In: J Neurochem (1984 Aug) 43(2):507-16 ISSN: 0022-3042 Cathepsin D (CD) was purified to homogeneity from postmortem human cerebral cortex. Incubation of CD with human neurofilament proteins (NFPs) prepared by axonal flotation led to the rapid degradation of the 200,000, 160,000, and 70,000 NFP subunits (200K, 160K, and 70K) which had been separated by one- or two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Degradation was appreciable at enzyme activity-to-substrate protein ratios that were two- to threefold lower than those in unfractionated homogenates from cerebral cortex. Quantitative measurements of NFPs separated by PAGE revealed that, at early stages of digestion, the 160K NFP was somewhat more rapidly degraded than the 70K subunit while the 200K NFP had an intermediate rate of degradation. At sufficiently high enzyme concentrations, all endogenous proteins in human NF preparations were susceptible to the action of CD. Human brain CD also degraded cytoskeletal proteins in NF preparations from mouse brain with a similar specificity. To identify specific NFP break-down products, antisera against each of the major NFPs were applied to nitrocellulose electroblots of NFPs separated by two-dimensional SDS- PAGE. In addition to detecting the 200K, 160K, and 70K NFP in human NF preparations, the antisera also detected nonoverlapping groups of polypeptides resembling those in NF preparations from fresh rat brain. When human NF preparations were incubated with CD, additional polypeptides were released in specific patterns from each NFP subunit. Some of the immuno-cross-reactive fragments generated from NFPs by CD comigrated on two-dimensional gels with polypeptides present in unincubated preparations. These results demonstrate that NFPs and other cytoskeletal proteins are substrates for CD. The physiological significance of these findings and the possible

usefulness of analyzing protein degradation products for establishing the action of proteinases in vivo are discussed. Registry Numbers: EC 3.4.-(Cathepsins) Registry Numbers: EC 3.4.23.5(Cathepsin D) (REFERENCE 35 OF 100) 83137726 Sajdel-Sulkowska E, Coughlin JF, Marotta CA In vitro synthesis of polypeptides of moderately large size by poly(A)-containing messenger RNA from postmortem human brain and mouse brain. In: J Neurochem (1983 Mar) 40(3):670-80 ISSN: 0022-3042 Studies were undertaken to optimize the conditions for isolation and in vitro translation of poly(A)-containing mRNA from human postmortem brain. The comparison of several methods for preparation of mRNA from frozen mouse brain indicated that although the yield of mRNA was increased using polysomes prepared in the presence of ribonucleoside vanadyl complexes and subsequently extracted with guanidinium thiocyanate, the translation products were indistinguishable from those synthesized by total cellular RNA directly extracted from tissue with guanidinium thiocyanate. The oligo d(T)-cellulose- purified poly(A)-containing mRNA preparations were translated in vitro in a rabbit reticulocyte lysate in the presence of L- [35S]methionine. Messenger RNA from frozen mouse brain stimulated protein synthesis from 9- to 20-fold over endogenous mRNA. Over 450 polypeptides were reproducibly synthesized and separated by two- dimensional polyacrylamide gel electrophoresis (PAGE); size classes up to 130,000 daltons were present. Direct extraction of RNA from frozen human cerebral cortex and cerebellum with guanidinium thiocyanate followed by oligo d(T)-cellulose chromatography yielded 1.8 micrograms/g and 2.0 micrograms/g, respectively, of poly(A)- containing mRNA; this represents a two- to fourfold increase over our earlier results. In the rabbit reticulocyte translation system human brain mRNA stimulated protein synthesis nearly threefold over endogenous mRNA. Compared with earlier studies, the number of newly synthesized polypeptides was increased by 30%. Over 300 species were separated by two-dimensional PAGE, and size classes up to 130,000 daltons were present, as compared to 70,000 in an earlier report. The polypeptides synthesized by human cerebral cortex and cerebellum were indistinguishable. However, several appeared to be uniquely human when compared with the products synthesized by mouse brain mRNA. The method described for the preparation of postmortem human brain mRNA eliminates the need to prepare polysomes, which are recovered in variable and low yield from the postmortem human brain. The procedure appears applicable to studies on the synthesis of moderately large human brain polypeptides and for investigations of brain protein polymorphism when relatively large numbers of products are required for analysis. Registry Numbers: 24937-83-5(Poly A)

(REFERENCE 36 OF 100) 83110996 Brown BA, Majocha RE, Staton DM, Marotta CA Axonal polypeptides cross-reactive with antibodies to neurofilament proteins. In: J Neurochem (1983 Feb) 40(2):299-308 ISSN: 0022-3042 Antibodies were prepared to mammalian CNS neurofilament proteins (NFPs) and the antibody specificities were compared using a sensitive immunoblotting method. This procedure was used to detect and characterize cross-reactive proteins and their degradation products in neurofilament preparations. NFPs were prepared by axon flotation. Rabbits were immunized with 200,000, 140,000, and 70,000 NFPs (200K, 140K, and 70K) that had been electrophoretically purified by polyacrylamide gel electrophoresis (PAGE). By immunohistofluorescence it was shown that all antisera stained similar filamentous structures in rat cerebellar neurons. By use of a horseradish peroxidase- conjugated indirect antibody procedure, however, differences were detected in the cross-reactivities of the antisera to rat NFPs, separated by PAGE and electrophoretically transferred to nitrocellulose membranes. Each antiserum exhibited strong binding to the homologous NFP and, thus, was suitable for the detection of cross- reactive polypeptides and proteolytic degradation products derived exclusively from the individual NFPs. Anti-200K, anti-140K, or anti- 70K was applied to overloaded two-dimensional nitrocellulose blots of NFPs prepared by axon flotation. Each of the three sera detected a group of unique nonoverlapping polypeptides, some of which were identified as NFP degradation products. A different group of polypeptides was cross-reactive with antiserum to purified glial fibrillary acidic protein. The immunostaining of polypeptides on nitrocellulose was far more sensitive for detecting NFP degradation products than was staining polyacrylamide gels with Coomassie blue. Titers for the antisera were two to three orders of magnitude higher with the immunoblotting procedure than with immunohistologic methods. The sensitivity and the specificity of the described methods suggest their usefulness for examining proteolytic cleavage products of NFPs under a variety of conditions. (REFERENCE 37 OF 100) 81144091 Marotta CA, Brown BA, Strocchi P, Bird ED, Gilbert JM In vitro synthesis of human brain proteins including tubulin and actin by purified postmortem polysomes. In: J Neurochem (1981 Mar) 36(3):966-75 ISSN: 0022-3042 Polysomes were prepared from human brain tissue 2-6 h postmortem; the polysomes were active in a cell-free protein synthesis system

containing rabbit reticulocyte factors. Protein synthesis was totally dependent upon added MgCl2, ATP, the reticulocyte factor fraction, and the human polysome fraction. Human brain proteins synthesized in the presence of L-[35S]methionine were analyzed by one- and two- dimensional polyacrylamide gel electrophoresis. Over 250 proteins were synthesized and they extended in size up to 250,000 d; many of the most abundant native human brain proteins were synthesized, including tubulin and actin. It was shown that human brain alpha and beta tubulin and actin isomers synthesized in vitro from human postmortem polysomes have the same apparent molecular weights and isoelectric points as the corresponding proteins synthesized by rat polysomes from fresh cortices. The corresponding tubulin and actin synthesized by human and rat brain polysomes also yield the same radioactive methionine-containing peptides after digestion with Staphylococcus aureus V8 protease. These analyses indicate that postmortem polysomes contain active messenger RNA which can direct the partial and/or complete synthesis of actin and tubulin subunits and other human brain proteins. (REFERENCE 38 OF 100) 81119016 Brown BA, Nixon RA, Strocchi P, Marotta CA Characterization and comparison of neurofilament proteins from rat and mouse CNS. In: J Neurochem (1981 Jan) 36(1):143-53 ISSN: 0022-3042 Rat and mouse CNS neurofilament proteins (NFPs) were characterized and compared, in terms of electrophoretic properties on polyacrylamide gels and by peptide mapping, with one another and with other co-purifying lower-molecular-weight CNS proteins, including alpha and beta tubulin. NFPs were partially purified by modification of the axon flotation procedure of Norton and co-workers and were demyelinated with Triton X-100. On one-dimensional SDS polyacrylamide gels the molecular weights of the triad of NFPs from both rat and mouse were approximately 200,000, 140,000, and 70,000. Prominent lower-molecular-weight proteins (63,000-16,000) as well as minor amounts of tubulin and actin were observed after gel electrophoresis. On two-dimensional gels (isoelectric focusing followed by SDS gel electrophoresis) each of the NFPs appeared to be composed of more than one component and the corresponding NFPs from rat and mouse had similar isoelectric points. Gel electrophoresis peptide mapping using Staphylococcus aureus V8 protease indicated the following: (1) the triad of NFPs of different sizes have different peptide maps; (2) alpha and beta tubulin have nonidentical digestion products, which are dissimilar to those of the NFPs; (3) other proteins that co- purify by the axon flotation procedure also have nonidentical peptide maps; and(4) the corresponding NFPs from rat and mouse have similar peptide maps. The co-purifying proteins examined in detail (63,000- 49,000) do not appear to be derived by proteolytic cleavage of NFPs and may represent other cytoskeletal constituents. (REFERENCE 39 OF 100) 81217478

Strocchi P, Gilbert JM, Marotta CA Variations in brain cortical polysome translation products among rats of the same strain. In: J Neurochem (1981 Jun) 36(6):2044-51 ISSN: 0022-3042 Studies were undertaken to determine whether there exist variations among the translation products of polysomes from different brains of animals of the same strain. Polysomes were prepared from individual rat cortices and translated in a reticulocyte protein-synthesizing system containing rabbit reticulocyte factors and L-[35S]methionine; he resulting radioactive proteins were analyzed by two-dimensional polyacrylamide gel electrophoresis autoradiography. Comparison of the autoradiographs revealed that two acidic proteins, A and B, of apparent 54,000 M. W. occur as three phenotype: A only, B only, or A plus B. These proteins were not detectable by Coomassie brilliant blue staining of two-dimensional electrophoretograms of brain protein preparations. Messenger RNA was extracted from pooled cortices and translated in a wheat germ extract, and both A and B proteins were detected among the products of translation. Cyclic AMP affinity chromatography of the translation products of cortical polysomes showed that both A and B proteins bind to cyclic AMP. Our data are consistent with the conclusion that there are qualitative differences in the polysome translation products that bind to cyclic AMP among individual cortices of rats of the same strain. Registry Numbers: 60-92-4(Cyclic AMP) Registry Numbers: 63-68-3(Methionine) (REFERENCE 40 OF 100) 81144092 Gilbert JM, Brown BA, Strocchi P, Bird ED, Marotta CA The preparation of biologically active messenger RNA from human postmortem brain tissue. In: J Neurochem (1981 Mar) 36(3):976-84 ISSN: 0022-3042 Messenger RNA (mRNA) was extracted from human postmortem brain tissue by alkaline phenol extraction of polysomes followed by oligo (dT)- cellulose chromatography. The mRNA preparations stimulated protein synthesis in a cell-free system containing wheat germ homogenate. The products of protein synthesis were analyzed by one- and two- dimensional gel electrophoresis. These analyses indicated that numerous polypeptides, including tubulin subunits and actin isomers, were synthesized by the human mRNA. The molecular weight range of polypeptides synthesized by human mRNA fractions from two brain specimens were identical, and analysis by two-dimensional gel electrophoresis indicated qualitatively similar products. The yield of mRNA extracted per gram of human tissue was less than the yield obtained with rat forebrains from animals sacrificed immediately

before brain removal and mRNA purification. A decrease in the amount of polysomes isolated from human tissue relative to rat brain tissue was a major factor contributing to the low yield. The molecular weight distribution of polypeptides synthesized by human and rat brain mRNA fractions in wheat germ homogenate was similar; thus, there was no indication for selective breakdown or inactivation of high molecular weight mRNA species in the human tissue. Our studies indicate that it is possible to utilize postmortem tissue for molecular biological investigations of human brain mRNA. (REFERENCE 41 OF 100) 81144073 Gilbert JM, Strocchi P, Brown BA, Marotta CA Tubulin synthesis in rat forebrain: studies with free and membrane- bound polysomes. In: J Neurochem (1981 Mar) 36(3):839-46 ISSN: 0022-3042 Free and membrane-bound polysomes were prepared from rat forebrain and added to a cell-free system containing rabbit reticulocyte factors and L-[35S]methionine. The translation products were analyzed by two-dimensional gel electrophoresis followed by autoradiography. The free polysomes synthesized actin and at least four major tubulin subunits (alpha 1, alpha 2, beta 1, and beta 2) that are found in rat forebrain cytoplasm. The membrane-bound polysomes synthesized predominantly one protein (MB) in the tubulin region of the two- dimensional gel. MB has a molecular weight and isoelectric point similar to alpha-tubulin. Only trace amounts of alpha- and beta- tubulin and action were synthesized by the membrane-bound polysomes. MB co-purified with cytoplasmic tubulin after two cycles of aggregation and disaggregation. MB synthesized in vitro (from membrane-bound polysomes) and alpha- and beta-tubulin and actin subunits (synthesized from free polysomes) were digested with Staphylococcus aureus V8 protease, and the resulting peptides were separated by slab gel electrophoresis followed by autoradiography. The peptide pattern of MB was similar but not identical to the peptide patterns of alpha- and beta-tubulin; MB yielded peptides not found in tubulin. We conclude that membrane-bound polysomes from rat forebrain do not synthesize significant amounts of the predominant tubulin subunits synthesized by free polysomes. A major protein (MB) is synthesized by membrane-bound polysomes and is similar, but not identical, to alpha-tubulin synthesized by free polysomes on the basis of molecular weight, isoelectric point, and peptide analysis. (REFERENCE 42 OF 100) 79218133 Marotta CA, Strocchi P, Gilbert JM Biosynthesis of heterogeneous forms of mammalian brain tubulin subunits by multiple messenger RNAs. In: J Neurochem (1979 Jul) 33(1):231-46 ISSN: 0022-3042

[No Abstract Available] (REFERENCE 43 OF 100) 78219787 Marotta CA, Strocchi P, Gilbert JM Microheterogeneity of brain cytoplasmic and synaptoplasmic actins. In: J Neurochem (1978 Jun) 30(6):1441-51 ISSN: 0022-3042 [No Abstract Available] (REFERENCE 44 OF 100) 78219786 Marotta CA, Harris JL, Gilbert JM Characterization of multiple forms of brain tubulin subunits. In: J Neurochem (1978 Jun) 30(6):1431-40 ISSN: 0022-3042 [No Abstract Available] *****JOURNAL OF NEUROSCIENCE***** (REFERENCE 45 OF 100) 87197498 Nixon RA, Lewis SE, Marotta CA Posttranslational modification of neurofilament proteins by phosphate during axoplasmic transport in retinal ganglion cell neurons. In: J Neurosci (1987 Apr) 7(4):1145-58 ISSN: 0270-6474 The progressive modification of newly synthesized neurofilament proteins (NFPs) during axoplasmic transport in mouse retinal ganglion cell (RGC) neurons was studied after RGC perikarya were pulse-labeled with 32P-orthophosphate or radiolabeled amino acids. The 3 NFP subunits, H(igh), M(iddle), and L(ow), were among a group of axonally transported proteins that incorporated high levels of 32P. Covalent addition of phosphate slowed the electrophoretic mobility of H and M on SDS polyacrylamide gels and shifted the charge of all 3 subunits toward more acidic pH values, thereby providing an index of the phosphorylation state of this radiolabeled population of NFPs. NFPs were extensively phosphorylated before they entered axons at the optic nerve level, and continued to be modified during transport along RGC axons at the optic nerve and tract level. H and M exhibited charge shifts of 0.2-0.6 units toward a more acidic pH during axoplasmic transport. The charge modifications became more prominent when NFPs reached distal axonal levels, which may indicate regional

differences in the activity of this modification process along axons. By contrast, the L subunit became more basic in charge, consistent with decreases in the phosphorylation state during transport. Additional observations (Nixon and Lewis, 1986) that a considerable proportion of phosphate groups initially added to L and M were later removed as neurofilaments advanced along RGC axons support the notion that the changing phosphorylation state of NFP subunits during axoplasmic transport reflects a dynamic equilibrium between phosphorylation and dephosphorylation events. Topographical remodeling of phosphate groups on NFPs during axoplasmic transport is proposed as a possible mechanism for coordinating interactions between neurofilaments and other constituents, as these elements are transported and integrated into the axonal cytoskeleton. Registry Numbers: 147-85-3(Proline) Registry Numbers: 7005-18-7(Methionine) *****JOURNAL OF NEUROSCIENCE METHODS***** (REFERENCE 46 OF 100) 88188764 Sajdel-Sulkowska EM, Majocha RE, Salim M, Zain SB, Marotta CA The postmortem Alzheimer brain is a source of structurally and functionally intact astrocytic messenger RNA. In: J Neurosci Methods (1988 Mar) 23(2):173-9 ISSN: 0165-0270 Although the precise role of astrocytes in the pathogenesis of Alzheimer's disease (AD) is currently undefined, studies carried out at the molecular level may lead to new insights into the functioning of this class of brain cells in dementia. In order to facilitate such investigations, methods are described that establish that structurally and functionally intact messenger RNA (mRNA) for an astrocytic marker, glial fibrillary acidic protein (GFAP), is present in the postmortem Alzheimer's disease brain after long postmortem intervals. Rapid preparative procedures were used to obtain poly(A)+ RNA from postmortem control and AD cortices. In vitro protein synthesis was carried out in a reticulocyte system. Relative to controls, AD mRNA synthesized a two-fold higher level of a 50,000 mol.wt. protein that was immunologically identified as GFAP. High levels of GFAP synthesis by purified mRNA from AD cortices was independent of age at death and postmortem interval up to 24 h. Northern blot hybridization using a cloned human GFAP riboprobe was used to evaluate postmortem GFAP mRNA stability. No appreciable degradation products of GFAP mRNA were observed on Northern blots for at least 10 h postmortem in poly(A)+ RNA extracted from the AD brain. The described methodology demonstrates that the postmortem AD brain is an excellent source of functionally and structurally intact astrocyte-specific mRNA. Institutional address: Neuroscience Program

Harvard Medical School Boston MA 02115. *****JOURNAL OF NEUROSCIENCE RESEARCH***** (REFERENCE 47 OF 100) 96301866 Nagy JI, Hossain MZ, Hertzberg EL, Marotta CA Induction of connexin43 and gap junctional communication in PC12 cells overexpressing the carboxy terminal region of amyloid precursor protein. In: J Neurosci Res (1996 Apr 15) 44(2):124-32 ISSN: 0360-4012 Previous studies have shown that PC12 cells overexpressing beta/A4 amyloid peptide display altered morphology characterized by pronounced membrane ruffling and extensive intercellular appositions. Having observed other cell types in which these features accompany increased connexin43 (Cx43) production and gap junctional communication, we examined Cx43 in normal and beta/A4-transfected PC12 cells. Studies of two beta/A4-transfected PC12 clones revealed an induction of Cx43 expression by Western blotting, intracellular and plasma membrane-associated Cx43 in some cells of cultures processed by immunofluorescence, dye-transfer between some cells microinjected with Lucifer Yellow, and gap junctions between cells examined by EM. Normal and vector-transfected PC12 cells exhibited none of these properties. Increased immunofluorescence in some clusters of beta/A4-transfected cells was also observed with a monoclonal antibody against connexin32. The results suggest that beta/A4 amyloid peptide may cause aberrant intercellular communication and gap junction formation through induction or increased expression of connexins in cells that are not normally coupled or only poorly coupled by gap junctions. Institutional address: Department of Physiology University of Manitoba Winnipeg Manitoba Canada. *****JOURNAL OF NUCLEAR MEDICINE***** (REFERENCE 48 OF 100) 93094922 Majocha RE, Reno JM, Friedland RP, VanHaight C, Lyle LR, Marotta CA Development of a monoclonal antibody specific for beta/A4 amyloid in Alzheimer's disease brain for application to in vivo imaging of amyloid angiopathy.

In: J Nucl Med (1992 Dec) 33(12):2184-9 ISSN: 0161-5505 We evaluated the efficacy of murine monoclonal antibodies (Mabs) targeted to beta/A4 amyloid for development of procedures for the in vivo identification of amyloid angiopathy (AA) in Alzheimer's disease (AD). Mabs to beta/A4 amyloid were prepared and screened for effectiveness in visualizing AA and senile plaques in postmortem AD brain sections. They were assessed again after enzymatic cleavage to produce Fab fragments and after labeling with 99mTc using a diamide dimercaptide ligand system. Modified and radiolabeled Fab fragments retained activity and specificity towards amyloid-laden blood vessels and senile plaques. A highly specific murine Mab, 10H3, was identified and characterized that fulfills criteria necessary for the development of a diagnostic imaging agent. Expansion and adaptation of these strategies may provide the methods and materials for the noninvasive analysis of AA in living patients, and permit assessment of the contribution of AA to the clinical and pathological features of AD. Registry Numbers: 7440-26-8(Technetium) Institutional address: Neurobiology Laboratory Massachusetts General Hospital Boston. *****JOURNAL OF PSYCHIATRIC RESEARCH***** (REFERENCE 49 OF 100) 90308771 Chou WG, Zain SB, Rehman S, Tate-Ostroff B, Majocha RE, Benes FM, Marotta CA Alzheimer cortical neurons containing abundant amyloid mRNA. Relationship to amyloid deposition and senile plaques. In: J Psychiatr Res (1990) 24(1):37-50 ISSN: 0022-3956 Since the detailed molecular events leading to the formation of amyloid-containing senile plaques of the Alzheimer's disease (AD) brain are incompletely understood, the present studies were undertaken to address this issue using a combination of molecular and cytochemical approaches. Amyloid precursor protein riboprobes containing the A4 (beta-amyloid) domain were applied to cortex using the in situ hybridization method to examine the distribution of neuronal amyloid mRNA in relation to the laminar pattern of amyloid deposition and the localization of plaques. The derived data indicated that high levels of amyloid mRNA can be synthesized by AD cortical neurons that appeared to be morphologically intact. The distribution of these cells was not coincident with the cortical laminar pattern that is typical of amyloid deposits observed after

immunostaining with anti-A4 monoclonal antibodies. Further, there was no obvious relationship between neurons containing abundant amyloid mRNA and the distribution of plaques identified by thioflavin S staining. While the neuronal synthesis of amyloid may be a significant factor at some point during plaque formation, it may not be the exclusive determinant. The possibility is raised that processes affecting secretion, diffusion, and/or transport of amyloid away from neuronal or non-neuronal cells of origin to sites of deposition may be meaningful aspects of the molecular pathology of Alzheimer's disease. Institutional address: Department of Biochemistry University of Rochester Medical School NY 14642. *****METHODS IN ENZYMOLOGY***** (REFERENCE 50 OF 100) 74305878 Marotta CA, Lebowitz P, Dhar R, Zain BS, Weissman SM Preparation of RNA transcripts of discrete segments of DNA. In: Methods Enzymol (1974) 29(0):254-72 ISSN: 0076-6879 [No Abstract Available] *****MOLECULAR AND CHEMICAL NEUROPATHOLOGY***** (REFERENCE 51 OF 100) 93221620 Majocha RE, Tate B, Marotta CA PC12 cells release stimulatory factors after transfection with beta/A4-C-terminal DNA of the Alzheimer amyloid precursor protein. In: Mol Chem Neuropathol (1993 Jan-Feb) 18(1-2):99-113 ISSN: 1044-7393 The beta/A4 region of the amyloid precursor protein (APP) accumulates in brains of victims of Alzheimer disease (AD) where it is a major component of senile plaques. We examined the pathophysiological consequences of overexpression of the beta/A4-C-terminal DNA in PC12 cells. Serum-free conditioned media (SFCM) from positive transfectants stimulated control PC12 cells to extend neurites and increase in size. Unlike the factor that affected cell size, neurite lengthening activity was significantly decreased after immunoabsorption with anti-beta/A4 monoclonal antibodies (MAb) and changes in pH. The data support the view that among the consequences of beta/A4-C-terminal DNA overexpression in PC12 cells is the release of factors that stimulate nontransfected cells to undergo

morphological transformations that include differentiation to a neuronal phenotype. It is hypothesized that similar activities that may contribute to the molecular pathophysiology of the disorder may be present in the AD brain. Registry Numbers: 9007-49-2(DNA) Institutional address: Massachusetts General Hospital Department of Psychiatry Boston. *****MOLECULAR NEUROBIOLOGY***** (REFERENCE 52 OF 100) 95194581 Friedland RP, Majocha RE, Reno JM, Lyle LR, Marotta CA Development of an anti-A beta monoclonal antibody for in vivo imaging of amyloid angiopathy in Alzheimer's disease. In: Mol Neurobiol (1994 Aug-Dec) 9(1-3):107-13 ISSN: 0893-7648 We evaluated the efficacy of murine monoclonal antibodies (MAbs) targeted to the A beta amyloid of Alzheimer's disease for development of procedures for the in vivo identification of amyloid angiopathy (AA). MAbs to A beta were prepared and screened for effectiveness in visualizing AA and neuritic plaques in postmortem AD brain sections. They were assessed again after enzymatic cleavage to produce Fab fragments and after labeling with technetium-99m (99mTc) using a diamide dimercaptide ligand system. Modified and radiolabeled Fab fragments retained activity and specificity toward amyloid-laden blood vessels and neuritic plaques. A highly specific murine MAb, 10H3, was identified and characterized that fulfills criteria necessary for the development of an in vivo diagnostic imaging agent. Toxicity studies in rats showed the MAb to be safe. Biodistribution studies in mice demonstrated desirable properties for use as an imaging agent. Expansion and adaptation of these strategies may provide the methods and materials for the noninvasive analysis of AA in living patients, and permit assessment of the contribution of AA to the clinical and pathological features of AD. Institutional address: Department of Neurology Case Western Reserve University Cleveland OH 44106. *****NEUROBIOLOGY OF AGING***** (REFERENCE 53 OF 100) 88233132

Salim M, Rehman S, Sajdel-Sulkowska EM, Chou WG, Majocha RE, Marotta CA, Zain SB Preparation of a recombinant cDNA library from poly(A+) RNA of the Alzheimer brain. Identification and characterization of a cDNA copy encoding a glial-specific protein. In: Neurobiol Aging (1988 Mar-Apr) 9(2):163-71 ISSN: 0197-4580 Studies were undertaken to assess the extent to which messenger RNA prepared from the postmortem Alzheimer's disease (AD) brain can be used for the successful preparation of a recombinant cDNA library. Initial experiments focused on the glial-specific marker glial fibrillary acidic protein (GFAP) since GFAP expression appeared to be a model for further studies on mRNAs that may continue to be expressed at high levels in the vicinity of lesioned sites in the AD brain. An AD cDNA library, prepared in the lambda gt11 expression vector system contained GFAP-specific recombinants. One of these was sequenced and the insert was shown to exhibit 88% homology with the similar sequence from mouse GFAP. As established by Northern blots, the size of the GFAP mRNA prepared from the routinely acquired postmortem AD cortex, approximately 2.7 kb, was the same as from a neurologically normal control brain. These results agree with earlier studies on GFAP mRNA from fresh mouse brain. The results demonstrate that in the postmortem AD brain, astroglial-specific mRNA remains sufficiently stable for molecular genetic analysis and may serve as a useful model for examining the genetic expression of mRNAs that may be related to the molecular pathogenesis and the etiology of AD. Registry Numbers: 9007-49-2(DNA) Institutional address: Cancer Center University of Rochester Medical School NY 14642. *****NEUROCHEMICAL RESEARCH***** (REFERENCE 54 OF 100) 92382695 Honda T, Marotta CA Arginine specific endopeptidases modify the aggregation properties of a synthetic peptide derived from Alzheimer beta/A4 amyloid. In: Neurochem Res (1992 Apr) 17(4):367-74 ISSN: 0364-3190 A synthetic peptide corresponding to the first 28 amino acids of the Alzheimer disease amyloid beta/A4 peptide (3.2 kDa) aggregated to a high molecular weight (15 kDa) on SDS/urea polyacrylamide gels. Proteinase K, V8 protease, trypsin, and endopeptidase Lys-C readily degraded the aggregate. By contrast, when digested by endopeptidase Arg-C, a new polypeptide aggregate of higher molecular weight (16

kDa) was observed on denaturing gels without degraded smaller products. The new aggregate was comprised of three peptides: an intact beta/A4(1-28) and partially degraded peptides beta/A4(1-5) plus beta/A4(6-28). The results were confirmed by treatment of beta/A4 with other arginine-specific proteases: the gamma subunit of nerve growth factor and clostripain. The results indicate that arginine-specific proteases, including a growth factor processing enzyme, can nick aggregated beta/A4(1-28) amyloid and alter the configuration to produce a more complex aggregated form. If similar highly specific proteolytic mechanisms occur in the Alzheimer disease brain, the processing may promote the formation of high molecular weight aggregates that contribute to the development of relatively insoluble senile plaque core protein. Registry Numbers: EC 3.4.-(Peptide Peptidohydrolases) Registry Numbers: EC 3.4.21(Serine Proteinases) Institutional address: Molecular Neurobiology Laboratory McLean Hospital Belmont Massachusetts. *****NEUROSCIENCE LETTERS***** (REFERENCE 55 OF 100) 96149986 Lynn BD, Marotta CA, Nagy JI Propagation of intercellular calcium waves in PC12 cells overexpressing a carboxy-terminal fragment of amyloid precursor protein. In: Neurosci Lett (1995 Oct 13) 199(1):21-4 ISSN: 0304-3940 We have recently found that PC12 cells overexpressing a C-terminal 97 amino acid fragment containing the beta/A4 amyloid peptide of amyloid precursor protein (APP) exhibit an induced production of the gap junctional protein connexin43 (Cx43) and an induction of gap junctional communication as assessed by dye-transfer. In studies of two beta/A4-transfected PC12 clones, we show here that these cells, unlike normal PC12 cells or those transfected with vector alone, have the capacity for intercellular transmission of calcium waves as determined by imaging of calcium with fura-2. Intercellular wave propagation occurred in the absence of extracellular calcium and was blocked by known inhibitors of gap junctional communication (octanol and 12-O-tetradecanoyl-phorbol-13-acetate), suggesting mediation by gap junctions. The results indicate a disruptive influence of the C- terminal region of APP or of beta/A4 amyloid peptide on intercellular signaling via gap junctions, which may be relevant to the normal functions of APP or to pathology in Alzheimer's disease. Registry Numbers: 7440-70-2(Calcium)

Registry Numbers: 96314-98-6(Fura-2) Institutional address: Department of Physiology University of Manitoba Winnipeg Canada. (REFERENCE 56 OF 100) 89071022 Shea TB, Majocha RE, Marotta CA, Nixon RA Soluble, phosphorylated forms of the high molecular weight neurofilament protein in perikarya of cultured neuronal cells. In: Neurosci Lett (1988 Oct 17) 92(3):291-7 ISSN: 0304-3940 The high molecular weight subunit of neurofilaments (NF-H) in mouse NB2a/d1 neuroblastoma cells is extensively phosphorylated and exhibits an apparent molecular weight of 200 kDa by SDS gel electrophoresis. In this study, we observed that extensively phosphorylated NF-H variants exist as both Triton-soluble and - insoluble forms, which display different cellular distributions. Perikarya and neurites of differentiated NB2a/d1 cells were immunostained by a polyclonal antiserum (anti-NF-H) that specifically recognizes the extensively phosphorylated NF-H forms and a monoclonal antibody (SMI-31) that recognizes phosphorylated epitopes of neurofilament proteins (NFPs). When cells were extracted with Triton X-100 to remove soluble proteins, however, only axonal neurites remained immunoreactive. Immunoblot analyses established the specificity of anti-NF-H and SMI-31 and demonstrated that both Triton- soluble and -insoluble NF-H subunits exhibit an apparent molecular weight of 200 kDa. Incorporation of radiolabeled phosphate into Triton-soluble NF-H following incubation of intact NB2a/d1 cells with 32P-orthophosphate confirmed that the Triton-soluble form of NF-H is a phosphoprotein. Most NF-H subunits in the Triton-soluble fraction sedimented after centrifugation at 100,000 g for 1 h, indicating that they may be present as oligomers. The implications of these data for the development of neurofibrillary pathology are discussed. Registry Numbers: 9002-93-1(Octoxynol) Institutional address: Ralph Lowell Laboratories Mailman Research Center McLean Hospital Belmont MA 02178. *****NEUROSCIENCE***** (REFERENCE 57 OF 100)

92066147 Benes FM, Farol PA, Majocha RE, Marotta CA, Bird ED Evidence for axonal loss in regions occupied by senile plaques in Alzheimer cortex. In: Neuroscience (1991) 42(3):651-60 ISSN: 0306-4522 The studies described have sought to determine what, if any, relationship exists between axons and the senile plaque, a hallmark histopathological feature of Alzheimer's disease. A double stain was performed on both early and late Alzheimer frontal cortex tissues in order to examine the interaction between axons stained with antibodies against the 200,000 mol. wt neurofilament subunit (NFP- 200) of the axon cytoskeleton and Thioflavin-S, a fluorescent dye that stains plaques. Serial photomicrographs of plaques were taken and axon and plaque profiles were three-dimensionally reconstructed. Analysis of computer-processed images revealed that there were fewer axons within plaques than in regions lying one and two plaque distances away. When axons were observed passing through plaques, swelling and disruption of normal morphology was frequently present. Statistical analyses of axon counts within and around placques showed a gradient of axon density, with increased numbers occurring at progressive distances from the placque. Similar patterns were seen for early and late stages of the disease. The results of this study indicate that disruption of the axonal cytoskeleton may occur within the regions occupied by plaques. Registry Numbers: 108688-71-7(neurofilament protein H) Registry Numbers: 2390-54-7(thioflavin T) Institutional address: Department of Psychiatry Harvard Medical School Belmont MA. (REFERENCE 58 OF 100) 91270507 Vickers JC, Costa M, Vitadello M, Dahl D, Marotta CA Neurofilament protein-triplet immunoreactivity in distinct subpopulations of peptide-containing neurons in the guinea-pig coeliac ganglion. In: Neuroscience (1990) 39(3):743-59 ISSN: 0306-4522 A battery of polyclonal and monoclonal antibodies raised against the triplet of identified neurofilament protein subunits was used to investigate neurofilament protein immunoreactivity in neurons of the guinea-pig coeliac ganglion. Using optimal conditions of fixation and tissue processing for each antibody we found that only 20% of the

postganglionic sympathetic neurons in the guinea-pig coeliac ganglion contain neurofilament protein-triplet immunoreactivity. Double labelling with neurofilament protein-triplet antibodies raised in different species demonstrated that all of these antibodies labelled the same population of neurons. Double labelling using mouse monoclonal antibodies against neurofilament proteins in combination with rabbit polyclonals to neuronal markers showed that neurofilament protein-triplet immunoreactivity is restricted to specific chemically coded subpopulations of noradrenergic neurons. Approximately 52% of neurons in the ganglion contain neuropeptide Y and are presumed vasomotor neurons projecting to blood vessels in the submucosa of the small intestine. Virtually none of the neuropeptide Y-containing neurons were labelled with neurofilament protein-triplet antibodies. Neurons that contain somatostatin (21%) project to the submucous ganglia of the small intestine. Approximately two-thirds of neurons containing somatostatin are immunoreactive for the neurofilament protein-triplet. The other postganglionic neurons in the ganglion (27%) project to the myenteric plexus of the small intestine and do not contain either neuropeptide Y or somatostatin. Approximately a quarter of these neurons were labelled with neurofilament protein- triplet antibodies. These results suggest that the neurofilament protein-triplet may not be an intrinsic component of the cytoskeleton of all neurons. Furthermore the idea of a chemical coding of neurons should be extended to cytoskeletal proteins. The finding that these neurofilament proteins are confined to specific neuronal subpopulations has important implications for the search for a role of the neurofilament protein-triplet in neurons, for the interpretation of classical neurohistological silver impregnation techniques which appear to stain only neurofilament protein-triplet- containing neurons, as well as for neuropathological conditions that may involve these proteins in disease processes. Registry Numbers: EC 1.14.16.2(Tyrosine 3-Monooxygenase) Registry Numbers: 37221-79-7(Vasoactive Intestinal Peptide) Institutional address: Department of Physiology Flinders University of South Australia Bedford Park. (REFERENCE 59 OF 100) 90265487 Benes FM, Reifel JL, Majocha RE, Marotta CA Evidence for a diffusional model of Alzheimer amyloid A4 (beta- amyloid) deposition during neuritic plaque formation. In: Neuroscience (1989) 33(3):483-8 ISSN: 0306-4522 A recent study reported that Alzheimer senile plaques immunostained with monoclonal antibodies against the A4 (beta-amyloid) region of the amyloid precursor protein show gradients of density (Majocha R. E., Benes F. M., Reifel R. L., Rodenrys A. M. and Marotta C. A., Proc. natn. Acad. Sci. U.S.A. 85, 6182-6186, 1988). Although more

than one explanation was suggested for this observation, the possible involvement of a diffusional process during plaque maturation was considered. In order to examine this hypothesis, specimens from prefrontal cortex, entorhinal area and hippocampal formation were immunoprocessed in a similar fashion and subjected to quantitative microdensitometric analyses of A4 amyloid reaction product. All plaques in the three brain areas examined showed a curvilinear relationship between the area of amyloid reaction product (expressed in pixel counts) and optical density (expressed as each of six grey scale levels). There was an increase in the area of amyloid at progressively lower density levels. When the area of amyloid reaction product at each density level was correlated with the overall size of individual plaques, it was found that there was a striking increase in the correlation coefficients at progressively lower grey scale levels, with r = 0.853 at the lowest level examined. When a second order derivation of these correlations was performed by expressing individual r-values with respect to an optical density index, an asymptotic relationship resulted with the lowest density levels showing an increasingly sharp rise toward unity. These data are consistent overall with a model for plaque maturation that involves diffusion of amyloid protein through the extracellular space from focal regions of high density where synthesis and/or release may occur. Institutional address: Department of Psychiatry Harvard Medical School Boston MA 02115. *****NUCLEIC ACIDS RESEARCH***** (REFERENCE 60 OF 100) 78011677 Wilson JT, deRiel JK, Forget BG, Marotta CA, Weissman SM Nucleotide sequence of 3' untranslated portion of human alpha globin mRNA. In: Nucleic Acids Res (1977 Jul) 4(7):2353-68 ISSN: 0305-1048 We have determined the nucleotide sequence of 75 nucleotides of the 3'-untranslated portion of normal human alpha globin mRNA which corresponds to the elongated amino acid sequence of the chain termination mutant Hb Constant Spring. This was accomplished by sequence analysis of cDNA fragments obtained by restriction endonuclease or T4 endonuclease IV cleavage of human globin cDNA synthesized from globin mRNA by use of viral reverse transcriptase. Analysis of cRNA synthesized from cDNA by use of RNA polymerase provided additional confirmatory sequence information. Possible polymorphism has been identified at one site of the sequence. Our sequence overlaps with, and extends the sequence of 43 nucleotides determined by Proudfood and coworkers for the very 3'-terminal portion of human alpha globin mRNA. The complete 3'-untranslated

sequence of human alpha globin mRNA (112 nucleotides including termination codon) shows little homology to that of the human or rabbit beta globin mRNAs except for the presence of the hexanucleotide sequence AAUAAA which is found in most eukaryotic mRNAs near the 3'-terminal poly (A). Registry Numbers: 9004-22-2(Globins) Registry Numbers: EC 2.7.1.78(Polynucleotide 5'-Hydroxyl-Kinase) *****PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES***** (REFERENCE 61 OF 100) 92357777 Tate B, Aboody-Guterman KS, Morris AM, Walcott EC, Majocha RE, Marotta CA Disruption of circadian regulation by brain grafts that overexpress Alzheimer beta/A4 amyloid. In: Proc Natl Acad Sci U S A (1992 Aug 1) 89(15):7090-4 ISSN: 0027-8424 Alzheimer disease patients exhibit irregularities in the patterns of normally circadian (daily) rhythms. Alzheimer-type pathology has been reported in the hypothalamus and in the suprachiasmatic nuclei, the putative site of the circadian oscillator. We examined the relationship between the neuropathology of Alzheimer disease, as modeled by an animal system, and circadian dysregulation by grafting genetically transformed cells that overexpress beta/A4 amyloid into the suprachiasmatic nuclei of adult rats. Grafts of beta/A4-positive cells, but not of control cells, significantly altered the pattern of activity of implanted rats. Although experimental conditions included light-dark cycles that normally tend to drive rats to 24-h rhythms, animals with grafts of beta/A4-positive cells showed abnormally high levels of activity during the light phase in addition to a disrupted circadian pattern. Periodogram analysis demonstrated significant rhythms outside of a circadian range. The body temperature rhythm of these animals was also weak 6 weeks after grafting; however, unlike activity patterns, body temperature regained a circadian period by 8 weeks after cell implantation. These data indicate that disruption of circadian activity is a behavioral measure of the consequences of beta/A4 accumulation in brain implants. Institutional address: Department of Psychiatry Harvard Medical School Boston MA 02115. (REFERENCE 62 OF 100) 89099008 Tate-Ostroff B, Majocha RE, Marotta CA

Identification of cellular and extracellular sites of amyloid precursor protein extracytoplasmic domain in normal and Alzheimer disease brains. In: Proc Natl Acad Sci U S A (1989 Jan) 86(2):745-9 ISSN: 0027-8424 Information concerning the distribution of various subdomains of the amyloid precursor protein (APP) in brain may illuminate aspects of the normal metabolism of this membrane-associated protein, as well as putative abnormal processing that may occur in Alzheimer disease (AD). We prepared affinity-purified antibody, P2, against an extracytoplasmic APP site and applied it, along with monoclonal antibodies to the beta-peptide, or A4 region, in conjunction with selective cytochemical staining methods, to control and AD tissues. The following was noted: (i) in contrast to A4 epitopes, which are easily demonstrable primarily in extracellular senile plaques of AD patients, the extracytoplasmic P2 antigen was found in association with neurons, glia, and blood vessels in both normal and AD prefrontal cortex; (ii) a subset of senile plaques contained both A4 and P2 antigens; (iii) in some instances, P2 antigen occurred as an extracellular deposit in the absence of A4; (iv) the P2 antigen, but not A4, was also associated with corpora amylacea. In addition to identifying the unique cellular distribution of the APP extracytoplasmic antigen, the results support the view that a segment of this domain undergoes processing and deposition at extracellular sites, including a subset of senile plaques. Institutional address: Department of Psychiatry Harvard Medical School Boston MA 02115. (REFERENCE 63 OF 100) 89098911 Marotta CA, Chou WG, Majocha RE, Watkins R, LaBonne C, Zain SB Overexpression of amyloid precursor protein A4 (beta-amyloid) immunoreactivity in genetically transformed cells: implications for a cellular model of Alzheimer amyloidosis. In: Proc Natl Acad Sci U S A (1989 Jan) 86(1):337-41 ISSN: 0027-8424 Among the major obstacles to clarifying molecular mechanisms involved in amyloid metabolism in Alzheimer disease has been the unavailability of laboratory models for this uniquely human disorder. The present studies were aimed at establishing genetically engineered cell lines that overexpress amyloid immunoreactivity and that may be relevant to amyloid accumulation in the Alzheimer disease brain. We used cloned amyloid cDNA that contains a region encoding A4 (beta- polypeptide) amino acids along with recently developed tumor virus vectors derived from simian virus 40 to prepare transformed cells. After transient and permanent transfection, a variety of cell types

overexpressed A4 immunoreactivity that was detected by highly specific monoclonal antibodies. We observed that the use of an amyloid subdomain containing the A4 region, but lacking the sequence of a Kunitz-type protease inhibitor found in amyloid precursor protein variants, was sufficient to obtain cells that overproduced an A4 epitope. The transformed cells were readily propagated in culture and may provide an experimental medium to elucidate aspects of the molecular pathogenesis of Alzheimer disease. The cellular models may also serve as tools for deriving potentially useful therapeutic agents. Institutional address: Cancer Center University of Rochester Medical School NY 14642. (REFERENCE 64 OF 100) 88124954 Zain SB, Salim M, Chou WG, Sajdel-Sulkowska EM, Majocha RE, Marotta CA Molecular cloning of amyloid cDNA derived from mRNA of the Alzheimer disease brain: coding and noncoding regions of the fetal precursor mRNA are expressed in the cortex. In: Proc Natl Acad Sci U S A (1988 Feb) 85(3):929-33 ISSN: 0027-8424 To gain insight into factors associated with the excessive accumulation of beta-amyloid in the Alzheimer disease (AD) brain, the present studies were initiated to distinguish between a unique primary structure of the AD-specific amyloid precursor mRNA vis a vis other determinants that may affect amyloid levels. Previous molecular cloning experiments focused on amyloid derived from sources other than AD cases. In the present work, we cloned and characterized amyloid cDNA derived directly from AD brain mRNA. Poly(A)+ RNA from AD cortices was used for the preparation of lambda gt11 recombinant cDNA libraries. An insert of 1564 nucleotides was isolated that included the beta-amyloid domain and corresponded to 75% of the coding region and approximately equal to 70% of the 3'-noncoding region of the fetal precursor amyloid cDNA reported by others. On RNA blots, the AD amyloid mRNA consisted of a doublet of 3.2 and 3.4 kilobases. In control and AD cases, the amyloid mRNA levels were nonuniform and were independent of glial-specific mRNA levels. Based on the sequence analysis data, we conclude that a segment of the amyloid gene is expressed in the AD cortex as a high molecular weight precursor mRNA with major coding and 3'-noncoding regions that are identical to the fetal brain gene product. Registry Numbers: 24937-83-5(Poly A) Registry Numbers: 9007-49-2(DNA) Institutional address: Cancer Center University of Rochester Medical School

NY 14642. (REFERENCE 65 OF 100) 88320405 Majocha RE, Benes FM, Reifel JL, Rodenrys AM, Marotta CA Laminar-specific distribution and infrastructural detail of amyloid in the Alzheimer disease cortex visualized by computer-enhanced imaging of epitopes recognized by monoclonal antibodies. In: Proc Natl Acad Sci U S A (1988 Aug) 85(16):6182-6 ISSN: 0027-8424 Monoclonal antibodies to the A4 amyloid polypeptide were used in immunocytochemical staining of the Alzheimer disease prefrontal cortex. Analysis of the resulting staining patterns allowed us to evaluate the amounts and distribution of amyloid-protein deposits exclusive of other senile-plaque components. Previously unappreciated infra-structural details of amyloid in the Alzheimer disease brain became accessible through computer-enhanced imaging procedures. Four discrete morphologic classes of amyloid deposits were observed and classified as punctate, macular, ring, and ring-with-core configurations. Computer imaging indicated that all four classes of immunostained deposits contain internal gradients of density. The classes were nonuniformly distributed with regard to size and location within cortical laminae. Our results support two separate but complementary hypotheses concerning the molecular neuropathology of Alzheimer disease in the prefrontal cortex. (i) Irrespective of cortical layer or morphology, density-gradient analyses suggest that amyloid deposits are elaborated through molecular and cellular events that may involve diffusion or coalescence of the A4 polypeptide. (ii) The distribution and morphology of prefrontal cortical amyloid deposits may be dependent upon underlying laminar-specific structures of the neocortex. Institutional address: Department of Psychiatry Harvard Medical School Boston MA 02115. (REFERENCE 66 OF 100) 76053173 Forget BG, Marotta CA, Weissman SM, Cohen-Solal M Nucleotide sequences of the 3'-terminal untranslated region of messenger RNA for human beta globin chain. In: Proc Natl Acad Sci U S A (1975 Sep) 72(9):3614-8 ISSN: 0027-8424 In normal messenger RNA for the human beta-globin chain, nucleotide sequences have been identified which can be matched to the amino-acid sequence of the abnormally long segment of the beta-chain of hemoglobin Cranston. The finding of these sequences strengthens the

hypothesis that the betaCranston chain arose by a frameshift mutation allowing the "readthrough" of the normal termination codon and translation of usually untranslated portions of the messenger RNA for the beta-globin chain. The oligonucleotides which match the amino- acid sequence of hemoglobin Cranston provide a sequence of 36 nucleotides which follows the normal beta-chain termination codon UAA. (REFERENCE 67 OF 100) 74275150 Marotta CA, Forget BG, Weissman SM, Verma IM, McCaffrey RP, Baltimore D Nucleotide sequences of human globin messenger RNA. In: Proc Natl Acad Sci U S A (1974 Jun) 71(6):2300-4 ISSN: 0027-8424 [No Abstract Available] (REFERENCE 68 OF 100) 77607803 Forget BG, Marotta CA, Weissman SM, Cohensolal M NUCLEOTIDE SEQUENCES OF THE 3'-TERMINAL UNTRANSLATED REGION OF MESSENGER RNA FOR HUMAN BETA GLOBIN CHAIN In: Proc Natl Acad Sci U S A (1975) 72(9):3614-3618 ISSN: 0027-8424 In normal messenger RNA for the human beta-globin chain, nucleotide sequences have been identified which can be matched to the amino-acid sequence of the abnormally long segment of the beta-chain of hemoglobin Cranston. The finding of these sequences strengthens the hypothesis that the beta**Cranston chain arose by a frameshift mutation allowing the 'readthrough' of the normal termination codon and translation of usually untranslated portions of the messenger RNA for the beta-globin chain. The oligonucleotides which match the amino- acid sequence of hemoglobin Cranston provide a sequence of 36 nucleotides which follows the normal beta-chain termination codon UAA. (Author Abstract) Institutional address: The Division of Hematology-Oncology of the Department of Medicine Children'S Hospital Medical Center Boston Massachusetts 02115 *****PROGRESS IN BRAIN RESEARCH***** (REFERENCE 69 OF 100) 87205135

Marotta CA, Majocha RE, Coughlin JF, Manz HJ, Davies P, Ventosa-Michelman M, Chou WG, Zain SB, Sajdel-Sulkowska EM Transcriptional and translational regulatory mechanisms during normal aging of the mammalian brain and in Alzheimer's disease. In: Prog Brain Res (1986) 70:303-20 ISSN: 0079-6123 [No Abstract Available] Registry Numbers: 63231-63-0(RNA) *****PROGRESS IN CLINICAL AND BIOLOGICAL RESEARCH***** (REFERENCE 70 OF 100) 90099547 Chou WG, Zhu W, Salim M, Rehman S, Tate-Ostroff B, Majocha RE, Sajdel-Sulkowska EM, Marotta CA, Zain SB Extracytoplasmic and A4 domains of the amyloid precursor protein: molecular cloning, genetically engineered cell lines and immunocytochemical investigations. In: Prog Clin Biol Res (1989) 317:991-9 ISSN: 0361-7742 [No Abstract Available] Institutional address: Cancer Center and Department of Biochemistry University of Rochester Medical School NY 14642. *****PROGRESS IN NUCLEIC ACID RESEARCH AND MOLECULAR BIOLOGY***** (REFERENCE 71 OF 100) 77126403 Marotta CA, Forget BG, Cohen-Solal M, Weissman SM Nucleotide sequence analysis of coding and noncoding regions of human beta-globin mRNA. In: Prog Nucleic Acid Res Mol Biol (1976) 19:165-75 ISSN: 0079-6603 [No Abstract Available] Registry Numbers: 9004-22-2(Globins) Registry Numbers: 9007-49-2(DNA)

*****SCANNING MICROSCOPY***** (REFERENCE 72 OF 100) 95215822 Maestre GE, Tate BA, Majocha RE, Maguire J, Marotta CA Intercellular interactions in PC12 cells overexpressing beta/A4 amyloid. In: Scanning Microsc (1994) 8(2):325-35; discussion 335-6 ISSN: 0891-7035 The amyloid precursor protein (APP) is an integral membrane component of eukaryotic cells. A variety of research approaches have addressed the contribution of the beta amyloid peptide region of the APP to neuritic plaque structure and formation in the Alzheimer disease brain as well as the relationship between beta amyloid accumulation and the occurrence of dementia. However, there is limited information available concerning the cellular consequences of amyloid deposition. The present studies were undertaken to investigate the relationship between beta amyloid and intercellular junctions. Transfected PC12 cell lines, that overexpress the beta amyloid peptide, exhibit structural and functional alterations at the cell surface and tend to form aggregates more readily than normal control cells. Intermediate junctions were the most common intercellular interactions of both normal and transfected cells. However, the control and transfected cells differed since areas of continuous and extensive junctions were readily seen in transfected cells and infrequently seen in control cells. The data suggest that excess accumulation of beta amyloid is associated with the junctional apparatus and may be related to increased intercellular adhesion. Institutional address: Department of Psychiatry Harvard Medical School Boston MA 02115. *****SCIENCE***** (REFERENCE 73 OF 100) 84300278 Sajdel-Sulkowska EM, Marotta CA Alzheimer's disease brain: alterations in RNA levels and in a ribonuclease-inhibitor complex. In: Science (1984 Aug 31) 225(4665):947-9 ISSN: 0036-8075 A macromolecular alteration occurs at the posttranscriptional level in the Alzheimer's disease (AD) brain. Compared with age-matched controls, total cellular RNA and polyadenylated RNA were substantially reduced in the AD cortex with many neuritic plaques and

neurofibrillary tangles. RNA changes are associated with a significant increase in alkaline ribonuclease activity due to an abnormality in the ribonuclease-inhibitor complex. The decrease in protein synthesis in the AD brain, previously observed in patients severely affected with AD, and in translation systems in vitro with AD cortical messenger RNA, may be partly related to an enzyme- inhibitor alteration that affects RNA levels and activity. Decreased protein synthesis therefore may contribute to the characteristic decline in certain neurotransmitter enzymes and to the loss of neurons in the AD brain. Registry Numbers: EC 3.1.-(Ribonucleases) Registry Numbers: 24937-83-5(Poly A) *****TRANSACTIONS OF THE ASSOCIATION OF AMERICAN PHYSICIANS***** (REFERENCE 74 OF 100) 78160788 Wilson JT, Marotta CA, Forget BG, Weissman SM Structure of human hemoglobin messenger RNA and its relation to hemoglobinopathies. In: Trans Assoc Am Physicians (1977) 90:117-26 ISSN: 0066-9458 1. One-fifth to 1/4 of globin mRNA is untranslated sequence other than polyadenylic acid. 2. The untranslated sequences of mRNA vary markedly in their sequence and in their length. 3. Globin mRNAs demonstrate a marked bias in codon selection. 4. Viral mRNA shows a quite different pattern of codon selection; therefore, the selection of codons is not uniform for all mRNAs functioning in animal cells. 5. Elongated hemoglobin chains can be accounted for by frame-shift mutations, or point mutations within the normal termination codon. The additional amino acids are then coded for by sequences that are normally untranslated. 6. Certain hemoglobin deletion mutants occur at sites where there are partially reiterated sequences within the heomoglobin messenger RNA. Registry Numbers: 56-45-1(Serine) Registry Numbers: 61-90-5(Leucine)

*********************************************************************** OTHER PUBLISHED RESEARCH REPORTS, MONOGRAPHS, and BOOKS 75 Marotta, C.A., Strocchi, P., Brown, B.A., Bonventre, J.A. and Gilbert, J.M.: Gel electrophoresis methods for the examination of human and rat brain fibrous proteins. Application to studies of human brain proteins from postmortem tissue. In: Genetic Research Strategies for Psychobiology and Psychiatry (E.S. Gershon, S. Matthysse, X.O. Breakefield and R.D. Ciaranello, Eds.), Boxwood Press, Pacific Grove, 1981, pp. 39-57 76 Selkoe, D.J., Salazar, F.J., Brown, B.A. and Marotta, C.A.: Characterization of altered neuronal proteins in Alzheimer disease. Trans. Amer. Neurol. Assoc. 105:1-4, 1980 77 Marotta, C.A., Strocchi, P., Brown, B.A. and Gilbert, J.M.: Genetic aspects of mammalian brain macromolecular complexity. In: Psychiatry and the Biology of the Human Brain (S.W. Matthysse, Ed.), Elsevier, Amsterdam, 1981, pp. 71-87 78 Sajdel-Sulkowska, E.M., Coughlin, J.F., Staton, D.M. and Marotta, C.A.: In vitro protein synthesis by messenger RNA from the Alzheimer's disease brain. Banbury Report 15: Biological Aspects of Alzheimer's Disease, Cold Spring Harbor, 1983, pp. 193-200. 79 Marotta, C.A., Ed., Neurofilaments. University of Minnesota Press, Minneapolis, 1983 80 Marotta, C.A.: Some aspects of current and future neurofilament research. In: Neurofilaments (C.A. Marotta, Ed.), University of Minnesota Press, Minneapolis, 1983, pp. 222-229 81 Marotta, C.A.: Neuronal Intermediate Filaments. In: Handbook of Neurochemistry, Vol. 7 (A. Lajtha, Ed.), Plenum, NY, 1984, pp. 281-314 82 Sajdel-Sulkowska, E.M. and Marotta, C.A.: Functional messenger RNA from the postmortem human brain: comparison of aged normal with Alzheimer's

disease. In: Molecular Biology of Aging, Gene Stability and Gene Expression (R.S. Sohal, L. Birnbaum and R.G. Cutler, Eds.), Raven Press, NY, 1985, pp. 243-256 83 Marotta, C.A. Majocha, R.E., Coughlin, J.F., Manz, H.J., Davies, P., Ventosa- Michelman, M., Chu, W.-G., Zain, S.B. and Sajdel-Sulkowska, E.M.: Transcriptional and translational regulatory mechanisms during normal aging of the mammalian brain and in Alzheimer's disease. Prog. Brain Res. 70: 303- 320, 1986 84 Marotta,C.A.: Postmortem human brain: functional messenger RNA. In: Encyclopedia of Neuroscience (G. Adelman, Ed.), Birkhauser, Boston, 1987, p. 148 85 Marotta, C.A.: Both genetic and non-genetic risk factors of Alzheimer's disease must continue to be evaluated. Neurobiol. Aging 7:481-482, 1987 86 Marotta C.A.: Alzheimer's disease brain investigated by postmortem RNA studies. In: Discussions in Neurosciences: Molecular Mechanisms of Pathogenesis of the Central Nervous System Disorders, FESN, III:83-88, 1986 87 Benes, F.M., Matthysse, S.W., Bird, E.D., Marotta, C.A. and Davidson, J.: Formulating strategies for studying cerebral cortical cytoarchitecture in schizophrenia. Biological Psychiatry (Proceedings of the IV World Congress on Biological Psychiatry), Elsevier, Amsterdam, 1986, pp. 1021-1023 88 Chou, W.-G., Majocha, R.E., Sajdel-Sulkowska, E.M., Benes, F.M., Salim, M., Stoler, M., Fulwiler, C.E., Ventosa-Michelman, M., Rodenrys, A.M., Moore, K.E., Webb, T., Zain, S.B. and Marotta, C.A.: Immunologic and molecular genetic studies on amyloid deposition in the Alzheimer's disease brain. Brain Dysfunction 1:133- 145, 1988 89 Salim, M., Zain, S.B., Chou, W.-G., Sajdel-Sulkowska, E.M., Majocha, R.E., Rehman, S., Benes, F.M. and Marotta, C.A.: Molecular cloning of amyloid cDNA from Alzheimer brain messenger RNA. Correlative neuro- immunologic and in situ hybridization studies. In: Familial Alzheimer's Disease: Molecular Genetics, Clinical Prospects and Societal Issues (J.P. Blass, G.D. Miner, L.A. Miner, R.W. Richter and J.L. Valentine, Eds.), Marcel Dekker, NY, 1988, pp. 153-165

90 Zain, S.B., Majocha, R.E., Chou, W.-G., Sajdel-Sulkowska, E.M., Salim, M., Tate-Ostroff, B., Rehman, S., Ventosa-Michelman, M., Zhu, W., Labonne, C., Ramakrishnan, A., Walcott, E., and Marotta, C.A.: Alzheimer amyloid: epitopic distribution, postmortem molecular cloning, and development of a cellular model for amyloidosis. Proceedings of the World Congress of Gerontology, Fourteenth Meeting, International Association of Gerontology, 1989 91 Sajdel-Sulkowska, E.M., Manz, H.J. and Marotta, C.A.: Postmortem stability of RNA metabolism in human brain: studies of the non-demented control and Alzheimer cases. In: Novel Approaches to the Treatment of Alzheimer's Disease. (E.M. Meyer, Ed.), Plenum, NY, 1989, pp. 349-356 92 Marotta, C.A.: Molecular and cellular biology of Alzheimer amyloid. J. Mol. Neurosci. 3: 111-125 1992 93 Tate, B.A., Majocha, R.E., Maestre, G.E., and Marotta, C.A.: Biological and behavioral alterations induced by ß/A4-amyloid. Bull. Clin. Neurosci. 56: 131-139, 1991 94 Marotta, C.A.: Alzheimer amyloid: models, membranes and memory. Proceedings of the American Neuropsychiatric Association, Sixth Annual Meeting, pp. 12-13, 1994 95 Marotta, C.A.: Molecular strategies for the treatment of Alzheimer's disease. In: Alzheimer's Disease. A Guide to Practical Management. (R. Richter and J.P.Blass, Eds.), Mosby, New York, Vol. IV, pp. 203-207, 1996 96 Binsack, D.P., Agrawal, S., and Marotta, C.A.: Behavioral assessment of antisense oligonucleotides targeted to messenger RNAs of genes associated with Alzheimer's disease. In: Application of Antisense Strategies for Investigation of Receptor Mechanisms In Vivo and In Vitro. (R.B. Raffa, Ed.), Landes, New York, 1996 *********************************************************************** DOCTORAL DISSERTAION, YALE UNIVERSITY 97 Marotta, C.A.: Sequence analysis of human globin messenger RNA. Doctoral Dissertation, Yale University (University Microfilms), 1975.

*********************************************************************** In preparation for publication: 98 Banks, J.L., Marotta, C.A.: Outcomes Validity and Reliability of the Modified Rankin Scale. Implications for Stroke Clinical Trials. A Literature Review and Synthesis 99 Banks, J.L., Salas, M., Marotta, C.A.: The National Institutes of Health Stroke Scale: Reliable Predictor of Disability and Mortality in Patients with Stroke 100 Banks, J.L., Salas, M., Marotta, C.A , et al: Meta-Analysis of Stroke State Utilities by Severity, Utility Elicitation Method and Sample Characteristics *********************************************************************** ABSTRACTS: >135 REPORTS

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