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Biomedical subjects

J W Jacobson

Publications and source records attributed to J W Jacobson.

At least 55 records · Page 3Linked to original sources

Problem behavior and psychiatric impairment within a developmentally disabled population I: behavior frequency.

This report summarizes information about problem behavior and psychiatric impairment in a population of over 30,000 individuals receiving developmental disabilities services. Teenage and adult individuals displayed more problem behaviors than young children or the elderly, more markedly mentally retarded individuals were more likely to display problem behavior, individuals in less restrictive residential settings displayed less problem behavior than those in more restrictive settings, and persons with psychiatric impairment displayed more frequent problems than persons without psychiatric impairment. The proportion of developmentally disabled persons with a psychiatric impairment was positively associated with presence of problem behaviors classified as affective or cognitive. Implications for community placement and further research are addressed.

Adolescent↗

The character of developmental disabilities in New York State: preliminary observations.

Preliminary information about the population characteristics of developmentally disabled persons in the State of New York is presented. The information was derived through a "rate-under-treatment" survey approach using the Developmental Disabilities Information Survey (DDIS). The results are based upon statewide data, derived from 36,334 cases, composed of 34% children and 66% adults. The findings revealed that the subject populations socio-demographics were consistent with those of the general population in the areas of age and ethnic distribution, but not in gender ratio. Developmentally disabled persons were located primarily in community settings; only 24% of the children and 45% of the adults were found to reside within various institutional settings. Most were mentally retarded, however more children than adults were identified as having a secondary condition such as autism, cerebral palsy, epilepsy or some other neurological impairment. About half were reported to also have some type of physical disability. The majority were free of ambulation difficulties. Only half had basic self-care skills, and a third were facile in communication skills. Learning capabilities were found to be affected by maturation and experience. It was suggested that a "rate-under-treatment" approach is effective, within certain constraints, in capturing reliable information about a population that could be used for planning and developing public policies.

Activities of Daily Living↗

Projected impact of the functional definition of developmental disabilities: the categorically disabled population and service eligibility.

Indicators of substantial limitation in seven life activity areas defined in P.L. 95-602 were generated from a comprehensive needs assessment/screening instrument used in New York State. These indicators were then applied to a data base of over 35,000 individuals having one or more of the categorical developmental disabilities of autism, cerebral palsy, epilepsy, mental retardation, or other neurological impairments. Results of this analysis suggest that potential inclusion within the functional developmental disabilities definition of P.L. 95-602 varies as a function of categorical disability, age, and level of intellectual functioning. Implications of these findings for access to services were discussed.

Activities of Daily Living↗

Expression in Escherichia coli K-12 of the structural gene for catabolic dehydroquinase of Neurospora crassa.

The inducible quinic acid catabolic pathway of Neurospora crassa is controlled by four genes, the qa cluster which includes structural genes qa-2, qa-3, qa-4 for three enzymes and a regulatory gene, qa-1. In this paper we report the molecular cloning of at least the qa-2 gene which encodes the catabolic dehydroquinase (5-dehydroquinate hydro-lyase, EC 4.2.1.10). Endo.R.HindIII restriction endonuclease fragments of N. crassa DNA from a qa-1(c) (constitutive) mutant and of Escherichia coli plasmid pBR322 DNA were ligated in vitro and used to transform an aroD6 (5-dehydroquinate hydrolyase deficient) strain of E. coli K12. The recombinant plasmid (pVK55) isolated from one AroD(+) transformant (SK1518) contained, in addition to pBR322, two N. crassa HindIII fragments with molecular weights of 2.3 x 10(6) and 1.9 x 10(6). Derivatives of SK1518 cured of plasmid DNA were phenotypically Amp(s) and AroD(-). These cured strains, retransformed with pVK55, were phenotypically Amp(R) and AroD(+). Strains transformed with pVK55 possessed 5-dehydroquinate hydrolyase activity but no activity was present in any AroD(-) strain. The enzyme extracted from strains containing the recombinant plasmid was identical to N. crassa catabolic dehydroquinase by the criteria of heat stability, ammonium sulfate fractionation, immunological crossreactivity, molecular weight, and purification characteristics. This identity demonstrates that the N. crassa qa-2(+) gene is carried by the recombinant plasmid and is apparently transcribed and translated with complete fidelity. Furthermore, subunit assembly of the N. crassa polypeptides also occurs in E. coli, because the catabolic dehydroquinase is a multimer composed of approximately 20 identical subunits.

DNA Restriction Enzymes↗

Isolation and characterization of nuclei from Neurospora crassa.

A procedure was developed for isolating nuclei from either the conidial or germinated conidial growth phase of Neurospora crassa. A frozen conidial suspension was lysed by passage through a French pressure cell, and the nuclei were freed from the broken cells by repeated homogenization in an Omni-Mixer. Pure nuclei were obtained from the crude nuclear fraction by density banding in a Ludox gradient. The final nuclear yield was 20 to 30%. The nuclei had a deoxyribonucleic acid (DNA):ribonucleic acid (RNA):protein ratio of 1:3.5:7 and were active in RNA synthesis. The nuclei, stained with the DNA stain 4,6-diamidino-2-phenylindole, appeared under fluorescence microscopy as bright blue spheres, 1 micron in diameter, essentially free from cytoplasmic attachments. Chromatin extracted from the nuclei in a 70 to 75% yield by dissociation with 2 M sodium chloride and 5 M urea had a DNA:RNA:protein ratio of 1:1.05:1.7. Chromatin reconstituted from this preparation exhibited a level of RNA polymerase template activity lower than that of pure Neurospora DNA, but the maximum level of reconstitution obtained was only 10%. Fractionation of Neurospora chromatin on hydroxylapatite separated the histones from the chromatin acidic proteins. The normal complement of histone proteins was present in both the reconstituted and dissociated chromatin preparations. The acidic protein fraction exhibited a variety of bands on sodium dodecyl sulfate gel electrophoresis ranging in molecular weight from 15,000 to 70,000. The gel pattern was much more complex for total dissociated chromatin than for reconstituted chromatin.

Cell Nucleus↗

Characterization and in vitro translation of polyadenylated messenger ribonucleic acid from Neurospora crassa.

Ribonucleic acid (RNA) extracted from Neurospora crassa has been fractionated by oligodeoxythymidylic acid [oligo(dT)]-cellulose chromatography into polyadenylated messenger RNA [poly(A) mRNA] and unbound RNA. The poly(A) mRNA, which comprises approximately 1.7% of the total cellular RNA, was further characterized by Sepharose 4B chromatography and polyacrylamide gel electrophoresis. Both techniques showed that the poly(A) mRNA was heterodisperse in size, with an average molecular weight similar to that of 17S ribosomal RNA (rRNA). The poly(A) segments isolated from the poly(A) mRNA were relatively short, with three major size classes of 30, 55, and 70 nucleotides. Gel electrophoresis of the non-poly(A) RNA indicated that it contained primarily rRNA and 4S RNA. The optimal conditions were determined for the translation of Neurospora mRNA in a cell-free wheat germ protein-synthesizing system. Poly(A) mRNA stimulated the incorporation of [14C]leucine into polypeptides ranging in size from 10,000 to 100,000 daltons. The RNA that did not bind to oligo(dT)-cellulose also stimulated the incorporation of [14C]leucine, indicating that this fraction contains a significant concentration of mRNA which has either no poly(A) or very short poly(A) segments. In addition, the translation of both poly(A) mRNA and unbound mRNA was inhibited by 7-methylguanosine-5'-monophosphate (m7G5'p). This is preliminary evidence for the existence of a 5'-RNA "cap" on Neurospora mRNA.

Chromatography, Agarose↗

Purification and characterization of catabolic dehydroquinase, an enzyme in the inducible quinic acid catabolic pathway of Neurospora crassa.

Catabolic dehydroquinase which functions in the inducible quinic acid catabolic pathway in Neurospora crassa has been purified 8000-fold. The enzyme was purified by two methods. One used heat denaturation of contaminating proteins; the other used antibody affinity chromatography. The preparations obtained by these two methods were identical by all criteria. The purified enzyme is extremely resistant to thermal denaturation as well as denaturation 0y urea and guanidine hydrochloride at 25 degrees. It is irreversibly inactivated, although not efficiently dissociated, by sodium dodecyl sulfate and guanidine hydrochloride at 55 degrees. At pH 3.0, the enzyme is reversibly dissociated into inactive subunits. At high concentrations catabolic dehydroquinase aggregates into an inactive, high molecular weight complex. The native enzyme, which has a very high specific activity, has a molecular weight of approximately 220,000 and is composed of identical subunits of 8,000 to 12,000 molecular weight each. The native enzyme and the subunit are both asymmetric.

Amino Acids↗

Effect of mutations in the qa gene cluster of Neurospora crassa on the enzyme catabolic dehydroquinase.

Catabolic dehydroquinase, which functions in the inducible quinic acid catabolic pathway of Neurospora crassa, has been purified from wild type (74-A) and three mutants in the qa gene cluster. The mutant strains were: 105c, a temperature-sensitive constitutive mutant in the qa-1 regulatory locus; M-16, a qa-3 mutant deficient in quinate dehydrogenase activity; and 237, a leaky qa-2 mutant which possess very low levels of catabolic dehydroquinase activity. The enzymes purified from strains 74-A, 105c, and M-16 are identical with respect to behavior during purification, specific activity, electrophoretic behavior, stability, molecular weight, subunit structure, immunological cross-reactivity, and amino acid content. The mutant enzyme from strain 237 is 1,500-fold less active and appears to have a slightly different amino acid content. It is identical by a number of the other criteria listed above and is presumed to be a mutant at or near the enzyme active site. These data demonstrate that the qa-1 gene product is not involved in the posttranslational expression of enzyme activity. The biochemical identity of catabolic dehydroquinase isolated from strains 105c and M-16 with that from wild type also demonstrates that neither the inducer, quinic acid, nor other enzymes encoded in the qa gene cluster are necessary for the expression of activity. Therefore the combined genetic and biochemical data on the qa system continue to support the hypothesis that the qa-1 regulatory protein acts as a positive initiator of qa enzyme synthesis.

Alcohol Oxidoreductases↗