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Biomedical subjects

J McManus

Publications and source records attributed to J McManus.

15 recordsLinked to original sources

Unusual chromosome structure of fission yeast DNA in mouse cells.

Chromosomes from the fission yeast Schizosaccharomyces pombe have been introduced into mouse cells by protoplast fusion. In most cell lines the yeast DNA integrates into a single site within a mouse chromosome and results in striking chromosome morphology at metaphase. Both light and electron microscopy show that the yeast chromosome region is narrower than the flanking mouse DNA. Regions of the yeast insert stain less intensely with propidium iodide than surrounding DNA and bear a morphological resemblance to fragile sites. We investigate the composition of the yeast transgenomes and the modification and chromatin structure of this yeast DNA in mouse cells. We suggest that the underlying basis for the structure we see lies above the level of DNA modification and nucleosome assembly, and may reflect the attachment of the yeast DNA to the rodent cell nucleoskeleton. The yeast integrant replicates late in S phase at a time when G bands of the mouse chromosomes are being replicated, and participates in sister chromatid exchanges at a high frequency. We discuss the implications of these studies to the understanding of how chromatin folding relates to metaphase chromosome morphology and how large stretches of foreign DNA behave when introduced into mammalian cells.

Anaphase

Mobile mission.

The "Big Blue" van of The Children's Aid Society brings much needed health services to homeless and underserved children of New York City.

Child

Fecal coproporphyrin isomers in hereditary coproporphyria.

To see whether the fecal coproporphyrin III:coproporphyrin I (CIII:CI) ratio (determined by HPLC) would be suitable for screening patients at risk of hereditary coproporphyria (HC), we compared such ratios with the lymphocyte coproporphyrinogen oxidase (EC 1.3.3.3) activities (COOX) in 38 subjects from one large family and two smaller families with HC. The CIII:CI ratio was normal (less than 1.3) in adults with normal COOX (greater than 180 nmol/g of protein per hour) and high (greater than 2) in those with low COOX. Results were difficult to interpret in six of 10 children, who had borderline or low COOX but normal fecal CIII:CI ratios. Five subjects with low COOX and abnormal fecal CIII:CI ratios had normal fecal total porphyrin, indicating that the latter investigation alone is inadequate for family studies. The sample for determining the fecal CIII:CI ratio is easier to obtain and the assay is technically less demanding than COOX. We found the fecal CIII:CI ratio suitable for investigation of adults in a family study, but its usefulness in children needs to be established.

Adolescent

An unusual case of variegate porphyria with possible homozygous inheritance.

We report an unusual case of variegate porphyria in a young girl with epilepsy, mental retardation and premature adrenarche. Symptoms of porphyria commenced about the age of 12 years and death occurred about 18 months later. The patient had very low protoporphyrinogen oxidase activity in her cultured fibroblasts. Both parents had half the normal activity of this enzyme in lymphocytes and are heterozygous for the abnormal gene for variegate porphyria. Therefore, it is possible that the patient was a homozygous variant. Anticonvulsant therapy and low hepatic 5 alpha reductase activity were probably other contributing factors to the severity of the condition in this patient.

Animals

An assay of uroporphyrinogen decarboxylase in erythrocytes.

Measurement of uroporphyrinogen decarboxylase (UROD; EC 4.1.1.37) activity in erythrocytes is useful in distinguishing between familial porphyria cutanea tarda (PCT), in which UROD activities are low, and acquired PCT, in which UROD activity is normal. In this method for measuring UROD, pentacarboxylic acid porphyrinogen I (PPI) is used as substrate. A sample of the patient's whole blood is incubated with PPI at 37 degrees C for 30 min at pH 6.0. The reaction is stopped by adding trichloroacetic acid/dimethyl sulfoxide containing mesoporphyrin (internal standard). The coproporphyrin so produced is measured directly by high-performance liquid chromatography, with fluorescence detection. Our values by this method for healthy subjects and non-PCT patients ranged from 1.8 to 4.0 U/L. The CV for the assay was 10% at 1.1 U/L and 9% at 2.4 U/L. Twelve of 42 patients with PCT had low erythrocyte UROD activities. In each of six families of patients with low UROD activity we found at least one other family member with a low UROD activity in erythrocytes.

Adult

Polymer support oligonucleotide synthesis XVIII: use of beta-cyanoethyl-N,N-dialkylamino-/N-morpholino phosphoramidite of deoxynucleosides for the synthesis of DNA fragments simplifying deprotection and isolation of the final product.

Various 5'O-N-protected deoxynucleoside-3'-O-beta-cyanoethyl-N,N-dialkylamino-/N- morpholinophosphoramidites were prepared from beta-cyanoethyl monochlorophosphoramidites of N,N-dimethylamine, N,N-diisopropylamine and N-morpholine. These active deoxynucleoside phosphates have successfully been used for oligodeoxynucleotide synthesis on controlled pore glass as polymer support and are very suitable for automated DNA-synthesis due to their stability in solution. The intermediate dichloro-beta- cyanoethoxyphosphine can easily be prepared free from any PC1(3) contamination. The active monomers obtained from beta-cyanoethyl monochloro N,N- diisopropylaminophosphoramidites are favoured. Cleavage of the oligonucleotide chain from the polymer support, N-deacylation and deprotection of beta-cyanoethyl group from the phosphate triester moiety can be performed in one step with concentrated aqueous ammonia. Mixed oligodeoxynucleotides are characterized by the sequencing method of Maxam and Gilbert.

DNA

Sensitive radioimmunoassay for somatostatin using N-[125I]-Tyr-somatostatin as labelled antigen.

A sensitive radioimmunoassay for somatostatin using N-[125I]-Tyr-somatostatin is described and compared with that using [125I]-Tyr1-somatostatin. The minimum detectable amount of somatostatin using N-[125I]-Tyr-somatostatin as tracer was 0.1 to 0.5 pg, which is approximately 10-fold lower than the lower detection limit of the RIA using [125I]-Tyr1-somatostatin. Moreover, it was found that the shelf-life of N[125I]Tyr-somatostatin was prolonged in comparison with labelled Tyr1-somatostatin. Human pancreatic and gastric extracts displayed immunological similarity to synthetic somatostatin tetradecapeptide.

Animals

Experience with hyperselective vagotomy in patients with duodenal ulcer.

Hyperselective vagotomy is a relatively recent surgical procedure used for the treatment of duodenal ulcer. We report excellent results in 98 consecutive patients during the last four years. After a mean follow-up period of 2.2 years, the results, according to the classification of Visick, are excellent in a large number of patients. There was a significant gain in body weight in a majority of the patients. The recurrence rate for an ulcer is only 4.5 per cent. There is a significant reduction of basal acid output of 63 per cent, maximal acid output of 46.8 per cent and postinsulin acid output of 73.7 per cent.

Adult

Medications not to be refrigerated.

A survey of selected drug labelers was conducted to generate a list of drug products that should not be refrigerated. Letters asking for information on products adversely affected by refrigeration were mailed to 109 drug product labelers. A second letter was sent to nonresponders and to labelers providing incomplete information. Responses were received from 97 labelers, 43 of whom stated that none of their products would be harmed by refrigeration. Eleven labelers were unable to provide conclusive data or a list of specific drug products that refrigeration would harm. Lists of drug products not to be refrigerated were provided by 43 labelers, some including explanations of the adverse effects of refrigeration. Pharmacists may find the survey's data useful in their patient education activities.

Drug Labeling

Prevention by specific perceptual remediation for vulnerable first-graders. Controlled study and follow-up of lasting effects.

In a pretest-posttest design with two matched control groups, 86 first-graders screened as vulnerable to academic failure and behavioral decompensation were each assigned to one of three groups: (1) channel-specific perceptual stimulation, (2) regular academic tutoring (contact controls), or (3) no contact. On most measures, including perceptual and achievement tests and behavior ratings by teachers, group 1 showed more improvement than either of the control groups. Several of these differences were significant (P less than .05, two-tailed). In general, the two control groups came closer to each other than to group 1. With no further intervention, follow-up one year later showed more dramatic significance. On every measure, group 1's improvement surpassed that of the control groups, and diverged from them even more than at posttest; this divergence was significant on most measures. Group 1 improved significantly (P less than .05 to P less than .01) in reading, IQ, and three behavior scales, while both control groups showed only deterioration on these measures. Group 1 gained five IQ points on the Wechsler Intelligence Scale for Children, while the controls lost five. On the Davids Scale, group 1 improved significantly (P less than .01) to "probably not hyperkinetic," while both control groups deteriorated to "probably hyperkinetic." The hypothesized mechanism of group 1's behavioral superiority at follow-up was its significant (P less than 0.1) reading improvement, with secondary emotional benefits.

Adolescent