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An Arabidopisis cDNA clone was isolated that encodes a protein similar to the antifungal chitin-binding protein hevein from rubber tree latex. This hevein-like (HEL) mRNA was inducible by either turnip crinkle virus infection or ethylene treatment. In addition, expression was moderately inducible by treatment with the resistance-inducing compounds salicylic acid and 2,6-dichlorisonicotinic acid. The 786-bp cDNA contains an open reading frame of 212 codons. The deduced amino acid sequence contains a putative signal sequence of 21 amino acids followed by a 43-amino-acid cysteine-rich lectin domain and a 129-amino-acid carboxy-terminal domain. The predicted protein is approximately 70% identical to hevein, to the wound-inducible WIN1 and WIN2 proteins from potato, and to PR-4, a pathogenesis-related protein from tobacco.

Data from the Partnership Attitude Tracking Study are analyzed to provide a greater understanding of the contributing factors to alcohol and other drug use among children and adolescents. The data were collected from children (ages 9 to 12) and teenagers (ages 13 to 17) in central locations (usually shopping malls) located in 100 primary sampling units across the United States. The analysis, which primarily employed stepwise multiple regression for model estimation, reinforced the importance of friends use, perceived risk, and tobacco and alcohol use in predicting marijuana use. An exposure model, which included antidrug advertisements, was a particularly powerful model. Implications of the findings are discussed.

The prevalence of smoking amongst a group of 77 diabetic teenagers aged 11-18 years attending two paediatric clinics was determined using a questionnaire and urine cotinine assay. Five individuals were identified as definite smokers with two further individuals being probable smokers, all of whom were aged 15 years or more. This low prevalence (9%) is in contrast to a 48% prevalence in a young adult diabetic clinic. Many diabetic teenagers appear to acquire a regular smoking habit after leaving the paediatric clinic so it is important that health education is targeted at this group.

The influence of exposure to tobacco smoke on maternal serum alpha-fetoprotein (AFP) levels at 16 wk gestation was examined. Urinary cotinine levels were used to quantify exposure to tobacco smoke. Significantly higher levels of maternal serum AFP were found in 101 women who had more than 1.0 microgram cotinine/mg urinary creatinine compared with 180 women whose urinary cotinine levels were below this level [(mean +/- SD) 1.23 +/- 0.64 and 1.06 +/- 0.54 respectively; 95 per cent CI of difference of means 0.01-0.31; P < 0.05]. There was a mild albeit statistically significant correlation between urinary cotinine levels and maternal serum AFB (r 0.099; P < 0.05). However, the difference in maternal serum AFP levels between the two groups was not found to be significant, when adjustments for maternal body mass index were made.

Complementary DNA clones encoding acidic and basic isoforms of the class III chitinase were isolated from Nicotiana tabacum. The clones share ca. 65% identity, are equally homologous to the class III chitinases from cucumber and Arabidopsis, and are members of small gene families in tobacco. An acidic class III chitinase was purified from the intercellular fluid of tobacco leaves infected with tobacco mosaic virus (TMV). Partial amino acid sequencing of the protein confirmed that it was encoded by one of the cDNA clones. The mRNAs of the class III chitinases are coordinately expressed in response to TMV infection, both in infected and uninfected tissue. The acidic and basic class III chitinases constitute previously undescribed pathogenesis-related proteins in tobacco.

The programmed senescence of carnation (Dianthus caryophyllus L.) petals requires active gene expression and is associated with the expression of several senescence-related mRNAs. Expression of the mRNA represented by the cDNA clone pSR12 has previously been shown to be transcriptionally activated by ethylene specifically in senescing flowers. We report in this paper the structural analysis of this cDNA and its corresponding gene. One cloned genomic DNA fragment, SR12-B, contained the entire transcription unit in 17 exons, interrupted by 16 introns. A second gene, SR12-A, was highly homologous to SR12-B with several nucleotide substitutions and a 489 bp deletion in the 5' flanking DNA sequence. The SR12 transcript has an open reading frame of 2193 bp sufficient to encode a protein of 82.8 kDa. No significant homology at the DNA or protein levels was found with other known genes. We have identified a DNA-binding factor which specifically interacts with two upstream fragments (-149 to -337 and -688 to -1055) of SR12-B. Both fragments apparently compete for the same binding factor. The DNA-binding activity was present in nuclear extracts from both presenescent and senescing carnation petals. The upstream DNA fragments that bind this factor have sequence homology with promoter sequences of other ethylene-regulated genes.

Ethylene plays a regulatory role in carnation (Dianthus caryophyllus L.) flower senescence. Petal senescence coincides with a burst of ethylene production, is induced prematurely in response to exogenous ethylene, and is delayed by inhibitors of ethylene biosynthesis or action. We have investigated the role of ethylene in the regulation of three senescence-related cDNA clones isolated from a senescent carnation petal library (KA Lawton et al. [1989] Plant Physiol 90: 690-696). Expression of two of the cloned mRNAs in response to ethylene is floral specific, while the expression of another mRNA can be induced in both leaves and flowers exposed to ethylene. Although ethylene induces expression of these mRNAs in petals, message abundance decreases when flowers are removed from ethylene unless an autoenhancement of ethylene production is induced. This indicates continued perception of ethylene is required for their expression. Interruption of ethylene action following the onset of natural senescence results in a substantial decrease in transcript abundance of two of these mRNAs. However, the abundance of another mRNA remains unaffected, indicating this gene responds to temporal cues as well as to ethylene. As flowers age the dosage of exogenous ethylene required to induce expression of the cloned mRNAs decreases, indicating sensitivity to ethylene changes as the tissue matures. Nuclear run-on transcription experiments indicate that relative transcription rates of cloned mRNAs increase in response to exogenous ethylene.

The smoking habits of 300 consecutive pregnant women and of other members of their household were obtained by interview at 16 and 32 weeks' gestation and after delivery. Urine samples for estimation of cotinine were collected at the same times during pregnancy and on admission in labour. Both self-reports of active maternal smoking and urinary cotinine levels were significantly associated with lighter babies but urinary cotinine correlated the better. Active maternal smoking was associated with a decrease in birth-weight of 12 g for every cigarette smoked in a day while there was a decrease of 25 g in birth-weight for every microgram of cotinine/mg of urinary creatinine. Passive maternal smoking was associated with a decrease of 66 g in mean birth-weight but this decrease was not statistically significant. Self-reports of active and passive smoking accounted for 43% of the variation in urinary cotinine levels suggesting that urinary cotinine assays can be used to assess and monitor exposure to tobacco smoke in pregnancy.

Biochemical kinetic analyses of the ATP stimulation of the A2 form of calf DNA alpha-polymerase show that when DNA or primer termini are the variable substrates, maximum reaction velocity is independent of ATP concentration. When dNTP concentration is the variable substrate, the apparent Km is invariant with ATP. Such results indicate that the increase in the synthetic rate caused by ATP results from an improvement in synthesis initiation at primer termini. The effect of ATP on the DNA binding affinity of alpha-A2-polymerase was examined by using column chromatography. Passage of the polymerase through native DNA-cellulose at 70 mM ionic strength resulted in 40% binding of the enzyme. In the presence of 4 mM ATP, binding increased to 80%. In both cases, the bound polymerase could be eluted by a 370 mM ionic strength wash. An elution profile similar to that observed in the absence of ATP was obtained with 0.1 mM ATP, 4 mM GTP, or 4 mM each of the nonhydrolyzable ATP analogues adenyl-5'-yl imidodiphosphate or adenosine 5'-O-(3-thiotriphosphate). These results suggest that hydrolysis of the gamma-phosphate occurs at millimolar levels of ATP and leads to a higher affinity of polymerase for DNA. To distinguish the effects of ATP on RNA priming from those on DNA synthesis, products synthesized processively by alpha-A2-polymerase were sized by gel filtration. Results indicate that essentially all products made on a gapped fd replicative form template in the presence of four dNTPs and 4 mM ATP result from the extension of preexisting DNA primers.(ABSTRACT TRUNCATED AT 250 WORDS)

A 70-year-old man developed severe immune intravascular hemolysis and renal failure following ingestion of fenoprofen, a nonsteroidal, anti-inflammatory drug. The patient's red blood cells were sensitized with both IgG and C3d. The serum reacted with normal red blood cells in the presence and absence of the drug. Addition of albumin to the serum inhibited the reactivity with both neat and drug-treated serum. These atypical serologic findings for drug-related immune hemolytic anemia were explained by (1) the measurement of fenoprofen by high performance liquid chromatography (HPLC) in the neat serum; and (2) solid-phase adsorption studies showing that albumin can bind drug, leading to the disappearance of agglutination when albumin is added. This case demonstrates the utility of drug levels and adsorption techniques to confirm the diagnosis of drug-induced immune hemolytic anemia despite the anomalous serologic results which obscured the diagnosis and management of the patient.

A panel of monoclonal antibodies raised against the hemagglutinin-neuraminidase glycoprotein (HN) of the Australia-Victoria strain of Newcastle disease virus has been used to compare that strain and eight other strains of the virus. The ability of the antibodies to neutralize infectivity, inhibit hemagglutination and neuraminidase, and bind to purified virions in solid-phase radioimmunoassays was determined for each strain. Of the four antigenic sites delineated by these antibodies on the HN of the homologous strain, site 1 (that with the greatest neutralizing susceptibility), is apparently conserved in all the strains tested as revealed by neutralization assays. The least neutralizing site, number 4, is also conserved in most of the strains tested. Site 2, which lies at or near the neuraminidase site, appears to be conserved in the avirulent strains but not in the virulent strains. An antibody to site 3 is unable to bind to a significant extent to any of the heterologous strains tested, and thus recognizes a strain-specific epitope. Inhibition of hemagglutination and neuraminidase by antibodies to each site were also examined and the results suggest that antibodies to sites 1 and 2 may distinguish virulent and avirulent strains at least with respect to these functions.

The senescence of carnation (Dianthus caryophyllus L.) flower petals is associated with increased production of ethylene which plays an important role in regulating this developmental event. Three senescence-related cDNA clones were isolated from a cDNA library prepared from mRNA isolated from senescing petals. These cDNAs are representative of two classes of mRNAs which increase in abundance in senescing petal tissue. The mRNA for one class is present at low levels during the early stages of development and begins to accumulate in mature petals prior to the increase in ethylene production. The accumulation of this mRNA is reduced, but not eliminated, in petals treated with aminooxyacetic acid, an inhibitor of ethylene biosynthesis, or silver thiosulfate, an ethylene action inhibitor. In contrast, expression of the second class of mRNAs appears to be highly regulated by ethylene. These mRNAs are not detectable prior to the rise in ethylene production and increase in abundance in parallel with the ethylene climacteric. Furthermore, expression of these mRNAs is significantly inhibited by both aminooxyacetic acid and silver thiosulfate. Expression of these mRNAs in vegetative and floral organs was limited to floral tissue, and predominantly to senescing petals.

Exposure of carnation (Dianthus caryophyllus L.) flowers to ethylene evokes the developmental program of petal senescence. The temporal relationship of several aspects of this developmental program following treatment with ethylene was investigated. Exposure of mature, presenescent flowers to 7.5 microliters per liter ethylene for at least 6 hours induced petal in-rolling and premature senescence. Autocatalytic ethylene production was induced in petals following treatment with ethylene for 12 or more hours. A number of changes in mRNA populations were noted in response to ethylene, as determined by in vitro translation of petal polyadenylated RNA. At least 6 mRNAs accumulated following ethylene exposure. The molecular weights of their in vitro translation products were 81, 58, 42, 38, 35, and 25 kilodaltons. Significant increases in abundance of most mRNAs were observed 3 hours following ethylene exposure. Ethylene exposure resulted in decreased abundance of another group of mRNAs. Treatment of flowers with competitive inhibitors of ethylene action largely prevented the induction of these ethylene responses in petals. An increase in flower age was accompanied by an increase in the capacity for ethylene to induce petal in-rolling, autocatalytic ethylene production, and changes in mRNA populations suggesting that these responses are regulated by both sensitivity to ethylene and ethylene concentration. These results indicate that changes in petal physiology resulting from exposure to ethylene may be the result of rapid changes in gene expression.

The various clinical types of depression may correspond to biochemically distinct forms of the condition. If these could be characterized, antidepressant treatment might be tailored to correction of the underlying biochemical change(s) in individual patients. In particular, the selective actions of clomipramine and maprotiline on the serotonergic and noradrenergic systems respectively might be exploited therapeutically. Studies of the clinical response to these two antidepressants included tests of urinary 5-hydroxy-indoleacetic acid (5-HIAA) excretion and of serum thyroxine levels. While thyroid stimulation tests may prove of value for investigating depressed patients, urinary 5-HIAA levels are inconvenient, subject to extraneous influences and hard to interpret in terms of brain chemistry. (An alternative is proposed in the next paper.)