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Biomedical subjects

M Rabinowitz

Publications and source records attributed to M Rabinowitz.

At least 37 records · Page 2Linked to original sources

Temperature sensitive pet mutants in yeast Saccharomyces cerevisiae that lose mitochondrial RNA.

This is a description of a new class of temperature sensitive pet mutants in Saccharomyces cereviase that lose all or part of their mitochondrial RNA at the restrictive temperature. These mutants fall into 8 different complementation groups, mna1 to mna8, and 2 different classes based on their phenotype. Class I mutations, mna1-1 through mna5-1, cause complete or partial loss of mitochondrial RNA at the restrictive temperature. The mutation, mna1-1, is especially interesting since it causes a loss of both mitochondrial DNA and RNA when the mutant is grown on a fermentable carbon source at the restrictive temperature. However, when this mutant is grown at the permissive temperature on a non-fermentable carbon source then shifted to the restrictive temperature, only the mitochondrial RNA is lost. This indicates that the primary cause for the pet phenotype is due to the loss of mitochondrial RNA and not DNA. Class II mutations, mna6-1 through mna8-1, cause complete loss of the 14S rRNA after growth at the restrictive temperature in a fermentable carbon source. This loss appears to be specific for the 14S rRNA, since all other transcripts probed by Northern analysis are normal.

DNA, Mitochondrial

A modified, solid phase radioimmunoassay for the differential diagnosis of acute and convalescent phases of hepatitis A infection.

The commercial assays for diagnosing the presence of hepatitis A antibodies (HAVAB; Abbott Laboratories, North Chicago, IL) or the presence of IgM class anti-hepatitis A virus antibodies (HAVAB-M; Abbott) do not provide precise information as to the timing of the acute infection. IgM class antibodies are detected as late as six months after the acute infection. In this study the authors describe a modified HAVAB test that inactivates the IgM class antibodies. It thus measures the proportion of IgG antibodies out of the total anti-hepatitis A virus antibodies. In a study of 139 patients with impaired liver function, the available and modified tests showed good agreement except for the convalescent phase of hepatitis A. During serial testing for three months after the acute infection, the commercial tests continuously detected IgM class antibodies. The modified test detected predominantly IgG class antibodies from four weeks on. By six weeks, 85% of the patients had predominantly IgG class antibodies. The modified test thus provides information on the timing of recent hepatitis A infection.

Acute Disease

Pregnancy hypertension, blood pressure during labor, and blood lead levels.

Pregnancy hypertension, blood pressure during labor, and the umbilical cord blood lead concentration were assessed in 3851 women for whom additional demographic, medical, and personal information was available. Lead levels correlated with both systolic (Pearson r = 0.081, p = 0.0001) and diastolic (r = 0.051, p = 0.002) blood pressures during labor. The incidence of pregnancy hypertension increased with lead level. Multivariate models of pregnancy hypertension and systolic blood pressure as a function of maternal age, parity, hematocrit, ponderal index, race, and diabetes were improved by including lead as a predictor variable. At these observed levels of exposure (mean blood lead, 6.9 +/- 3.3 [SD] micrograms/dl), lead appears to have a small but demonstrable association with pregnancy hypertension and blood pressure at the time of delivery, but not with preeclampsia.

Adult

Correlates of low-level lead exposure in urban children at 2 years of age.

The blood lead levels of a large number of US preschool children approach the value regarded as the upper limit of normal. To reduce the number of children whose levels increase into the range thought to be toxic, the antecedents and correlates of levels in the 0- to 25-micrograms/dL range must be identified. In a large longitudinal study of middle and upper-middle class children living in metropolitan Boston, we evaluated how well five sets of variables predicted children's blood lead levels at 2 years of age: environmental lead sources, mouthing activity, home environment/care giving, prior developmental status, and sociodemographic characteristics. A series of bivariate and multivariate analyses indicated that only environmental lead sources and, to a lesser extent, mouthing activity accounted for significant portions of the variance in blood lead levels. Environmental lead sources were not significantly related to the home environment/care-giving variables or to sociodemographic characteristics. The most promising approach for achieving community-wide reductions in children's blood lead levels is reduction in the amount of lead in the proximate environment.

Analysis of Variance

Expression of myosin heavy chains during thyroid hormone-induced cardiac growth.

The expression of mRNAs for two cardiac myosins has been studied in the ventricles of hypo- and hyperthyroid rabbits by using cloned cDNA sequences corresponding to the mRNAs of the alpha- and beta-myosin heavy chains (HCs). The temporal change in relative levels of the alpha and beta HC mRNAs after triiodothyronine (T3) treatment of hypothyroid rabbits was determined by nuclease S1 mapping. In the hypothyroid state, only NC beta-mRNA was expressed in the ventricles. The HC alpha-mRNA was first detectable 4 h after administration of T3 (200 micrograms/kg) to hypothyroid animals. By 12, 24, and 72 h, HC alpha-mRNA represented 20, 50, and 90% of total myosin mRNA. The relationship between the relative mRNA levels and relative synthesis rates of myosin HCs was evaluated in 5- to 6-wk-old normal and thyrotoxic rabbits. Myosin synthesis rates were determined by labeling of protein in vivo with [2H]leucine. The V1 (HC alpha) and V3 (HC beta) isomyosins were separated by immune affinity chromatography and the HCs were isolated electrophoretically. In a normal euthyroid group of animals and in animals 12 and 24 h after administration of 200 micrograms of thyroxine, the relative mRNA levels and relative synthesis rates of the alpha and beta HCs were not significantly different. Our results show that, first, thyroid hormone causes a rapid accumulation of HC alpha-mRNA and loss of HC alpha-mRNA, and second, in normal and thyrotoxic rabbits, the relative synthesis rates of HC alpha and HC beta reflect the relative abundance of their respective mRNAs. These data are consistent with the thyroid hormones regulating synthesis of ventricular myosin at steps that precede translation of its message.

Animals

Environmental correlates of infant blood lead levels in Boston.

From a blood lead survey of 11,837 births, 249 newborns were enrolled in a 2-year, longitudinal study. Their blood leads (PbB) were measured semiannually, and their homes were visited for repeated collections of dust, soil, indoor air, tap water, and paint. Recent refinishing activity and the sizes of nearby streets were recorded. Overall mean PbB was 7.2 micrograms/dl (SD = 5.3) at birth. PbB did not vary systematically with age. Each subject's average postnatal PbB correlated highly with the amount of lead in dust (r = 0.4, P less than 0.0001) and soil (r = 0.3, P less than 0.001), and with the lead in paint (r = 0.2, P less than 0.01). Dust, soil and air lead levels correlated with one another. Refinishing activity in the presence of lead paint was associated with elevations of PbB. Water lead, proximate traffic, weight of recovered dust, race, maternal age and education, and sex were not predictive of PbB. Multivariate models of PbB were constructed that become increasingly predictive with age (r2 = 20 to 37%). Indoor dust lead, lead in soil, refinishing activity, and season were the independent variables.

Air Pollutants

Characterization of a yeast mitochondrial promoter by deletion mutagenesis.

We have generated collections of mutants of the promoter for the small rRNA gene from the mitochondria of yeast deleted from either the 3' or 5' end. Plasmids containing the partially deleted promoter were assayed for their ability to direct correct transcriptional initiation in a homologous in vitro system. We find that the region required for high-efficiency promoter function lies between positions -10 and +2. Our methods detected no effect of flanking sequences on the strength of this promoter.

Base Sequence

Nuclear-cytoplasmic interactions affecting DNA synthesis during induced cardiac muscle growth in the rat.

Nuclear-cytoplasmic interactions affecting DNA synthesis during induced cardiac muscle growth were examined in 29 to 46 day old rats. DNA synthesis was examined in vitro using isolated nuclei from rat heart and adult X. laevis spleen. Cytoplasmic extract (CE) was obtained from a 105 000 g supernatant of rat heart and fetal liver homogenates. To measure DNA synthesis we utilised DNA within the isolated quiescent nucleus as the template and measured the effect of CE on the incorporation of 3H-TTP into an acid precipitable product. In a homologous system of rat heart nuclei from weanling rats and CE from cardiac muscle undergoing induced growth, no stimulation of 3H-TTP incorporation was observed. Cardiac muscle CE however, did possess stimulatory factor(s) since quiescent X. laevis nuclei could be stimulated with the rat heart CE. Furthermore, CE from hearts undergoing induced growth had greater activity than extract from control hearts. While cardiac muscle nuclei were not stimulated by heart CE, they showed substantial stimulation by CE from fetal rat liver, which contains a large population of proliferating cells. Stimulation by fetal rat liver was greater with nuclei obtained from hearts undergoing induced growth than from control hearts. Stimulatory factor(s) in CE was distinct from DNA polymerase-alpha activity, as shown by separation of the two activities on a 5 to 15% glycerol gradient.

Animals

Home refinishing, lead paint, and infant blood lead levels.

We measured the blood lead levels of 249 infants semi-annually from birth to two years of age; we sampled the home paint and recorded any recent home refinishing activity. Mean blood lead from birth to age 2 years did not vary systematically with age but did correlate significantly with the amount of lead in the indoor paint (p less than .01). Refinishing activity in homes with high lead paint was associated with elevations of blood lead averaging 69 per cent.

Adult

The relationship between prenatal exposure to lead and congenital anomalies.

We obtained umbilical cord blood from 5,183 consecutive deliveries of at least 20 weeks' gestation and analyzed them for lead concentration. Those demographic and socioeconomic variables, including lead, which were shown on univariate analysis to be associated with increased risk for congenital anomalies were evaluated and controlled by entering them into a stepwise logistic-regression model with malformation as the outcome. Coffee, alcohol, tobacco, and marijuana use, which were associated with lead level, but not risk of malformation, were also controlled. The model was reduced in steps by eliminating the variables with the highest P value, until the most parsimonious model was created. The relative risk for anomalies associated with lead was then calculated while holding other covariates constant. Lead was found to be associated, in a dose-related fashion, with an increased risk for minor anomalies.

Abnormalities, Drug-Induced

Expression of rabbit ventricular alpha-myosin heavy chain messenger RNA sequences in atrial muscle.

We have constructed and isolated a cardiac myosin heavy chain (HC) cDNA clone, pMHC alpha 81, with mRNA from ventricular heart muscle of hyperthyroid rabbits. The clone encodes approximately 480 amino acids of the COOH terminus of light meromyosin and all of the 3' nontranslated region of the corresponding mRNA. Nuclease S1 analyses indicated that the clone is transcribed in hyperthyroid, but not in hypothyroid ventricles and, therefore, corresponds to ventricular alpha-HC mRNA. With probes from the more divergent 3' non-translated region of pMHC alpha 81 and also from selected portions of two previously characterized rabbit cDNA clones ( pMHC alpha 252 and pMHC beta 174), we analyzed the myosin HC mRNAs of atrial, fast skeletal, and slow skeletal muscles by nuclease S1 mapping. In atrial muscle, only one major transcript was detected. The sequence of this transcript was indistinguishable from ventricular alpha-HC mRNA in the 3' nontranslated region and in two coding segments. In contrast, the sequence divergence between the ventricular alpha-HC mRNA and the mRNAs of ventricular beta, fast skeletal, and slow skeletal myosin HCs was clearly detected. There appeared to be, however, considerable homology between coding sequences of ventricular beta and slow skeletal myosin HC mRNAs. The results strongly suggest that rabbit atrial and ventricular alpha-HCs are encoded by the same gene.

Amino Acid Sequence

Regulation of myosin synthesis by thyroid hormone: relative change in the alpha- and beta-myosin heavy chain mRNA levels in rabbit heart.

The expression of mRNAs for two cardiac myosins has been examined in the ventricles of hypo- and hyperthyroid rabbits by means of cloned cDNA sequences corresponding to the mRNAs of the alpha- and beta-myosin heavy chains (HCs). The temporal change in the relative levels of the alpha- and beta-HC mRNAs after 3,5,3'-triiodothyronine (T3) treatment of hypothyroid rabbits was determined by nuclease S1 mapping. In the hypothyroid state, only HC beta-mRNA was expressed in the ventricles. The HC alpha-mRNA was first detectable 4 h after administration of T3 (200 micrograms/kg) to hypothyroid animals. By 12 h, HC alpha-mRNA represented 20% of total myosin mRNA, increasing to 50% by 24 h and to about 90% by 72 h. The relationship between the relative mRNA levels and relative synthesis rates of the myosin HCs was evaluated in 5-6-week-old normal and thyrotoxic rabbits. Myosin synthesis rates were determined by labeling of protein in vivo with [3H]leucine. The V1 (HC alpha) and V3 (HC beta) isomyosins were separated by affinity chromatography with monoclonal antibodies, and the HCs were isolated electrophoretically. In a normal euthyroid group of animals and in animals 12 and 24 h after administration of 200 micrograms of 3,5,3',5'-tetraiodothyronine/kg, the relative mRNA levels and relative synthesis rates of the alpha- and beta-HCs were not significantly different. Our results show that, first, thyroid hormone causes a rapid accumulation of HC alpha-mRNA and loss of HC beta-mRNA and, second, in normal and thyrotoxic rabbits, the relative synthesis rates of HC alpha and HC beta reflect the relative abundance of the alpha- and beta-HC mRNAs.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Analysis of cloned mRNA sequences encoding subfragment 2 and part of subfragment 1 of alpha- and beta-myosin heavy chains of rabbit heart.

Two cardiac myosin heavy chain cDNA clones, pMHC alpha 252 and pMHC beta 174, were constructed using rabbit ventricular mRNA isolated from adult thyrotoxic and normal hearts, respectively. The complete DNA sequences of the 2.2- and 1.4-kilobase inserts of pMHC beta 174 and pMHC alpha 252, respectively, were obtained. The 736 amino acids specified by pMHC beta 174 begin 439 (1.3 kilobases) residues from the heavy chain NH2 terminus and include a 400-amino acid segment of subfragment 1 and the entire subfragment 2 region. Clone pMHC alpha 252 encodes 465 amino acids encompassing all of subfragment 2 and a portion of light meromyosin. Comparison of these two clones revealed extensive sequence overlap which included 1107 nucleotides specifying a 369-amino acid segment corresponding to subfragment 2. Within this region 78 (7%) base and 32 (8.7%) amino acid mismatches were noted. These differences were clustered within discrete regions, with the subfragment 1/subfragment 2 junctional region being particularly divergent. Structural differences between pMHC alpha 252 and pMHC beta 174 indicate that these two clones represent two similar but distinct myosin heavy chain genes whose expression is responsible for ventricular myosin heavy chain isoforms alpha and beta, respectively. The derived amino acid sequences of both clones exhibit extensive homology (greater than 81%) with sequences obtained by direct analysis of adult rabbit skeletal muscle myosin heavy chain protein. The sequences corresponding to the subfragment 2 region are consistent with an alpha-helical conformation with a characteristic 7-residue periodicity in the linear distribution of nonpolar amino acids. Conversely, subfragment 1 sequences specified by pMHC beta 174 suggest a folded highly irregular structure.

Amino Acid Sequence

Cadmium content of umbilical cord blood.

Cadmium was measured in the umbilical cord blood at birth from 94 healthy babies. Samples were dried and ashed at low temperatures with an oxygen plasma prior to atomic absorption spectrometry. The concentration of cadmium ranged from 0.003 to 0.210 microgram/dl, with a mean of 0.045 +/- 0.063 (SD). Blood lead, maternal smoking, and proximity of residence to automobile traffic were not statistically related to cadmium levels.

Automobiles

Characterization of genomic clones specifying rabbit alpha- and beta-ventricular myosin heavy chains.

We have isolated gene sequences coding for the alpha- and beta-myosin heavy chains (HC) of rabbit ventricular muscle. A rabbit genomic library was screened with previously characterized cDNA clones specifying part of the light meromyosin and the entire subfragment 2 portion of alpha- and beta-myosin HCs, as well as with a clone containing the 3' nontranslated sequences of the alpha-myosin HC mRNA. Seven strongly hybridizing clones were analyzed in detail. One genomic clone encoded all of the 3' nontranslated sequences of an alpha-cDNA clone and, therefore, contained the 3' end of the alpha-myosin HC gene. Electron microscopic heteroduplex analysis and DNA sequence analysis showed that this clone overlapped a second genomic clone providing more than 25 kilobase pairs of the alpha-myosin HC gene. The exons within this region corresponded to approximately equal to 85% of the mRNA and were separated by at least 28 introns. A clone for the beta-myosin HC gene was also identified by Southern blot hybridization, by heteroduplex mapping, and by comparing the DNA sequence of a subfragment 2 exon to sequences of the alpha- and beta-cDNA clones. The introns of the alpha- and beta-myosin HC genes were in the same position but showed marked variation in length. These results conclusively showed that the alpha- and beta-myosin HCs are products of separate genes.

Animals